Evaluation of the effects of insufficient blood volume samples on the performance of blood glucose self-test meters.

PubMed

Pfützner, Andreas; Schipper, Christina; Ramljak, Sanja; Flacke, Frank; Sieber, Jochen; Forst, Thomas; Musholt, Petra B

2013-11-01

Accuracy of blood glucose readings is (among other things) dependent on the test strip being completely filled with sufficient sample volume. The devices are supposed to display an error message in case of incomplete filling. This laboratory study was performed to test the performance of 31 commercially available devices in case of incomplete strip filling. Samples with two different glucose levels (60-90 and 300-350 mg/dl) were used to generate three different sample volumes: 0.20 µl (too low volume for any device), 0.32 µl (borderline volume), and 1.20 µl (low but supposedly sufficient volume for all devices). After a point-of-care capillary reference measurement (StatStrip, NovaBiomedical), the meter strip was filled (6x) with the respective volume, and the response of the meters (two devices) was documented (72 determinations/meter type). Correct response was defined as either an error message indicating incomplete filling or a correct reading (±20% compared with reference reading). Only five meters showed 100% correct responses [BGStar and iBGStar (both Sanofi), ACCU-CHEK Compact+ and ACCU-CHEK Mobile (both Roche Diagnostics), OneTouch Verio (LifeScan)]. The majority of the meters (17) had up to 10% incorrect reactions [predominantly incorrect readings with sufficient volume; Precision Xceed and Xtra, FreeStyle Lite, and Freedom Lite (all Abbott); GlucoCard+ and GlucoMen GM (both Menarini); Contour, Contour USB, and Breeze2 (all Bayer); OneTouch Ultra Easy, Ultra 2, and Ultra Smart (all LifeScan); Wellion Dialog and Premium (both MedTrust); FineTouch (Terumo); ACCU-CHEK Aviva (Roche); and GlucoTalk (Axis-Shield)]. Ten percent to 20% incorrect reactions were seen with OneTouch Vita (LifeScan), ACCU-CHEK Aviva Nano (Roche), OmniTest+ (BBraun), and AlphaChek+ (Berger Med). More than 20% incorrect reactions were obtained with Pura (Ypsomed), GlucoCard Meter and GlucoMen LX (both Menarini), Elite (Bayer), and MediTouch (Medisana). In summary, partial and incomplete blood filling of glucose meter strips is often associated with inaccurate reading. These findings underline the importance of appropriate patient education on this aspect of blood glucose self-monitoring. © 2013 Diabetes Technology Society.

Miniaturized blood sampling techniques to benefit reduction in mice and refinement in nonhuman primates: applications to bioanalysis in toxicity studies with antibody-drug conjugates.

PubMed

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

2015-03-01

Minimizing the number of animals in regulatory toxicity studies while achieving study objectives to support the development of future medicines contributes to good scientific and ethical practices. Recent advances in technology have enabled the development of miniaturized blood sampling methods (including microsampling and dried blood spots) applicable to toxicokinetic determinations of small-molecule drugs. Implementation of miniaturized blood sampling methods in the context of biotherapeutic drugs is desirable because a limitation to this type of medicine remains the total blood volume needed from a single animal to support toxicokinetic determinations of several analytes (parent drug, metabolites[s], antidrug antibodies, and so forth). We describe here the technical details, applicability, and relevance of new miniaturized blood sampling procedures in mice and nonhuman primates in the context of the toxicologic evaluation of biotherapeutic drugs consisting of antibody-drug conjugates developed for oncology indications. These examples illustrate how these techniques can benefit the reduction of animal usage in mouse toxicity studies by decreasing the number of animals dedicated to toxicokinetic determinations and the refinement of practices in nonhuman primate toxicity studies by decreasing the blood volume repeatedly drawn for toxicokinetic determinations.

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

PubMed Central

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

2015-01-01

Minimizing the number of animals in regulatory toxicity studies while achieving study objectives to support the development of future medicines contributes to good scientific and ethical practices. Recent advances in technology have enabled the development of miniaturized blood sampling methods (including microsampling and dried blood spots) applicable to toxicokinetic determinations of small-molecule drugs. Implementation of miniaturized blood sampling methods in the context of biotherapeutic drugs is desirable because a limitation to this type of medicine remains the total blood volume needed from a single animal to support toxicokinetic determinations of several analytes (parent drug, metabolites[s], antidrug antibodies, and so forth). We describe here the technical details, applicability, and relevance of new miniaturized blood sampling procedures in mice and nonhuman primates in the context of the toxicologic evaluation of biotherapeutic drugs consisting of antibody–drug conjugates developed for oncology indications. These examples illustrate how these techniques can benefit the reduction of animal usage in mouse toxicity studies by decreasing the number of animals dedicated to toxicokinetic determinations and the refinement of practices in nonhuman primate toxicity studies by decreasing the blood volume repeatedly drawn for toxicokinetic determinations. PMID:25836960

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

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

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

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

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

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

DOE PAGES

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

2016-04-07

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

Some physiologic blood values of wild diving ducks

USGS Publications Warehouse

Kocan, R.M.

1972-01-01

Blood samples were obtained from 54 canvasbacks (Aythya valisineria), 30 lesser scaup (A. affinis), 3 ring-necks (A. collaris), and 3 buffleheads (Bucephala albeola), which were wintering on Chesapeake Bay. These blood samples were used for: red blood cell counts (cans. 2.56 x 106/mm3; scaup 2.45 x 106/mm3; ring-necks 2.50 x 106mm3; buffiehead 2.64 x lO6/mm3), packed cell volume (cans. 52.2%; scaup 57.1%; ring-neck 49.1%; buffieheads 54.3%), total protein (cans. 4.4 g/100 ml; scaup 4.4 g/100 mi; ring-neck 3.5 g/100 ml; buffiehead 3.8 g/100 ml), erythrocyte measurements, and electrophoretic analysis of plasma and serum. Variations between individuals of the same species and sex were slight but a possible difference between male and female erythrocyte numbers and packed cell volumes was evident in the canvasback and scaup samples.

Effect of Increased Plasma Osmolality on Cold-Induced Thirst Attenuation

DTIC Science & Technology

2008-08-01

Xuid intake. Oropharyngeal and gastric stim- uli, as well as blood volume and plasma osmolality (Posm) are important factors modifying thirst (Greenleaf...reported to result in an increase in the osmotic thresh- old for the stimulation of thirst due to an increase in central blood volume (Sobocinska and...model 5877, Abbott Hospital, Inc., Chicago, IL, USA) was inserted into the catheter port for acquisition of subsequent blood samples. The catheter

Ultrasensitive Detection of Shigella Species in Blood and Stool.

PubMed

Luo, Jieling; Wang, Jiapeng; Mathew, Anup S; Yau, Siu-Tung

2016-02-16

A modified immunosensing system with voltage-controlled signal amplification was used to detect Shigella in stool and blood matrixes at the single-digit CFU level. Inactivated Shigella was spiked in these matrixes and detected directly. The detection was completed in 78 min. Detection limits of 21 CFU/mL and 18 CFU/mL were achieved in stool and blood, respectively, corresponding to 2-7 CFUs immobilized on the detecting electrode. The outcome of the detection of extremely low bacterium concentration, i.e., below 100 CFU/mL, blood samples show a random nature. An analysis of the detection probabilities indicates the correlation between the sample volume and the success of detection and suggests that sample volume is critical for ultrasensitive detection of bacteria. The calculated detection limit is qualitatively in agreement with the empirically determined detection limit. The demonstrated ultrasensitive detection of Shigella on the single-digit CFU level suggests the feasibility of the direct detection of the bacterium in the samples without performing a culture.

Comparison of the image-derived radioactivity and blood-sample radioactivity for estimating the clinical indicators of the efficacy of boron neutron capture therapy (BNCT): 4-borono-2-18F-fluoro-phenylalanine (FBPA) PET study.

PubMed

Isohashi, Kayako; Shimosegawa, Eku; Naka, Sadahiro; Kanai, Yasukazu; Horitsugi, Genki; Mochida, Ikuko; Matsunaga, Keiko; Watabe, Tadashi; Kato, Hiroki; Tatsumi, Mitsuaki; Hatazawa, Jun

2016-12-01

In boron neutron capture therapy (BNCT), positron emission tomography (PET) with 4-borono-2- 18 F-fluoro-phenylalanine (FBPA) is the only method to estimate an accumulation of 10 B to target tumor and surrounding normal tissue after administering 10 B carrier of L-paraboronophenylalanine and to search the indication of BNCT for individual patient. Absolute concentration of 10 B in tumor has been estimated by multiplying 10 B concentration in blood during BNCT by tumor to blood radioactivity (T/B) ratio derived from FBPA PET. However, the method to measure blood radioactivity either by blood sampling or image data has not been standardized. We compared image-derived blood radioactivity of FBPA with blood sampling data and studied appropriate timing and location for measuring image-derived blood counts. We obtained 7 repeated whole-body PET scans in five healthy subjects. Arterialized venous blood samples were obtained from the antecubital vein, heated in a heating blanket. Time-activity curves (TACs) of image-derived blood radioactivity were obtained using volumes of interest (VOIs) over ascending aorta, aortic arch, pulmonary artery, left and right ventricles, inferior vena cava, and abdominal aorta. Image-derived blood radioactivity was compared with those measured by blood sampling data in each location. Both the TACs of blood sampling radioactivity in each subject, and the TACs of image-derived blood radioactivity showed a peak within 5 min after the tracer injection, and promptly decreased soon thereafter. Linear relationship was found between blood sampling radioactivity and image-derived blood radioactivity in all the VOIs at any timing of data sampling (p < 0.001). Image-derived radioactivity measured in the left and right ventricles 30 min after injection showed high correlation with blood radioactivity. Image-derived blood radioactivity was lower than blood sampling radioactivity data by 20 %. Reduction of blood radioactivity of FBPA in left ventricle after 30 min of FBPA injection was minimal. We conclude that the image-derived T/B ratio can be reliably used by setting the VOI on the left ventricle at 30 min after FBPA administration and correcting for underestimation due to partial volume effect and reduction of FBPA blood radioactivity.

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

An affordable method to obtain cultured endothelial cells from peripheral blood

PubMed Central

Bueno-Betí, Carlos; Novella, Susana; Lázaro-Franco, Macarena; Pérez-Cremades, Daniel; Heras, Magda; Sanchís, Juan; Hermenegildo, Carlos

2013-01-01

The culture of endothelial progenitor cells (EPC) provides an excellent tool to research on EPC biology and vascular regeneration and vasculogenesis. The use of different protocols to obtain EPC cultures makes it difficult to obtain comparable results in different groups. This work offers a systematic comparison of the main variables of most commonly used protocols for EPC isolation, culture and functional evaluation. Peripheral blood samples from healthy individuals were recovered and mononuclear cells were cultured. Different recovery and culture conditions were tested: blood volume, blood anticoagulant, coating matrix and percentage of foetal bovine serum (FBS) in culture media. The success of culture procedure, first colonies of endothelial cells appearance time, correlation with number of circulating EPC (cEPC) and functional comparison with human umbilical vein endothelial cells (HUVEC) were studied. The use of heparin, a minimum blood volume of 30 ml, fibronectin as a coating matrix and endothelial growing media-2 supplemented with 20% FBS increased the success of obtaining EPC cultures up to 80% of the processed samples while reducing EPC colony appearance mean time to a minimum of 13 days. Blood samples exhibiting higher cEPC numbers resulted in reduced EPC colony appearance mean time. Cells isolated by using this combination were endothelial cell-like EPCs morphological and phenotypically. Functionally, cultured EPC showed decreased growing and vasculogenic capacity when compared to HUVEC. Thus, above-mentioned conditions allow the isolation and culture of EPC with smaller blood volumes and shorter times than currently used protocols. PMID:24118735

Evacuated blood-collection tubes for haematological tests - a quality evaluation prior to their intended use for specimen collection.

PubMed

Gros, Nataša

2013-05-01

An inappropriate anticoagulant concentration in a blood sample can cause cell shrinkage and affect the haematocrit and mean corpuscular volume (MCV). In evacuated blood-collection tubes there are two parameters affecting the quality of the product: the anticoagulant amount introduced into the tube during its production and the internal under-pressure at the instant of the blood-specimen collection affecting the draw-volume. No testing procedures that would give an insight into the anticoagulant concentration that can be expected for blood samples after specimen collection have been available up until now. The methodology suggested here combines the draw-volume test performed with deionised water using a laboratory made measuring device, and a conductivity measurement. The corrections taking into account the air pressure and ambient temperature provide an insight into the anticoagulant concentration that can be expected for blood samples. Results presented in the form of a nomogram facilitate the routine use of the suggested methodology. Our 338-day study confirmed significant differences and variations in the quality and the anticoagulant concentrations of the K₃EDTA and K2EDTA tubes of different producers and identified different examples of non-compliance with the norms during the shelf life of the tubes. The quality evaluation of the evacuated blood-collection tubes prior to their intended use as suggested here can, in everyday laboratory practice, ensure that the tubes are used only if, and only until, their quality is adequate.

Role of cardiac volume receptors in the control of ADH release during acute simulated weightlessness in man

NASA Technical Reports Server (NTRS)

Convertino, V. A.; Benjamin, B. A.; Keil, L. C.; Sandler, H.

1984-01-01

Hemodynamic responses and antidiuretic hormone (ADH) were measured during body position changes, designed to induce central blood volume shifts in ten cardiac and one heart-lung transplant recipients, to assess the contribution of cardiac volume receptors in the control of ADH release during the initial acute phase of exposure to weightlessness. Each subject underwent 15 min of a sitting-control period (C) followed by 30 min of 6 deg headdown tilt (T) and 30 min of resumed sitting (S). Venous blood samples and cardiac dimensions were taken at 0 and 15 min of C; 5, 15, and 30 min of T; and 5, 15, and 30 min of S. Blood samples were analyzed for hematocrit, plasma osmolality, plasma renin activity (PRA), and ADH. Heart rate and blood pressure were recorded every two min. Plasma osmolality was not altered by posture changes. Mean left ventricular end-diastolic volume increased (P less than 0.05) from 90 ml in C to 106 ml in T and returned to 87 ml in S. Plasma ADH was reduced by 20 percent (P less than 0.05) with T, and returned to control levels with S. These responses were similar in six normal cardiac-innervated control subjects. These data may suggest that cardiac volume receptors are not the primary mechanism for the control of ADH release during acute central volume shifts in man.

Method and Apparatus for the Collection, Storage, and Real Time Analysis of Blood and Other Bodily Fluids

NASA Technical Reports Server (NTRS)

Whiitson, Peggy A. (Inventor); Clift, Vaughan L. (Inventor)

1999-01-01

The present invention provides a method and apparatus for separating a blood sample having a volume of up to about 20 milliliters into cellular and acellular fractions. The apparatus includes a housing divided by a fibrous filter into a blood sample collection chamber having a volume of at least about 1 milliliter and a serum sample collection chamber. The fibrous filter has a pore size of less than about 3 microns, and is coated with a mixture including between about 1-40 wt/vol % mannitol and between about 0.1-15 wt/vol % of plasma fraction protein (or an animal or vegetable equivalent thereof). The coating causes the cellular fraction to be trapped by the small pores, leaving the cellular fraction intact on the fibrous filter while the acellular fraction passes through the filter for collection in unaltered form from the serum sample collection chamber.

Clinical biochemistry laboratory rejection rates due to various types of preanalytical errors.

PubMed

Atay, Aysenur; Demir, Leyla; Cuhadar, Serap; Saglam, Gulcan; Unal, Hulya; Aksun, Saliha; Arslan, Banu; Ozkan, Asuman; Sutcu, Recep

2014-01-01

Preanalytical errors, along the process from the beginning of test requests to the admissions of the specimens to the laboratory, cause the rejection of samples. The aim of this study was to better explain the reasons of rejected samples, regarding to their rates in certain test groups in our laboratory. This preliminary study was designed on the rejected samples in one-year period, based on the rates and types of inappropriateness. Test requests and blood samples of clinical chemistry, immunoassay, hematology, glycated hemoglobin, coagulation and erythrocyte sedimentation rate test units were evaluated. Types of inappropriateness were evaluated as follows: improperly labelled samples, hemolysed, clotted specimen, insufficient volume of specimen and total request errors. A total of 5,183,582 test requests from 1,035,743 blood collection tubes were considered. The total rejection rate was 0.65 %. The rejection rate of coagulation group was significantly higher (2.28%) than the other test groups (P < 0.001) including insufficient volume of specimen error rate as 1.38%. Rejection rates of hemolysis, clotted specimen and insufficient volume of sample error were found to be 8%, 24% and 34%, respectively. Total request errors, particularly, for unintelligible requests were 32% of the total for inpatients. The errors were especially attributable to unintelligible requests of inappropriate test requests, improperly labelled samples for inpatients and blood drawing errors especially due to insufficient volume of specimens in a coagulation test group. Further studies should be performed after corrective and preventive actions to detect a possible decrease in rejecting samples.

Peripheral venous vs. capillary microfilariaemia in a dog co-infected with Dirofilaria repens and D. immitis: A comparative approach using triatomine bugs for blood collection.

PubMed

Păstrav, Ioana Raluca; Ionică, Angela Monica; Peştean, Cosmin; Novakova, Eva; Modrý, David; Mihalca, Andrei Daniel

2018-06-15

Dirofilaria immitis and D. repens are mosquito-borne nematodes, primarily infecting dogs, but also other species of carnivores and even humans. Given their impact on animal and human health, the transmission of these filarioids has been widely studied. The microfilariaemia has been shown to have a circadian variation for both Dirofilaria species infecting dogs. Due to methodological difficulties, the periodicity was only studied using venous blood samples, while the mosquitoes feed, in fact, on capillary blood. In this context, the present study aimed to test the feasibility of using triatomine bugs for the collection of capillary blood and to comparatively evaluate the level of microfilariaemia and its circadian variation in capillary blood vs. peripheral venous blood in a dog naturally co-infected with D. immitis and D. repens. The results showed a feeding success of 50%, with variations in the blood meal volume that the bugs ingested. The relative values of microfilariaemia (mf/bug) were strongly correlated with the volume of blood recovered: the more blood recovered from each bug, the higher values of microfilariaemia in the evening samples while the opposite results were obtained for the morning samples. The counting of microfilariae revealed a dominance of D. immitis in all the samples, but with significantly higher microfilariaemia in the venous blood. Meanwhile, for D. repens, the situation was opposite, with higher counts in the capillary blood samples. Our study showed that triatomine bugs can be used as a model for the collection and study of microfilariaemia in the capillary blood in mammals. Copyright © 2018 Elsevier B.V. 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.

OPTIMIZED DETERMINATION OF TRACE JET FUEL VOLATILE ORGANIC COMPOUNDS IN HUMAN BLOOD USING IN-FIELD LIQUID-LIQUID EXTRACTION WITH SUBSEQUENT LABORATORY GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS AND ON-COLUMN LARGE VOLUME INJECTION

EPA Science Inventory

A practical and sensitive method to assess volatile organic compounds (VOCs) from JP-8 jet fuel in human whole blood was developed by modifying previously established liquid-liquid extraction procedures, optimizing extraction times, solvent volume, specific sample processing te...

Adhesive blood microsampling systems for steroid measurement via LC-MS/MS in the rat.

PubMed

Heussner, Kirsten; Rauh, Manfred; Cordasic, Nada; Menendez-Castro, Carlos; Huebner, Hanna; Ruebner, Matthias; Schmidt, Marius; Hartner, Andrea; Rascher, Wolfgang; Fahlbusch, Fabian B

2017-04-01

Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) allows for the direct analysis of multiple hormones in a single probe with minimal sample volume. Rodent-based animal studies strongly rely on microsampling, such as the dry blood spot (DBS) method. However, DBS suffers the drawback of hematocrit-dependence (non-volumetric). Hence, novel volumetric microsampling techniques were introduced recently, allowing sampling of fixed accurate volumes. We compared these methods for steroid analysis in the rat to improve inter-system comparability. We analyzed steroid levels in blood using the absorptive microsampling devices Whatman® 903 Protein Saver Cards, Noviplex™ Plasma Prep Cards and the Mitra™ Microsampling device and compared the obtained results to the respective EDTA plasma levels. Quantitative steroid analysis was performed via LC-MS/MS. For the determination of the plasma volume factor for each steroid, their levels in pooled blood samples from each human adults and rats (18weeks) were compared and the transferability of these factors was evaluated in a new set of juvenile (21days) and adult (18weeks) rats. Hematocrit was determined concomitantly. Using these approaches, we were unable to apply one single volume factor for each steroid. Instead, plasma volume factors had to be adjusted for the recovery rate of each steroid and device individually. The tested microsampling systems did not allow the use of one single volume factor for adult and juvenile rats based on an unexpectedly strong hematocrit-dependency and other steroid specific (pre-analytic) factors. Our study provides correction factors for LC-MS/MS steroid analysis of volumetric and non-volumetric microsampling systems in comparison to plasma. It argues for thorough analysis of chromatographic effects before the use of novel volumetric systems for steroid analysis. Copyright © 2017 Elsevier Inc. All rights reserved.

Design of portable ultraminiature flow cytometers for medical diagnostics

NASA Astrophysics Data System (ADS)

Leary, James F.

2018-02-01

Design of portable microfluidic flow/image cytometry devices for measurements in the field (e.g. initial medical diagnostics) requires careful design in terms of power requirements and weight to allow for realistic portability. True portability with high-throughput microfluidic systems also requires sampling systems without the need for sheath hydrodynamic focusing both to avoid the need for sheath fluid and to enable higher volumes of actual sample, rather than sheath/sample combinations. Weight/power requirements dictate use of super-bright LEDs with top-hat excitation beam architectures and very small silicon photodiodes or nanophotonic sensors that can both be powered by small batteries. Signal-to-noise characteristics can be greatly improved by appropriately pulsing the LED excitation sources and sampling and subtracting noise in between excitation pulses. Microfluidic cytometry also requires judicious use of small sample volumes and appropriate statistical sampling by microfluidic cytometry or imaging for adequate statistical significance to permit real-time (typically in less than 15 minutes) initial medical decisions for patients in the field. This is not something conventional cytometry traditionally worries about, but is very important for development of small, portable microfluidic devices with small-volume throughputs. It also provides a more reasonable alternative to conventional tubes of blood when sampling geriatric and newborn patients for whom a conventional peripheral blood draw can be problematical. Instead one or two drops of blood obtained by pin-prick should be able to provide statistically meaningful results for use in making real-time medical decisions without the need for blood fractionation, which is not realistic in the doctor's office or field.

Method and Apparatus for the Collection Storage and Real Time Analysis of Blood and Other Bodily Fluids

NASA Technical Reports Server (NTRS)

Whitson, Peggy A. (Inventor); Clift, Vaughan L. (Inventor)

1997-01-01

The present invention provides an apparatus for separating a relatively large volume of blood into cellular and acellular fractions without the need for centrifugation. The apparatus comprises a housing divided by a fibrous filter into a blood sample collection chamber having a volume of at least about 1 milliliter and a serum sample collection chamber. The fibrous filter has a pore size of less than about 3 microns, and is coated with a mixture of mannitol and plasma fraction protein (or an animal or vegetable equivalent thereof). The coating causes the cellular fraction to be trapped by the small pores, leaving the cellular fraction intact on the fibrous filter while the acellular fraction passes through the filter for collection in unaltered form from the serum sample collection chamber.

[Transfusion supply optimization in multiple-discipline surgical hospital].

PubMed

Solov'eva, I N; Trekova, N A; Krapivkin, I A

2016-01-01

To define optimal variant of transfusion supply of hospital by blood components and to decrease donor blood expense via application of blood preserving technologies. Donor blood components expense, volume of hemotransfusions and their proportion for the period 2012-2014 were analyzed. Number of recipients of packed red cells, fresh-frozen plasma and packed platelets reduced 18.5%, 25% and 80% respectively. Need for donor plasma decreased 35%. Expense of autologous plasma in cardiac surgery was 76% of overall volume. Preoperative plasma sampling is introduced in patients with aortic aneurysm. Number of cardiac interventions performed without donor blood is increased 7-31% depending on its complexity.

Biomonitoring of perfluorinated compounds in a drop of blood.

PubMed

Mao, Pan; Wang, Daojing

2015-06-02

Biomonitoring of pollutants and their metabolites and derivatives using biofluids provides new opportunities for spatiotemporal assessment of human risks to environmental exposures. Perfluorinated compounds (PFCs) have been used widely in industry and pose significant environmental concerns due to their stability and bioaccumulation in humans and animals. However, current methods for extraction and measurement of PFCs require relatively large volumes (over one hundred microliters) of blood samples, and therefore, are not suitable for frequent blood sampling and longitudinal biomonitoring of PFCs. We have developed a new microassay, enabled by our silicon microfluidic chip platform, for analyzing PFCs in small volumes (less than five microliters) of blood. Our assay integrates on-chip solid-phase extraction (SPE) with online nanoflow liquid chromatography-electrospray ionization-mass spectrometry (nanoLC-ESI-MS) detection. We demonstrated high sample recovery, excellent interday and intraday accuracy and precision, and a limit of detection down to 50 femtogram of PFCs, in one microliter of human plasma. We validated our assay performance using pooled human plasma and NIST SRM 1950 samples. Our microfluidic chip-based assay may enable frequent longitudinal biomonitoring of PFCs and other environmental toxins using a finger prick of blood, thereby providing new insights into their bioaccumulation, bioavailability, and toxicity.

Coagulation measurement from whole blood using vibrating optical fiber in a disposable cartridge

NASA Astrophysics Data System (ADS)

Yaraş, Yusuf Samet; Gündüz, Ali Bars; Saǧlam, Gökhan; Ölçer, Selim; Civitçi, Fehmi; Baris, İbrahim; Yaralioǧlu, Göksenin; Urey, Hakan

2017-11-01

In clinics, blood coagulation time measurements are performed using mechanical measurements with blood plasma. Such measurements are challenging to do in a lab-on-a-chip (LoC) system using a small volume of whole blood. Existing LoC systems use indirect measurement principles employing optical or electrochemical methods. We developed an LoC system using mechanical measurements with a small volume of whole blood without requiring sample preparation. The measurement is performed in a microfluidic channel where two fibers are placed inline with a small gap in between. The first fiber operates near its mechanical resonance using remote magnetic actuation and immersed in the sample. The second fiber is a pick-up fiber acting as an optical sensor. The microfluidic channel is engineered innovatively such that the blood does not block the gap between the vibrating fiber and the pick-up fiber, resulting in high signal-to-noise ratio optical output. The control plasma test results matched well with the plasma manufacturer's datasheet. Activated-partial-thromboplastin-time tests were successfully performed also with human whole blood samples, and the method is proven to be effective. Simplicity of the cartridge design and cost of readily available materials enable a low-cost point-of-care device for blood coagulation measurements.

Coagulation measurement from whole blood using vibrating optical fiber in a disposable cartridge.

PubMed

Yaraş, Yusuf Samet; Gündüz, Ali Bars; Sağlam, Gökhan; Ölçer, Selim; Civitçi, Fehmi; Baris, İbrahim; Yaralioğlu, Göksenin; Urey, Hakan

2017-11-01

In clinics, blood coagulation time measurements are performed using mechanical measurements with blood plasma. Such measurements are challenging to do in a lab-on-a-chip (LoC) system using a small volume of whole blood. Existing LoC systems use indirect measurement principles employing optical or electrochemical methods. We developed an LoC system using mechanical measurements with a small volume of whole blood without requiring sample preparation. The measurement is performed in a microfluidic channel where two fibers are placed inline with a small gap in between. The first fiber operates near its mechanical resonance using remote magnetic actuation and immersed in the sample. The second fiber is a pick-up fiber acting as an optical sensor. The microfluidic channel is engineered innovatively such that the blood does not block the gap between the vibrating fiber and the pick-up fiber, resulting in high signal-to-noise ratio optical output. The control plasma test results matched well with the plasma manufacturer's datasheet. Activated-partial-thromboplastin-time tests were successfully performed also with human whole blood samples, and the method is proven to be effective. Simplicity of the cartridge design and cost of readily available materials enable a low-cost point-of-care device for blood coagulation measurements. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

[Research of bleeding volume and method in blood-letting acupuncture therapy based on data mining].

PubMed

Liu, Xin; Jia, Chun-Sheng; Wang, Jian-Ling; Du, Yu-Zhu; Zhang, Xiao-Xu; Shi, Jing; Li, Xiao-Feng; Sun, Yan-Hui; Zhang, Shen; Zhang, Xuan-Ping; Gang, Wei-Juan

2014-03-01

Through computer-based technology and data mining method, with treatment in cases of bloodletting acupuncture therapy in collected literature as sample data, the association rule in data mining was applied. According to self-built database platform, the data was input, arranged and summarized, and eventually required data was acquired to perform the data mining of bleeding volume and method in blood-letting acupuncture therapy, which summarized its application rules and clinical values to provide better guide for clinical practice. There were 9 kinds of blood-letting tools in the literature, in which the frequency of three-edge needle was the highest, accounting for 84.4% (1239/1468). The bleeding volume was classified into six levels, in which less volume (less than 0.1 mL) had the highest frequency (401 times). According to the results of the data mining, blood-letting acupuncture therapy was widely applied in clinical practice of acupuncture, in which use of three-edge needle and less volume (less than 0.1 mL) of blood were the most common, however, there was no central tendency in general.

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

The application of capillary microsampling in GLP toxicology studies.

PubMed

Verhaeghe, Tom; Dillen, Lieve; Stieltjes, Hans; Zwart, Loeckie de; Feyen, Bianca; Diels, Luc; Vroman, Ann; Timmerman, Philip

2017-04-01

Capillary microsampling (CMS) to collect microplasma volumes is gradually replacing traditional, larger volume sampling from rats in GLP toxicology studies. About 32 µl of blood is collected with a capillary, processed to plasma and stored in a 10- or 4-µl capillary which is washed out further downstream in the laboratory. CMS has been standardized with respect to materials, assay validation experiments and application for sample analysis. The implementation of CMS has resulted in blood volume reductions in the rat from 300 to 32 µl per time point and the elimination of toxicokinetic satellite groups in the majority of the rat GLP toxicology studies. The technique has been successfully applied in 26 GLP studies for 12 different projects thus far.

The use of biomarkers to describe plasma-, red cell-, and blood volume from a simple blood test.

PubMed

Lobigs, Louisa Margit; Sottas, Pierre-Edouard; Bourdon, Pitre Collier; Nikolovski, Zoran; El-Gingo, Mohamed; Varamenti, Evdokia; Peeling, Peter; Dawson, Brian; Schumacher, Yorck Olaf

2017-01-01

Plasma volume and red cell mass are key health markers used to monitor numerous disease states, such as heart failure, kidney disease, or sepsis. Nevertheless, there is currently no practically applicable method to easily measure absolute plasma or red cell volumes in a clinical setting. Here, a novel marker for plasma volume and red cell mass was developed through analysis of the observed variability caused by plasma volume shifts in common biochemical measures, selected based on their propensity to present with low variations over time. Once a month for 6 months, serum and whole blood samples were collected from 33 active males. Concurrently, the CO-rebreathing method was applied to determine target levels of hemoglobin mass (HbM) and blood volumes. The variability of 18 common chemistry markers and 27 Full Blood Count variables was investigated and matched to the observed plasma volume variation. After the removal of between-subject variations using a Bayesian model, multivariate analysis identified two sets of 8 and 15 biomarkers explaining 68% and 69% of plasma volume variance, respectively. The final multiparametric model contains a weighting function to allow for isolated abnormalities in single biomarkers. This proof-of-concept investigation describes a novel approach to estimate absolute vascular volumes, with a simple blood test. Despite the physiological instability of critically ill patients, it is hypothesized the model, with its multiparametric approach and weighting function, maintains the capacity to describe vascular volumes. This model has potential to transform volume management in clinical settings. Am. J. Hematol. 92:62-67, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

In Vitro Effect of Activated Recombinant Factor VII (rFVIIa) on Coagulation Properties of Human Blood at Hypothermic Temperatures

DTIC Science & Technology

2007-11-01

purified soy phosphatides in ellagic acid (Actin FS Activated PTT Reagent, Dade Behring, Marburg, Germany) was added to plasma samples in a 1:1 volume...acetylsali- cylic acid or any other nonsteroidal anti-inflammatory drugs for the 7 days before blood sampling. A smooth cubital venipuncture was

Erythrocyte migration and gap formation in rabbit blood clots in vitro.

PubMed

Ueki, T; Yazama, F; Horiuchi, T; Yamada, M

2008-04-01

Thrombolytic agents must be carried by the blood circulation to thrombi to exert their functions. Structural gaps exist between blood vessels and thrombi or in the area surrounding thrombi. Therefore, information about fundamental gap formation at thrombotic areas is critically important for thrombolytic therapy. We previously reported that t-PA accelerates the activities of bovine erythrocytes and hemoglobin (Hb) towards bovine plasminogen activation. Here, we examined gap generation by observing morphological changes during thrombolytic processes in rabbit blood clots deformation of erythrocytes from blood clots and Hb transfer from erythrocytes to serum in vitro. Rabbit venous blood samples (1 ml) were stored under sterile conditions in glass tubes at 37 degrees C for 2, 24, 48 h, 1, and 2 weeks. We examined clot diameter, erythrocyte diameter and number as well as Hb volume in the serum, as well as histological changes in the clots. The diameter of blood clots did not change until 2 weeks after sampling. Erythrocyte diameter decreased within 48 h and at 2 weeks after sampling at the clot surface (p < 0.001) and interior (p < 0.001). The number of erythrocytes in the serum started to increase starting from 24 h after sampling (p < 0.01). Serum Hb volume also gradually increased from 24 h until 2 weeks after sampling (p < 0.01). The erythrocyte envelope became disrupted and cytoplasm started to flow through pores into the serum at 24 h. The results indicated that blood clots are reduced due to clot retraction, erythrocyte dissociation and cytoplasm leakage without a distinct fibrinolytic reaction. These results indicated that gaps start to form between 2 and 24 h after blood clotting.

Smart fast blood counting of trace volumes of body fluids from various mammalian species using a compact custom-built microscope cytometer (Conference Presentation)

NASA Astrophysics Data System (ADS)

Smith, Zachary J.; Gao, Tingjuan; Lin, Tzu-Yin; Carrade-Holt, Danielle; Lane, Stephen M.; Matthews, Dennis L.; Dwyre, Denis M.; Wachsmann-Hogiu, Sebastian

2016-03-01

Cell counting in human body fluids such as blood, urine, and CSF is a critical step in the diagnostic process for many diseases. Current automated methods for cell counting are based on flow cytometry systems. However, these automated methods are bulky, costly, require significant user expertise, and are not well suited to counting cells in fluids other than blood. Therefore, their use is limited to large central laboratories that process enough volume of blood to recoup the significant capital investment these instruments require. We present in this talk a combination of a (1) low-cost microscope system, (2) simple sample preparation method, and (3) fully automated analysis designed for providing cell counts in blood and body fluids. We show results on both humans and companion and farm animals, showing that accurate red cell, white cell, and platelet counts, as well as hemoglobin concentration, can be accurately obtained in blood, as well as a 3-part white cell differential in human samples. We can also accurately count red and white cells in body fluids with a limit of detection ~3 orders of magnitude smaller than current automated instruments. This method uses less than 1 microliter of blood, and less than 5 microliters of body fluids to make its measurements, making it highly compatible with finger-stick style collections, as well as appropriate for small animals such as laboratory mice where larger volume blood collections are dangerous to the animal's health.

Simplifications in analyzing positron emission tomography data: effects on outcome measures.

PubMed

Logan, Jean; Alexoff, David; Kriplani, Aarti

2007-10-01

Initial validation studies of new radiotracers generally involve kinetic models that require a measured arterial input function. This allows for the separation of tissue binding from delivery and blood flow effects. However, when using a tracer in a clinical setting, it is necessary to eliminate arterial blood sampling due to its invasiveness and the extra burden of counting and analyzing the blood samples for metabolites. In some cases, it may also be necessary to replace dynamic scanning with a shortened scanning period some time after tracer injection, as is done with FDG (F-18 fluorodeoxyglucose). These approximations represent loss of information. In this work, we considered several questions related to this: (1) Do differences in experimental conditions (drug treatments) or populations affect the input function, and what effect, if any, does this have on the final outcome measure? (2) How do errors in metabolite measurements enter into results? (3) What errors are incurred if the uptake ratio is used in place of the distribution volume ratio? (4) Is one- or two-point blood sampling any better for FDG data than the standardized uptake value? and (5) If blood sampling is necessary, what alternatives are there to arterial blood sampling? The first three questions were considered in terms of data from human dynamic positron emission tomography (PET) studies under conditions of baseline and drug pretreatment. Data from [11C]raclopride studies and those from the norepinephrine transporter tracer (S,S)-[11C]O-methyl reboxetine were used. Calculation of a metabolic rate for FDG using the operational equation requires a measured input function. We tested a procedure based on two blood samples to estimate the plasma integral and convolution that occur in the operational equation. There are some tracers for which blood sampling is necessary. Strategies for brain studies involve using the internal carotids in estimating the radioactivity after correcting for partial volume and spillover in order to eliminate arterial sampling. Some venous blood samples are still required for metabolite measurements. The ultimate solution to the problem of arterial sampling may be a wrist scanner, which acts as a small PET camera for imaging the arteries in the wrist. This is currently under development.

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

PubMed

Aristov, Alexander; Nosova, Ekaterina

2017-04-01

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

Long-Term Bioeffects of 435MHz Radiofrequency Radiation on Selected Blood-Borne Endpoints in Cannulated Rats. Volume 2. Plasma ACTH (adrenocorticotropic Hormone) and Plasma Corticosterone

DTIC Science & Technology

1987-08-01

B.- and Soutas. Dpmptrio; I 13a. TYPE OF REPORT 13b. TIME COVERED 114. DATE OF REPORT (Year, Month, Day) S. PAGE COUNT Final FROM84/8/20 TO 86/2/16...cages and placed in sampling boxes. At this moment a first 0.2 mL sample of blood was withdrawn. At 0 time (20 min later) another 0.2 mL sample of blood...concentration data scatter diagram (exposure group) ............. . . . . . . . . . . . . . . . 18 10. Mean plasma ACTH concentrations versus time ....... 19

Comparison of 4 different types of surgical gloves used for preventing blood contact.

PubMed

Wittmann, Andreas; Kralj, Nenad; Köver, Jan; Gasthaus, Klaus; Lerch, Hartmut; Hofmann, Friedrich

2010-05-01

Needlestick injuries are always associated with a risk of infection, because these types of punctures may expose healthcare workers to a patient's blood and/or body fluids. To compare the efficacy of 4 different types of surgical gloves for preventing exposure to blood as a result of needlestick injury. For simulation of needlestick injury, a circular sample of pork skin was tightened onto a bracket, and a single finger from a medical glove was stretched over the sample. First, a powder-free surgical glove with a gel coating was used to test blood contact. Second, a glove with a patented puncture indication system was used to test blood contact with a double-gloved hand. Third, 2 powder-free latex medical gloves of the same size and hand were combined for double gloving, again to test blood contact. Finally, we tested a glove with an integrated disinfectant on the inside. The punctures were carried out using diverse sharp surgical devices that were contaminated with (99)Tc-marked blood. The amount of blood contact was determined from the transmitted radioactivity. For the powder-free surgical glove with a gel coating, a mean volume of 0.048 microL of blood (standard error of the mean [SEM], 0.077 microL) was transferred in punctures with an automated lancet at a depth of 2.4 mm through 1 layer of latex. For the glove with an integrated disinfectant on the inside, the mean volume of blood transferred was 0.030 microL (SEM, 0.0056 microL) with a single glove and was 0.024 microL (SEM, 0.003 microL) with 2 gloves. For the glove with the patented puncture indication system, a mean volume of 0.024 microL (SEM, 0.003 microL) of blood was transferred. Double gloving or the use of a glove with disinfectant can result in a decrease in the volume of blood transferred. Therefore, the use of either of these gloving systems could help to minimize the risk of bloodborne infections for medical staff.

Comparison of a Point-of-Care Glucometer and a Laboratory Autoanalyzer for Measurement of Blood Glucose Concentrations in Domestic Pigeons ( Columba livia domestica).

PubMed

Mohsenzadeh, Mahdieh Sadat; Zaeemi, Mahdieh; Razmyar, Jamshid; Azizzadeh, Mohammad

2015-09-01

Biochemical analysis is necessary for diagnosis and monitoring of diseases in birds; however, the small volume of blood that can be safely obtained from small avian species often limits laboratory diagnostic testing. Consequently, a suitable methodology requiring only a small volume of blood must be used. This study was designed to compare blood glucose concentrations in domestic pigeons ( Columba livia domestica) as measured by a commercial, handheld, human glucometer and a standard autoanalyzer. During the first phase of the study, whole blood samples obtained from 30 domestic pigeons were used to measure the blood glucose concentration with a glucometer, the packed cell volume (PCV), and the total erythrocyte count (nRBC). Plasma separated from the each sample was then used to obtain the plasma glucose concentration with the autoanalyzer. During the second phase of the study, 30 pigeons were assigned to 2 equal groups (n = 15). Hypoglycemia or hyperglycemia was induced in each group by intravenous injection of insulin or glucose, respectively. Blood was collected and processed, and glucose concentrations, PCV, and nRBC were measured as previously described. Linear-regression models demonstrated a significant relationship between results measured by the glucometer and autoanalyzer results from normoglycemic (correlation coefficient [R] = 0.43, P = .02), hypoglycemic (R = 0.95; P < .001), and hyperglycemic (R = 0.81; P < .001) birds. The results of this study suggest that we can predict the real blood-glucose concentration of pigeons by using results obtained by a glucometer.

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

Kinetic Modeling of PET Data Without Blood Sampling

NASA Astrophysics Data System (ADS)

Bentourkia, M.

2005-06-01

In positron emission tomography (PET) imaging, application of kinetic modeling always requires an input curve (IC) together with the PET data. The IC can be obtained by means of external blood sampling or, in the case of cardiac studies, by means of a region-of-interest (ROI) drawn on the blood pool. It is, however, very unsuitable to withdraw and to analyze blood samples, and in small animals, these operations become difficult, while ICs determined from ROIs are generally contaminated by emissions from neighboring sites, or they are underestimated because of partial volume effect. In this paper, we report a new method to extract kinetic parameters from dynamic PET studies without a priori knowledge of the IC. The method is applied in human brain data measured with fluorodeoxyglucose (FDG) human-brain and in cardiac-rat perfusion studies with /sup 13/N-ammonia and /sup 11/C-acetate. The tissue blood volume (TBV), usually fitted together with the rate constants, is extracted simultaneously with the tissue time activity curves for cardiac studies, while for brain gray matter, TBV is known to be about 4% to 7%. The shape of IC is obtained by means of factor analysis from an ROI drawn around a cardiac tissue or a brain artery. The results show a good correlation (p<0.05) between the cerebral metabolic rate of glucose, myocardial blood flow, and oxygen consumption obtained with the new method in comparison to the usual method. In conclusion, it is possible to apply kinetic modeling without any blood sampling, which significantly simplifies PET acquisition and data analysis.

Diagnosis of neonatal group B Streptococcus sepsis by nested-PCR of residual urine samples

PubMed Central

Cezarino, Bruno Nicolino; Yamamoto, Lidia; Del Negro, Gilda Maria Barbaro; Rocha, Daisy; Okay, Thelma Suely

2008-01-01

Group B streptococcus (GBS) remains the most common cause of early-onset sepsis in newborns. Laboratory gold-standard, broth culture methods are highly specific, but lack sensitivity. The aim of this study was to validate a nested-PCR and to determine whether residue volumes of urine samples obtained by non invasive, non sterile methods could be used to confirm neonatal GBS sepsis. The nested-PCR was performed with primers of the major GBS surface antigen. Unavailability of biological samples to perform life supporting exams, as well as others to elucidate the etiology of infections is a frequent problem concerning newborn patients. Nevertheless, we decided to include cases according to strict criteria: newborns had to present with signs and symptoms compatible with GBS infection; at least one of the following biological samples had to be sent for culture: blood, urine, or cerebrospinal fluid; availability of residue volumes of the samples sent for cultures, or of others collected on the day of hospitalization, prior to antibiotic therapy prescription, to be analyzed by PCR; favorable outcome after GBS empiric treatment. In only one newborn GBS infection was confirmed by cultures, while infection was only presumptive in the other three patients (they fulfilled inclusion criteria but were GBS-culture negative). From a total of 12 biological samples (5 blood, 3 CSF and 4 urine specimen), eight were tested by culture methods (2/8 were positive), and 8 were tested by PCR (7/8 were positive), and only 4 samples were simultaneously tested by both methods (1 positive by culture and 3 by PCR). In conclusion, although based on a restricted number of neonates and samples, our results suggest that the proposed nested-PCR might be used to diagnose GBS sepsis as it has successfully amplified the three types of biological samples analyzed (blood, urine and cerebrospinal fluid), and was more sensitive than culture methods as PCR in urine confirmed diagnosis in all four patients. Moreover, PCR has enabled us to use residue volumes of urine samples collected by non invasive, non sterile methods, what is technically adequate as GBS is not part of the normal urine flora, thus avoiding invasive procedures such as suprapubic bladder punction or transurethral catheterization. At the same time, the use of urine instead of blood samples could help preventing newborns blood spoliation. PMID:24031170

Reducing the risk of fatal and disabling hypoglycaemia: a comparison of arterial blood sampling systems.

PubMed

Brennan, K A; Eapen, G; Turnbull, D

2010-04-01

In 2008, the National Patient Safety Agency (NPSA) published a report after 42 incidents and two deaths where glucose-containing flush solutions were attached to the arterial line. The molar concentration of 5% glucose is 277 mmol litre(-1). Only a tiny amount of sample contamination will lead to an artificially high glucose. As the NPSA sought a solution, a bench model was constructed to compare the performance of three open and three closed arterial line systems in limiting sample contamination. All arterial line systems were set up in a standard manner and pressurized to 300 mm Hg with 5% glucose used as the flush solution. This was connected to the 'radial artery' using an 18 G needle representing the radial cannula. The radial artery was simulated using a wide-bore extension set with 'blood' flow at 60 ml min(-1). Blood was simulated by the addition of red dye to Hartmann's solution. Increasing multiples of arterial line dead space were aspirated and discarded. Blood samples were then obtained and glucose concentration was measured. Significant glucose contamination (3 mmol litre(-1) +/-3.4) was detected in all open arterial line systems up to an aspiration volume of five times the dead space. No samples from the closed systems recorded glucose concentration >1 mmol litre(-1). Recommended minimal discard volumes are inadequate in the presence of glucose as the flush solution and can lead to high blood glucose readings, inappropriate insulin use, and iatrogenic neuroglycopaenia. Our study demonstrates that the closed-loop arterial sampling system could be the universal solution sought by the NPSA.

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood.

PubMed

Park, Yang-Seok; Sunkara, Vijaya; Kim, Yubin; Lee, Won Seok; Han, Ja-Ryoung; Cho, Yoon-Kyoung

2016-04-16

Enzyme-linked immunosorbent assay (ELISA) is a promising method to detect small amount of proteins in biological samples. The devices providing a platform for reduced sample volume and assay time as well as full automation are required for potential use in point-of-care-diagnostics. Recently, we have demonstrated ultrasensitive detection of serum proteins, C-reactive protein (CRP) and cardiac troponin I (cTnI), utilizing a lab-on-a-disc composed of TiO2 nanofibrous (NF) mats. It showed a large dynamic range with femto molar (fM) detection sensitivity, from a small volume of whole blood in 30 min. The device consists of several components for blood separation, metering, mixing, and washing that are automated for improved sensitivity from low sample volumes. Here, in the video demonstration, we show the experimental protocols and know-how for the fabrication of NFs as well as the disc, their integration and the operation in the following order: processes for preparing TiO2 NF mat; transfer-printing of TiO2 NF mat onto the disc; surface modification for immune-reactions, disc assembly and operation; on-disc detection and representative results for immunoassay. Use of this device enables multiplexed analysis with minimal consumption of samples and reagents. Given the advantages, the device should find use in a wide variety of applications, and prove beneficial in facilitating the analysis of low abundant proteins.

Long-Term Bioeffects of 435-MHz Radiofrequency Radiation on Selected Blood-Borne Endpoints in Cannulated Rats. Volume 4. Plasma Catecholamines.

DTIC Science & Technology

1987-08-01

out. To use each animal as its own control , arterial blood was sampled by means of chronically implanted aortic cannulas 112,13,14]. This simple...APPENDIX B STATISTICAL METHODOLOGY 37 APPENDIX B STATISTICAL METHODOLOGY The balanced design of this experiment (requiring that 25 animals from each...protoccl in that, in numerous cases, samples were collected at odd intervals (invalidating the orthogonality of the design ) and the number of samples’taken

Increased mean platelet volume in patients with idiopathic subjective tinnitus.

PubMed

Sarıkaya, Yasin; Bayraktar, Cem; Karataş, Mehmet; Doğan, Sedat; Olt, Serdar; Kaskalan, Emin; Türkbeyler, İbrahim Halil

2016-11-01

Tinnitus is the perception of sound with no external stimulus and idiopathic subjective tinnitus is the most common type in adults. Mean platelet volume (MPV) alterations were shown in some inflammatory diseases and were evaluated as a clinically useful marker. Our aim was to investigate MPV alterations in idiopathic subjective tinnitus patients. A total of 101 patients and 54 age- and sex-matched healthy control subjects were enrolled in the study. Patients included in the study had complaints of tinnitus for at least 3 months. All patients underwent detailed otolaryngologic examination, blood sampling, pure tone audiometry, magnetic resonance imaging of ear, and vertebrobasilar artery Doppler ultrasonography to make the differential diagnosis of tinnitus. Blood sampling consisted of renal-liver-thyroid function tests, lipid profile, and complete blood count. All tests and examinations except the imaging modalities were also performed for the control group. There were no differences in age and sex distribution of groups. Mean platelet volume values were significantly increased in tinnitus patients when compared with controls (p = 0.001). We think that MPV can be qualified as a useful marker in tinnitus patients.

Automated processing of whole blood samples into microliter aliquots of plasma.

PubMed

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

1988-01-01

A rotor that accepts and automatically processes a bulk aliquot of a single blood sample into multiple aliquots of plasma has been designed and built. The rotor consists of a central processing unit, which includes a disk containing eight precision-bore capillaries. By varying the internal diameters of the capillaries, aliquot volumes ranging 1 to 10 mul can be prepared. In practice, an unmeasured volume of blood is placed in a centre well, and, as the rotor begins to spin, is moved radially into a central annular ring where it is distributed into a series of processing chambers. The rotor is then spun at 3000 rpm for 10 min. When the centrifugal field is removed by slowly decreasing the rotor speed, an aliquot of plasma is withdrawn by capillary action into each of the capillary tubes. The disk containing the eight measured aliquots of plasma is subsequently removed and placed in a modifed rotor for conventional centrifugal analysis. Initial evaluation of the new rotor indicates that it is capable of producing discrete, microliter volumes of plasma with a degree of accuracy and precision approaching that of mechanical pipettes.

[Susceptibility of induced sickle in samples of heterozygous hemoglobin S patients (sickle cell trait) suffering diabetes mellitus type 2].

PubMed

Díaz-Piedra, Pablo; Cervantes-Villagrana, Alberto Rafael; Ramos-Jiménez, Raúl; Presno-Bernal, José Miguel; Cervantes-Villagrana, Rodolfo Daniel

2015-01-01

Hemoglobin S is an abnormal protein that induces morphological changes in erythrocyte in low-oxygen conditions. In Mexico, it is reported that up to 13.7% of the population with mutation in one allele are considered asymptomatic (sickle cell trait). The sickle cell trait and diabetes mellitus are conditions that occur together in more than one million patients worldwide. Both diseases possibly produce microvascular changes in retinopathy and acute chest syndrome. The aim of this study was to evaluate the induction of sickle cells in samples of diabetic patients with sickle cell trait to identify altered red cell parameters. We obtained samples of diabetic patients to determine hemoglobin A1c and S; furthermore, red blood cell biometrics data were analyzed. We found that older men with diabetes were susceptible to generate sickle cells and this correlated with reduced red blood cell count and an increase in media cell volume. In samples of women diabetes, there were no differences. We conclude that samples from patients with sickle cell trait and diabetes can cause sickle cells with high frequency in men, with lower red blood cells count and increased mean corpuscular volume as susceptibility parameters.

Measuring Blood Glucose Concentrations in Photometric Glucometers Requiring Very Small Sample Volumes.

PubMed

Demitri, Nevine; Zoubir, Abdelhak M

2017-01-01

Glucometers present an important self-monitoring tool for diabetes patients and, therefore, must exhibit high accuracy as well as good usability features. Based on an invasive photometric measurement principle that drastically reduces the volume of the blood sample needed from the patient, we present a framework that is capable of dealing with small blood samples, while maintaining the required accuracy. The framework consists of two major parts: 1) image segmentation; and 2) convergence detection. Step 1 is based on iterative mode-seeking methods to estimate the intensity value of the region of interest. We present several variations of these methods and give theoretical proofs of their convergence. Our approach is able to deal with changes in the number and position of clusters without any prior knowledge. Furthermore, we propose a method based on sparse approximation to decrease the computational load, while maintaining accuracy. Step 2 is achieved by employing temporal tracking and prediction, herewith decreasing the measurement time, and, thus, improving usability. Our framework is tested on several real datasets with different characteristics. We show that we are able to estimate the underlying glucose concentration from much smaller blood samples than is currently state of the art with sufficient accuracy according to the most recent ISO standards and reduce measurement time significantly compared to state-of-the-art methods.

Techniques for Nonterminal Blood Sampling in Black-Tailed Prairie Dogs (Cynomys ludovicianus).

PubMed

Head, Valerie; Eshar, David; Nau, Melissa R

2017-03-01

Black-tailed prairie dogs (Cynomys ludovicianus) are used as an animal model for research on gallbladder stones and several infectious diseases. A comprehensive, instructive resource regarding the appropriate techniques for venipuncture and collection of nonterminal blood samples in this species has not yet been published. Blood samples (1 mL or larger) were readily obtained from the jugular vein, femoral vein, or cranial vena cava, whereas peripheral sites, such as the cephalic vein, saphenous vein, and tarsal vein, mainly were useful for obtaining smaller volumes. The detailed and illustrated information presented here can aid clinicians and researchers in performing venipuncture, anesthesia, and handling of this species.

Blood transfer devices for malaria rapid diagnostic tests: evaluation of accuracy, safety and ease of use.

PubMed

Hopkins, Heidi; Oyibo, Wellington; Luchavez, Jennifer; Mationg, Mary Lorraine; Asiimwe, Caroline; Albertini, Audrey; González, Iveth J; Gatton, Michelle L; Bell, David

2011-02-08

Malaria rapid diagnostic tests (RDTs) are increasingly used by remote health personnel with minimal training in laboratory techniques. RDTs must, therefore, be as simple, safe and reliable as possible. Transfer of blood from the patient to the RDT is critical to safety and accuracy, and poses a significant challenge to many users. Blood transfer devices were evaluated for accuracy and precision of volume transferred, safety and ease of use, to identify the most appropriate devices for use with RDTs in routine clinical care. Five devices, a loop, straw-pipette, calibrated pipette, glass capillary tube, and a new inverted cup device, were evaluated in Nigeria, the Philippines and Uganda. The 227 participating health workers used each device to transfer blood from a simulated finger-prick site to filter paper. For each transfer, the number of attempts required to collect and deposit blood and any spilling of blood during transfer were recorded. Perceptions of ease of use and safety of each device were recorded for each participant. Blood volume transferred was calculated from the area of blood spots deposited on filter paper. The overall mean volumes transferred by devices differed significantly from the target volume of 5 microliters (p < 0.001). The inverted cup (4.6 microliters) most closely approximated the target volume. The glass capillary was excluded from volume analysis as the estimation method used is not compatible with this device. The calibrated pipette accounted for the largest proportion of blood exposures (23/225, 10%); exposures ranged from 2% to 6% for the other four devices. The inverted cup was considered easiest to use in blood collection (206/226, 91%); the straw-pipette and calibrated pipette were rated lowest (143/225 [64%] and 135/225 [60%] respectively). Overall, the inverted cup was the most preferred device (72%, 163/227), followed by the loop (61%, 138/227). The performance of blood transfer devices varied in this evaluation of accuracy, blood safety, ease of use, and user preference. The inverted cup design achieved the highest overall performance, while the loop also performed well. These findings have relevance for any point-of-care diagnostics that require blood sampling.

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

PubMed Central

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

2015-01-01

Capillary blood sampling is a medical procedure aimed at assisting in patient diagnosis, management and treatment, and is increasingly used worldwide, in part because of the increasing availability of point-of-care testing. It is also frequently used to obtain small blood volumes for laboratory testing because it minimizes pain. The capillary blood sampling procedure can influence the quality of the sample as well as the accuracy of test results, highlighting the need for immediate, widespread standardization. A recent nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia has shown that capillary sampling procedures are not standardized and that only a small proportion of Croatian laboratories comply with guidelines from the Clinical Laboratory Standards Institute (CLSI) or the World Health Organization (WHO). The aim of this document is to provide recommendations for capillary blood sampling. This document has been produced by the Working Group for Capillary Blood Sampling within the Croatian Society of Medical Biochemistry and Laboratory Medicine. Our recommendations are based on existing available standards and recommendations (WHO Best Practices in Phlebotomy, CLSI GP42-A6 and CLSI C46-A2), which have been modified based on local logistical, cultural, legal and regulatory requirements. We hope that these recommendations will be a useful contribution to the standardization of capillary blood sampling in Croatia. PMID:26524965

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

PubMed

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

2015-01-01

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

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

PubMed

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

2016-10-01

In this study, a fast and simple blood sampling and sample pre-treatment method based on the use of the dried blood spot (DBS) cards and ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for the quantification of olive oil phenolic metabolites in human blood was developed and validated. After validation, the method was applied to determine hydroxytyrosol metabolites in human blood samples after the acute intake of an olive oil phenolic extract. Using the FTA DMPK-A DBS card under optimum conditions, with 20µL as the blood solution volume, 100µL of methanol/Milli-Q water (50/50, v/v) as the extraction solvent and 7 disks punched out from the card, the main hydroxytyrosol metabolites (hydroxytyrosol-3-O-sulphate and hydroxytyrosol acetate sulphate) were identified and quantified. The developed methodology allowed detecting and quantifying the generated metabolites at low μM levels. The proposed method is a significant improvement over existing methods to determine phenolic metabolites circulating in blood and plasma samples, thus making blood sampling possible with the volunteer pricking their own finger, and the subsequent storage of the blood in the DBS cards prior to chromatographic analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

"Center punch" and "whole spot" bioanalysis of apixaban in human dried blood spot samples by UHPLC-MS/MS.

PubMed

Zheng, Naiyu; Yuan, Long; Ji, Qin C; Mangus, Heidi; Song, Yan; Frost, Charles; Zeng, Jianing; Aubry, Anne-Françoise; Arnold, Mark E

2015-04-15

Apixaban (Eliquis™) was developed by Bristol-Myers Squibb (BMS) and Pfizer to use as an antithrombotic/anticoagulant agent and has been recently approved for the prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation. A clinical study of apixaban, sponsored by BMS and Pfizer, included a pilot exploratory portion to evaluate the potential for future drug concentration monitoring using dried blood spot (DBS) sample collection. For DBS sample collection, a fixed blood volume was dispensed onto a DBS card by either regular volumetric pipette (venous blood collection) or capillary dispenser (finger prick blood collection). A 96-well semi-automated liquid-liquid extraction sample preparation procedure was developed to provide clean extracts for UHPLC-MS/MS quantitation. Assays using both partial-spot center punch and whole spot punch were developed and validated. The linear dynamic ranges for all the analyses were from 0.5 to 500 ng/mL. The coefficient of determination (r(2)) values was >0.9944 for all the validation runs. For the center punch approach, the intra-assay precision (%CV) was within 4.4% and inter-assay precision was within 2.6%. The assay accuracy, expressed as %Dev., was within ± 5.4% of the nominal concentrations. One accuracy and precision run was performed using the whole spot approach, the intra-assay precision (%CV) was within 7.1% and the accuracy was within ± 8.0% of the nominal concentrations. In contrast to the center punch approach, the whole spot approach eliminated the effect of hematocrit and high lipids on the analysis of apixaban in human DBS when an accurate sample blood volume was collected on DBS cards. Copyright © 2015 Elsevier B.V. All rights reserved.

Non-terminal blood sampling techniques in guinea pigs.

PubMed

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

2014-10-11

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

21 CFR 606.122 - Instruction circular.

Code of Federal Regulations, 2012 CFR

2012-04-01

...: (1) Instructions to administer a suitable plasma volume expander if Red Blood Cells are substituted... approximate volume of plasma from which a sample unit of Platelets is prepared. (2) Instructions to begin administration as soon as possible, but not more than 4 hours after entering the container. (m) For Plasma, the...

Study of Umbilical Cord Blood Culture in Diagnosis of Early-onset Sepsis Among Newborns with High-risk Factors.

PubMed

Kalathia, Mitul Babubhai; Shingala, Prakash Ashokbhai; Parmar, Parin Niranjanbhai; Parikh, Yogesh Narenedrabhai; Kalathia, Ila Mitulkumar

2013-10-01

Blood culture is gold standard for diagnosis of neonatal sepsis. Low sensitivity of blood culture is usually due to small volume of blood sample, intrapartum antibiotics, and antibiotics given to newborn before sampling. We evaluated use of Umbilical cord blood culture (UCBC) in diagnosis of neonatal sepsis as compared to peripheral venous blood culture. This study was done in tertiary care teaching hospital during May-June 2012. A total of 45 newborns with presence of two or more risk factors of sepsis were included. Blood sample from placental end of umbilical cord was collected and cultured. Primary outcome was diagnosis of neonatal sepsis by use of umbilical cord blood sample as compared with venous blood sample. Secondary outcome was to compare organisms identified by UCBC and venous blood culture. Sensitivity, specificity, positive and negative predictive values of UCBC were calculated. A total of 24.44% (11 out of 45) high-risk newborns had positive UCBC. A total of 17.8% (8 out of 45) newborns had positive blood culture report. Organisms grown in UCBC were Pseudomonas (45%, 5 out of 11), Acinetobacter (27.27%, 3 out of 11), Escherichia coli (18.18%, 2 out of 11), and Klebsiella (9%, 1 out of 11). UCBC is a good method for diagnosis of neonatal sepsis among high-risk newborns as compared to venous blood culture with a sensitivity of 80% and specificity of 91.43%. Organisms grown are comparable to blood culture samples.

Customization of Advia 120 thresholds for canine erythrocyte volume and hemoglobin concentration, and effects on morphology flagging results.

PubMed

Grimes, Carolyn N; Fry, Michael M

2014-12-01

This study sought to develop customized morphology flagging thresholds for canine erythrocyte volume and hemoglobin concentration [Hgb] on the ADVIA 120 hematology analyzer; compare automated morphology flagging with results of microscopic blood smear evaluation; and examine effects of customized thresholds on morphology flagging results. Customized thresholds were determined using data from 52 clinically healthy dogs. Blood smear evaluation and automated morphology flagging results were correlated with mean cell volume (MCV) and cellular hemoglobin concentration mean (CHCM) in 26 dogs. Customized thresholds were applied retroactively to complete blood (cell) count (CBC) data from 5 groups of dogs, including a reference sample group, clinical cases, and animals with experimentally induced iron deficiency anemia. Automated morphology flagging correlated more highly with MCV or CHCM than did blood smear evaluation; correlation with MCV was highest using customized thresholds. Customized morphology flagging thresholds resulted in more sensitive detection of microcytosis, macrocytosis, and hypochromasia than default thresholds.

IMPROVED DERIVATION OF INPUT FUNCTION IN DYNAMIC MOUSE [18F]FDG PET USING BLADDER RADIOACTIVITY KINETICS

PubMed Central

Wong, Koon-Pong; Zhang, Xiaoli; Huang, Sung-Cheng

2013-01-01

Purpose Accurate determination of the plasma input function (IF) is essential for absolute quantification of physiological parameters in positron emission tomography (PET). However, it requires an invasive and tedious procedure of arterial blood sampling that is challenging in mice because of the limited blood volume. In this study, a hybrid modeling approach is proposed to estimate the plasma IF of 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) in mice using accumulated radioactivity in urinary bladder together with a single late-time blood sample measurement. Methods Dynamic PET scans were performed on nine isoflurane-anesthetized male C57BL/6 mice after a bolus injection of [18F]FDG at the lateral caudal vein. During a 60- or 90-min scan, serial blood samples were taken from the femoral artery. Image data were reconstructed using filtered backprojection with CT-based attenuation correction. Total accumulated radioactivity in the urinary bladder was fitted to a renal compartmental model with the last blood sample and a 1-exponential function that described the [18F]FDG clearance in blood. Multiple late-time blood sample estimates were calculated by the blood [18F]FDG clearance equation. A sum of 4-exponentials was assumed for the plasma IF that served as a forcing function to all tissues. The estimated plasma IF was obtained by simultaneously fitting the [18F]FDG model to the time-activity curves (TACs) of liver and muscle and the forcing function to early (0–1 min) left-ventricle data (corrected for delay, dispersion, partial-volume effects and erythrocytes uptake) and the late-time blood estimates. Using only the blood sample acquired at the end of the study to estimate the IF and the use of liver TAC as an alternative IF were also investigated. Results The area under the plasma TACs calculated for all studies using the hybrid approach was not significantly different from that using all blood samples. [18F]FDG uptake constants in brain, myocardium, skeletal muscle and liver computed by the Patlak analysis using estimated and measured plasma TACs were in excellent agreement (slope ~ 1; R2 > 0.938). The IF estimated using only the last blood sample acquired at the end of the study and the use of liver TAC as plasma IF provided less reliable results. Conclusions The estimated plasma IFs obtained with the hybrid model agreed well with those derived from arterial blood sampling. Importantly, the proposed method obviates the need of arterial catheterization, making it possible to perform repeated dynamic [18F]FDG PET studies on the same animal. Liver TAC is unsuitable as an input function for absolute quantification of [18F]FDG PET data. PMID:23322346

Capillary blood sampling as an alternative to venipuncture in the assessment of serum 25 hydroxyvitamin D levels.

PubMed

Dayre McNally, J; Matheson, Loren A; Sankaran, Koravangattu; Rosenberg, Alan M

2008-11-01

This study compared 25-hydroxyvitamin D [25(OH)D] measurements in capillary and venous blood samples collected, respectively by fingerprick and venipuncture. Capillary blood for measuring 25(OH)D has potential advantages by reducing blood volume required (2mL versus 0.3mL for venipuncture and capillary sampling, respectively), facilitating blood collection for those populations in whom venipuncture is difficult (e.g. infants and children), improving patient convenience and reducing costs associated with phlebotomy. The results demonstrated a highly significant relationship between 25(OH)D levels in serum derived from venous and capillary blood samples (r(2)=0.901). Despite statistically higher 25(OH)D levels in fingerprick samples (108+/-9nmol/L) compared with venipuncture samples (90+/-7nmol/L), the correlation between venous and capillary samples provides support for this approach as a practical alternative to venipuncture for vitamin D determination. However, clinical application may require the incorporation of a correction factor for the assessment of insufficiency, and research studies should avoid using the two methods interchangeably. Studying vitamin D's role in health and disease requires collection techniques and measurement methods that are reliable, reproducible, easily accessible, inexpensive and minimally burdensome to the patient. The option to collect patient samples by fingerprick may facilitate the collection process.

Technical considerations and precautions in in situ bronchoalveolar lavage and alveolar infiltrating cells isolation in rats.

PubMed

Qamar, Wajhul

2015-01-01

Bronchoalveolar lavage fluid (BALF) is an important tool in experimental toxicology and pharmacology. A number of researchers have utilized BAL in studies involving rodents. However, the detailed procedure of performing BAL in rodents has rarely been reported. In the present article, in situ BAL in rats has been described with technical points that ensure the quality and validity of BALF samples. BAL was performed in rats of Wistar strain. Flow cytometry and microscopy were utilized to analyze the BAL cells. The collected volume of BALF was 84.32 ± 2.7% of instilled volume. Alveolar macrophages were found to be prevalent in normal lungs when analyzed under microscope and by flow cytometry. The main problem that one may face is the accidental contamination of BALF samples with blood of the animal itself. Presence of blood in BALF certainly adds a significant number of cells and other biochemical variables. No blood contamination was detected in BALF. Here, a simple procedure for BAL and collection of alveolar cells (macrophages in the present study) is explained with an emphasis on technical steps and precautions, which ensures the quality of the BALF samples. Data exhibit that there is no blood contamination in the BALF and provide evidence that the technical points considered in the procedure here are successful in maintaining the quality and validity of BALF samples.

Acute effects of 30 minutes of exposure to a smartphone call on in vitro platelet function

PubMed Central

Lippi, Giuseppe; Danese, Elisa; Brocco, Giorgio; Gelati, Matteo; Salvagno, Gian Luca; Montagnana, Martina

2017-01-01

Background Significant concerns are now regularly raised about the safety of excessive mobile phone use. This study was aimed to assess the acute effects of radiofrequency waves emitted by a commercial smartphone on platelet function. Materials and methods Two sequential citrated blood samples were collected from 16 healthy volunteers recruited from laboratory staff. The first sample was placed in a plastic rack, 1 cm distant from a commercial smartphone receiving a 30-min call and emitting 900 MHz radiofrequency waves. The second sample was placed in another plastic rack, isolated from radiofrequency wave sources, for the same period. The platelet count and the mean platelet volume were then assessed in all blood samples, whereas platelet function was evaluated using the platelet function analyser-100 (PFA-100). Results A 30-min exposure of citrated blood to smartphone radiofrequency waves induced significant prolongation of collagen-epinephrine aggregation (median increase, 10%) and a considerable increase of mean platelet volume (median increase, 5%), whereas collagen-adenosine diphosphate aggregation and platelet count remained unchanged. Discussion This study demonstrates that smartphone radiofrequency waves induce significant perturbation of platelet structure and function, thus providing further support to concerns regarding excessive use of mobile phones. Caution should also be taken with regards to blood products containing platelets, which should be kept far away from mobile phones and smartphones throughout the production pipeline and storage period. PMID:27177410

Acute effects of 30 minutes of exposure to a smartphone call on in vitro platelet function.

PubMed

Lippi, Giuseppe; Danese, Elisa; Brocco, Giorgio; Gelati, Matteo; Salvagno, Gian Luca; Montagnana, Martina

2017-05-01

Significant concerns are now regularly raised about the safety of excessive mobile phone use. This study was aimed to assess the acute effects of radiofrequency waves emitted by a commercial smartphone on platelet function. Two sequential citrated blood samples were collected from 16 healthy volunteers recruited from laboratory staff. The first sample was placed in a plastic rack, 1 cm distant from a commercial smartphone receiving a 30-min call and emitting 900 MHz radiofrequency waves. The second sample was placed in another plastic rack, isolated from radiofrequency wave sources, for the same period. The platelet count and the mean platelet volume were then assessed in all blood samples, whereas platelet function was evaluated using the platelet function analyser-100 (PFA-100). A 30-min exposure of citrated blood to smartphone radiofrequency waves induced significant prolongation of collagen-epinephrine aggregation (median increase, 10%) and a considerable increase of mean platelet volume (median increase, 5%), whereas collagen-adenosine diphosphate aggregation and platelet count remained unchanged. This study demonstrates that smartphone radiofrequency waves induce significant perturbation of platelet structure and function, thus providing further support to concerns regarding excessive use of mobile phones. Caution should also be taken with regards to blood products containing platelets, which should be kept far away from mobile phones and smartphones throughout the production pipeline and storage period.

A method for the direct injection and analysis of small volume human blood spots and plasma extracts containing high concentrations of organic solvents using revered-phase 2D UPLC/MS.

PubMed

Rainville, Paul D; Simeone, Jennifer L; Root, Dan S; Mallet, Claude R; Wilson, Ian D; Plumb, Robert S

2015-03-21

The emergence of micro sampling techniques holds great potential to improve pharmacokinetic data quality, reduce animal usage, and save costs in safety assessment studies. The analysis of these samples presents new challenges for bioanalytical scientists, both in terms of sample processing and analytical sensitivity. The use of two dimensional LC/MS with, at-column-dilution for the direct analysis of highly organic extracts prepared from biological fluids such as dried blood spots and plasma is demonstrated. This technique negated the need to dry down and reconstitute, or dilute samples with water/aqueous buffer solutions, prior to injection onto a reversed-phase LC system. A mixture of model drugs, including bromhexine, triprolidine, enrofloxacin, and procaine were used to test the feasibility of the method. Finally an LC/MS assay for the probe pharmaceutical rosuvastatin was developed from dried blood spots and protein-precipitated plasma. The assays showed acceptable recovery, accuracy and precision according to US FDA guidelines. The resulting analytical method showed an increase in assay sensitivity of up to forty fold as compared to conventional methods by maximizing the amount loaded onto the system and the MS response for the probe pharmaceutical rosuvastatin from small volume samples.

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

PubMed

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

2016-10-01

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

Hormonal and electrolyte responses to acute isohemic volume expansion in unanesthetized rats

NASA Technical Reports Server (NTRS)

Chenault, V. M.; Morris, M.; Lynch, C. D.; Maultsby, S. J.; Hutchins, P. M.

1993-01-01

This study was undertaken to explore the time course of the metabolic response to isohemic blood volume expansion (30%) in normotensive, unanesthetized Sprague-Dawley rats. Whole blood, drawn from a femoral artery catheter of conscious donor rats, was infused into the jugular vein of recipient rats. Blood samples were drawn from a carotid artery of recipient rats at time points beginning immediately post-volume expansion (IPVE) up through 5 days post-volume expansion (PVE). To characterize the attendant compensatory mechanisms, the plasma concentrations of electrolytes and fluid regulatory hormones were determined. Hematocrit began to raise IPVE and was significantly elevated above control IPVE 20, 30, 40, 60, and 90 min, and 2, 4, 6, 8, 12, and 24 hr PVE. Consistent with our current understanding of the hormonal response to excess volume, atrial natriuretic factor was significantly increased above the prevolume expansion (control) values 0-30 min PVE. Surprisingly, plasma aldosterone levels were significantly increased above control at 20 and 30 min and 6 hr PVE, whereas plasma renin activity was significantly decreased 30-40 min PVE. Plasma sodium was not changed from control values except for a significant increase at 6 hr post-volume expansion. Plasma potassium, osmolality, and arginine vasopressin levels were not altered by the volume expansion. These studies delineate the physiologic time scheme operative in the regulation of fluid volume during acute ischemic volume expansion.

Comparison of human umbilical cord blood processing with or without hydroxyethyl starch.

PubMed

Souri, Milad; Nikougoftar Zarif, Mahin; Rasouli, Mahboobeh; Golzadeh, Khadijeh; Nakhlestani Hagh, Mozhdeh; Ezzati, Nasim; Atarodi, Kamran

2017-11-01

Umbilical cord blood (UCB) processing with hydroxyethyl starch (HES) is the most common protocol in the cord blood banks. The quality of UCB volume reduction was guaranteed by minimum manipulation of cord blood samples in the closed system. This study aimed to analyze and compare cell recovery and viability of UCB processed using the Sepax automated system in the presence and absence of HES. Thirty UCB bags with a total nucleated cell (TNC) count of more than 2.5 × 10 9 were divided in two bags with equal volume. HES solution was added to one bag and another was intact. Both bags were processed with the Sepax. To determine cell recovery, viability, and potential of colony-forming cells (CFCs), preprocessing, postprocessing, and thawing samples were analyzed. The mean TNC recovery after processing and after thaw was significantly better with the HES method (p < 0.01 for the postprocessing step and p < 0.05 for the postthaw step). There were no significant differences to mononucleated cells (MNCs) and CD34+ cell recovery between the two methods after processing and after thaw. TNC and MNC viability was significantly higher without HES after processing and after thaw (p < 0.01). The results of the CFC assay were similar for both methods after processing and after thaw. These results showed that processing of UCB using the Sepax system with the without-HES protocol due to the lower manipulation of samples could be used as an eligible protocol to reduce the volume of UCB. © 2017 AABB.

Effect of a commercial anion dietary supplement on acid-base balance, urine volume, and urinary ion excretion in male goats fed oat or grass hay diets.

PubMed

Stratton-Phelps, Meri; House, John K

2004-10-01

To determine whether feeding a commercial anionic dietary supplement as a urinary acidifier to male goats may be useful for management of urolithiasis. 8 adult sexually intact male Toggenburg, Saanen, and Nubian goats. Goats were randomly assigned by age-, breed-, and weight-matched pairs to an oat or grass hay diet that was fed for 12 days. On days 13 to 14 (early sample collection time before supplementation), measurements were made of blood and urine sodium, potassium, calcium, magnesium, chloride, phosphorus, and sulfur concentrations; blood and urine pH; urine production; and water consumption. During the next 28 days, the anionic dietary supplement was added to the oat and grass hay diets to achieve a dietary cation-anion difference of 0 mEq/100g of dry matter. Blood and urine samples were analyzed during dietary supplementation on days 12 to 13 (middle sample collection time) and 27 to 28 (late sample collection time). Blood bicarbonate, pH, and urine pH of goats fed grass hay and goats fed oat hay were significantly decreased during the middle and late sample collection times, compared with the early sample collection time. Water consumption and urine production in all goats increased significantly during the late sample collection time, compared with the early sample collection time. The anionic dietary supplement used in our study increases urine volume, alters urine ion concentrations, and is an efficacious urinary acidifier in goats. Goats treated with prolonged anionic dietary supplementation should be monitored for secondary osteoporosis from chronic urinary calcium loss.

Evaluation of two portable meters for determination of blood triglyceride concentration in dogs.

PubMed

Kluger, Elissa K; Dhand, Navneet K; Malik, Richard; Ilkin, William J; Snow, David H; Govendir, Merran

2010-02-01

To evaluate agreement between 2 portable triglyceride meters and a veterinary laboratory for measurement of blood triglyceride concentrations in dogs and evaluate effects of Hct and blood volume analyzed. 97 blood samples collected from 60 dogs. Triglyceride concentrations were measured in blood by use of 2 meters and compared with serum triglyceride concentrations determined by a veterinary laboratory. Within- and between-day precision, accuracy, and effects of blood volume and Hct were analyzed. Accuracy of both meters varied with triglyceride concentration, although both accurately delineated dogs with triglyceride concentrations < 180 mg/dL versus > or = 180 mg/dL. One meter had results with excellent overall correlation with results of the standard laboratory method, with a concordance correlation coefficient of 0.94 and mean difference of 20.3 mg/dL. The other meter had a good overall concordance correlation coefficient of 0.86 with a higher absolute mean difference of -27.7 mg/dL. Results were only affected by blood volume; triglyceride concentrations determined via both meters were significantly lower when 7 microL of EDTA-anticoagulated blood was used, compared with larger volumes. 1 meter had greater accuracy in the range of 140 to 400 mg/dL and was therefore well suited to detect hypertriglyceridemia. The other meter was accurate with triglyceride values < 140 mg/dL and yielded results similar to those of the veterinary laboratory in the range of 140 to 400 mg/dL, therefore being suitable for determination of triglyceride concentrations in nonfed dogs and dogs with mildly high concentrations.

Dried blood spot analysis of an iron chelator--deferasirox and its potential application to therapeutic drug monitoring.

PubMed

Nirogi, Ramakrishna; Ajjala, Devender Reddy; Kandikere, Vishwottam; Aleti, Raghupathi; Srikakolapu, SuryaRao; Vurimindi, Himabindu

2012-10-15

Deferasirox is an iron chelating agent for the treatment of transfusional iron over load in patients with chronic anemia. These anemic patients require close monitoring of the deferasirox exposures for ensuring its therapeutic efficacy. Dried blood spot (DBS) sampling methodology has the advantages of low volume of blood withdrawal and ease of transportation and storage over liquid blood methods. A LC-MS/MS based analytical method was developed using reversed phase column with gradient elution program and quantitated in MRM mode. Linearity range for the liquid blood was 1-1000 ng/mL and for DBS was 5-5000 ng/mL under similar mass spectrometry conditions. The method was validated with respective (M-H)(-) ions, m/z 372→118 for deferasirox and m/z 410→348 for fluvastatin (internal standard). The validated method was applied for the analysis of DBS samples from a rat pharmacokinetic study and results were compared against liquid blood samples from the same animal. The mean C(max) from DBS sample (1121 ng/mL) was comparable to mean C(max) found in blood samples (1015 ng/mL) at 2h after oral dose of deferasirox. All the other calculated pharmacokinetic parameters were quite comparable for both liquid blood and DBS samples. Copyright © 2012. Published by Elsevier B.V.

Quantification of 18F-fluorocholine kinetics in patients with prostate cancer.

PubMed

Verwer, Eline E; Oprea-Lager, Daniela E; van den Eertwegh, Alfons J M; van Moorselaar, Reindert J A; Windhorst, Albert D; Schwarte, Lothar A; Hendrikse, N Harry; Schuit, Robert C; Hoekstra, Otto S; Lammertsma, Adriaan A; Boellaard, Ronald

2015-03-01

Choline kinase is upregulated in prostate cancer, resulting in increased (18)F-fluoromethylcholine uptake. This study used pharmacokinetic modeling to validate the use of simplified methods for quantification of (18)F-fluoromethylcholine uptake in a routine clinical setting. Forty-minute dynamic PET/CT scans were acquired after injection of 204 ± 9 MBq of (18)F-fluoromethylcholine, from 8 patients with histologically proven metastasized prostate cancer. Plasma input functions were obtained using continuous arterial blood-sampling as well as using image-derived methods. Manual arterial blood samples were used for calibration and correction for plasma-to-blood ratio and metabolites. Time-activity curves were derived from volumes of interest in all visually detectable lymph node metastases. (18)F-fluoromethylcholine kinetics were studied by nonlinear regression fitting of several single- and 2-tissue plasma input models to the time-activity curves. Model selection was based on the Akaike information criterion and measures of robustness. In addition, the performance of several simplified methods, such as standardized uptake value (SUV), was assessed. Best fits were obtained using an irreversible compartment model with blood volume parameter. Parent fractions were 0.12 ± 0.4 after 20 min, necessitating individual metabolite corrections. Correspondence between venous and arterial parent fractions was low as determined by the intraclass correlation coefficient (0.61). Results for image-derived input functions that were obtained from volumes of interest in blood-pool structures distant from tissues of high (18)F-fluoromethylcholine uptake yielded good correlation to those for the blood-sampling input functions (R(2) = 0.83). SUV showed poor correlation to parameters derived from full quantitative kinetic analysis (R(2) < 0.34). In contrast, lesion activity concentration normalized to the integral of the blood activity concentration over time (SUVAUC) showed good correlation (R(2) = 0.92 for metabolite-corrected plasma; 0.65 for whole-blood activity concentrations). SUV cannot be used to quantify (18)F-fluoromethylcholine uptake. A clinical compromise could be SUVAUC derived from 2 consecutive static PET scans, one centered on a large blood-pool structure during 0-30 min after injection to obtain the blood activity concentrations and the other a whole-body scan at 30 min after injection to obtain lymph node activity concentrations. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

Bacterial and fungal DNA extraction from blood samples: automated protocols.

PubMed

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

2015-01-01

Automation in DNA isolation is a necessity for routine practice employing molecular diagnosis of infectious agents. To this end, the development of automated systems for the molecular diagnosis of microorganisms directly in blood samples is at its beginning. Important characteristics of systems demanded for routine use include high recovery of microbial DNA, DNA-free containment for the reduction of DNA contamination from exogenous sources, DNA-free reagents and consumables, ideally a walkaway system, and economical pricing of the equipment and consumables. Such full automation of DNA extraction evaluated and in use for sepsis diagnostics is yet not available. Here, we present protocols for the semiautomated isolation of microbial DNA from blood culture and low- and high-volume blood samples. The protocols include a manual pretreatment step followed by automated extraction and purification of microbial DNA.

21 CFR 862.1130 - Blood volume test system.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Blood volume test system. 862.1130 Section 862....1130 Blood volume test system. (a) Identification. A blood volume test system is a device intended to measure the circulating blood volume. Blood volume measurements are used in the diagnosis and treatment of...

Menstrual blood loss measurement: validation of the alkaline hematin technique for feminine hygiene products containing superabsorbent polymers.

PubMed

Magnay, Julia L; Nevatte, Tracy M; Dhingra, Vandana; O'Brien, Shaughn

2010-12-01

To validate the alkaline hematin technique for measurement of menstrual blood loss using ultra-thin sanitary towels that contain superabsorbent polymer granules as the absorptive agent. Laboratory study using simulated menstrual fluid (SMF) and Always Ultra Normal, Long, and Night "with wings" sanitary towels. Keele Menstrual Disorders Laboratory. None. None. Recovery of blood, linearity, and interassay variation over a range of SMF volumes applied to towels. Because of the variable percentage of blood in menstrual fluid, blood recovery was assessed from SMF constituted as 10%, 25%, 50%, and 100% blood. The lower limit of reliable detection and the effect of storing soiled towels for up to 4 weeks at 15°C-20°C, 4°C, and -20°C before analysis were determined. Ninety percent recovery was reproducibly achieved up to 30 mL applied volume at all tested SMF compositions, except at low volume or high dilution equivalent to <2 mL whole blood. Samples could be stored for 3 weeks at all tested temperatures without loss of recovery. The technique was suitable for processing towels individually or in batches. The alkaline hematin technique is a suitable and validated method for measuring menstrual blood loss from Always Ultra sanitary towels that contain superabsorbent polymers. Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Effect of increasing the sampling interval to 2 seconds on the radiation dose and accuracy of CT perfusion of the head and neck.

PubMed

Tawfik, Ahmed M; Razek, Ahmed A; Elhawary, Galal; Batouty, Nihal M

2014-01-01

To evaluate the effect of increasing the sampling interval from 1 second (1 image per second) to 2 seconds (1 image every 2 seconds) on computed tomographic (CT) perfusion (CTP) of head and neck tumors. Twenty patients underwent CTP studies of head and neck tumors with images acquired in cine mode for 50 seconds using sampling interval of 1 second. Using deconvolution-based software, analysis of CTP was done with sampling interval of 1 second and then 2 seconds. Perfusion maps representing blood flow, blood volume, mean transit time, and permeability surface area product (PS) were obtained. Quantitative tumor CTP values were compared between the 2 sampling intervals. Two blinded radiologists compared the subjective quality of CTP maps using a 3-point scale between the 2 sampling intervals. Radiation dose parameters were recorded for the 2 sampling interval rates. No significant differences were observed between the means of the 4 perfusion parameters generated using both sampling intervals; all P >0.05. The 95% limits of agreement between the 2 sampling intervals were -65.9 to 48.1) mL/min per 100 g for blood flow, -3.6 to 3.1 mL/100 g for blood volume, -2.9 to 3.8 seconds for mean transit time, and -10.0 to 12.5 mL/min per 100 g for PS. There was no significant difference between the subjective quality scores of CTP maps obtained using the 2 sampling intervals; all P > 0.05. Radiation dose was halved when sampling interval increased from 1 to 2 seconds. Increasing the sampling interval rate to 1 image every 2 seconds does not compromise the image quality and has no significant effect on quantitative perfusion parameters of head and neck tumors. The radiation dose is halved.

Clinical and anatomic pathology effects of serial blood sampling in rat toxicology studies, using conventional or microsampling methods.

PubMed

Caron, Alexis; Lelong, Christine; Bartels, T; Dorchies, O; Gury, T; Chalier, Catherine; Benning, Véronique

2015-08-01

As a general practice in rodent toxicology studies, satellite animals are used for toxicokinetic determinations, because of the potential impact of serial blood sampling on toxicological endpoints. Besides toxicological and toxicokinetic determinations, blood samples obtained longitudinally from a same animal may be used for the assessment of additional parameters (e.g., metabolism, pharmacodynamics, safety biomarkers) to maximize information that can be deduced from rodents. We investigated whether removal of up to 6 × 200 μL of blood over 24h can be applied in GLP rat toxicology studies without affecting the scientific outcome. 8 week-old female rats (200-300 g) were dosed for up to 1 month with a standard vehicle and subjected or not (controls) to serial blood sampling for sham toxicokinetic/ancillary determinations, using miniaturized methods allowing collection of 6 × 50, 100 or 200 μL over 24h. In-life endpoints, clinical pathology parameters and histopathology of organs sensitive to blood volume reduction were evaluated at several time points after completion of sampling. In sampled rats, minimal and reversible changes in red blood cell mass (maximally 15%) and subtle variations in liver enzymes, fibrinogen and neutrophils were not associated with any organ/tissue macroscopic or microscopic correlate. Serial blood sampling (up to 6 × 200 μL over 24h) is compatible with the assessment of standard toxicity endpoints in adult rats. Copyright © 2015 Elsevier Inc. All rights reserved.

Specimen rejection in laboratory medicine: Necessary for patient safety?

PubMed

Dikmen, Zeliha Gunnur; Pinar, Asli; Akbiyik, Filiz

2015-01-01

The emergency laboratory in Hacettepe University Hospitals receives specimens from emergency departments (EDs), inpatient services and intensive care units (ICUs). The samples are accepted according to the rejection criteria of the laboratory. In this study, we aimed to evaluate the sample rejection ratios according to the types of pre-preanalytical errors and collection areas. The samples sent to the emergency laboratory were recorded during 12 months between January to December, 2013 in which 453,171 samples were received and 27,067 specimens were rejected. Rejection ratios was 2.5% for biochemistry tests, 3.2% for complete blood count (CBC), 9.8% for blood gases, 9.2% for urine analysis, 13.3% for coagulation tests, 12.8% for therapeutic drug monitoring, 3.5% for cardiac markers and 12% for hormone tests. The most frequent rejection reasons were fibrin clots (28%) and inadequate volume (9%) for biochemical tests. Clotted samples (35%) and inadequate volume (13%) were the major causes for coagulation tests, blood gas analyses and CBC. The ratio of rejected specimens was higher in the EDs (40%) compared to ICUs (30%) and inpatient services (28%). The highest rejection ratio was observed in neurology ICU (14%) among the ICUs and internal medicine inpatient service (10%) within inpatient clinics. We detected an overall specimen rejection rate of 6% in emergency laboratory. By documentation of rejected samples and periodic training of healthcare personnel, we expect to decrease sample rejection ratios below 2%, improve total quality management of the emergency laboratory and promote patient safety.

Circulating blood volume determination using electronic spin resonance spectroscopy.

PubMed

Facorro, Graciela; Bianchin, Ana; Boccio, José; Hager, Alfredo

2006-09-01

There have been numerous methods proposed to measure the circulating blood volume (CBV). Nevertheless, none of them have been massively and routinely accepted in clinical diagnosis. This study describes a simple and rapid method, on a rabbit model, using the dilution of autologous red cells labeled with a nitroxide radical (Iodoacetamide-TEMPO), which can be detected by electronic spin resonance (ESR) spectroscopy. Blood samples were withdrawn and re-injected using the ears' marginal veins. The average CBV measured by the new method/body weight (CBV(IAT)/BW) was 59 +/- 7 mL/kg (n = 33). Simultaneously, blood volume determinations using the nitroxide radical and (51)Cr (CBV(Cr)) were performed. In the plot of the difference between the methods (CBV(IAT) - CBV(Cr)) against the average (CBV(IAT) + CBV(Cr))/2, the mean of the bias was -1.1 +/- 6.9 mL and the limits of agreement (mean difference +/-2 SD) were -14.9 and 12.7 mL. Lin's concordance correlation coefficient p(c) = 0.988. Thus, both methods are in close agreement. The development of a new method that allows a correct estimation of the CBV without using radioactivity, avoiding blood manipulation, and decreasing the possibility of blood contamination with similar accuracy and precision of that of the "gold standard method" is an innovative proposal.

Blood volume analysis: a new technique and new clinical interest reinvigorate a classic study.

PubMed

Manzone, Timothy A; Dam, Hung Q; Soltis, Daniel; Sagar, Vidya V

2007-06-01

Blood volume studies using the indicator dilution technique and radioactive tracers have been performed in nuclear medicine departments for over 50 y. A nuclear medicine study is the gold standard for blood volume measurement, but the classic dual-isotope blood volume study is time-consuming and can be prone to technical errors. Moreover, a lack of normal values and a rubric for interpretation made volume status measurement of limited interest to most clinicians other than some hematologists. A new semiautomated system for blood volume analysis is now available and provides highly accurate results for blood volume analysis within only 90 min. The availability of rapid, accurate blood volume analysis has brought about a surge of clinical interest in using blood volume data for clinical management. Blood volume analysis, long a low-volume nuclear medicine study all but abandoned in some laboratories, is poised to enter the clinical mainstream. This article will first present the fundamental principles of fluid balance and the clinical means of volume status assessment. We will then review the indicator dilution technique and how it is used in nuclear medicine blood volume studies. We will present an overview of the new semiautomated blood volume analysis technique, showing how the study is done, how it works, what results are provided, and how those results are interpreted. Finally, we will look at some of the emerging areas in which data from blood volume analysis can improve patient care. The reader will gain an understanding of the principles underlying blood volume assessment, know how current nuclear medicine blood volume analysis studies are performed, and appreciate their potential clinical impact.

Left Atrial 4D Blood Flow Dynamics and Hemostasis following Electrical Cardioversion of Atrial Fibrillation.

PubMed

Cibis, Merih; Lindahl, Tomas L; Ebbers, Tino; Karlsson, Lars O; Carlhäll, Carl-Johan

2017-01-01

Background: Electrical cardioversion in patients with atrial fibrillation is followed by a transiently impaired atrial mechanical function, termed atrial stunning. During atrial stunning, a retained risk of left atrial thrombus formation exists, which may be attributed to abnormal left atrial blood flow patterns. 4D Flow cardiovascular magnetic resonance (CMR) enables blood flow assessment from the entire three-dimensional atrial volume throughout the cardiac cycle. We sought to investigate left atrial 4D blood flow patterns and hemostasis during left atrial stunning and after left atrial mechanical function was restored. Methods: 4D Flow and morphological CMR data as well as blood samples were collected in fourteen patients at two time-points: 2-3 h (Time-1) and 4 weeks (Time-2) following cardioversion. The volume of blood stasis and duration of blood stasis were calculated. In addition, hemostasis markers were analyzed. Results: From Time-1 to Time-2: Heart rate decreased (61 ± 7 vs. 56 ± 8 bpm, p = 0.01); Maximum change in left atrial volume increased (8 ± 4 vs. 22 ± 15%, p = 0.009); The duration of stasis (68 ± 11 vs. 57 ± 8%, p = 0.002) and the volume of stasis (14 ± 9 vs. 9 ± 7%, p = 0.04) decreased; Thrombin-antithrombin complex (TAT) decreased (5.2 ± 3.3 vs. 3.3 ± 2.2 μg/L, p = 0.008). A significant correlation was found between TAT and the volume of stasis ( r 2 = 0.69, p < 0.001) at Time-1 and between TAT and the duration of stasis ( r 2 = 0.34, p = 0.04) at Time-2. Conclusion: In this longitudinal study, left atrial multidimensional blood flow was altered and blood stasis was elevated during left atrial stunning compared to the restored left atrial mechanical function. The coagulability of blood was also elevated during atrial stunning. The association between blood stasis and hypercoagulability proposes that assessment of left atrial 4D flow can add to the pathophysiological understanding of thrombus formation during atrial fibrillation related atrial stunning.

Performance evaluation of the Abbott CELL-DYN Ruby and the Sysmex XT-2000i haematology analysers.

PubMed

Leers, M P G; Goertz, H; Feller, A; Hoffmann, J J M L

2011-02-01

Two mid-range haematology analysers (Abbott CELL-DYN Ruby and Sysmex XT-2000i) were evaluated to determine their analytical performance and workflow efficiency in the haematology laboratory. In total 418 samples were processed for determining equivalence of complete blood count (CBC) measurements, and 100 for reticulocyte comparison. Blood smears served for assessing the agreement of the differential counts. Inter-instrument agreement for most parameters was good although small numbers of discrepancies were observed. Systematic biases were found for mean cell volume, reticulocytes, platelets and mean platelet volume. CELL-DYN Ruby WBC differentials were obtained with all samples while the XT-2000i suppressed differentials partially or completely in 13 samples (3.1%). WBC subpopulation counts were otherwise in good agreement with no major outliers. Following first-pass CBC/differential analysis, 88 (21%) of XT-2000i samples required further analyser processing compared to 18 (4.3%) for the CELL-DYN Ruby. Smear referrals for suspected WBC/nucleated red blood cells and platelet abnormalities were indicated for 106 (25.4%) and 95 (22.7%) of the XT-2000i and CELL-DYN Ruby samples respectively. Flagging efficiencies for both analysers were found to be similar. The Sysmex XT-2000i and Abbott CELL-DYN Ruby analysers have broadly comparable analytical performance, but the CELL-DYN Ruby showed superior first-pass efficiency. © 2010 Blackwell Publishing Ltd.

Utility of capillary microsampling for rat pharmacokinetic studies: Comparison of tail-vein bleed to jugular vein cannula sampling.

PubMed

Korfmacher, Walter; Luo, Yongyi; Ho, Stacy; Sun, Wei; Shen, Liduo; Wang, Jie; Wu, Zhongtao; Guo, Yang; Snow, Gregory; O'Shea, Thomas

2015-01-01

Serial sampling methods have been used for rat pharmacokinetic (PK) studies for over 20 years. Currently, it is still common to take 200-250 μL of blood at each timepoint when performing a PK study in rats and using serial sampling. While several techniques have been employed for collecting blood samples from rats, there is only limited published data to compare these methods. Recently, microsampling (≤ 50 μL) techniques have been reported as an alternative process for collecting blood samples from rats. In this report, five compounds were dosed orally into rats. For three proprietary compounds, jugular vein cannula (JVC) sampling was used to collect whole blood and plasma samples and capillary microsampling (CMS) was used to collect blood samples from the tail vein of the same animal. For the two other compounds, marketed drugs fluoxetine and glipizide, JVC sampling was used to collect both whole blood and blood CMS samples while tail-vein sampling from the same rats was also used to collect both whole blood and blood CMS samples. For the three proprietary compounds, the blood AUC as well as the blood concentration-time profile that were obtained from the tail vein were different from those obtained via JVC sampling. For fluoxetine, the blood total exposure (AUC) was not statistically different when comparing tail-vein sampling to JVC sampling, however the blood concentration-time profile that was obtained from the tail vein was different than the one obtained from JVC sampling. For glipizide, the blood AUC and concentration-time profile were not statistically different when comparing the tail-vein sampling to the JVC sampling. For both fluoxetine and glipizide, the blood concentration profiles obtained from CMS were equivalent to the blood concentration profiles obtained from the standard whole blood sampling, collected at the same sampling site. The data in this report provide strong evidence that blood CMS is a valuable small volume blood sampling approach for rats and that it provides results for test compound concentrations that are equivalent to those obtained from traditional whole blood sampling. The data also suggest that for some compounds, the concentration-time profile that is obtained for a test compound based on sampling from a rat tail vein may be different from that obtained from rat JVC sampling. In some cases, this shift in the concentration-time profile will result in different PK parameters for the test compound. Based on these observations, it is recommended that a consistent blood sampling method should be used for serial microsampling in discovery rat PK studies when testing multiple new chemical entities. If the rat tail vein sampling method is selected for PK screening, then conducting a bridging study on the lead compound is recommended to confirm that the rat PK obtained from JVC sampling is comparable to the tail-vein sampling. Copyright © 2015 Elsevier Inc. All rights reserved.

Fetal blood drawing.

PubMed

Hobbins, J C; Mahoney, M J

1975-07-19

A small sample of fetal blood suitable for studies of haemoglobin synthesis was obtained from a placental vessel under endoscopic visualisation in 23 of 26 patients in whom the procedure was attempted prior to second-trimester abortion. Fetal blood loss, calculated in 23 cases, was between 0-2 ml. and 2-5 ml., and fetal blood-volume depletion varied from 0-5% to 15%. No short-term ill-effects were demonstrated in mother or fetus in any of 16 patients in whom the injection of aborti-facient was postponed for between 16 and 24 hours after the procedure.

Use of self-collected capillary blood samples for islet autoantibody screening in relatives: a feasibility and acceptability study.

PubMed

Liu, Y; Rafkin, L E; Matheson, D; Henderson, C; Boulware, D; Besser, R E J; Ferrara, C; Yu, L; Steck, A K; Bingley, P J

2017-07-01

To evaluate the feasibility of using self-collected capillary blood samples for islet autoantibody testing to identify risk in relatives of people with Type 1 diabetes. Participants were recruited via the observational TrialNet Pathway to Prevention study, which screens and monitors relatives of people with Type 1 diabetes for islet autoantibodies. Relatives were sent kits for capillary blood collection, with written instructions, an online instructional video link and a questionnaire. Sera from capillary blood samples were tested for autoantibodies to glutamic acid decarboxylase, islet antigen-2, insulin and zinc transporter 8. 'Successful' sample collection was defined as obtaining sufficient volume and quality to provide definitive autoantibody results, including confirmation of positive results by repeat assay. In 240 relatives who returned samples, the median (range) age was 15.5 (1-49) years and 51% were male. Of these samples, 98% were sufficient for glutamic acid decarboxylase, islet antigen-2 and zinc transporter 8 autoantibody testing and 84% for insulin autoantibody testing and complete autoantibody screen. The upper 90% confidence bound for unsuccessful collection was 4.4% for glutamic acid decarboxylase, islet antigen-2 and/or zinc transporter 8 autoantibody assays, and 19.3% for insulin autoantibodies. Despite 43% of 220 questionnaire respondents finding capillary blood collection uncomfortable or painful, 82% preferred home self-collection of capillary blood samples compared with outpatient venepuncture (90% of those aged <8 years, 83% of those aged 9-18 years and 73% of those aged >18 years). The perceived difficulty of collecting capillary blood samples did not affect success rate. Self-collected capillary blood sampling offers a feasible alternative to venous sampling, with the potential to facilitate autoantibody screening for Type 1 diabetes risk. © 2017 Diabetes UK.

Comparison of haematology, coagulation and clinical chemistry parameters in blood samples from the sublingual vein and vena cava in Sprague-Dawley rats.

PubMed

Seibel, J; Bodié, K; Weber, S; Bury, D; Kron, M; Blaich, G

2010-10-01

The investigation of clinical pathology parameters (haematology, clinical chemistry and coagulation) is an important part of the preclinical evaluation of drug safety. However, the blood sampling method employed should avoid or minimize stress and injury in laboratory animals. In the present study, we compared the clinical pathology results from blood samples collected terminally from the vena cava (VC) immediately before necropsy with samples taken from the sublingual vein (VS) also prior to necropsy in order to determine whether the sampling method has an influence on clinical pathology parameters. Forty-six 12-week-old male Sprague-Dawley rats were assigned to two groups (VC or VS; n = 23 each). All rats were anaesthetized with isoflurane prior to sampling. In the VC group, blood was withdrawn from the inferior VC. For VS sampling, the tongue was gently pulled out and the VS was punctured. The haematology, coagulation and clinical chemistry parameters were compared. Equivalence was established for 13 parameters, such as mean corpuscular volume, white blood cells and calcium. No equivalence was found for the remaining 26 parameters, although they were considered to be similar when compared with the historical data and normal ranges. The most conspicuous finding was that activated prothrombin time was 30.3% less in blood taken from the VC (16.6 ± 0.89 s) than that in the VS samples (23.8 ± 1.58 s). Summing up, blood sampling from the inferior VC prior to necropsy appears to be a suitable and reliable method for terminal blood sampling that reduces stress and injury to laboratory rats in preclinical drug safety studies.

Servo-control of water and sodium homeostasis during renal clearance measurements in conscious rats.

PubMed

Thomsen, Klaus; Shirley, David G

2007-01-01

Servo-controlled fluid and sodium replacement during clearance studies is used in order to prevent loss of body fluid and sodium following diuretic/natriuretic procedures. However, even under control conditions, the use of this technique is sometimes associated with increases in proximal tubular fluid output (assessed by lithium clearance) and excretion rates. The present study examined the reason for these increases. The first series of experiments showed that one cause is volume overloading. This can occur if the servo system is activated from the start, i.e., during the establishment of a suitably high urine flow rate by constant infusion of hypotonic glucose solution. The second series of experiments showed that replacement of blood samples with donor blood can also lead to increases in fractional lithium excretion and accompanying increases in water and sodium excretion, a problem not seen when blood samples are replaced with the animal's own red blood cells resuspended in isotonic saline. When these pitfalls are avoided, servo-controlled sodium and fluid replacement is a reliable technique that makes it possible to study the effects of natriuretic and/or diuretic stimuli without interference from unwanted changes in extracellular volume. 2007 S. Karger AG, Basel

A novel instrument for studying the flow behaviour of erythrocytes through microchannels simulating human blood capillaries.

PubMed

Sutton, N; Tracey, M C; Johnston, I D; Greenaway, R S; Rampling, M W

1997-05-01

A novel instrument has been developed to study the microrheology of erythrocytes as they flow through channels of dimensions similar to human blood capillaries. The channels are produced in silicon substrates using microengineering technology. Accurately defined, physiological driving pressures and temperatures are employed whilst precise, real-time image processing allows individual cells to be monitored continuously during their transit. The instrument characterises each cell in a sample of ca. 1000 in terms of its volume and flow velocity profile during its transit through a channel. The unique representation of the data in volume/velocity space provides new insight into the microrheological behaviour of blood. The image processing and subsequent data analysis enable the system to reject anomalous events such as multiple cell transits, thereby ensuring integrity of the resulting data. By employing an array of microfluidic flow channels we can integrate a number of different but precise and highly reproducible channel sizes and geometries within one array, thereby allowing multiple, concurrent isobaric measurements on one sample. As an illustration of the performance of the system, volume/velocity data sets recorded in a microfluidic device incorporating multiple channels of 100 microns length and individual widths ranging between 3.0 and 4.0 microns are presented.

Quantification of myo-inositol, 1,5-anhydro-D-sorbitol, and D-chiro-inositol using high-performance liquid chromatography with electrochemical detection in very small volume clinical samples

PubMed Central

Schimpf, Karen J.; Meek, Claudia C.; Leff, Richard D.; Phelps, Dale L.; Schmitz, Daniel J.; Cordle, Christopher T.

2015-01-01

Inositol is a six-carbon sugar alcohol and is one of nine biologically significant isomers of hexahydroxycyclohexane. Myo-inositol is the primary biologically active form and is present in higher concentrations in the fetus and newborn than in adults. It is currently being examined for the prevention of retinopathy of prematurity in newborn preterm infants. A robust method for quantifying myo-inositol (MI), D-chiro-inositol (DCI) and 1,5-anhydro-D-sorbitol (ADS) in very small-volume (25 μL) urine, blood serum and/or plasma samples was developed. Using a multiple-column, multiple mobile phase liquid chromatographic system with electrochemical detection, the method was validated with respect to (a) selectivity, (b) accuracy/recovery, (c) precision/reproducibility, (d) sensitivity, (e) stability and (f) ruggedness. The standard curve was linear and ranged from 0.5 to 30 mg/L for each of the three analytes. Above-mentioned performance measures were within acceptable limits described in the Food and Drug Administration’s Guidance for Industry: Bioanalytical Method Validation. The method was validated using blood serum and plasma collected using four common anticoagulants, and also by quantifying the accuracy and sensitivity of MI measured in simulated urine samples recovered from preterm infant diaper systems. The method performs satisfactorily measuring the three most common inositol isomers on 25 μL clinical samples of serum, plasma milk, and/or urine. Similar performance is seen testing larger volume samples of infant formulas and infant formula ingredients. MI, ADS and DCI may be accurately tested in urine samples collected from five different preterm infant diapers if the urine volume is greater than 2–5 mL. PMID:26010453

Levonorgestrel-releasing intrauterine system use in premenopausal women with symptomatic uterine leiomyoma: a systematic review.

PubMed

Jiang, Wenxiao; Shen, Qi; Chen, Miaomiao; Wang, Ying; Zhou, Qingfeng; Zhu, Xuejie; Zhu, Xueqiong

2014-08-01

A systematic review is done to determine the efficacy and safety of levonorgestrel-releasing intrauterine systems as a treatment using in premenopausal women with symptomatic uterine leiomyoma. We searched the Medline, Central and ICTRP databases for all articles published from inception through July 2013 that examined the following outcomes: uterine volume, uterine leiomyoma volume, endometrial thickness, then menstrual blood loss, blood haemoglobin, ferritin and hematocrit levels, treatment failure rate, device expulsion rate, hysterectomy rate and side effects. From 645 studies, a total of 11 studies met our inclusion criteria with sample sizes ranging from 10 to 104. Evidence suggested that levonorgestrel-releasing intrauterine systems could decrease uterine volume and endometrial thickness, significantly reduce menstrual blood loss, and increase blood haemoglobin, ferritin and hematocrit levels. There was no evidence for decreasing uterine leiomyoma volume. There were no adverse effects on the ovarian function except for ovarian cysts. Device expulsion rates were low, which associated with leiomyoma size (larger than 3cm) but not with leiomyoma location. Irregular bleeding/spotting was observed at the beginning of the follow-up period and then decreased progressively. Results of this systematic review indicate that levonorgestrel-releasing intrauterine systems may be effective and safe treatment for symptomatic uterine leiomyoma in premenopausal women. Copyright © 2014 Elsevier Inc. All rights reserved.

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.

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

PubMed

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

2015-01-01

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

Comparison of the effects of intravenous administration of isotonic and hypertonic sodium bicarbonate solutions on venous acid-base status in dehydrated calves with strong ion acidosis.

PubMed

Coskun, Alparslan; Sen, Ismail; Guzelbektes, Hasan; Ok, Mahmut; Turgut, Kursat; Canikli, Sebnem

2010-05-15

OBJECTIVE-To compare the effects of IV administration of isotonic (1.3%) and hypertonic (8.4%) sodium bicarbonate (NaHCO(3)) solutions on acid-base status in dehydrated calves with strong ion (metabolic) acidosis. DESIGN-Randomized controlled clinical trial. ANIMALS-50 calves with diarrhea and severe dehydration. PROCEDURES-Calves were randomly assigned to receive isotonic NaHCO(3) solution (65 mL/kg [29.5 mL/lb], IV) over 3 hours (n = 30) or hypertonic NaHCO(3) solution (10 mL/kg [4.5 mL/lb], IV) over 20 minutes (20). Blood samples were collected at 0 hours (immediately prior to solution administration) and at 0.5, 1, 2, and 4 hours after administration began. Samples were submitted for blood gas analysis, serum biochemical analysis, and determination of blood Na(+), K(+), and Cl(-) concentrations and percentage change in plasma volume. RESULTS-Calves that received isotonic NaHCO(3) solution had an increase in venous blood pH, HCO(3) concentration, and base excess; a small, transient increase in Po(2); and no change in Pco(2) within 4 hours after administration began. Calves that received hypertonic NaHCO(3) solution had an immediate increase in venous blood pH, HCO(3) concentration, and base excess; a small, transient increase Pco(2); and no change in Po(2) within 0.5 hours after treatment began. Plasma volume increased to a greater extent following administration of isotonic solution than after administration of hypertonic solution. CONCLUSIONS AND CLINICAL RELEVANCE-IV administration of 8.4% NaHCO(3) solution in small volumes provided fast and effective improvement of severe acid-base abnormalities in calves with severe strong ion acidosis but did not improve hydration status as well as administration of a larger volume of isotonic NaHCO(3) solution.

Comparison of Pre- and Postoperative Hemoglobin and Hematocrit Levels in Hip Arthroscopy

PubMed Central

Seijas, Roberto; Espinosa, Wenceslao; Sallent, Andrea; Cuscó, Xavier; Cugat, Ramón; Ares, Oscar

2015-01-01

Purpose : to assess the loss in hematocrit and hemoglobin, if any, 24 hours after hip arthroscopy. Methods : thirty-five patients were included. Laboratory tests including complete blood count and white blood cells were performed one week prior to surgery and 24 hours after. Surgical time, volume of saline perfusion and pump perfusion was also recorded. Results : mean preoperative hematocrit was 42.01% (4.63 SD), whereas mean postoperative hematocrit at 24 h decreased to 36.78% (SD 5.11) (p <0.021.). Mean preoperative hemoglobin was 14.23 g/dL (1.73 SD), and mean postoperative hemoglobin at 24 h decreased to 12.40 g/dL (SD 1.92) (p =0.03.). Platelets and white blood cells, as well as the remaining biochemical parameters showed no significant difference between preoperative and postoperative samples. Lost blood volume worked out with the logarithmic method for estimated blood loss was which 0.78 liters (SD 0.45). Lost blood volume taking into account, the red blood cell mass was also 0.78 liters (SD 0.45). Conclusion : a significant decrease in hemoglobin and hematocrit after hip arthroscopy was observed. Although patients did not show clinical signs of anemia or bleeding, blood loss should be considered when planning a hip arthroscopy, especially in patients at risk of anemia. According to our results, we recommend a postoperative control analysis at 24 h. Level of Evidence : level II, Diagnostic Study. PMID:26401169

Global transcriptomic profiling using small volumes of whole blood: a cost-effective method for translational genomic biomarker identification in small animals.

PubMed

Fricano, Meagan M; Ditewig, Amy C; Jung, Paul M; Liguori, Michael J; Blomme, Eric A G; Yang, Yi

2011-01-01

Blood is an ideal tissue for the identification of novel genomic biomarkers for toxicity or efficacy. However, using blood for transcriptomic profiling presents significant technical challenges due to the transcriptomic changes induced by ex vivo handling and the interference of highly abundant globin mRNA. Most whole blood RNA stabilization and isolation methods also require significant volumes of blood, limiting their effective use in small animal species, such as rodents. To overcome these challenges, a QIAzol-based RNA stabilization and isolation method (QSI) was developed to isolate sufficient amounts of high quality total RNA from 25 to 500 μL of rat whole blood. The method was compared to the standard PAXgene Blood RNA System using blood collected from rats exposed to saline or lipopolysaccharide (LPS). The QSI method yielded an average of 54 ng total RNA per μL of rat whole blood with an average RNA Integrity Number (RIN) of 9, a performance comparable with the standard PAXgene method. Total RNA samples were further processed using the NuGEN Ovation Whole Blood Solution system and cDNA was hybridized to Affymetrix Rat Genome 230 2.0 Arrays. The microarray QC parameters using RNA isolated with the QSI method were within the acceptable range for microarray analysis. The transcriptomic profiles were highly correlated with those using RNA isolated with the PAXgene method and were consistent with expected LPS-induced inflammatory responses. The present study demonstrated that the QSI method coupled with NuGEN Ovation Whole Blood Solution system is cost-effective and particularly suitable for transcriptomic profiling of minimal volumes of whole blood, typical of those obtained with small animal species.

Performance of Identifiler Direct and PowerPlex 16 HS on the Applied Biosystems 3730 DNA Analyzer for processing biological samples archived on FTA cards.

PubMed

Laurin, Nancy; DeMoors, Anick; Frégeau, Chantal

2012-09-01

Direct amplification of STR loci from biological samples collected on FTA cards without prior DNA purification was evaluated using Identifiler Direct and PowerPlex 16 HS in conjunction with the use of a high throughput Applied Biosystems 3730 DNA Analyzer. In order to reduce the overall sample processing cost, reduced PCR volumes combined with various FTA disk sizes were tested. Optimized STR profiles were obtained using a 0.53 mm disk size in 10 μL PCR volume for both STR systems. These protocols proved effective in generating high quality profiles on the 3730 DNA Analyzer from both blood and buccal FTA samples. Reproducibility, concordance, robustness, sample stability and profile quality were assessed using a collection of blood and buccal samples on FTA cards from volunteer donors as well as from convicted offenders. The new developed protocols offer enhanced throughput capability and cost effectiveness without compromising the robustness and quality of the STR profiles obtained. These results support the use of these protocols for processing convicted offender samples submitted to the National DNA Data Bank of Canada. Similar protocols could be applied to the processing of casework reference samples or in paternity or family relationship testing. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Dried Blood Spot Analysis Suitable for Therapeutic Drug Monitoring of Voriconazole, Fluconazole, and Posaconazole

PubMed Central

van der Elst, Kim C. M.; Span, Lambert F. R.; van Hateren, Kai; Vermeulen, Karin M.; van der Werf, Tjip S.; Greijdanus, Ben; Kosterink, Jos G. W.; Uges, Donald R. A.

2013-01-01

Invasive aspergillosis and candidemia are important causes of morbidity and mortality in immunocompromised and critically ill patients. The triazoles voriconazole, fluconazole, and posaconazole are widely used for the treatment and prophylaxis of these fungal infections. Due to the variability of the pharmacokinetics of the triazoles among and within individual patients, therapeutic drug monitoring is important for optimizing the efficacy and safety of antifungal treatment. A dried blood spot (DBS) analysis was developed and was clinically validated for voriconazole, fluconazole, and posaconazole in 28 patients. Furthermore, a questionnaire was administered to evaluate the patients' opinions of the sampling method. The DBS analytical method showed linearity over the concentration range measured for all triazoles. Results for accuracy and precision were within accepted ranges; samples were stable at room temperature for at least 12 days; and different hematocrit values and blood spot volumes had no significant influence. The ratio of the drug concentration in DBS samples to that in plasma was 1.0 for voriconazole and fluconazole and 0.9 for posaconazole. Sixty percent of the patients preferred DBS analysis as a sampling method; 15% preferred venous blood sampling; and 25% had no preferred method. There was significantly less perception of pain with the DBS sampling method (P = 0.021). In conclusion, DBS analysis is a reliable alternative to venous blood sampling and can be used for therapeutic drug monitoring of voriconazole, fluconazole, and posaconazole. Patients were satisfied with DBS sampling and had less pain than with venous sampling. Most patients preferred DBS sampling to venous blood sampling. PMID:23896473

High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots.

PubMed

Holmberg, Rebecca C; Gindlesperger, Alissa; Stokes, Tinsley; Brady, Dane; Thakore, Nitu; Belgrader, Philip; Cooney, Christopher G; Chandler, Darrell P

2013-06-11

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).

Techniques for immobilizing and bleeding marmots and woodrats.

PubMed

Frase, B A; Van Vuren, D

1989-07-01

Blood samples were obtained in the field by femoral vein puncture in bushy-tailed woodrats (Neotoma cinerea) and yellow-bellied marmots (Marmota flaviventris) that had been injected intramuscularly with ketamine hydrochloride. Dosages ranged from 50 mg/kg for marmots to 30 to 110 mg/kg for woodrats. Sedated animals were handled easily, and a volume of blood sufficient for hematological assays or electrophoresis could be collected.

Amylase, lipase, and volume of drainage fluid in gastrectomy for the early detection of complications caused by pancreatic leakage.

PubMed

Seo, Kyung Won; Yoon, Ki Young; Lee, Sang Ho; Shin, Yeon Myung; Choi, Kyung Hyun; Hwang, Hyun Yong

2011-12-01

Pancreatic leakage is a serious complication of gastrectomy due to stomach cancer. Therefore, we analyzed amylase and lipase concentrations in blood and drainage fluid, and evaluated the volume of drainage fluid to discern their usefulness as markers for the early detection of serious pancreatic leakage requiring reoperation after gastrectomy. From January 2001 to December 2007, we retrospectively analyzed data from 24,072 patient samples. We divided patients into two groups; 1) complications with pancreatic leakage (CG), and 2) no complications associated with pancreatic leakage (NCG). Values of amylase and lipase in the blood and drainage fluid, volume of the drainage fluid, and relationships among the volumes, amylase values, and lipase values in the drainage fluid were evaluated, respectively in the two groups. The mean amylase values of CG were significantly higher than those of NCG in blood and drainage fluid (P < 0.05). For lipase, statistically significant differences were observed in drainage fluid (P < 0.05). The mean volume (standard deviation) of the drained fluid through the tube between CG (n = 22) and NCG (n = 236) on postoperative day 1 were 368.41 (266.25) and 299.26 (300.28), respectively. There were no statistically significant differences between the groups (P = 0.298). There was a correlation between the amylase and lipase values in the drainage fluid (r = 0.812, P = 0.000). Among postoperative amylase and lipase values in blood and drainage fluid, and the volume of drainage fluid, the amylase in drainage fluid was better differentiated between CG and NCG than other markers. The volume of the drainage fluid did not differ significantly between groups.

A randomized, controlled, double-blind crossover study on the effects of 1-L infusions of 6% hydroxyethyl starch suspended in 0.9% saline (voluven) and a balanced solution (Plasma Volume Redibag) on blood volume, renal blood flow velocity, and renal cortical tissue perfusion in healthy volunteers.

PubMed

Chowdhury, Abeed H; Cox, Eleanor F; Francis, Susan T; Lobo, Dileep N

2014-05-01

We compared the effects of intravenous administration of 6% hydroxyethyl starch (maize-derived) in 0.9% saline (Voluven; Fresenius Kabi, Runcorn, United Kingdom) and a "balanced" preparation of 6% hydroxyethyl starch (potato-derived) [Plasma Volume Redibag (PVR); Baxter Healthcare, Thetford, United Kingdom] on renal blood flow velocity and renal cortical tissue perfusion in humans using magnetic resonance imaging. Hyperchloremia resulting from 0.9% saline infusion may adversely affect renal hemodynamics when compared with balanced crystalloids. This phenomenon has not been studied with colloids. Twelve healthy adult male subjects received 1-L intravenous infusions of Voluven or PVR over 30 minutes in a randomized, double-blind manner, with crossover studies 7 to 10 days later. Magnetic resonance imaging proceeded for 60 minutes after commencement of infusion to measure renal artery blood flow velocity and renal cortical perfusion. Blood was sampled, and weight was recorded at 0, 30, 60, 120, 180, and 240 minutes. Mean peak serum chloride concentrations were 108 and 106 mmol/L, respectively, after Voluven and PVR infusion (P = 0.032). Changes in blood volume (P = 0.867), strong ion difference (P = 0.219), and mean renal artery flow velocity (P = 0.319) were similar. However, there was a significant increase in mean renal cortical tissue perfusion after PVR when compared with Voluven (P = 0.033). There was no difference in urinary neutrophil gelatinase-associated liopcalin to creatinine ratios after the infusion (P = 0.164). There was no difference in the blood volume-expanding properties of the 2 preparations of 6% hydroxyethyl starch. The balanced starch produced an increase in renal cortical tissue perfusion, a phenomenon not seen with starch in 0.9% saline.

Specimen rejection in laboratory medicine: Necessary for patient safety?

PubMed Central

Dikmen, Zeliha Gunnur; Pinar, Asli; Akbiyik, Filiz

2015-01-01

Introduction The emergency laboratory in Hacettepe University Hospitals receives specimens from emergency departments (EDs), inpatient services and intensive care units (ICUs). The samples are accepted according to the rejection criteria of the laboratory. In this study, we aimed to evaluate the sample rejection ratios according to the types of pre-preanalytical errors and collection areas. Materials and methods The samples sent to the emergency laboratory were recorded during 12 months between January to December, 2013 in which 453,171 samples were received and 27,067 specimens were rejected. Results Rejection ratios was 2.5% for biochemistry tests, 3.2% for complete blood count (CBC), 9.8% for blood gases, 9.2% for urine analysis, 13.3% for coagulation tests, 12.8% for therapeutic drug monitoring, 3.5% for cardiac markers and 12% for hormone tests. The most frequent rejection reasons were fibrin clots (28%) and inadequate volume (9%) for biochemical tests. Clotted samples (35%) and inadequate volume (13%) were the major causes for coagulation tests, blood gas analyses and CBC. The ratio of rejected specimens was higher in the EDs (40%) compared to ICUs (30%) and inpatient services (28%). The highest rejection ratio was observed in neurology ICU (14%) among the ICUs and internal medicine inpatient service (10%) within inpatient clinics. Conclusions We detected an overall specimen rejection rate of 6% in emergency laboratory. By documentation of rejected samples and periodic training of healthcare personnel, we expect to decrease sample rejection ratios below 2%, improve total quality management of the emergency laboratory and promote patient safety. PMID:26527231

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

PubMed

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

2011-03-01

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

Quasi-static acoustic tweezing thromboelastometry.

PubMed

Holt, R G; Luo, D; Gruver, N; Khismatullin, D B

2017-07-01

Essentials Blood coagulation measurement during contact with an artificial surface leads to unreliable data. Acoustic tweezing thromboelastometry is a novel non-contact method for coagulation monitoring. This method detects differences in the blood coagulation state within 10 min. Coagulation data were obtained using a much smaller sample volume (4 μL) than currently used. Background Thromboelastography is widely used as a tool to assess the coagulation status of critical care patients. It allows observation of changes in material properties of whole blood, beginning with early stages of clot formation and ending with clot lysis. However, the contact activation of the coagulation cascade at surfaces of thromboelastographic systems leads to inherent variability and unreliability in predicting bleeding or thrombosis risks. Objectives To develop acoustic tweezing thromboelastometry as a non-contact method for perioperative assessment of blood coagulation. Methods Acoustic tweezing is used to levitate microliter drops of biopolymer and human blood samples. By quasi-statically changing the acoustic pressure we control the sample drop location and deformation. Sample size, deformation and location are determined by digital imaging at each pressure. Results Simple Newtonian liquid solutions maintain a constant, reversible location vs. deformation curve. In contrast, the location/deformation curves for gelatin, alginate, whole blood and blood plasma uniquely change as the samples solidify. Increasing elasticity causes the sample to deform less, leading to steeper stress/strain curves. By extracting a linear regime slope, we show that whole blood or blood plasma exhibits a unique slope profile as it begins to clot. By exposing blood samples to pro- or antithrombotic agents, the slope profile changes, allowing detection of hyper- or hypocoagulable states. Conclusions We demonstrate that quasi-static acoustic tweezing can yield information about clotting onset, maturation and strength. The advantages of small sample size, non-contact and rapid measurement make this technique desirable for real-time monitoring of blood coagulation. © 2017 International Society on Thrombosis and Haemostasis.

Anemia in patients with coinherited thalassemia and glucose-6-phosphate dehydrogenase deficiency.

PubMed

Pornprasert, Sakorn; Phanthong, Siratcha

2013-01-01

Thalassemia and glucose-6-phosphate dehydrogenase (G-6-PD) deficiency are genetic disorders that cause hemolytic anemia. In areas with high frequencies of both hematological disorders, coinheritance of G-6-PD deficiency with thalassemia can be found. Whether G-6-PD deficiency, coinherited with thalassemia, enhances severe anemia is still unclear. Hematological parameters between thalassemia carriers with G-6-PD deficiency and those without G-6-PD deficiency were compared. The G-6-PD deficiency was diagnosed in 410 blood samples from thalassemia patients using a fluorescent spot test. The levels of hemoglobin (Hb), packed cell volume (PCV), mean corpuscular volume (MCV) and Hb A2/Hb E [β26(B8)Glu→Lys; HBB: c.79G>A] were measured using an automated blood counter and high performance liquid chromatography (HPLC), respectively. The G-6-PD deficiency was found in 37 samples (9.02%). Mean levels of Hb, PCV, MCV and Hb A2/E were similar between the two groups. Thus, G-6-PD deficiency did not enhance red blood cell pathology or induce more anemic severity in thalassemia patients.

Detection limits of 405 nm and 633 nm excited PpIX fluorescence for brain tumor detection during stereotactic biopsy

NASA Astrophysics Data System (ADS)

Markwardt, Niklas; Götz, Marcus; Haj-Hosseini, Neda; Hollnburger, Bastian; Sroka, Ronald; Stepp, Herbert; Zelenkov, Petr; Rühm, Adrian

2016-04-01

5-aminolevulinic-acid-(5-ALA)-induced protoporphyrin IX (PpIX) fluorescence may be used to improve stereotactic brain tumor biopsies. In this study, the sensitivity of PpIX-based tumor detection has been investigated for two potential excitation wavelengths (405 nm, 633 nm). Using a 200 μm fiber in contact with semi-infinite optical phantoms containing ink and Lipovenös, PpIX detection limits of 4.0 nM and 200 nM (relating to 1 mW excitation power) were determined for 405 nm and 633 nm excitation, respectively. Hence, typical PpIX concentrations in glioblastomas of a few μM should be well detectable with both wavelengths. Additionally, blood layers of selected thicknesses were placed between fiber and phantom. Red excitation was shown to be considerably less affected by blood interference: A 50 μm blood layer, for instance, blocked the 405- nm-excited fluorescence completely, but reduced the 633-nm-excited signal by less than 50%. Ray tracing simulations demonstrated that - without blood layer - the sensitivity advantage of 405 nm rises for decreasing fluorescent volume from 50-fold to a maximum of 100-fold. However, at a tumor volume of 1 mm3, which is a typical biopsy sample size, the 633-nm-excited fluorescence signal is only reduced by about 10%. Further simulations revealed that with increasing fiber-tumor distance, the signal drops faster for 405 nm. This reduces the risk of detecting tumor tissue outside the needle's coverage, but diminishes the overlap between optically and mechanically sampled volumes. While 405 nm generally offers a higher sensitivity, 633 nm is more sensitive to distant tumors and considerably superior in case of blood-covered tumor tissue.

What a drop can do: dried blood spots as a minimally invasive method for integrating biomarkers into population-based research.

PubMed

McDade, Thomas W; Williams, Sharon; Snodgrass, J Josh

2007-11-01

Logistical constraints associated with the collection and analysis of biological samples in community-based settings have been a significant impediment to integrative, multilevel bio-demographic and biobehavioral research. However recent methodological developments have overcome many of these constraints and have also expanded the options for incorporating biomarkers into population-based health research in international as well as domestic contexts. In particular using dried blood spot (DBS) samples-drops of whole blood collected on filter paper from a simple finger prick-provides a minimally invasive method for collecting blood samples in nonclinical settings. After a brief discussion of biomarkers more generally, we review procedures for collecting, handling, and analyzing DBS samples. Advantages of using DBS samples-compared with venipuncture include the relative ease and low cost of sample collection, transport, and storage. Disadvantages include requirements for assay development and validation as well as the relatively small volumes of sample. We present the results of a comprehensive literature review of published protocols for analysis of DBS samples, and we provide more detailed analysis of protocols for 45 analytes likely to be of particular relevance to population-level health research. Our objective is to provide investigators with the information they need to make informed decisions regarding the appropriateness of blood spot methods for their research interests.

Emergency Physician Estimation of Blood Loss

PubMed Central

Ashburn, Jeffery C.; Harrison, Tamara; Ham, James J.; Strote, Jared

2012-01-01

Introduction Emergency physicians (EP) frequently estimate blood loss, which can have implications for clinical care. The objectives of this study were to examine EP accuracy in estimating blood loss on different surfaces and compare attending physician and resident performance. Methods A sample of 56 emergency department (ED) physicians (30 attending physicians and 26 residents) were asked to estimate the amount of moulage blood present in 4 scenarios: 500 mL spilled onto an ED cot; 25 mL spilled onto a 10-pack of 4 × 4-inch gauze; 100 mL on a T-shirt; and 150 mL in a commode filled with water. Standard estimate error (the absolute value of (estimated volume − actual volume)/actual volume × 100) was calculated for each estimate. Results The mean standard error for all estimates was 116% with a range of 0% to 1233%. Only 8% of estimates were within 20% of the true value. Estimates were most accurate for the sheet scenario and worst for the commode scenario. Residents and attending physicians did not perform significantly differently (P > 0.05). Conclusion Emergency department physicians do not estimate blood loss well in a variety of scenarios. Such estimates could potentially be misleading if used in clinical decision making. Clinical experience does not appear to improve estimation ability in this limited study. PMID:22942938

Blood indicators of seasonal metabolic patterns in captive adult gray wolves

USGS Publications Warehouse

Seal, U.S.; Mech, L.D.

1983-01-01

Blood samples and physical data were collected weekly from a colony of gray wolves (Canis lupus) maintained under natural weather arid light conditions. Sampling over 33 continuous months indicated that hemoglobin, hematocrit, red blood cells, mean corpuscular hemoglobin concentration (MCHC), and thyroxine exhibited consistent circannual patterns of variation in both males and females. Hemoglobin levels peaked at 15-16 g/dl in January in females and at 16-17 g/dl in February in males, and were lowest in August at 10.5-11.5 g/dl (P < 0.00001). The cyclic patterns of hematocrit, red blood cells, and MCHC were similarly timed. Females also had a cyclic pattern of white blood cell counts and body weight; their weight peaked in early February and was lowest in August (P < 0.001). Body temperature, urea nitrogen, mean corpuscular volume (MCV), serum glucose, and cortisol did not follow a consistent seasonal pattern.

Handheld Universal Diagnostic Sensor

NASA Technical Reports Server (NTRS)

Chan, Eugene

2012-01-01

The rHEALTH technology is designed to shrink an entire hospital testing laboratory onto a handheld device. A physician or healthcare provider performs the test by collecting a fingerstick of blood from a patient. The tiny volume of blood is inserted into the rHEALTH device. Inside the device is a microfluidic chip that contains small channels about the width of a human hair. These channels help move the blood and analyze the blood sample. The rHEALTH sensor uses proprietary reagents called nanostrips, which are nanoscale test strips that enable the clinical assays. The readout is performed by laser-induced fluorescence. Overall, the time from blood collection through analysis is less than a minute.

Electrospun biocompatible Chitosan/MIL-101 (Fe) composite nanofibers for solid-phase extraction of Δ9-tetrahydrocannabinol in whole blood samples using Box-Behnken experimental design.

PubMed

Asiabi, Mina; Mehdinia, Ali; Jabbari, Ali

2017-01-06

The nanofibers of biocompatible Chitosan/MIL-101 (Fe) composite were synthesized by a simple, cheap and accessible electrospining method and applied for mat-based extraction of trace amount of Δ9-tetrahydrocannabinol (THC) from human whole blood sample following its combination by high performance liquid chromatography-ultraviolet detection. The composite nanofibres were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and N 2 adsorption-desorption experiments. The volume of eluting solvent, sorbent amount, pH and% NaCl (w/v) influencing on the responses were investigated using factorial experimental design. The optimum point was achieved by analysis of the results according to design expert (DX) software. The volume of eluting solvent, sorbent amount and pH were significant variables, and 150μL, 7mg and 7.0 were respectively chosen for obtaining the best extraction response. Under the optimum conditions, the method was exhibited a linear range of 0.1-100μgL -1 (R 2 =0.9943) for THC with a detection limit of 0.04μgL -1 . Acceptable values for intra-day (3.2%) and inter-day (4.8%) relative standard deviations were obtained. The high preconcentration factor (970) and satisfactory recoveries (88.2%-92.4%) in whole blood samples were achieved which proved the capability of the method for trace determination of THC in the human whole blood samples. Copyright © 2016 Elsevier B.V. All rights reserved.

21 CFR 864.5950 - Blood volume measuring device.

Code of Federal Regulations, 2011 CFR

2011-04-01

... § 864.5950 Blood volume measuring device. (a) Identification. A blood volume measuring device is a manual, semiautomated, or automated system that is used to calculate the red cell mass, plasma volume... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Blood volume measuring device. 864.5950 Section...

21 CFR 864.5950 - Blood volume measuring device.

Code of Federal Regulations, 2010 CFR

2010-04-01

... § 864.5950 Blood volume measuring device. (a) Identification. A blood volume measuring device is a manual, semiautomated, or automated system that is used to calculate the red cell mass, plasma volume... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Blood volume measuring device. 864.5950 Section...

21 CFR 864.5950 - Blood volume measuring device.

Code of Federal Regulations, 2014 CFR

2014-04-01

... § 864.5950 Blood volume measuring device. (a) Identification. A blood volume measuring device is a manual, semiautomated, or automated system that is used to calculate the red cell mass, plasma volume... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Blood volume measuring device. 864.5950 Section...

21 CFR 864.5950 - Blood volume measuring device.

Code of Federal Regulations, 2013 CFR

2013-04-01

... § 864.5950 Blood volume measuring device. (a) Identification. A blood volume measuring device is a manual, semiautomated, or automated system that is used to calculate the red cell mass, plasma volume... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Blood volume measuring device. 864.5950 Section...

21 CFR 864.5950 - Blood volume measuring device.

Code of Federal Regulations, 2012 CFR

2012-04-01

... § 864.5950 Blood volume measuring device. (a) Identification. A blood volume measuring device is a manual, semiautomated, or automated system that is used to calculate the red cell mass, plasma volume... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Blood volume measuring device. 864.5950 Section...

Vascular capacitance and cardiac output in pacing-induced canine models of acute and chronic heart failure.

PubMed

Ogilvie, R I; Zborowska-Sluis, D

1995-11-01

The relationship between stressed and total blood volume, total vascular capacitance, central blood volume, cardiac output (CO), and pulmonary capillary wedge pressure (Ppcw) was investigated in pacing-induced acute and chronic heart failure. Acute heart failure was induced in anesthetized splenectomized dogs by a volume load (20 mL/kg over 10 min) during rapid right ventricular pacing at 250 beats/min (RRVP) for 60 min. Chronic heart failure was induced by continuous RRVP for 2-6 weeks (average 24 +/- 2 days). Total vascular compliance and capacitance were calculated from the mean circulatory filling pressure (Pmcf) during transient circulatory arrest after acetylcholine at three different circulating volumes. Stressed blood volume was calculated as a product of compliance and Pmcf, with the total blood volume measured by a dye dilution. Central blood volume (CBV) and CO were measured by thermodilution. Central (heart and lung) vascular capacitance was estimated from the plot of Ppcw against CBV. Acute volume loading without RRVP increased capacitance and CO, whereas after volume loading with RRVP, capacitance and CO were unaltered from baseline. Chronic RRVP reduced capacitance and CO. All interventions, volume +/- RRVP or chronic RRVP, increased stressed and central blood volumes and Ppcw. Acute or chronic RRVP reduced central vascular capacitance. Cardiac output was increased when stressed and unstressed blood volumes increased proportionately as during volume loading alone. When CO was reduced and Ppcw increased, as during chronic RRVP or acute RRVP plus a volume load, stressed blood volume was increased and unstressed blood volume was decreased. Thus, interventions that reduced CO and increased Ppcw also increased stressed and reduced unstressed blood volume and total vascular capacitance.

Simple, miniaturized blood plasma extraction method.

PubMed

Kim, Jin-Hee; Woenker, Timothy; Adamec, Jiri; Regnier, Fred E

2013-12-03

A rapid plasma extraction technology that collects a 2.5 μL aliquot of plasma within three minutes from a finger-stick derived drop of blood was evaluated. The utility of the plasma extraction cards used was that a paper collection disc bearing plasma was produced that could be air-dried in fifteen minutes and placed in a mailing envelop for transport to an analytical laboratory. This circumvents the need for venipuncture and blood collection in specialized vials by a phlebotomist along with centrifugation and refrigerated storage. Plasma extraction was achieved by applying a blood drop to a membrane stack through which plasma was drawn by capillary action. During the course of plasma migration to a collection disc at the bottom of the membrane stack blood cells were removed by a combination of adsorption and filtration. After the collection disc filled with an aliquot of plasma the upper membranes were stripped from the collection card and the collection disc was air-dried. Intercard differences in the volume of plasma collected varied approximately 1% while volume variations of less than 2% were seen with hematocrit levels ranging from 20% to 71%. Dried samples bearing metabolites and proteins were then extracted from the disc and analyzed. 25-Hydroxy vitamin D was quantified by LC-MS/MS analysis following derivatization with a secosteroid signal enhancing tag that imparted a permanent positive charge to the vitamin and reduced the limit of quantification (LOQ) to 1 pg of collected vitamin on the disc; comparable to values observed with liquid-liquid extraction (LLE) of a venipuncture sample. A similar study using conventional proteomics methods and spectral counting for quantification was conducted with yeast enolase added to serum as an internal standard. The LOQ with extracted serum samples for enolase was 1 μM, linear from 1 to 40 μM, the highest concentration examined. In all respects protein quantification with extracted serum samples was comparable to that observed with serum samples obtained by venipuncture.

Finger Blood Volume Responses of Counselors, Counselor Trainees, and Non Counselors to Stimuli from an Empathy Test

ERIC Educational Resources Information Center

Gellen, Murray I.

1970-01-01

The major finding was that Counselors in this sample displayed higher arousal than the sample of scientists during the presentation of dialogs depicting human interactions. This finding is consistent with the widely held view that counselors reflect a greater degree of humanistic interpersonal orientation than noncounselors, and offers support for…

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

PubMed Central

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

2015-01-01

Purpose. The gravimetric method of weighing surgical sponges is used to quantify intraoperative blood loss. The dry mass minus the wet mass of the gauze equals the volume of blood lost. This method assumes that the density of blood is equivalent to water (1 gm/mL). This study's purpose was to validate the assumption that the density of blood is equivalent to water and to correlate density with hematocrit. Methods. 50 µL of whole blood was weighed from eighteen rats. A distilled water control was weighed for each blood sample. The averages of the blood and water were compared utilizing a Student's unpaired, one-tailed t-test. The masses of the blood samples and the hematocrits were compared using a linear regression. Results. The average mass of the eighteen blood samples was 0.0489 g and that of the distilled water controls was 0.0492 g. The t-test showed P = 0.2269 and R 2 = 0.03154. The hematocrit values ranged from 24% to 48%. The linear regression R 2 value was 0.1767. Conclusions. The R 2 value comparing the blood and distilled water masses suggests high correlation between the two populations. Linear regression showed the hematocrit was not proportional to the mass of the blood. The study confirmed that the measured density of blood is similar to water. PMID:26464949

Metabolic liver function measured in vivo by dynamic (18)F-FDGal PET/CT without arterial blood sampling.

PubMed

Horsager, Jacob; Munk, Ole Lajord; Sørensen, Michael

2015-01-01

Metabolic liver function can be measured by dynamic PET/CT with the radio-labelled galactose-analogue 2-[(18)F]fluoro-2-deoxy-D-galactose ((18)F-FDGal) in terms of hepatic systemic clearance of (18)F-FDGal (K, ml blood/ml liver tissue/min). The method requires arterial blood sampling from a radial artery (arterial input function), and the aim of this study was to develop a method for extracting an image-derived, non-invasive input function from a volume of interest (VOI). Dynamic (18)F-FDGal PET/CT data from 16 subjects without liver disease (healthy subjects) and 16 patients with liver cirrhosis were included in the study. Five different input VOIs were tested: four in the abdominal aorta and one in the left ventricle of the heart. Arterial input function from manual blood sampling was available for all subjects. K*-values were calculated using time-activity curves (TACs) from each VOI as input and compared to the K-value calculated using arterial blood samples as input. Each input VOI was tested on PET data reconstructed with and without resolution modelling. All five image-derived input VOIs yielded K*-values that correlated significantly with K calculated using arterial blood samples. Furthermore, TACs from two different VOIs yielded K*-values that did not statistically deviate from K calculated using arterial blood samples. A semicircle drawn in the posterior part of the abdominal aorta was the only VOI that was successful for both healthy subjects and patients as well as for PET data reconstructed with and without resolution modelling. Metabolic liver function using (18)F-FDGal PET/CT can be measured without arterial blood samples by using input data from a semicircle VOI drawn in the posterior part of the abdominal aorta.

Clinical application of microsampling versus conventional sampling techniques in the quantitative bioanalysis of antibiotics: a systematic review.

PubMed

Guerra Valero, Yarmarly C; Wallis, Steven C; Lipman, Jeffrey; Stove, Christophe; Roberts, Jason A; Parker, Suzanne L

2018-03-01

Conventional sampling techniques for clinical pharmacokinetic studies often require the removal of large blood volumes from patients. This can result in a physiological or emotional burden, particularly for neonates or pediatric patients. Antibiotic pharmacokinetic studies are typically performed on healthy adults or general ward patients. These may not account for alterations to a patient's pathophysiology and can lead to suboptimal treatment. Microsampling offers an important opportunity for clinical pharmacokinetic studies in vulnerable patient populations, where smaller sample volumes can be collected. This systematic review provides a description of currently available microsampling techniques and an overview of studies reporting the quantitation and validation of antibiotics using microsampling. A comparison of microsampling to conventional sampling in clinical studies is included.

Heart rate sensitive optical coherence angiography

NASA Astrophysics Data System (ADS)

Alvarez, Karl; Lopez-Tremoleda, Jordi; Donnan, Rob; Michael-Titus, Adina T.; Tomlins, Peter H.

2018-02-01

Optical coherence angiography (OCA) enables visualisation of three-dimensional micro-vasculature from optical coherence tomography data volumes. Typically, various statistical methods are used to discriminate static tissue from blood flow within vessels. In this paper, we introduce a new method that relies upon the beating heart frequency to isolate blood vessels from the surrounding tissue. Vascular blood flow is assumed to be more strongly modulated by the heart-beat compared to surrounding tissue and therefore short-time Fourier transform of sequential measurements can discriminate the two. Furthermore, it is demonstrated that adjacent B-Scans within an OCT data volume can provide the required sampling frequency. As such, the technique can be considered to be a spatially mapped variation of photoplethysmography (PPG), whereby each image voxel operates as a PPG detector. This principle is demonstrated using both a model system and in vivo for monitoring the vascular changes effected by traumatic brain injury in mice. In vivo measurements were acquired at an A-Scan rate of 10kHz to form a 500x500x512 (lateral x lateral x axial) pixel volume, enabling sequential sampling of the mouse heart rate in an expected range of 300-600 bpm. One of the advantages of this new OCA processing method is that it can be used in conjunction with existing algorithms as an additional filter for signal to noise enhancement.

Knowledge of Good Blood Culture Sampling Practice among Healthcare Staffs in An Emergency Department - Are We Getting It Right?

PubMed

Chew, K S; Mohd Hashairi, F; Jusoh, A F; Aziz, A A; Nik Hisamuddin, N A R; Siti Asma, H

2013-08-01

Although a vital test, blood culture is often plagued with the problem of contamination and false results, especially in a chaotic emergency department setting. The objectives of this pilot study is to find out the level of understanding among healthcare staffs in emergency department, Hospital Universiti Sains Malaysia (HUSM) regarding good blood culture sampling practice. All healthcare staffs in emergency department, HUSM who consented to this study were given a set of selfadministered anonymous questionnaire to fill. More than half (53.1%) of the 64 participants are emergency medicine residents. Majority of them (75%) have been working in the emergency medicine, HUSM for more than 2 years. More than half of them were able to answer correctly the amount of blood volume needed for culture in adult and pediatric patients. When asked what are the factors required to improve the true yield as well as to reduce the risk of culture contamination, the four commonest answers given were observing proper aseptic technique during blood sampling, donning sterile glove, proper hand scrubbing as well as ensuring the sterility of the equipments. This study suggests that there is a lack of proper knowledge of good blood culture sampling practice among our healthcare staffs in emergency department.

Detection and capture of single circulating melanoma cells using photoacoustic flowmetry

NASA Astrophysics Data System (ADS)

O'Brien, Christine; Mosley, Jeffrey; Goldschmidt, Benjamin S.; Viator, John A.

2010-02-01

Photoacoustic flowmetry has been used to detect single circulating melanoma cells in vitro. Circulating melanoma cells are those cells that travel in the blood and lymph systems to create secondary tumors and are the hallmark of metastasis. This technique involves taking blood samples from patients, separating the white blood and melanoma cells from whole blood and irradiating them with a pulsed laser in a flowmetry set up. Rapid, visible wavelength laser pulses on the order of 5 ns can induce photoacoustic waves in melanoma cells due to their melanin content, while surrounding white blood cells remain acoustically passive. We have developed a system that identifies rare melanoma cells and captures them in 50 microliter volumes using suction applied near the photoacoustic detection chamber. The 50 microliter sample is then diluted and the experiment is repeated using the new sample until only a melanoma cell remains. We have tested this system on dyed microspheres ranging in size from 300 to 500 microns. Capture of circulating melanoma cells may provide the opportunity to study metastatic cells for basic understanding of the spread of cancer and to optimize patient specific therapies.

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

A novel method for blood volume estimation using trivalent chromium in rabbit models.

PubMed

Baby, Prathap Moothamadathil; Kumar, Pramod; Kumar, Rajesh; Jacob, Sanu S; Rawat, Dinesh; Binu, V S; Karun, Kalesh M

2014-05-01

Blood volume measurement though important in management of critically ill-patients is not routinely estimated in clinical practice owing to labour intensive, intricate and time consuming nature of existing methods. The aim was to compare blood volume estimations using trivalent chromium [(51)Cr(III)] and standard Evans blue dye (EBD) method in New Zealand white rabbit models and establish correction-factor (CF). Blood volume estimation in 33 rabbits was carried out using EBD method and concentration determined using spectrophotometric assay followed by blood volume estimation using direct injection of (51)Cr(III). Twenty out of 33 rabbits were used to find CF by dividing blood volume estimation using EBD with blood volume estimation using (51)Cr(III). CF is validated in 13 rabbits by multiplying it with blood volume estimation values obtained using (51)Cr(III). The mean circulating blood volume of 33 rabbits using EBD was 142.02 ± 22.77 ml or 65.76 ± 9.31 ml/kg and using (51)Cr(III) was estimated to be 195.66 ± 47.30 ml or 89.81 ± 17.88 ml/kg. The CF was found to be 0.77. The mean blood volume of 13 rabbits measured using EBD was 139.54 ± 27.19 ml or 66.33 ± 8.26 ml/kg and using (51)Cr(III) with CF was 152.73 ± 46.25 ml or 71.87 ± 13.81 ml/kg (P = 0.11). The estimation of blood volume using (51)Cr(III) was comparable to standard EBD method using CF. With further research in this direction, we envisage human blood volume estimation using (51)Cr(III) to find its application in acute clinical settings.

Integrated hollow microneedle-optofluidic biosensor for therapeutic drug monitoring in sub-nanoliter volumes

NASA Astrophysics Data System (ADS)

Ranamukhaarachchi, Sahan A.; Padeste, Celestino; Dübner, Matthias; Häfeli, Urs O.; Stoeber, Boris; Cadarso, Victor J.

2016-07-01

Therapeutic drug monitoring (TDM) typically requires painful blood drawn from patients. We propose a painless and minimally-invasive alternative for TDM using hollow microneedles suitable to extract extremely small volumes (<1 nL) of interstitial fluid to measure drug concentrations. The inner lumen of a microneedle is functionalized to be used as a micro-reactor during sample collection to trap and bind target drug candidates during extraction, without requirements of sample transfer. An optofluidic device is integrated with this microneedle to rapidly quantify drug analytes with high sensitivity using a straightforward absorbance scheme. Vancomycin is currently detected by using volumes ranging between 50-100 μL with a limit of detection (LoD) of 1.35 μM. The proposed microneedle-optofluidic biosensor can detect vancomycin with a sample volume of 0.6 nL and a LoD of <100 nM, validating this painless point of care system with significant potential to reduce healthcare costs and patients suffering.

Integrated hollow microneedle-optofluidic biosensor for therapeutic drug monitoring in sub-nanoliter volumes

PubMed Central

Ranamukhaarachchi, Sahan A.; Padeste, Celestino; Dübner, Matthias; Häfeli, Urs O.; Stoeber, Boris; Cadarso, Victor J.

2016-01-01

Therapeutic drug monitoring (TDM) typically requires painful blood drawn from patients. We propose a painless and minimally-invasive alternative for TDM using hollow microneedles suitable to extract extremely small volumes (<1 nL) of interstitial fluid to measure drug concentrations. The inner lumen of a microneedle is functionalized to be used as a micro-reactor during sample collection to trap and bind target drug candidates during extraction, without requirements of sample transfer. An optofluidic device is integrated with this microneedle to rapidly quantify drug analytes with high sensitivity using a straightforward absorbance scheme. Vancomycin is currently detected by using volumes ranging between 50–100 μL with a limit of detection (LoD) of 1.35 μM. The proposed microneedle-optofluidic biosensor can detect vancomycin with a sample volume of 0.6 nL and a LoD of <100 nM, validating this painless point of care system with significant potential to reduce healthcare costs and patients suffering. PMID:27380889

Muscle damage produced during a simulated badminton match in competitive male players.

PubMed

Abián, Pablo; Del Coso, Juan; Salinero, Juan José; Gallo-Salazar, César; Areces, Francisco; Ruiz-Vicente, Diana; Lara, Beatriz; Soriano, Lidón; Muñoz, Victor; Lorenzo-Capella, Irma; Abián-Vicén, Javier

2016-01-01

The purpose of the study was to assess the occurrence of muscle damage after a simulated badminton match and its influence on physical and haematological parameters. Sixteen competitive male badminton players participated in the study. Before and just after a 45-min simulated badminton match, maximal isometric force and badminton-specific running/movement velocity were measured to assess muscle fatigue. Blood samples were also obtained before and after the match. The badminton match did not affect maximal isometric force or badminton-specific velocity. Blood volume and plasma volume were significantly reduced during the match and consequently haematite, leucocyte, and platelet counts significantly increased. Blood myoglobin and creatine kinase concentrations increased from 26.5 ± 11.6 to 197.3 ± 70.2 µg·L(-1) and from 258.6 ± 192.2 to 466.0 ± 296.5 U·L(-1), respectively. In conclusion, a simulated badminton match modified haematological parameters of whole blood and serum blood that indicate the occurrence of muscle fibre damage. However, the level of muscle damage did not produce decreased muscle performance.

Microanalyzer for Biomonitoring of Lead (Pb) in Blood and Urine

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

Yantasee, Wassana; Timchalk, Chuck; Lin, Yuehe

2007-01-01

Biomonitoring of lead (Pb) in blood and urine enables quantitative evaluation of human occupational and environmental exposures to Pb. The state-of-the-art ICP-MS instruments analyze metals in laboratories, resulting in lengthy turn around time, and are expensive. In response to the growing need for metal analyzer for on-site, real-time monitoring of trace metals in individuals, we developed a portable microanalyzer based on flow-injection/adsorptive stripping voltammetry and used it to analyze Pb in rat blood and urine. Fouling of electrodes by proteins often prevents the effective use of electrochemical sensors in biological matrices. Minimization of such fouling was accomplished with the suitablemore » sample pretreatment and the turbulent flowing of Pb contained blood and urine onto the glassy electrode inside the microanalyzer, which resulted in no apparent electrode fouling even when the samples contained 50% urine or 10% blood by volume. There was no matrix effect on the voltammetric Pb signals even when the samples contained 10% blood or 10% urine. The microanalyzer offered linear concentration range relevant to Pb exposure levels in human (0-20 ppb in 10%-blood samples, 0-50 ppb in 50%-urine samples). The device had excellent sensitivity and reproducibility; Pb detection limits were 0.54 ppb and 0.42 ppb, and % RSDs were 4.9 and 2.4 in 50%-urine and 10%-blood samples, respectively. It offered a high throughput (3 min per sample) and had economical use of samples (60 ?L per measurement), making the collection of blood being less invasive especially to children, and had low reagent consumption (1 ?g of Hg per measurement), thus minimizing the health concerns of mercury use. Being miniaturized in size, the microanalyzer is portable and field-deployable. Thus, it has a great potential to be the next-generation analyzer for biomonitoring of toxic metals.« less

A microfluidic device integrating dual CMOS polysilicon nanowire sensors for on-chip whole blood processing and simultaneous detection of multiple analytes.

PubMed

Kuan, Da-Han; Wang, I-Shun; Lin, Jiun-Rue; Yang, Chao-Han; Huang, Chi-Hsien; Lin, Yen-Hung; Lin, Chih-Ting; Huang, Nien-Tsu

2016-08-02

The hemoglobin-A1c test, measuring the ratio of glycated hemoglobin (HbA1c) to hemoglobin (Hb) levels, has been a standard assay in diabetes diagnosis that removes the day-to-day glucose level variation. Currently, the HbA1c test is restricted to hospitals and central laboratories due to the laborious, time-consuming whole blood processing and bulky instruments. In this paper, we have developed a microfluidic device integrating dual CMOS polysilicon nanowire sensors (MINS) for on-chip whole blood processing and simultaneous detection of multiple analytes. The micromachined polymethylmethacrylate (PMMA) microfluidic device consisted of a serpentine microchannel with multiple dam structures designed for non-lysed cells or debris trapping, uniform plasma/buffer mixing and dilution. The CMOS-fabricated polysilicon nanowire sensors integrated with the microfluidic device were designed for the simultaneous, label-free electrical detection of multiple analytes. Our study first measured the Hb and HbA1c levels in 11 clinical samples via these nanowire sensors. The results were compared with those of standard Hb and HbA1c measurement methods (Hb: the sodium lauryl sulfate hemoglobin detection method; HbA1c: cation-exchange high-performance liquid chromatography) and showed comparable outcomes. Finally, we successfully demonstrated the efficacy of the MINS device's on-chip whole blood processing followed by simultaneous Hb and HbA1c measurement in a clinical sample. Compared to current Hb and HbA1c sensing instruments, the MINS platform is compact and can simultaneously detect two analytes with only 5 μL of whole blood, which corresponds to a 300-fold blood volume reduction. The total assay time, including the in situ sample processing and analyte detection, was just 30 minutes. Based on its on-chip whole blood processing and simultaneous multiple analyte detection functionalities with a lower sample volume requirement and shorter process time, the MINS device can be effectively applied to real-time diabetes diagnostics and monitoring in point-of-care settings.

[Blood volume for biochemistry determinations--laboratory needs and everyday practice].

PubMed

Sztefko, Krystyna; Mamica, Katarzyna; Bugajska, Jolanta; Maziarz, Barbara; Tomasik, Przemysław

2014-01-01

Blood loss due to diagnostic phlebotomy jest a very serious problem, especially for newborn, infants and critically ill patients on intensive care units. Although single blood loss can be easily tolerated in adults, in small babies and in patients who are frequently monitored based on laboratory tests iatrogenic anaemia can occur. To evaluate the blood volume drawn for routine biochemistry tests in relation to patient age and the number of parameters requested. Blood volume drawn for routine biochemistry measurements from patients hospitalized in University Children's Hospital (N = 2980, children age from one day to 18 years) and in University Hospital (N = 859, adults, aged > 1.8 years) in Cracow has been analyzed. Blood volume was calculated based on regular tube diameter and blood heights in the tube. In case of microvettes the blood volume was 0.2 ml. Statistical analysis has been performed by using PRISM 5.0. The statistical significance was set at p < 0.05. The mean values of blood volume were 3.02 +/- 0.92 ml and 4.12 +/- 0.68 ml in children and adults, respectively. Analyzing blood volume drawn in children using both microvettes and regular tubes, significant correlation between blood volume and patient age (p < 0.001) as well the number of requested parameters (p < 0.001). The latest relationship was true only for up to five parameters. However, analyzing the blood volume drawn into only into regular tubes blood volume was not related to patients age and number of laboratory tests requested. The proportion of microvettes used for blood collection was highest for newborns and infants, and in all cases where only one to three laboratory tests were requested. 1. All educational programs for nurses and doctors should include the information about current laboratory automation and methods miniaturization; 2) The amount of blood volume needed by laboratory for the requested number of tests should always be taken into account when diagnostic phlebotomy is necessary.

Destruction of newly released red blood cells in space flight

NASA Technical Reports Server (NTRS)

Alfrey, C. P.; Udden, M. M.; Huntoon, C. L.; Driscoll, T.

1996-01-01

Space flight results in a rapid change in total blood volume, plasma volume, and red blood cell mass because the space to contain blood is decreased. The plasma volume and total blood volume decreases during the first hours in space and remain at a decreased level for the remainder of the flight. During the first several hours following return to earth, plasma volume and total blood volume increase to preflight levels. During the first few days in space recently produced red blood cells disappear from the blood resulting in a decrease in red blood cell mass of 10-15%. Red cells 12 d old or older survive normally and production of new cells continues at near preflight levels. After the first few days in space, the red cell mass is stable at the decreased level. Following return to earth the hemoglobin and red blood cell mass concentrations decrease reflecting the increase in plasma volume. The erythropoietin levels increase responding to "postflight anemia"; red cell production increases, and the red cell mass is restored to preflight levels after several weeks.

Prediction of Packed Cell Volume after Whole Blood Transfusion in Small Ruminants and South American Camelids: 80 Cases (2006-2016).

PubMed

Luethy, D; Stefanovski, D; Salber, R; Sweeney, R W

2017-11-01

Calculation of desired whole blood transfusion volume relies on an estimate of an animal's circulating blood volume, generally accepted to be 0.08 L/kg or 8% of the animal's body weight in kilograms. To use packed cell volume before and after whole blood transfusion to evaluate the accuracy of a commonly used equation to predict packed cell volume after transfusion in small ruminants and South American camelids; to determine the nature and frequency of adverse transfusion reactions in small ruminants and camelids after whole blood transfusion. Fifty-eight small ruminants and 22 alpacas that received whole blood transfusions for anemia. Retrospective case series; medical record review for small ruminants and camelids that received whole blood transfusions during hospitalization. Mean volume of distribution of blood as a fraction of body weight in sheep (0.075 L/kg, 7.5% BW) and goats (0.076 L/kg, 7.6% BW) differed significantly (P < 0.01) from alpacas (0.103 L/kg, 10.3% BW). Mild transfusion reactions were noted in 16% of transfusions. The generally accepted value of 8% for circulating blood volume (volume of distribution of blood) is adequate for calculation of transfusion volumes; however, use of the species-specific circulating blood volume can improve calculation of transfusion volume to predict and achieve desired packed cell volume. The incidence of transfusion reactions in small ruminants and camelids is low. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Newborn screening blood spot analysis in the UK: influence of spot size, punch location and haematocrit.

PubMed

Lawson, A J; Bernstone, L; Hall, S K

2016-03-01

In dried blood spot analysis, punch location and variations in applied sample volume and haematocrit can produce different measured concentrations of analytes. We investigated the magnitude of these effects in newborn screening in the UK. Heparinized blood spiked with thyroid stimulating hormone (TSH), phenylalanine, tyrosine, leucine, methionine, octanoyl carnitine (C8), and immunoreactive trypsinogen (IRT) was spotted onto filter paper: (i) at a constant haematocrit of 50% at various volumes, and (ii) at a range of haematocrits using a constant volume. Subpunches (3.2 mm) of the dried blood spots were then analysed. Compared with a central punch from a 50 µL blood spot with 50% haematocrit, 10 µL spots can have significantly lower measured concentrations of all analytes, with decreases of 15% or more observed for leucine, methionine, phenylalanine, and tyrosine. Punching at the edge of a spot can increase measured concentrations up to 35%. Higher haematocrit decreased measured TSH and C8 yet increased amino acids and IRT by 15% compared with 50% haematocrit. Lower haematocrits had the opposite effect, but only with higher concentrations of some analytes. Differences in blood spot size, haematocrit and punch location substantially affect measured concentrations for analytes used in the UK newborn screening programme, and this could affect false positive and negative rates. To minimize analytical bias, these variables should be controlled or adjusted for where possible. © The Author(s) 2015.

The measurement of peripheral blood volume reactions to tilt test by the electrical impedance technique after exercise in athletes

NASA Astrophysics Data System (ADS)

Melnikov, A. A.; Popov, S. G.; Nikolaev, D. V.; Vikulov, A. D.

2013-04-01

We have investigated the distribution of peripheral blood volumes in different regions of the body in response to the tilt-test in endurance trained athletes after aerobic exercise. Distribution of peripheral blood volumes (ml/beat) simultaneously in six regions of the body (two legs, two hands, abdomen, neck and ECG) was assessed in response to the tilt-test using the impedance method (the impedance change rate (dZ/dT). Before and after exercise session cardiac stroke (CSV) and blood volumes in legs, arms and neck were higher in athletes both in lying and standing positions. Before exercise the increase of heart rate and the decrease of a neck blood volume in response to tilting was lower (p <0.05) but the decrease of leg blood volumes was higher (p<0.001) in athletes. The reactions in arms and abdomen blood volumes were similar. Also, the neck blood volumes as percentage of CSV (%/CSV) did not change in the control but increased in athletes (p <0.05) in response to the tilt test. After (10 min recovery) the aerobic bicycle exercise (mean HR = 156±8 beat/min, duration 30 min) blood volumes in neck and arms in response to the tilting were reduced equally, but abdomen (p<0.05) and leg blood volumes (p <0.001) were lowered more significantly in athletes. The neck blood flow (%/CSV) did not change in athletes but decreased in control (p<0.01), which was offset by higher tachycardia in response to tilt-test in controls after exercise. The data demonstrate greater orthostatic tolerance in athletes both before and after exercise during fatigue which is due to effective distribution of blood flows aimed at maintaining cerebral blood flow.

Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

PubMed Central

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

2015-01-01

Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample. PMID:25675242

Mean platelet volume (MPV) predicts middle distance running performance.

PubMed

Lippi, Giuseppe; Salvagno, Gian Luca; Danese, Elisa; Skafidas, Spyros; Tarperi, Cantor; Guidi, Gian Cesare; Schena, Federico

2014-01-01

Running economy and performance in middle distance running depend on several physiological factors, which include anthropometric variables, functional characteristics, training volume and intensity. Since little information is available about hematological predictors of middle distance running time, we investigated whether some hematological parameters may be associated with middle distance running performance in a large sample of recreational runners. The study population consisted in 43 amateur runners (15 females, 28 males; median age 47 years), who successfully concluded a 21.1 km half-marathon at 75-85% of their maximal aerobic power (VO2max). Whole blood was collected 10 min before the run started and immediately thereafter, and hematological testing was completed within 2 hours after sample collection. The values of lymphocytes and eosinophils exhibited a significant decrease compared to pre-run values, whereas those of mean corpuscular volume (MCV), platelets, mean platelet volume (MPV), white blood cells (WBCs), neutrophils and monocytes were significantly increased after the run. In univariate analysis, significant associations with running time were found for pre-run values of hematocrit, hemoglobin, mean corpuscular hemoglobin (MCH), red blood cell distribution width (RDW), MPV, reticulocyte hemoglobin concentration (RetCHR), and post-run values of MCH, RDW, MPV, monocytes and RetCHR. In multivariate analysis, in which running time was entered as dependent variable whereas age, sex, blood lactate, body mass index, VO2max, mean training regimen and the hematological parameters significantly associated with running performance in univariate analysis were entered as independent variables, only MPV values before and after the trial remained significantly associated with running time. After adjustment for platelet count, the MPV value before the run (p = 0.042), but not thereafter (p = 0.247), remained significantly associated with running performance. The significant association between baseline MPV and running time suggest that hyperactive platelets may exert some pleiotropic effects on endurance performance.

Use of artificial intelligence to identify cardiovascular compromise in a model of hemorrhagic shock.

PubMed

Glass, Todd F; Knapp, Jason; Amburn, Philip; Clay, Bruce A; Kabrisky, Matt; Rogers, Steven K; Garcia, Victor F

2004-02-01

To determine whether a prototype artificial intelligence system can identify volume of hemorrhage in a porcine model of controlled hemorrhagic shock. Prospective in vivo animal model of hemorrhagic shock. Research foundation animal surgical suite; computer laboratories of collaborating industry partner. Nineteen, juvenile, 25- to 35-kg, male and female swine. Anesthetized animals were instrumented for arterial and systemic venous pressure monitoring and blood sampling, and a splenectomy was performed. Following a 1-hr stabilization period, animals were hemorrhaged in aliquots to 10, 20, 30, 35, 40, 45, and 50% of total blood volume with a 10-min recovery between each aliquot. Data were downloaded directly from a commercial monitoring system into a proprietary PC-based software package for analysis. Arterial and venous blood gas values, glucose, and cardiac output were collected at specified intervals. Electrocardiogram, electroencephalogram, mixed venous oxygen saturation, temperature (core and blood), mean arterial pressure, pulmonary artery pressure, central venous pressure, pulse oximetry, and end-tidal CO(2) were continuously monitored and downloaded. Seventeen of 19 animals (89%) died as a direct result of hemorrhage. Stored data streams were analyzed by the prototype artificial intelligence system. For this project, the artificial intelligence system identified and compared three electrocardiographic features (R-R interval, QRS amplitude, and R-S interval) from each of nine unknown samples of the QRS complex. We found that the artificial intelligence system, trained on only three electrocardiographic features, identified hemorrhage volume with an average accuracy of 91% (95% confidence interval, 84-96%). These experiments demonstrate that an artificial intelligence system, based solely on the analysis of QRS amplitude, R-R interval, and R-S interval of an electrocardiogram, is able to accurately identify hemorrhage volume in a porcine model of lethal hemorrhagic shock. We suggest that this technology may represent a noninvasive means of assessing the physiologic state during and immediately following hemorrhage. Point of care application of this technology may improve outcomes with earlier diagnosis and better titration of therapy of shock.

Development of Metabolic Function Biomarkers in the Common Marmoset, Callithrix jacchus

PubMed Central

Ziegler, Toni E.; Colman, Ricki J.; Tardif, Suzette D.; Sosa, Megan E.; Wegner, Fredrick H.; Wittwer, Daniel J.; Shrestha, Hemanta

2013-01-01

Metabolic assessment of a nonhuman primate model of metabolic syndrome and obesity requires the necessary biomarkers specific to the species. While the rhesus monkey has a number of specific assays for assessing metabolic syndrome, the marmoset does not. Furthermore, the common marmoset (Callithrix jacchus) has a small blood volume that necessitates using a single blood volume for multiple analyses. The common marmoset holds a great potential as an alternative primate model for the study of human disease but assay methods need to be developed and validated for the biomarkers of metabolic syndrome. Here we report on the adaptation, development and validation of commercially available immunoassays for common marmoset samples in small volumes. We have performed biological validations for insulin, adiponectin, leptin, and ghrelin to demonstrate the use of these biomarkers in examining metabolic syndrome and other related diseases in the common marmoset. PMID:23447060

Deformability measurement of red blood cells using a microfluidic channel array and an air cavity in a driving syringe with high throughput and precise detection of subpopulations.

PubMed

Kang, Yang Jun; Ha, Young-Ran; Lee, Sang-Joon

2016-01-07

Red blood cell (RBC) deformability has been considered a potential biomarker for monitoring pathological disorders. High throughput and detection of subpopulations in RBCs are essential in the measurement of RBC deformability. In this paper, we propose a new method to measure RBC deformability by evaluating temporal variations in the average velocity of blood flow and image intensity of successively clogged RBCs in the microfluidic channel array for specific time durations. In addition, to effectively detect differences in subpopulations of RBCs, an air compliance effect is employed by adding an air cavity into a disposable syringe. The syringe was equally filled with a blood sample (V(blood) = 0.3 mL, hematocrit = 50%) and air (V(air) = 0.3 mL). Owing to the air compliance effect, blood flow in the microfluidic device behaved transiently depending on the fluidic resistance in the microfluidic device. Based on the transient behaviors of blood flows, the deformability of RBCs is quantified by evaluating three representative parameters, namely, minimum value of the average velocity of blood flow, clogging index, and delivered blood volume. The proposed method was applied to measure the deformability of blood samples consisting of homogeneous RBCs fixed with four different concentrations of glutaraldehyde solution (0%-0.23%). The proposed method was also employed to evaluate the deformability of blood samples partially mixed with normal RBCs and hardened RBCs. Thereafter, the deformability of RBCs infected by human malaria parasite Plasmodium falciparum was measured. As a result, the three parameters significantly varied, depending on the degree of deformability. In addition, the deformability measurement of blood samples was successfully completed in a short time (∼10 min). Therefore, the proposed method has significant potential in deformability measurement of blood samples containing hematological diseases with high throughput and precise detection of subpopulations in RBCs.

Haematological alterations in Rattus norvegicus (Wistar) experimentally infected with Echinostoma paraensei (Trematoda: Echinostomatidae).

PubMed

Garcia, J S; Pinheiro, J; Hooper, C S; Simões, R O; Ferraz, J S; Maldonado, A

2012-07-01

Laboratory rats (Rattus norvegicus) were infected with Echinostoma paraensei (Trematoda: Echinostomatidae). The rodents received 150 metacercariae each and blood samples were collected weekly until the fifth week of infection. The blood samples were analyzed for determination of haematocrit, total red blood cells with their dimensions, haemoglobin and haematimetric index (mean corpuscular volume, MCV; mean corpuscular haemoglobin, MCH; and mean corpuscular haemoglobin concentration, MCHC) and platelets. Red blood cells, haematocrit and haemoglobin in the first week had significantly lower levels than those of uninfected (control) rats, suggesting the development of normocytic and normocromic anaemia with anisocytic alteration. The number of eosinophils did not increase significantly among the groups. We concluded that E. paraensei produces haematological alterations in R. norvegicus, causing regenerative anaemia. This system can therefore be a useful model to study the direct and indirect effects of gastrointestinal infections. Copyright © 2012 Elsevier Inc. All rights reserved.

Accelerated Brain DCE-MRI Using Iterative Reconstruction With Total Generalized Variation Penalty for Quantitative Pharmacokinetic Analysis: A Feasibility Study.

PubMed

Wang, Chunhao; Yin, Fang-Fang; Kirkpatrick, John P; Chang, Zheng

2017-08-01

To investigate the feasibility of using undersampled k-space data and an iterative image reconstruction method with total generalized variation penalty in the quantitative pharmacokinetic analysis for clinical brain dynamic contrast-enhanced magnetic resonance imaging. Eight brain dynamic contrast-enhanced magnetic resonance imaging scans were retrospectively studied. Two k-space sparse sampling strategies were designed to achieve a simulated image acquisition acceleration factor of 4. They are (1) a golden ratio-optimized 32-ray radial sampling profile and (2) a Cartesian-based random sampling profile with spatiotemporal-regularized sampling density constraints. The undersampled data were reconstructed to yield images using the investigated reconstruction technique. In quantitative pharmacokinetic analysis on a voxel-by-voxel basis, the rate constant K trans in the extended Tofts model and blood flow F B and blood volume V B from the 2-compartment exchange model were analyzed. Finally, the quantitative pharmacokinetic parameters calculated from the undersampled data were compared with the corresponding calculated values from the fully sampled data. To quantify each parameter's accuracy calculated using the undersampled data, error in volume mean, total relative error, and cross-correlation were calculated. The pharmacokinetic parameter maps generated from the undersampled data appeared comparable to the ones generated from the original full sampling data. Within the region of interest, most derived error in volume mean values in the region of interest was about 5% or lower, and the average error in volume mean of all parameter maps generated through either sampling strategy was about 3.54%. The average total relative error value of all parameter maps in region of interest was about 0.115, and the average cross-correlation of all parameter maps in region of interest was about 0.962. All investigated pharmacokinetic parameters had no significant differences between the result from original data and the reduced sampling data. With sparsely sampled k-space data in simulation of accelerated acquisition by a factor of 4, the investigated dynamic contrast-enhanced magnetic resonance imaging pharmacokinetic parameters can accurately estimate the total generalized variation-based iterative image reconstruction method for reliable clinical application.

Differential effects of lower body negative pressure and upright tilt on splanchnic blood volume

PubMed Central

Taneja, Indu; Moran, Christopher; Medow, Marvin S.; Glover, June L.; Montgomery, Leslie D.; Stewart, Julian M.

2015-01-01

Upright posture and lower body negative pressure (LBNP) both induce reductions in central blood volume. However, regional circulatory responses to postural changes and LBNP may differ. Therefore, we studied regional blood flow and blood volume changes in 10 healthy subjects undergoing graded lower-body negative pressure (−10 to −50 mmHg) and 8 subjects undergoing incremental head-up tilt (HUT; 20°, 40°, and 70°) on separate days. We continuously measured blood pressure (BP), heart rate, and regional blood volumes and blood flows in the thoracic, splanchnic, pelvic, and leg segments by impedance plethysmography and calculated regional arterial resistances. Neither LBNP nor HUT altered systolic BP, whereas pulse pressure decreased significantly. Blood flow decreased in all segments, whereas peripheral resistances uniformly and significantly increased with both HUT and LBNP. Thoracic volume decreased while pelvic and leg volumes increased with HUT and LBNP. However, splanchnic volume changes were directionally opposite with stepwise decreases in splanchnic volume with LBNP and stepwise increases in splanchnic volume during HUT. Splanchnic emptying in LBNP models regional vascular changes during hemorrhage. Splanchnic filling may limit the ability of the splanchnic bed to respond to thoracic hypovolemia during upright posture. PMID:17085534

( sup 99m Tc)diphosphonate uptake and hemodynamics in arthritis of the immature dog knee

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

Hansen, E.S.; Soballe, K.; Henriksen, T.B.

1991-03-01

The relationship between (99mTc)diphosphonate uptake and bone hemodynamics was studied in canine carrageenan-induced juvenile chronic arthritis. Blood flow was determined with microspheres, plasma and red cell volumes were measured by labeled fibrinogen and red cells, and the microvascular volume and mean transit time of blood were calculated. Normal femoral epiphyses had lower central and higher subchondral blood flow and diphosphonate uptake values. Epiphyseal vascular volume was uniform, resulting in a greater transit time of blood centrally. In arthritis, blood flow and diphosphonate uptake were increased subchondrally and unaffected centrally, while epiphyseal vascular volume was increased throughout, leading to prolonged transitmore » time centrally. The normal metaphyses had low blood flow and diphosphonate uptake values in cancellous bone and very high values in growth plates, but a large vascular volume throughout. The mean transit time therefore was low in growth plates and high in adjacent cancellous bone. Arthritis caused decreased blood flow and diphosphonate uptake in growth plates but increased vascular volume and transit time of blood. Diphosphonate uptake correlated positively with blood flow and plasma volume and negatively with red cell volume in a nonlinear fashion. Thus, changes in diphosphonate uptake and microvascular hemodynamics occur in both epiphyseal and metaphyseal bone in chronic synovitis of the immature knee. The (99mTc)diphosphonate bone scan seems to reflect blood flow, plasma volume, and red cell volume of bone.« less

Real-Time Microfluidic Blood-Counting System for PET and SPECT Preclinical Pharmacokinetic Studies.

PubMed

Convert, Laurence; Lebel, Réjean; Gascon, Suzanne; Fontaine, Réjean; Pratte, Jean-François; Charette, Paul; Aimez, Vincent; Lecomte, Roger

2016-09-01

Small-animal nuclear imaging modalities have become essential tools in the development process of new drugs, diagnostic procedures, and therapies. Quantification of metabolic or physiologic parameters is based on pharmacokinetic modeling of radiotracer biodistribution, which requires the blood input function in addition to tissue images. Such measurements are challenging in small animals because of their small blood volume. In this work, we propose a microfluidic counting system to monitor rodent blood radioactivity in real time, with high efficiency and small detection volume (∼1 μL). A microfluidic channel is built directly above unpackaged p-i-n photodiodes to detect β-particles with maximum efficiency. The device is embedded in a compact system comprising dedicated electronics, shielding, and pumping unit controlled by custom firmware to enable measurements next to small-animal scanners. Data corrections required to use the input function in pharmacokinetic models were established using calibrated solutions of the most common PET and SPECT radiotracers. Sensitivity, dead time, propagation delay, dispersion, background sensitivity, and the effect of sample temperature were characterized. The system was tested for pharmacokinetic studies in mice by quantifying myocardial perfusion and oxygen consumption with (11)C-acetate (PET) and by measuring the arterial input function using (99m)TcO4 (-) (SPECT). Sensitivity for PET isotopes reached 20%-47%, a 2- to 10-fold improvement relative to conventional catheter-based geometries. Furthermore, the system detected (99m)Tc-based SPECT tracers with an efficiency of 4%, an outcome not possible through a catheter. Correction for dead time was found to be unnecessary for small-animal experiments, whereas propagation delay and dispersion within the microfluidic channel were accurately corrected. Background activity and sample temperature were shown to have no influence on measurements. Finally, the system was successfully used in animal studies. A fully operational microfluidic blood-counting system for preclinical pharmacokinetic studies was developed. Microfluidics enabled reliable and high-efficiency measurement of the blood concentration of most common PET and SPECT radiotracers with high temporal resolution in small blood volume. © 2016 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

Physiological responses and evaluation of effects of BMI, smoking and drinking in high altitude acclimatization: a cohort study in Chinese Han young males.

PubMed

Peng, Qian-Qian; Basang, Zhuoma; Cui, Chao-Ying; Li, Lei; Qian, Ji; Gesang, Quzhen; Yang, La; La, Zong; De, Yang; Dawa, Puchi; Qu, Ni; Suo, Qu; Dan, Zhen; Xiao, Duoji; Wang, Xiao-Feng; Jin, Li

2013-01-01

High altitude acclimatization is a series of physiological responses taking places when subjects go to altitude. Many factors could influence these processes, such as altitude, ascending speed and individual characteristics. In this study, based on a repeated measurement design of three sequential measurements at baseline, acute phase and chronic phase, we evaluated the effect of BMI, smoking and drinking on a number of physiological responses in high altitude acclimatization by using mixed model and partial least square path model on a sample of 755 Han Chinese young males. We found that subjects with higher BMI responses were reluctant to hypoxia. The effect of smoking was not significant at acute phase. But at chronic phase, red blood cell volume increased less while respiratory function increased more for smoking subjects compared with nonsmokers. For drinking subjects, red blood cell volume increased less than nondrinkers at both acute and chronic phases, while blood pressures increased more than nondrinkers at acute phase and respiratory function, red blood cell volume and oxygen saturation increased more than nondrinkers at chronic phase. The heavy and long-term effect of smoking, drinking and other factors in high altitude acclimatization needed to be further studied.

Physiological Responses and Evaluation of Effects of BMI, Smoking and Drinking in High Altitude Acclimatization: A Cohort Study in Chinese Han Young Males

PubMed Central

Cui, Chao-ying; Li, Lei; Qian, Ji; Gesang, Quzhen; Yang, La; La, Zong; De, Yang; Dawa, Puchi; Qu, Ni; Suo, Qu; Dan, Zhen; Xiao, Duoji; Wang, Xiao-feng; Jin, Li

2013-01-01

High altitude acclimatization is a series of physiological responses taking places when subjects go to altitude. Many factors could influence these processes, such as altitude, ascending speed and individual characteristics. In this study, based on a repeated measurement design of three sequential measurements at baseline, acute phase and chronic phase, we evaluated the effect of BMI, smoking and drinking on a number of physiological responses in high altitude acclimatization by using mixed model and partial least square path model on a sample of 755 Han Chinese young males. We found that subjects with higher BMI responses were reluctant to hypoxia. The effect of smoking was not significant at acute phase. But at chronic phase, red blood cell volume increased less while respiratory function increased more for smoking subjects compared with nonsmokers. For drinking subjects, red blood cell volume increased less than nondrinkers at both acute and chronic phases, while blood pressures increased more than nondrinkers at acute phase and respiratory function, red blood cell volume and oxygen saturation increased more than nondrinkers at chronic phase. The heavy and long-term effect of smoking, drinking and other factors in high altitude acclimatization needed to be further studied. PMID:24260204

Dynamic Neurovascular Coupling and Uncoupling during Ictal Onset, Propagation, and Termination Revealed by Simultaneous In Vivo Optical Imaging of Neural Activity and Local Blood Volume

PubMed Central

Zhao, Mingrui; Schwartz, Theodore H.

2013-01-01

Traditional models of ictal propagation involve the concept of an initiation site and a progressive outward march of activation. The process of neurovascular coupling, whereby the brain supplies oxygenated blood to metabolically active neurons presumably results in a similar outward cascade of hyperemia. However, ictal neurovascular coupling has never been assessed in vivo using simultaneous measurements of membrane potential change and hyperemia with wide spatial sampling. In an acute rat ictal model, using simultaneous intrinsic optical signal (IOS) and voltage-sensitive dye (VSD) imaging of cerebral blood volume and membrane potential changes, we demonstrate that seizures consist of multiple dynamic multidirectional waves of membrane potential change with variable onset sites that spread through a widespread network. Local blood volume evolves on a much slower spatiotemporal scale. At seizure onset, the VSD waves extend beyond the IOS signal. During evolution, spatial correlation with hemodynamic signal only exists briefly at the maximal spread of the VSD signal. At termination, the IOS signal extends spatially and temporally beyond the VSD waves. Hence, vascular reactivity evolves in a separate but parallel fashion to membrane potential changes resulting in a mechanism of neurovascular coupling and uncoupling, which is as dynamic as the seizure itself. PMID:22499798

The Effect of Intra-Dialytic Exercise on Inflammation and Blood Endotoxin Levels.

PubMed

Wong, Jonathan; Davis, Philip; Patidar, Ashish; Zhang, Yonglong; Vilar, Enric; Finkelman, Malcolm; Farrington, Ken

2017-01-01

In healthy individuals, an acute inflammatory response occurs after intense exercise due to gut ischaemia and intestinal bacterial endotoxin translocation into the bloodstream. This process maybe exacerbated in patients who exercise during dialysis due to large volume shifts experienced by many during haemodialysis (HD). The acute effect of intra-dialytic exercise on blood endotoxins and inflammation is not known. The effect of intra-dialytic exercise on blood endotoxin and inflammation was investigated in 10 patients and compared with resting haemodialysis. Blood was measured for endotoxin and inflammatory biomarkers before and after dialysis. With the exception of one sample, all samples tested negative for endotoxin. Intra-dialytic exercise attenuated the rise of interleukin-6, tumour necrosis factor-α and high-sensitivity C-reactive protein after the HD procedure. Intra-dialytic exercise was not associated with an observable rise in blood endotoxin, although it may ameliorate the inflammatory effects of the HD procedure. Larger studies are needed to confirm this finding. © 2017 S. Karger AG, Basel.

An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood*

PubMed Central

Li, Siyang; Plouffe, Brian D.; Belov, Arseniy M.; Ray, Somak; Wang, Xianzhe; Murthy, Shashi K.; Karger, Barry L.; Ivanov, Alexander R.

2015-01-01

Isolation and molecular characterization of rare cells (e.g. circulating tumor and stem cells) within biological fluids and tissues has significant potential in clinical diagnostics and personalized medicine. The present work describes an integrated platform of sample procurement, preparation, and analysis for deep proteomic profiling of rare cells in blood. Microfluidic magnetophoretic isolation of target cells spiked into 1 ml of blood at the level of 1000–2000 cells/ml, followed by focused acoustics-assisted sample preparation has been coupled with one-dimensional PLOT-LC-MS methodology. The resulting zeptomole detection sensitivity enabled identification of ∼4000 proteins with injection of the equivalent of only 100–200 cells per analysis. The characterization of rare cells in limited volumes of physiological fluids is shown by the isolation and quantitative proteomic profiling of first MCF-7 cells spiked into whole blood as a model system and then two CD133+ endothelial progenitor and hematopoietic cells in whole blood from volunteers. PMID:25755294

[Participation of leucocytes in pathogenesis of primary forms of lower limb chronic venous disease].

PubMed

Bogachev, V Iu; Golovanova, O V; Sergeeva, N A; Kuznetsov, A N

2011-01-01

The purpose of the study was to test the hypothesis on participation of WBCs in damaging the venous wall in patients presenting with primary forms of lower limb chronic venous diseases LLCVD . The study included a total of fifteen consecutively selected patients (13 women and 2 men) diagnosed as having grade C2-C-4 LLCVD according to the CEAP classification. Static loading (30 minutes in the sitting position) was followed by simultaneous sampling of blood from the varicose vein of the cms and ulnar vein. The total blood count including determination of both the absolute values and percentage of blood formed elements was performed using the automated haematological counter «Advia 7» («Bayer», USA). The obtained findings were statistically processed using the Microsoft Office Excel software by means of the pared two-sample τ-test for the average values. The number of leukocytes and their subpopulations in blood samples obtained from the crural varicose veins turned out to be significantly less as compared with that in blood sampled from the ulnar vein. Thus, blood sampled from the crural varicose veins demonstrated a decrease in the counts of WBC by 9.6% in fourteen (93.3%) patients, that of neutrophils by 4.9% in twelve (80%) patients, that of lymphocytes by 16,8% in fifteen (100%) patients, and that oi monocytes by 24% in twelve (80%) patients. The mentioned differences were statistically significant at a = 0.05. The eosinophilic counts in blood sampled from the upper and lower extremities appeared similar in 66.7% of the examined subjects. In 33.3% of cases the eosinophilic count in blood samples from crural varicose vein was by 16.7% lower than that for blood samples form the ulnar vein. No differences for the rest parameters of the clinical blood count were revealed. The absolute lymphocytic count in the blood samples taken after the 30-minute static loading from the crural varicose veins was significantly lower as compared with that in blood sampled form the cubital vein. The counts for RBCs and blood platelets, as well as other qualitative haematological indices (haemoglobin, haematocrit, average volume of the RBC, erythrocytic diameter, etc.) in blood sampled form crural and ulnar veins in the same patient were identical, thus strongly suggesting the lack of either haemodynamic or haemorheological phenomena capable of leading to redistribution of the blood formed elements in varicose veins. Hence a decrease in the counts of leukocytes and their subpopulations in blood sampled from crural varicose veins might be associated with the «leukocytic trap» phenomenon.

Continuing education: online monitoring of haemodialysis dose.

PubMed

Vartia, Aarne

2018-01-25

Kt/V urea reflects the efficacy of haemodialysis scaled to patient size (urea distribution volume). The guidelines recommend monthly Kt/V measurements based on blood samples. Modern haemodialysis machines are equipped with accessories monitoring the dose online at every session without extra costs, blood samples and computers. To describe the principles, devices, benefits and shortcomings of online monitoring of haemodialysis dose. A critical literature overview and discussion. UV absorbance methods measure Kt/V, ionic dialysance Kt (product of clearance and treatment time; cleared volume without scaling). Both are easy and useful methods, but comparison is difficult due to problems in scaling of the dialysis dose to the patient's size. The best dose estimation method is the one which predicts the quality of life and survival most accurately. There is some evidence on the predictive value of ionic dialysance Kt, but more documentation is required on the UV method. Online monitoring is a useful tool in everyday quality assurance, but blood samples are still required for more accurate kinetic modelling. After reading this article the reader should be able to: Understand the elements of the Kt/V equation for dialysis dose. Compare and contrast different methods of measurement of dialysis dose. Reflect on the importance of adequate dialysis dose for patient survival and life quality. © 2018 European Dialysis and Transplant Nurses Association/European Renal Care Association.

Development of a Novel Multipenicillin Assay and Assessment of the Impact of Analyte Degradation: Lessons for Scavenged Sampling in Antimicrobial Pharmacokinetic Study Design.

PubMed

Kipper, Karin; Barker, Charlotte I S; Standing, Joseph F; Sharland, Mike; Johnston, Atholl

2018-01-01

Penicillins are widely used to treat infections in children; however, the evidence is continuing to evolve in defining the optimal dosing. Modern pediatric pharmacokinetic study protocols frequently favor opportunistic, "scavenged" sampling. This study aimed to develop a small-volume single assay for five major penicillins and to assess the influence of sample degradation on inferences made using pharmacokinetic modeling, to investigate the suitability of scavenged sampling strategies. Using a rapid ultrahigh-performance liquid chromatographic-tandem mass spectrometric method, an assay for five penicillins (amoxicillin, ampicillin, benzylpenicillin, piperacillin, and flucloxacillin) in blood plasma was developed and validated. Penicillin stabilities were evaluated under different conditions. Using these data, the impact of drug degradation on inferences made during pharmacokinetic modeling was evaluated. All evaluated penicillins indicated good stability at room temperature (23 ± 2°C) over 1 h, remaining in the range of 98 to 103% of the original concentration. More-rapid analyte degradation had already occurred after 4 h, with stability ranging from 68% to 99%. Stability over longer periods declined: degradation of up to 60% was observed with delayed sample processing of up to 24 h. Modeling showed that analyte degradation can lead to a 30% and 28% bias in clearance and volume of distribution, respectively, and falsely show nonlinearity in clearance. Five common penicillins can now be measured in a single low-volume blood sample. Beta-lactam chemical instability in plasma can cause misleading pharmacokinetic modeling results, which could impact upon model-based dosing recommendations and the forthcoming era of beta-lactam therapeutic drug monitoring. Copyright © 2017 American Society for Microbiology.

[Optimization of isolation of the concentrate of stem cells from the umbilical blood].

PubMed

Tiumina, O V; Savchenko, V G; Gusarova, G I; Pavlov, V V; Zharkov, M N; Volchkov, S E; Rossiev, V A; Gridasov, G N

2005-01-01

To study correlations between body mass and height of the newborn, Apgar scale estimates, gestation time, volume of the obtained umbilical blood (UB), number of nucleated cells (NC); to compare manual and automatic modes of UB processing. 330 procurements of UB were made, 230 (69.7%) samples were frozen. Comparison of 2 techniques of UB processing was made in 73 cases of double centrifugation with hydroxyethylstarch (HES) and 47 cases of using separator Sepax (Biosafe, Switzerland). Blood cell count before and after UB processing and number of CD34+ cells were estimated. A correlation analysis was made of dependence of the volume of 102 samples of UB on the weight (r = 0.268, p < 0.01) and height of the fetus (r = 0.203, p < 0.05), estimation by Apgar scale (r = -0.092, p < 0.1) and gestation term (r = -0.003, p > 0.1); analysis of the number of NC dependence on the volume of UB (r = 0.102 p < 0.1), mass (r = 0.073 p > 0.1) and fetus height (r = 0.121 p > 0.1), gestation time (r = 0.159 p > 0.1), Apgar scale assessment (r = -0.174 p > 0.1). In manual UB management NC yield made up 71.9 +/- 6.7%, in automatic--81 +/- 8.0% (p < 0.05). Percent of erythrocytes removal was 73 +/- 5.7% and 80.5 +/- 6.1% (p < 0.05), respectively. A weak correlation was found between UB volume, mass and height of the fetus. The number of NC in UB depends on none of the parameters. Automatic processing of UB provides a greater release of NC and better elimination of erythrocytes in minimal risk of contamination.

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.

Development of Skylab medical equipment and flight preparations

NASA Technical Reports Server (NTRS)

Johnston, R. S.; Stonesifer, J. C.; Hawkins, W. R.

1975-01-01

The major medical systems in the Skylab orbital workshop are described. They comprise the food system, the waste management system, operational bioinstrumentation, personal hygiene, gas sampling, an inflight medical support system, and a cardiovascular counterpressure garment. Life sciences experiments carried out aboard Skylab are also reviewed; these include an ergometer and metabolic analyzer, a lower-body negative pressure device, an electrode harness and body temperature probe, a blood pressure cuff, a leg volume measuring band, sleep studies, a body-mass measuring device, a rotating litter chair, a blood sample processor, and small-mass measuring apparatus. All performance requirements were met with the equipment, and no failures were encountered.

Dielectrophoretic isolation and detection of cancer-related circulating cell-free DNA biomarkers from blood and plasma

PubMed Central

Sonnenberg, Avery; Marciniak, Jennifer Y.; Skowronski, Elaine A.; Manouchehri, Sareh; Rassenti, Laura; Ghia, Emanuela M.; Widhopf, George F.; Kipps, Thomas J.; Heller, Michael J.

2014-01-01

Conventional methods for the isolation of cancer-related circulating cell-free (ccf) DNA from patient blood (plasma) are time consuming and laborious. A DEP approach utilizing a microarray device now allows rapid isolation of ccf-DNA directly from a small volume of unprocessed blood. In this study, the DEP device is used to compare the ccf-DNA isolated directly from whole blood and plasma from 11 chronic lymphocytic leukemia (CLL) patients and one normal individual. Ccf-DNA from both blood and plasma samples was separated into DEP high-field regions, after which cells (blood), proteins, and other biomolecules were removed by a fluidic wash. The concentrated ccf-DNA was detected on-chip by fluorescence, and then eluted for PCR and DNA sequencing. The complete process from blood to PCR required less than 10 min; an additional 15 min was required to obtain plasma from whole blood. Ccf-DNA from the equivalent of 5 µL of CLL blood and 5 µL of plasma was amplified by PCR using Ig heavy-chain variable (IGHV) specific primers to identify the unique IGHV gene expressed by the leukemic B-cell clone. The PCR and DNA sequencing results obtained by DEP from all 11 CLL blood samples and from 8 of the 11 CLL plasma samples were exactly comparable to the DNA sequencing results obtained from genomic DNA isolated from CLL patient leukemic B cells (gold standard). PMID:24723219

Dielectrophoretic isolation and detection of cancer-related circulating cell-free DNA biomarkers from blood and plasma.

PubMed

Sonnenberg, Avery; Marciniak, Jennifer Y; Skowronski, Elaine A; Manouchehri, Sareh; Rassenti, Laura; Ghia, Emanuela M; Widhopf, George F; Kipps, Thomas J; Heller, Michael J

2014-07-01

Conventional methods for the isolation of cancer-related circulating cell-free (ccf) DNA from patient blood (plasma) are time consuming and laborious. A DEP approach utilizing a microarray device now allows rapid isolation of ccf-DNA directly from a small volume of unprocessed blood. In this study, the DEP device is used to compare the ccf-DNA isolated directly from whole blood and plasma from 11 chronic lymphocytic leukemia (CLL) patients and one normal individual. Ccf-DNA from both blood and plasma samples was separated into DEP high-field regions, after which cells (blood), proteins, and other biomolecules were removed by a fluidic wash. The concentrated ccf-DNA was detected on-chip by fluorescence, and then eluted for PCR and DNA sequencing. The complete process from blood to PCR required less than 10 min; an additional 15 min was required to obtain plasma from whole blood. Ccf-DNA from the equivalent of 5 μL of CLL blood and 5 μL of plasma was amplified by PCR using Ig heavy-chain variable (IGHV) specific primers to identify the unique IGHV gene expressed by the leukemic B-cell clone. The PCR and DNA sequencing results obtained by DEP from all 11 CLL blood samples and from 8 of the 11 CLL plasma samples were exactly comparable to the DNA sequencing results obtained from genomic DNA isolated from CLL patient leukemic B cells (gold standard). © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cerebral edema, mass effects, and regional blood volume in man.

PubMed

Penn, R D; Kurtz, D

1977-03-01

The authors conducted quantitative analysis of computerized tomography (CT) scans to measure tumor size, cerebral edema, and regional blood volume in man. Mass lesions without edema caused a local reduction in blood volume. Cerebral edema also reduced blood volume in proportion to its severity. Consideration of the electrolyte changes and water shifts in white-matter edema suggested that the decrease in absorption coefficient seen in CT scans was due to the increase in water content. Thus, in cerebral edema separation of blood vessels as well as increased interstitial pressure decrease blood volume, and the regional differences in turn reflect pressure gradients within the brain.

Blood Volume Response to Physical Activity and Inactivity

DTIC Science & Technology

2007-07-01

feedback to promote volume excretion and maintain blood volume at its normal baseline level. Physical activity does not alter either arterial pressure or...although this is less clear. The increase in total blood volume then increases the ability to maintain a high stroke volume and a lower heart rate for...compelling evidence that reduced blood vol- ume with age may be a result of a sedentary, high caloric lifestyle rather than the aging process. There

Development of a magnetic lab-on-a-chip for point-of-care sepsis diagnosis

NASA Astrophysics Data System (ADS)

Schotter, Joerg; Shoshi, Astrit; Brueckl, Hubert

2009-05-01

We present design criteria, operation principles and experimental examples of magnetic marker manipulation for our magnetic lab-on-a-chip prototype. It incorporates both magnetic sample preparation and detection by embedded GMR-type magnetoresistive sensors and is optimized for the automated point-of-care detection of four different sepsis-indicative cytokines directly from about 5 μl of whole blood. The sample volume, magnetic particle size and cytokine concentration determine the microfluidic volume, sensor size and dimensioning of the magnetic gradient field generators. By optimizing these parameters to the specific diagnostic task, best performance is expected with respect to sensitivity, analysis time and reproducibility.

Biochemical profile of African American women during three trimesters of pregnancy and at delivery.

PubMed

Knight, E M; Spurlock, B G; Edwards, C H; Johnson, A A; Oyemade, U J; Cole, O J; West, W L; Manning, M; James, H; Laryea, H

1994-06-01

The data presented are the results from a prospective observational study which was conducted to investigate the effects of nutrition and other related factors on the outcome of pregnancy in nulliparous African American women 16-35 years old. Fasting blood samples were collected from the women during the first, second and third trimesters of pregnancy. At delivery, both maternal and cord samples were collected. Biochemical variables such as, serum folate, vitamin B12, ascorbic acid, vitamin E, ferritin, selected minerals as well as complete blood count (CBC) and red cell folate were analyzed in the blood samples. The concentrations of hematocrit, hemoglobin, white blood cells, red blood cells and vitamin B12 were below the reference non-pregnant ranges throughout gestation. Maternal concentrations of folate and vitamin E increased sequentially with increased gestational age. Serum ferritin, during the third trimester, declined to 58% of the first trimester concentration. Maternal levels of ferritin at delivery were one third of the values found in the infant (cord) sample. Cord levels of folate, ascorbic acid and vitamin B12 were higher than the concentrations in the maternal delivery samples. The data suggest that among this group of pregnant women, major physiological changes, such as plasma volume expansion which alters blood chemistry and maternal to fetal transfer of nutrients, were similar to the findings of other investigators. In this population however, the findings for serum and whole blood folate are contrary to those reported by other researchers, and the sequential increase in the maternal concentration of the vitamin during pregnancy could be attributed to the use of vitamin supplements.

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma

DOE PAGES

Jacobs, Anne C.; Fair, Jeanne Marie

2015-10-09

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. Here, we tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throatedmore » flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. Finally, we caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis.« less

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma.

PubMed

Jacobs, Anne C; Fair, Jeanne M

2016-01-01

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. We tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throated flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. We caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis. Copyright © 2015 Elsevier Inc. All rights reserved.

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma

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

Jacobs, Anne C.; Fair, Jeanne Marie

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. Here, we tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throatedmore » flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. Finally, we caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis.« less

SU-E-I-79: Source Geometry Dependence of Gamma Well-Counter Measurements

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

Park, M; Belanger, A; Kijewski, M

Purpose: To determine the effect of liquid sample volume and geometry on counting efficiency in a gamma well-counter, and to assess the relative contributions of sample geometry and self-attenuation. Gamma wellcounters are standard equipment in clinical and preclinical studies, for measuring patient blood radioactivity and quantifying animal tissue uptake for tracer development and other purposes. Accurate measurements are crucial. Methods: Count rates were measured for aqueous solutions of 99m- Tc at four liquid volume values in a 1-cm-diam tube and at six volume values in a 2.2-cm-diam vial. Total activity was constant for all volumes, and data were corrected formore » decay. Count rates from a point source in air, supported by a filter paper, were measured at seven heights between 1.3 and 5.7 cm from the bottom of a tube. Results: Sample volume effects were larger for the tube than for the vial. For the tube, count efficiency relative to a 1-cc volume ranged from 1.05 at 0.05 cc to 0.84 at 3 cc. For the vial, relative count efficiency ranged from 1.02 at 0.05 cc to 0.87 at 15 cc. For the point source, count efficiency relative to 1.3 cm from the tube bottom ranged from 0.98 at 1.8 cm to 0.34 at 5.7 cm. The relative efficiency of a 3-cc liquid sample in a tube compared to a 1-cc sample is 0.84; the average relative efficiency for the solid sample in air between heights in the tube corresponding to the surfaces of those volumes (1.3 and 4.8 cm) is 0.81, implying that the major contribution to efficiency loss is geometry, rather than attenuation. Conclusion: Volume-dependent correction factors should be used for accurate quantitation radioactive of liquid samples. Solid samples should be positioned at the bottom of the tube for maximum count efficiency.« less

Salting-out homogenous extraction followed by ionic liquid/ionic liquid liquid-liquid micro-extraction for determination of sulfonamides in blood by high performance liquid chromatography.

PubMed

Liu, Zhongling; Yu, Wei; Zhang, Hanqi; Gu, Fanbin; Jin, Xiangqun

2016-12-01

Salting-out homogenous extraction followed by ionic liquid/ionic liquid dispersive liquid-liquid micro-extraction system was developed and applied to the extraction of sulfonamides in blood. High-performance liquid chromatography was applied to the determination of the analytes. The blood sample was centrifuged to obtain the serum. After the proteins in the serum were removed in the presence of acetonitrile, ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate, dipotassium hydrogen phosphate, ionic liquid 1-Hexyl-3-methylimidazolium hexafluorophosphate were added into the resulting solution. After the resulting mixture was ultrasonically shaken and centrifuged, the precipitate was separated. The acetonitrile was added in the precipitate and the analytes were extracted into the acetonitrile phase. The parameters affecting the extraction efficiency, such as volume of ionic liquid, amount of dipotassium hydrogen phosphate, volume of dispersant, extraction time and temperature were investigated. The limits of detection of sulfamethizole (STZ), sulfachlorpyridazine (SCP), sulfamethoxazole (SMX) and Sulfisoxazole (SSZ) were 4.78, 3.99, 5.21 and 3.77μgL -1 , respectively. When the present method was applied to the analysis of real blood samples, the recoveries of analytes ranged from 90.0% to 113.0% and relative standard deviations were lower than 7.2%. Copyright © 2016 Elsevier B.V. All rights reserved.

An assessment of the potential of laser-induced breakdown spectroscopy (LIBS) for the analysis of cesium in liquid samples of biological origin.

PubMed

Metzinger, Anikó; Kovács-Széles, Eva; Almási, István; Galbács, Gábor

2014-01-01

The present study describes the development of an analytical method for the determination of cesium in biological fluid samples (human urine and blood samples) by laser-induced breakdown spectroscopy (LIBS). The developed method is based on sample presentation by liquid-to-solid conversion, enhancing the emission signal by drying the liquid into small "pockets" created in a metal support (zinc plate), and allows the analysis to be carried out on as little as 1 μL of sample volume, in a closed sample cell. Absolute detection limits on the Cs I 852.1 nm spectral line were calculated by the IUPAC 3σ method to be 6 ng in the urine sample and 27 ng in the blood serum sample. It is estimated that LIBS may be used to detect highly elevated concentration levels of Cs in fluid samples taken from people potentially exposed to surges of Cs from non-natural sources.

A novel approach for quantitation of glucosylceramide in human dried blood spot using LC-MS/MS.

PubMed

Ji, Allena Ji; Wang, Haixing; Ziso-Qejvanaj, Enida; Zheng, Kefei; Chung, Lee Lee; Foley, Timothy; Chuang, Wei-Lien; Richards, Susan; Sung, Crystal

2015-01-01

Glucosylceramide, an efficacy biomarker for Gaucher Type 1 disease, exhibits poor solubility in polar solvents and whole blood which makes it difficult to prepare a homogenous blood standard. We developed a novel method using standard addition approach by spiking a small volume of analyte solution on the surface of prespotted dried blood spot. The whole spots were punched out for subsequent extraction and LC-MS/MS analysis. The assay performance met all validation acceptance criteria. Glucosylceramide concentrations in 50 paired plasma and dry blood spot samples obtained from Gaucher Type 1 patients were tested and the results demonstrated the feasibility of using the DBS method for clinical biomarker monitoring. The new approach greatly improves assay precision and accuracy.

Reductions in Central Venous Pressure by Lower Body Negative Pressure or Blood Loss Elicit Similar Hemodynamic Responses

DTIC Science & Technology

2014-05-29

echolamines, hematocrit, hemoglobin, arginine vasopressin, and blood gases. LBNP consisted of 5-min stages at 0, 15, 30, and 45 mmHg of suction. BL consisted...release of volume-regulating hormones such as arginine vasopressin (1, 2, 11, 18, 24, 37). Therefore, it is plausible that blood analyte responses to LBNP...regression analyses. A sample size of 8 subjects (age 32 3 yr; height 185.3 1.8 cm; weight 91.3 3.4 kg; BMI 26.6 0.8 kg/m2) was used for ANOVA

[Influence of postural changes on the variations of blood volume in hemodialysis and reincidence post-dialysis blood volume].

PubMed

López Ramón, I; Muro, B; Azcona, M; Moleres, M; Sagüés, C; Maeztu, B; Zubía, A; Martínez de Irujo, S

1997-01-01

The aim of the present work is to study the possible increment of blood volume, in the seated as well as the lying position, and to analyse the influence of changes of posture on the changes of blood volume during the hemodialysis. 17 patients were studied, evaluating the variations of the hematocrit in the extracorporeal circuit on entry into the dialyzer, by means of a photometric system (Crit-Line Instrument, Izasa). Every 15 minutes the values of the hematocrit were taken and heir repercussion in the blood volume in different changes of posture. Initially all of the patients improved their blood volume, both seated and lying down. During the session of hemodialysis and ultrafiltration, with the patient in a seated position, an important drop in the blood volume was noticed; this was the case in the 1st, 2nd and 3rd hours. This drop improved when the patient adopted a lying position and there was a "reincidence of post-dialysis blood volume" at the end. According to these results, the supine posture improves the refill, confirming that the patients position influences the vascular refill.

Alterations in hematologic indices during long-duration spaceflight.

PubMed

Kunz, Hawley; Quiriarte, Heather; Simpson, Richard J; Ploutz-Snyder, Robert; McMonigal, Kathleen; Sams, Clarence; Crucian, Brian

2017-01-01

Although a state of anemia is perceived to be associated with spaceflight, to date a peripheral blood hematologic assessment of red blood cell (RBC) indices has not been performed during long-duration space missions. This investigation collected whole blood samples from astronauts participating in up to 6-months orbital spaceflight, and returned those samples (ambient storage) to Earth for analysis. As samples were always collected near undock of a returning vehicle, the delay from collection to analysis never exceeded 48 h. As a subset of a larger immunologic investigation, a complete blood count was performed. A parallel stability study of the effect of a 48 h delay on these parameters assisted interpretation of the in-flight data. We report that the RBC and hemoglobin were significantly elevated during flight, both parameters deemed stable through the delay of sample return. Although the stability data showed hematocrit to be mildly elevated at +48 h, there was an in-flight increase in hematocrit that was ~3-fold higher in magnitude than the anticipated increase due to the delay in processing. While susceptible to the possible influence of dehydration or plasma volume alterations, these results suggest astronauts do not develop persistent anemia during spaceflight.

Comparison of human whole blood, plasma, and serum matrices for the determination of perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and other fluorochemicals

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

Ehresman, David J.; Froehlich, John W.; Olsen, Geary W.

2007-02-15

Interest in human exposure to perfluorinated acids, including perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), perfluorooctanesulfonate (PFOS), and perfluorooctanoate (PFOA) has led to their measurement in whole blood, plasma and serum. Comparison of measurements in these different blood-based matrices, however, has not been rigorously investigated to allow for across-matrix comparisons. This research evaluated concentrations of PFBS, PFHS, PFOS, and PFOA in whole blood collected in heparin (lithium) and ethylenediamine tetraacetic acid (EDTA), plasma samples collected in heparin and EDTA, and serum (from whole blood allowed to clot). Blood samples were collected from 18 voluntary participants employed at 3M Company. Solid phase extraction methodsmore » were used for all analytical sample preparations, and analyses were completed using high-pressure liquid chromatography/tandem mass spectrometry methods. Serum concentrations ranged from: limit of quantitation (LOQ, 5 ng/mL) to 25 ng/mL for PFBS; LOQ (5 ng/mL) to 75 ng/mL for PFHS; LOQ (5 ng/mL) to 880 ng/mL for PFOS; and LOQ (5 or 10 ng/mL) to 7320 ng/mL for PFOA. Values less than the LOQ were not included in the statistical analyses of the mean of the ratios of individual values for the matrices. PFBS was not quantifiable in most samples. Serum to plasma ratios for PFHS, PFOS, and PFOA were 1:1 and this ratio was independent of the level of concentrations measured. Serum or plasma to whole blood ratios, regardless of the anticoagulant used, approximated 2:1. The difference between plasma and serum and whole blood corresponded to volume displacement by red blood cells, suggesting that the fluorochemicals are not found intracellularly or attached to the red blood cells.« less

Application of Zeeman graphite furnace atomic absorption spectrometry with high-frequency modulation polarization for the direct determination of aluminum, beryllium, cadmium, chromium, mercury, manganese, nickel, lead, and thallium in human blood.

PubMed

Ivanenko, Natalya B; Solovyev, Nikolay D; Ivanenko, Anatoly A; Ganeev, Alexander A

2012-10-01

Determination of aluminum (Al), beryllium (Be), cadmium (Cd), chromium (Cr), mercury (Hg), manganese (Mn), nickel (Ni), lead (Pb), and thallium (Tl) concentrations in human blood using high-frequency modulation polarization Zeeman graphite furnace atomic absorption spectrometry (GFAAS) was performed. No sample digestion was used in the current study. Blood samples were diluted with deionized water or 0.1 % (m/v) Triton X-100 solution for Tl. Dilution factors ranged from 1/5 per volume for Be and Tl to 1/20 per volume for Cd and Pb. For Tl, Cd, and Hg, noble metals (gold, platinum, rhodium, etc.) were applied as surface modifiers. To mitigate chloride interference, 2 % (m/v) solution of NH(4)NO(3) was used as matrix modifier for Tl and Ni assessment. The use of Pd(NO(3))(2) as oxidative modifier was necessary for blood Hg and Tl measurement. Validation of the methods was performed by analyzing two-level reference material Seronorm. The precision of the designed methods as relative SD was between 4 and 12 % (middle of a dynamic range) depending on the element. For additional validation, spiked blood samples were analyzed. Limits of detection (LoDs, 3σ, n = 10) for undiluted blood samples were 2.0 μg L(-1) for Al, 0.08 μg L(-1) for Be, 0.10 μg L(-1) for Cd, 2.2 μg L(-1) for Cr, 7 μg L(-1) for Hg, 0.4 μg L(-1) for Mn, 2.3 μg L(-1) for Ni, 3.4 μg L(-1) for Pb, and 0.5 μg L(-1) for Tl. The LoDs achieved allowed determination of Al, Cd, Cr, Mn, Ni, and Pb at both toxic and background levels. Be, Hg, and Tl could be reliably measured at toxic levels only. The methods developed are used for clinical diagnostics and biological monitoring of work-related exposure.

Analysis of human serum and whole blood for mineral content by ICP-MS and ICP-OES: development of a mineralomics method.

PubMed

Harrington, James M; Young, Daniel J; Essader, Amal S; Sumner, Susan J; Levine, Keith E

2014-07-01

Minerals are inorganic compounds that are essential to the support of a variety of biological functions. Understanding the range and variability of the content of these minerals in biological samples can provide insight into the relationships between mineral content and the health of individuals. In particular, abnormal mineral content may serve as an indicator of illness. The development of robust, reliable analytical methods for the determination of the mineral content of biological samples is essential to developing biological models for understanding the relationship between minerals and illnesses. This paper describes a method for the analysis of the mineral content of small volumes of serum and whole blood samples from healthy individuals. Interday and intraday precision for the mineral content of the blood (250 μL) and serum (250 μL) samples was measured for eight essential minerals--sodium (Na), calcium (Ca), magnesium (Mg), potassium (K), iron (Fe), zinc (Zn), copper (Cu), and selenium (Se)--by plasma spectrometric methods and ranged from 0.635 to 10.1% relative standard deviation (RSD) for serum and 0.348-5.98% for whole blood. A comparison of the determined ranges for ten serum samples and six whole blood samples provided good agreement with literature reference ranges. The results demonstrate that the digestion and analysis methods can be used to reliably measure the content of these minerals and potentially of other minerals.

Magnetic measurements on human erythrocytes: Normal, beta thalassemia major, and sickle

NASA Astrophysics Data System (ADS)

Sakhnini, Lama

2003-05-01

In this article magnetic measurements were made on human erythrocytes at different hemoglobin states (normal and reduced hemoglobin). Different blood samples: normal, beta thalassemia major, and sickle were studied. Beta thalassemia major and sickle samples were taken from patients receiving lifelong blood transfusion treatment. All samples examined exhibited diamagnetic behavior. Beta thalassemia major and sickle samples showed higher diamagnetic susceptibilities than that for the normal, which was attributed to the increase of membrane to hemoglobin volume ratio of the abnormal cells. Magnetic measurements showed that the erythrocytes in the reduced state showed less diamagnetic response in comparison with erythrocytes in the normal state. Analysis of the paramagnetic component of magnetization curves gave an effective magnetic moment of μeff=7.6 μB per reduced hemoglobin molecule. The same procedure was applied to sickle and beta thalassemia major samples and values for μeff were found to be comparable to that of the normal erythrocytes.

Cyclic volatile methylsiloxanes in human blood as markers for ruptured silicone gel-filled breast implants.

PubMed

Rosendahl, Pia; Hippler, Joerg; Schmitz, Oliver J; Hoffmann, Oliver; Rusch, Peter

2016-05-01

The replacement of medical-grade silicone with industrial-grade silicone material in some silicone gel-filled breast implants (SBI) manufactured by Poly Implant Prothèse and Rofil Medical Nederland B.V., reported in 2010, which resulted in a higher rupture tendency of these SBI, demonstrates the need for non-invasive, sensitive monitoring and screening methods. Therefore a sensitive method based on large volume injection-gas chromatography coupled to mass spectrometry (LVI-GC/MS) was developed to determine octamethylcyclotetrasiloxane (D4), decamethylcyclopentasiloxane (D5), and dodecamethylcyclo-hexasiloxane (D6) in blood samples from women with intact (n = 13) and ruptured SBI (n = 11). With dichloromethane extraction, sample cooling during preparation, and analysis extraction efficiencies up to 100 % and limits of detection of 0.03-0.05 ng D4-D6/g blood were achieved. Blood samples from women with SBI were investigated. In contrast to women with intact SBI, in blood from women with ruptured SBI higher D4 and D6 concentrations up to 0.57 ng D4/g blood and 0.16 ng D6/g blood were detected. With concentrations above 0.18 D4 ng/blood and 0.10 ng D6/g blood as significant criteria for ruptured SBI, this developed analytical preoperative diagnostic method shows a significant increase of the recognition rate. Finally a higher precision (error rate 17%) than the commonly used clinical diagnostic method, mamma sonography (error rate 46%), was achieved.

Detection of prions in blood from patients with variant Creutzfeldt-Jakob disease

PubMed Central

Concha-Marambio, Luis; Pritzkow, Sandra; Moda, Fabio; Tagliavini, Fabrizio; Ironside, James W.; Schulz, Paul E.; Soto, Claudio

2017-01-01

Human prion diseases are infectious and invariably fatal neurodegenerative diseases. They include sporadic Creutzfeldt-Jakob disease (sCJD), the most common form, and variant CJD (vCJD), which is caused by interspecies transmission of prions from cattle infected by bovine spongiform encephalopathy. Development of a biochemical assay for the sensitive, specific, early, and noninvasive detection of prions (PrPSc) in the blood of patients affected by prion disease is a top medical priority to increase the safety of the blood supply. vCJD has already been transmitted from human to human by blood transfusion, and the number of asymptomatic carriers of vCJD in the U.K. alone is estimated to be 1 in 2000 people. We used the protein misfolding cyclic amplification (PMCA) technique to analyze blood samples from 14 cases of vCJD and 153 controls, including patients affected by sCJD and other neurodegenerative or neurological disorders as well as healthy subjects. Our results showed that PrPSc could be detected with 100% sensitivity and specificity in blood samples from vCJD patients. Detection was possible in any of the blood fractions analyzed and could be done with as little as a few microliters of sample volume. The PrPSc concentration in blood was estimated to be ~0.5 pg/ml. Our findings suggest that PMCA may be useful for premortem noninvasive diagnosis of vCJD and to identify prion contamination of the blood supply. Further studies are needed to fully validate the technology. PMID:28003548

The role of alcohol, marijuana, and other drugs in the accidents of injured drivers. Volume 1, Findings

DOT National Transportation Integrated Search

1982-01-01

Using samples of blood obtained from 497 injured drivers at a Rochester, NY hospital, this study determined the incidence rates of alcohol, THC (marijuana agent), and other drugs. Accident data (police reports, driver interviews) were also collected,...

The role of alcohol, marijuana, and other drugs in the accidents of injured drivers. Volume 2, Appendices

DOT National Transportation Integrated Search

1982-01-01

Using samples of blood obtained from 497 injured drivers at a Rochester, NY hospital, this study determined the incidence rates of alcohol, THC (marijuana agent), and other drugs. Accident data (police reports, driver interviews) were also collected,...

21 CFR 864.5600 - Automated hematocrit instrument.

Code of Federal Regulations, 2012 CFR

2012-04-01

... measures the packed red cell volume of a blood sample to distinguish normal from abnormal states, such as anemia and erythrocytosis (an increase in the number of red cells). (b) Classification. Class II... § 864.5600 Automated hematocrit instrument. (a) Identification. An automated hematocrit instrument is a...

21 CFR 864.5600 - Automated hematocrit instrument.

Code of Federal Regulations, 2011 CFR

2011-04-01

... measures the packed red cell volume of a blood sample to distinguish normal from abnormal states, such as anemia and erythrocytosis (an increase in the number of red cells). (b) Classification. Class II... § 864.5600 Automated hematocrit instrument. (a) Identification. An automated hematocrit instrument is a...

21 CFR 864.5600 - Automated hematocrit instrument.

Code of Federal Regulations, 2014 CFR

2014-04-01

... measures the packed red cell volume of a blood sample to distinguish normal from abnormal states, such as anemia and erythrocytosis (an increase in the number of red cells). (b) Classification. Class II... § 864.5600 Automated hematocrit instrument. (a) Identification. An automated hematocrit instrument is a...

21 CFR 864.5600 - Automated hematocrit instrument.

Code of Federal Regulations, 2013 CFR

2013-04-01

... measures the packed red cell volume of a blood sample to distinguish normal from abnormal states, such as anemia and erythrocytosis (an increase in the number of red cells). (b) Classification. Class II... § 864.5600 Automated hematocrit instrument. (a) Identification. An automated hematocrit instrument is a...

21 CFR 864.5600 - Automated hematocrit instrument.

Code of Federal Regulations, 2010 CFR

2010-04-01

... measures the packed red cell volume of a blood sample to distinguish normal from abnormal states, such as anemia and erythrocytosis (an increase in the number of red cells). (b) Classification. Class II... § 864.5600 Automated hematocrit instrument. (a) Identification. An automated hematocrit instrument is a...

Comparison of hematologic values in blood samples with lithium heparin or dipotassium ethylenediaminetetraacetic acid anticoagulants in Hispaniolan Amazon parrots (Amazona ventralis).

PubMed

Guzman, David Sanchez-Migallon; Mitchell, Mark A; Gaunt, Stephen D; Beaufrère, Hugues; Tully, Thomas N

2008-06-01

Blood samples were collected from 20 Hispaniolan Amazon parrots (Amazona ventralis) and were divided into tubes that contained dipotassium ethylenediaminetetraacetic acid (K2EDTA) and lithium heparin. Complete blood cell counts were determined in each sample within 2 hours of collection. The level of agreement in results was moderate for plasma protein, packed cell volume (PCV), and leukocyte, monocyte, and lymphocyte counts between the anticoagulants. Plasma protein and PCV values were significantly lower in samples with lithium heparin than in those with K2EDTA, whereas lymphocyte numbers were significantly higher in lithium heparin samples than in K2EDTA samples. The level of agreement was good for the other cell types (heterophils, eosinophils, and basophils) when comparing the different anticoagulants. The poor level of agreement between anticoagulants with the increase in thrombocyte clumping in lithium heparin samples indicates that the use of lithium heparin as anticoagulant may affect thrombocyte count. No negative effects on morphology and staining of blood cells were apparent in smears from heparin samples compared with K2EDTA samples. Within the different values compared, the limits of agreement are small enough to be confident that lithium heparin can be used for routine CBC counts in a clinical setting. The use of the same anticoagulant should be recommended to follow trends within the same patient, especially when considering plasma protein concentration, PCV, and lymphocyte count.

Evaluation of plasma chemistry and haematological studies on chickens infected with Eimeria tenella and E acervulina.

PubMed

Fukata, T; Komba, Y; Sasai, K; Baba, E; Arakawa, A

1997-07-12

Plasma chemistry and haematological studies were conducted on chickens with coccidiosis. Male White Leghorn chickens, of two weeks old, were inoculated with 5 x 10(4) Eimeria tenella sporulated oocysts or with 1 x 10(6) E acervulina sporulated oocysts. Blood samples were taken four, seven and 11 days after inoculation. A wet chemistry system was applied to measure the plasma activities of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyltransferase, creatine kinase, amylase and lactate dehydrogenase and the concentrations of creatine, total bilirubin, urate, total cholesterol, total protein, albumin, glucose and triglycerides. A dry chemistry system was applied to measure sodium, potassium, chloride and calcium. The number of red blood cells and packed cell volume were determined by a micro cell counter and blood pH was measured with a blood gas analyser. The erythrocyte count, packed cell volume, sodium and chloride levels in the chickens infected with E tenella were significantly (P < 0.05) lower than those of the uninfected controls. The significant decrease in blood pH of the chickens infected with E acervulina suggests malabsorption associated with duodenal lesions induced by the infection.

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

Preliminary Measures of Instructor Learning in Teaching Junctional Tourniquet Users.

PubMed

Kragh, John F; Aden, James K; Shackelford, Stacy; Dubick, Michael A

2016-01-01

The objective of the present study was to assess the effect of instructor learning on student performance in use of junctional tourniquets. From a convenience sample of data available after another study, we used a manikin for assessment of control of bleeding from a right groin gunshot wound. Blood loss was measured by the instructor while training users. The data set represented a group of 30 persons taught one at a time. The first measure was a plot of mean blood loss volumes for the sequential users. The second measure was a plot of the cumulative sum (CUSUM) of mean blood loss (BL) volumes for users. Mean blood loss trended down as the instructor gained experience with each newly instructed user. User performance continually improved as the instructor gained more experience with teaching. No plateau effect was observed within the 30 users. The CUSUM plot illustrated a turning point or cusp at the seventh user. The prior portion of the plot (users 1-7) had the greatest improvement; performance did not improve as much thereafter. The improvement after the seventh user was the only change detected in the instructor's trend of performance. The instructor's teaching experience appeared to directly affect user performance; in a model of junctional hemorrhage, the volume of blood loss from the manikin during junctional tourniquet placement was a useful metric of instructor learning. The CUSUM technique detected a small but meaningful change in trend where the instructor learning curve was greatest while working with the first seven users. 2016.

Quantification of Dynamic 11C-Phenytoin PET Studies.

PubMed

Mansor, Syahir; Boellaard, Ronald; Froklage, Femke E; Bakker, Esther D M; Yaqub, Maqsood; Voskuyl, Rob A; Schwarte, Lothar A; Verbeek, Joost; Windhorst, Albert D; Lammertsma, Adriaan

2015-09-01

The overexpression of P-glycoprotein (Pgp) is thought to be an important mechanism of pharmacoresistance in epilepsy. Recently, (11)C-phenytoin has been evaluated preclinically as a tracer for Pgp. The aim of the present study was to assess the optimal plasma kinetic model for quantification of (11)C-phenytoin studies in humans. Dynamic (11)C-phenytoin PET scans of 6 healthy volunteers with arterial sampling were acquired twice on the same day and analyzed using single- and 2-tissue-compartment models with and without a blood volume parameter. Global and regional test-retest (TRT) variability was determined for both plasma to tissue rate constant (K1) and volume of distribution (VT). According to the Akaike information criterion, the reversible single-tissue-compartment model with blood volume parameter was the preferred plasma input model. Mean TRT variability ranged from 1.5% to 16.9% for K1 and from 0.5% to 5.8% for VT. Larger volumes of interest showed better repeatabilities than smaller regions. A 45-min scan provided essentially the same K1 and VT values as a 60-min scan. A reversible single-tissue-compartment model with blood volume seems to be a good candidate model for quantification of dynamic (11)C-phenytoin studies. Scan duration may be reduced to 45 min without notable loss of accuracy and precision of both K1 and VT, although this still needs to be confirmed under pathologic conditions. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

Effect of carbohydrate composition on fluid balance, gastric emptying, and exercise performance.

PubMed

Cole, K J; Grandjean, P W; Sobszak, R J; Mitchell, J B

1993-12-01

This study examined the effects of serial feedings of different carbohydrate (CHO) solutions on plasma volume, gastric emptying (GE), and performance during prolonged cycling exercise. Solutions containing 6 g% glucose-sucrose (CHO-6GS), 8.3 g% high fructose corn syrup (CHO-8HF), 6.3 g% high fructose corn syrup + 2 g% glucose polymer (CHO-8HP), and a water placebo (WP) were compared. Ten trained male cyclists performed four cycling trials consisting of 105 min at 70% VO2max followed by a 15-min all-out, self-paced performance ride. Every 15 min the men consumed one of the four test solutions. Blood samples were taken before, during, and after exercise to determine blood glucose and plasma volume changes. There were no significant differences in performance, GE, or plasma volume changes between trials. Blood glucose was significantly elevated at the 105-min time-point in all CHO trials when compared to WP. The CHO-8HF and CHO-8HP drinks resulted in a significantly higher delivery of CHO to the intestine. Higher rates of CHO oxidation during the steady-state ride were observed only with the CHO-6GS drink.

Whole body and tissue blood volumes of two strains of rainbow trout (Oncorhynchus mykiss)

USGS Publications Warehouse

Gingerich, W.H.; Pityer, R.A.; Rach, J.J.

1990-01-01

1. Estimates of apparent packed cell, plasma and total blood volumes for the whole body and for 13 selected tissues were compared between Kamloops and Wytheville strains of rainbow trout (Oncorhynchus mykiss) by the simultaneous injection of two vascular tracers, radiolabeled trout erythrocytes (51Cr-RBC) and radioiodated bovine serum albumin (125I-BSA).2. Whole body total blood volume, plasma volume and packed cell volume were slightly, but not significantly greater in the Wytheville trout, whereas, the apparent plasma volumes and total blood volumes in 4 of 13 tissues were significantly greater in the Kamloops strain.3. Differences were most pronounced in highly perfused organs, such as the liver and kidney and in organs of digestion such as the stomach and intestines.4. Differences in blood volumes between the two strains may be related to the greater permeability of the vascular membranes in the Kamloops strain fish.

Resuscitative therapy with erythropoietin reduces oxidative stress and inflammatory responses of vital organs in a rat severe fixed-volume hemorrhagic shock model.

PubMed

Ranjbaran, Mina; Kadkhodaee, Mehri; Seifi, Behjat; Mirzaei, Reza; Ahghari, Parisa

2018-01-01

Hemorrhagic shock (HS) still has a high mortality rate and none of the known resuscitative regimens completely reverse its adverse outcomes. This study investigated the effects of different models of resuscitative therapy on the healing of organ damage in a HS model. Male Wistar rats were randomized into six groups: Sham, without HS induction; HS, without resuscitation; HS+Blood, resuscitation with the shed blood; HS+Blood+NS, resuscitation with blood and normal saline; HS+Blood+RL, resuscitation with blood and Ringer's lactate; EPO, erythropoietin was added to the blood and RL. Blood and urine samples were obtained 3 h after resuscitation. Kidney, liver and brain tissue samples were harvested for multiple organ failure evaluation. Survival rate was the highest in the Sham, EPO and HS+Blood+RL groups compared to others. Plasma creatinine concentration, ALT, AST, urinary NAG activity and renal NGAL mRNA expression significantly increased in the HS+Blood+RL group compared to the Sham group. There was a significant increase in tissue oxidative stress markers and pro-inflammatory cytokines in HS+Blood+RL group compared to the Sham rats. EPO had more protective effects on multiple organ failure compared to the HS+Blood+RL group. EPO, as a resuscitative treatment, attenuated HS-induced organ damage. It seems that it has a potential to be attractive for clinical trials.

Acute Traumatic Coagulopathy: Whole Blood Thrombelastography Measures the Tip of the Iceberg

DTIC Science & Technology

2015-01-01

with the thromboelastography analyser. Blood Coagul Fibrinolysis. 2009;20(6):436Y439. 14. Engstrom M , Schott U, Romner B , Reinstrup P. Acidosis impairs...PAGES 7 19a. NAME OF RESPONSIBLE PERSON a. REPORT unclassified b . ABSTRACT unclassified c. THIS PAGE unclassified Standard Form 298 (Rev. 8-98...and 10KL) of 1- M 2-[morpholino]ethanesulfonic acidYbuffered saline (MBS) to a 300-KL volume sample in the TEG cup (Table 1). MBS was chosen for its

Blood volume and red cell life span (M113), part C

NASA Technical Reports Server (NTRS)

Johnson, P. C., Jr.

1973-01-01

Prechamber, in-chamber, and postchamber blood samples taken from Skylab simulation crewmembers did not indicate significant shortening of the red cell life span during the mission. This does not suggest that the space simulation environment could not be associated with red cell enzyme changes. It does show that any changes in enzymes were not sufficiently great to significantly shorten red cell survival. There was no evidence of bone marrow erythropoetic suppression nor was there any evidence of increased red cell destruction.

Ultraviolet and visible light spectrophotometric approach to blood typing: objective analysis by agglutination index.

PubMed

Narayanan, S; Orton, S; Leparc, G F; Garcia-Rubio, L H; Potter, R L

1999-10-01

A new blood typing technology based on ultraviolet (UV) and visible light spectroscopy (UV/visible spectroscopy) has been developed. Blood groups and types are determined by quantifying reproducible changes in the UV and visible light spectra of blood in the presence of agglutinating antibodies. Samples of red cells in the presence and absence of agglutinating antibodies were examined by UV/visible spectroscopy. Blood groups and types were determined by comparing the optical density spectra obtained between 665 and 1000 nm. These comparisons generate numbers (agglutination index) ranging from 0 to 100, with smaller numbers corresponding to lack of agglutination and larger numbers corresponding to agglutination. The optical density of agglutinated blood is dramatically different from that of unagglutinated blood. The agglutination index derived from the relative slopes of the spectra is an objective indicator of agglutination strength. An agglutination index greater than 17 consistently and accurately established blood group- and type-specific agglutination. The method accurately predicted A, B, and O blood groups, and D type in over 275 samples. Scattering theory-based calculations of relative volumes of red cells before and after agglutination show a direct correlation with the agglutination index and provide the theoretical basis of the analysis. This quantitative technique is reproducible and has the potential for automation.

Image-derived input function with factor analysis and a-priori information.

PubMed

Simončič, Urban; Zanotti-Fregonara, Paolo

2015-02-01

Quantitative PET studies often require the cumbersome and invasive procedure of arterial cannulation to measure the input function. This study sought to minimize the number of necessary blood samples by developing a factor-analysis-based image-derived input function (IDIF) methodology for dynamic PET brain studies. IDIF estimation was performed as follows: (a) carotid and background regions were segmented manually on an early PET time frame; (b) blood-weighted and tissue-weighted time-activity curves (TACs) were extracted with factor analysis; (c) factor analysis results were denoised and scaled using the voxels with the highest blood signal; (d) using population data and one blood sample at 40 min, whole-blood TAC was estimated from postprocessed factor analysis results; and (e) the parent concentration was finally estimated by correcting the whole-blood curve with measured radiometabolite concentrations. The methodology was tested using data from 10 healthy individuals imaged with [(11)C](R)-rolipram. The accuracy of IDIFs was assessed against full arterial sampling by comparing the area under the curve of the input functions and by calculating the total distribution volume (VT). The shape of the image-derived whole-blood TAC matched the reference arterial curves well, and the whole-blood area under the curves were accurately estimated (mean error 1.0±4.3%). The relative Logan-V(T) error was -4.1±6.4%. Compartmental modeling and spectral analysis gave less accurate V(T) results compared with Logan. A factor-analysis-based IDIF for [(11)C](R)-rolipram brain PET studies that relies on a single blood sample and population data can be used for accurate quantification of Logan-V(T) values.

Dehydration, hemodynamics and fluid volume optimization after induction of general anesthesia.

PubMed

Li, Yuhong; He, Rui; Ying, Xiaojiang; Hahn, Robert G

2014-01-01

Fluid volume optimization guided by stroke volume measurements reduces complications of colorectal and high-risk surgeries. We studied whether dehydration or a strong hemodynamic response to general anesthesia increases the probability of fluid responsiveness before surgery begins. Cardiac output, stroke volume, central venous pressure and arterial pressures were measured in 111 patients before general anesthesia (baseline), after induction and stepwise after three bolus infusions of 3 ml/kg of 6% hydroxyethyl starch 130/0.4 (n=86) or Ringer's lactate (n=25). A subgroup of 30 patients who received starch were preloaded with 500 ml of Ringer's lactate. Blood volume changes were estimated from the hemoglobin concentration and dehydration was estimated from evidence of renal water conservation in urine samples. Induction of anesthesia decreased the stroke volume to 62% of baseline (mean); administration of fluids restored this value to 84% (starch) and 68% (Ringer's). The optimized stroke volume index was clustered around 35-40 ml/m2/beat. Additional fluid boluses increased the stroke volume by ≥10% (a sign of fluid responsiveness) in patients with dehydration, as suggested by a low cardiac index and central venous pressure at baseline and by high urinary osmolality, creatinine concentration and specific gravity. Preloading and the hemodynamic response to induction did not correlate with fluid responsiveness. The blood volume expanded 2.3 (starch) and 1.8 (Ringer's) times over the infused volume. Fluid volume optimization did not induce a hyperkinetic state but ameliorated the decrease in stroke volume caused by anesthesia. Dehydration, but not the hemodynamic response to the induction, was correlated with fluid responsiveness.

Prenatal dioxin exposure estimated from dioxins in breast milk and sex hormone levels in umbilical cord blood in Vietnamese newborn infants.

PubMed

Boda, Hitomi; Nghi, Tran Ngoc; Nishijo, Muneko; Thao, Pham Ngoc; Tai, Pham The; Van Luong, Hoang; Anh, Tran Hai; Morikawa, Yuko; Nishino, Yoshikazu; Nishijo, Hisao

2018-02-15

Dioxin concentrations remain elevated in the environment and humans residing near the former US Air Force base in Bien Hoa city, South Vietnam. We recruited 210 mother-infant pairs for whom breast milk dioxin levels were reported in our previous study. Cord blood samples were collected from 162 mother-infant pairs. We selected 16 cord blood samples with a volume over 20mL and fat content of ≥0.03g. Toxic equivalent levels of polychlorinated dibenzodioxins and polychlorinated dibenzofurans (TEQ-PCDD/Fs) and concentrations of 17 congeners, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), in cord blood were measured and compared with levels in breast milk (Study 1). Levels of 2,3,7,8-TCDD and TEQ-PCDD/Fs in cord blood samples were highly and significantly correlated with those in breast milk samples in the same pairs. This suggests dioxins in breast milk reflect prenatal dioxin exposure. Estradiol (E2) and testosterone (TS) were measured in cord blood serum from 162 samples. Associations between dioxins in breast milk and cord blood sex hormones were analyzed by infant sex, after adjusting for confounding factors (Study 2). Increased levels of TEQ-PCDD/Fs in breast milk were associated with decreased cord blood TS in girls. In boys, a significant reduction of cord blood TS was observed in those exposed to 2,3,7,8-TCDD at high levels (≥5.5pg/g lipid). There was no significant association between E2 and dioxins in breast milk in either sex. These results suggest increased prenatal dioxin exposure is associated with decreased cord TS, but in boys, only high level of 2,3,7,8-TCDD influence cord blood TS. Copyright © 2017. Published by Elsevier B.V.

Accuracy of predicted haemoglobin concentration on cardiopulmonary bypass in paediatric cardiac surgery: effect of different formulae for estimating patient blood volume.

PubMed

Redlin, Matthias; Boettcher, Wolfgang; Dehmel, Frank; Cho, Mi-Young; Kukucka, Marian; Habazettl, Helmut

2017-11-01

When applying a blood-conserving approach in paediatric cardiac surgery with the aim of reducing the transfusion of homologous blood products, the decision to use blood or blood-free priming of the cardiopulmonary bypass (CPB) circuit is often based on the predicted haemoglobin concentration (Hb) as derived from the pre-CPB Hb, the prime volume and the estimated blood volume. We assessed the accuracy of this approach and whether it may be improved by using more sophisticated methods of estimating the blood volume. Data from 522 paediatric cardiac surgery patients treated with CPB with blood-free priming in a 2-year period from May 2013 to May 2015 were collected. Inclusion criteria were body weight <15 kg and available Hb data immediately prior to and after the onset of CPB. The Hb on CPB was predicted according to Fick's principle from the pre-CPB Hb, the prime volume and the patient blood volume. Linear regression analyses and Bland-Altman plots were used to assess the accuracy of the Hb prediction. Different methods to estimate the blood volume were assessed and compared. The initial Hb on CPB correlated well with the predicted Hb (R 2 =0.87, p<0.001). A Bland-Altman plot revealed little bias at 0.07 g/dL and an area of agreement from -1.35 to 1.48 g/dL. More sophisticated methods of estimating blood volume from lean body mass did not improve the Hb prediction, but rather increased bias. Hb prediction is reasonably accurate, with the best result obtained with the simplest method of estimating the blood volume at 80 mL/kg body weight. When deciding for or against blood-free priming, caution is necessary when the predicted Hb lies in a range of ± 2 g/dL around the transfusion trigger.

Relation of blood volume and blood pressure in orthostatic intolerance

NASA Technical Reports Server (NTRS)

Jacob, G.; Biaggioni, I.; Mosqueda-Garcia, R.; Robertson, R. M.; Robertson, D.

1998-01-01

A complex but crucial relationship exists between blood volume and blood pressure in human subjects; it has been recognized that in essential hypertension, renovascular hypertension, and pheochromocytoma, the relationship between plasma volume and diastolic blood pressure is an inverse one. This phenomenon has not been studied in individuals with low normal and reduced blood pressures. Orthostatic intolerance is a commonly encountered abnormality in blood pressure regulation often associated with tachycardia in the standing position. Most of these patients have varying degrees of reduced blood volume. We tested the hypothesis that the relationship previously found between plasma volume and diastolic blood pressure in pressor states would also hold in orthostatic intolerance. We studied 16 patients with a history of symptomatic orthostatic intolerance associated with an elevation in plasma norepinephrine in the upright posture and hypovolemia in 9 patients and normovolemia in 7 patients. Our studies demonstrate an inverse relationship between plasma volume and diastolic blood pressure in patients with orthostatic intolerance. This finding also holds for the change in diastolic blood pressure in response to upright posture. In this relationship, patients with orthostatic intolerance with high plasma norepinephrine resemble those with essential hypertension, renovascular hypertension, and pheochromocytoma. We conclude that in a variety of conditions at both ends of the blood pressure spectrum, the seemingly paradoxical association of hypovolemia and diastolic blood pressure is preserved.

[The Contribution of Vascular Capacity and Blood Volume to Maintain Stable Blood Circulation during General Anesthesia].

PubMed

Shigemi, Kenji

2016-05-01

To maintain proper cardiac preload is one of the most effective procedures for the systemic circulation remaining stable. In particular, the balance between vascular capacity and total blood volume must be maintained within appropriate range by the administration of fluids, blood and/or vasoactive drugs with mean circulatory filling pressure (Pmcf), central venous pressure (CVP) or stroke volume variation (SVV). End-diastolic left ventricular volume (Ved) is theoretically the best index of cardiac preload; however, without transesophageal echocardbalanceiogram we cannot directly monitor Ved during anesthesia. The infused fluid volume remaining in intravascular space, the vascular capacity controlled by autonomic nervous system and/or vasoactive agents, and the unstressed blood volume properly mobilized to excess blood volume are the crucial factors to maintain cardiac output The knowledge of vascular physiology contribute the decision making to manipulate such factors to control blood circulation during general anesthesia. For example, CVP is usually maintained in the narrow range and seems to be stable; however, it must be changed just after the circulatory disturbances, such as acute bleeding, blood transfusion, and fluid infusion, and followed by gradual returning to initial value, because of the solid mechanism to preserve cardiac output

An automated system for chromosome analysis. Volume 1: Goals, system design, and performance

NASA Technical Reports Server (NTRS)

Castleman, K. R.; Melnyk, J. H.

1975-01-01

The design, construction, and testing of a complete system to produce karyotypes and chromosome measurement data from human blood samples, and a basis for statistical analysis of quantitative chromosome measurement data is described. The prototype was assembled, tested, and evaluated on clinical material and thoroughly documented.

Hormonal changes in antiorthostatic rats

NASA Technical Reports Server (NTRS)

Popovic, V.; Popovic, P.; Honeycutt, C.

1982-01-01

Hypokinesia, especially hypokinesia with negative tilt ('antiorthostatic hypokinesia'), mimics some of the effects of weightlessness. It is shown that cardiac output is increased during early exposure of rats to antiorthostatic hypokinesia. The increase of the stroke volume and of the cardiac output observed in the antiorthostatic hypokinetic rats is probably the consequence of a blood volume shift toward the chest brought forth by head-down positioning of the animals. It is also possible that struggling of the animals to escape from the harness and an increased metabolism contribute to the elevation of cardiac output. In order to study this hypothesis 'stress hormones' were measured in the antiorthostatic rats. Plasma levels of ACTH, corticosterone and prolactin were measured in the arterial blood (0.3 ml) sampled before, during and after hypokinesia from chronic aortic cannulas of the rats.

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

PubMed

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

2014-10-01

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

Specific binding of magnetic nanoparticle probes to platelets in whole blood detected by magnetorelaxometry

NASA Astrophysics Data System (ADS)

Eberbeck, Dietmar; Wiekhorst, Frank; Steinhoff, Uwe; Schwarz, Kay Oliver; Kummrow, Andreas; Kammel, Martin; Neukammer, Jörg; Trahms, Lutz

2009-05-01

The binding of monoclonal antibodies labelled with magnetic nanoparticles to CD61 surface proteins expressed by platelets in whole blood samples was measured by magnetorelaxometry. This technique is sensitive to immobilization of the magnetic labels upon binding. Control experiments with previous saturation of the epitopes on the platelet surfaces demonstrated the specificity of the binding. The kinetics of the antibody antigen reaction is accessible with a temporal resolution of 12 s. The minimal detectable platelet concentration is about 2000 μL -1 (sample volume 150 μL). The proportionality of the magnetic relaxation amplitude to the number of bound labels allows a quantification of the antibody binding capacity.

Blood values of the canvasback duck by age, sex and season

USGS Publications Warehouse

Kocan, R.M.; Pitts, S.M.

1976-01-01

Blood samples were obtained from canvasback ducklings from Manitoba and Saskatchewan and from immature and adult canvasbacks on the Mississippi River near LaCrosse, Wisconsin and the Chesapeake Bay. These samples were used to determine baseline data on red cell counts, hematocrit, total protein, glucose, cholesterol, hemoglobin and distribution of plasma proteins. Calculations were also made to determine mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. The major differences noted were between ducklings and adults. The former having higher total protein and lower hematocrit, glucose and cholesterol values. These hematologic values were collected in order to provide baseline information on apparently healthy canvasbacks, thereby providing disease investigators with a standard of comparison

Sample Preparation and Extraction in Small Sample Volumes Suitable for Pediatric Clinical Studies: Challenges, Advances, and Experiences of a Bioanalytical HPLC-MS/MS Method Validation Using Enalapril and Enalaprilat

PubMed Central

Burckhardt, Bjoern B.; Laeer, Stephanie

2015-01-01

In USA and Europe, medicines agencies force the development of child-appropriate medications and intend to increase the availability of information on the pediatric use. This asks for bioanalytical methods which are able to deal with small sample volumes as the trial-related blood lost is very restricted in children. Broadly used HPLC-MS/MS, being able to cope with small volumes, is susceptible to matrix effects. The latter restrains the precise drug quantification through, for example, causing signal suppression. Sophisticated sample preparation and purification utilizing solid-phase extraction was applied to reduce and control matrix effects. A scale-up from vacuum manifold to positive pressure manifold was conducted to meet the demands of high-throughput within a clinical setting. Faced challenges, advances, and experiences in solid-phase extraction are exemplarily presented on the basis of the bioanalytical method development and validation of low-volume samples (50 μL serum). Enalapril, enalaprilat, and benazepril served as sample drugs. The applied sample preparation and extraction successfully reduced the absolute and relative matrix effect to comply with international guidelines. Recoveries ranged from 77 to 104% for enalapril and from 93 to 118% for enalaprilat. The bioanalytical method comprising sample extraction by solid-phase extraction was fully validated according to FDA and EMA bioanalytical guidelines and was used in a Phase I study in 24 volunteers. PMID:25873972

The endocrine system in space flight

NASA Astrophysics Data System (ADS)

Leach, C. S.; Johnson, P. C.; Cintron, N. M.

Hormones are important effectors of the body's response to microgravity in the areas of fluid and electrolyte metabolism, erythropoiesis, and calcium metabolism. For many years antidiuretic hormone, cortisol and aldosterone have been considered the hormones most important for regulation of body fluid volume and blood levels of electrolytes, but they cannot account totally for losses of fluid and electrolytes during space flight. We have now measured atrial natriuretic factor (ANF), a hormone recently shown to regulate sodium and water excretion, in blood specimens obtained during flight. After 30 or 42 h of weightlessness, mean ANF was elevated. After 175 or 180 h, ANF had decreased by 59%, and it changed little between that time and soon after landing. There is probably an increase in ANF early inflight associated with the fluid shift, followed by a compensatory decrease in blood volume. Increased renal blood flow may cause the later ANF decrease. Erythropoietin (Ep), a hormone involved in the control of red blood cell production, was measured in blood samples taken during the first Spacelab mission and was significantly decreased on the second day of flight, suggesting also an increase in renal blood flow. Spacelab-2 investigators report that the active vitamin D metabolite 1α, 25-dihydroxyvitamin D 3 increased early in the flight, indicating that a stimulus for increased bone resorption occurs by 30 h after launch.

Computer simulation of preflight blood volume reduction as a countermeasure to fluid shifts in space flight

NASA Technical Reports Server (NTRS)

Simanonok, K. E.; Srinivasan, R.; Charles, J. B.

1992-01-01

Fluid shifts in weightlessness may cause a central volume expansion, activating reflexes to reduce the blood volume. Computer simulation was used to test the hypothesis that preadaptation of the blood volume prior to exposure to weightlessness could counteract the central volume expansion due to fluid shifts and thereby attenuate the circulatory and renal responses resulting in large losses of fluid from body water compartments. The Guyton Model of Fluid, Electrolyte, and Circulatory Regulation was modified to simulate the six degree head down tilt that is frequently use as an experimental analog of weightlessness in bedrest studies. Simulation results show that preadaptation of the blood volume by a procedure resembling a blood donation immediately before head down bedrest is beneficial in damping the physiologic responses to fluid shifts and reducing body fluid losses. After ten hours of head down tilt, blood volume after preadaptation is higher than control for 20 to 30 days of bedrest. Preadaptation also produces potentially beneficial higher extracellular volume and total body water for 20 to 30 days of bedrest.

Sensitive and quantitative measurement of gene expression directly from a small amount of whole blood.

PubMed

Zheng, Zhi; Luo, Yuling; McMaster, Gary K

2006-07-01

Accurate and precise quantification of mRNA in whole blood is made difficult by gene expression changes during blood processing, and by variations and biases introduced by sample preparations. We sought to develop a quantitative whole-blood mRNA assay that eliminates blood purification, RNA isolation, reverse transcription, and target amplification while providing high-quality data in an easy assay format. We performed single- and multiplex gene expression analysis with multiple hybridization probes to capture mRNA directly from blood lysate and used branched DNA to amplify the signal. The 96-well plate singleplex assay uses chemiluminescence detection, and the multiplex assay combines Luminex-encoded beads with fluorescent detection. The single- and multiplex assays could quantitatively measure as few as 6000 and 24,000 mRNA target molecules (0.01 and 0.04 amoles), respectively, in up to 25 microL of whole blood. Both formats had CVs < 10% and dynamic ranges of 3-4 logs. Assay sensitivities allowed quantitative measurement of gene expression in the minority of cells in whole blood. The signals from whole-blood lysate correlated well with signals from purified RNA of the same sample, and absolute mRNA quantification results from the assay were similar to those obtained by quantitative reverse transcription-PCR. Both single- and multiplex assay formats were compatible with common anticoagulants and PAXgene-treated samples; however, PAXgene preparations induced expression of known antiapoptotic genes in whole blood. Both the singleplex and the multiplex branched DNA assays can quantitatively measure mRNA expression directly from small volumes of whole blood. The assay offers an alternative to current technologies that depend on RNA isolation and is amenable to high-throughput gene expression analysis of whole blood.

The End-Diastolic Velocity of Thyroid Arteries Is Strongly Correlated with the Peak Systolic Velocity and Gland Volume in Patients with Autoimmune Thyroiditis

PubMed Central

Marui, Suemi; Buchpiguel, Carlos Alberto; Cerri, Giovanni Guido; Chammas, Maria Cristina

2017-01-01

Background The end-diastolic velocity (EDV) of thyroid arteries reflects peripheral blood flow resistance. Objective The aim was to evaluate EDV correlations with other Doppler sonography parameters and with clinical and biochemical variables in a sample of patients with hypothyroidism caused by chronic autoimmune thyroiditis (CAT). Methods A sample of 48 CAT hypothyroid patients receiving treatment with stable doses of levothyroxine was selected. The participants underwent clinical evaluation and measurement of serum thyrotropin (TSH), total triiodothyronine (T3), total thyroxine (T4), free T4, thyroid peroxidase antibodies (anti-TPO), and antithyroglobulin antibodies (anti-Tg) and Doppler sonography. Results The EDV of the inferior thyroid arteries (ITA-EDV) was strongly and positively correlated with the peak systolic velocity of the inferior thyroid arteries (ITA-PSV, r = 0.919), thyroid volume (r = 0.711), and thyroid visual vascularization pattern (TVP, r = 0.687). There was no correlation between ITA-EDV and the clinical variables, hormones, anti-TPO, or anti-Tg. Conclusion The strong correlation of ITA-EDV with ITA-PSV, TVP, and volume suggests that increased vascularization in CAT may be associated with a reduction in thyroid blood flow resistance, possibly due to an angiogenesis-induced increase in the total vascular cross-sectional area of the parenchyma. PMID:29062583

Determination of dioxin-like polychlorinated biphenyls in 1 mL whole blood using programmable temperature vaporization large volume injection coupled to gas chromatogram and high-resolution mass spectrometry.

PubMed

Shen, Haitao; Guan, Rongfa; Li, Jingguang; Zhang, Lei; Ren, Yiping; Xu, Xiaomin; Song, Yang; Zhao, Yunfeng; Han, Jianlong; Wu, Yongning

2013-03-12

A sensitive method based on programmable temperature vaporization large volume injection coupled to gas chromatogram and high-resolution mass spectrometry (PTV-GC-HRMS) has been developed for the determination of ultra trace levels of dioxin-like polychlorinated biphenyls (DL PCBs) in small amounts of human blood. Blood samples (1mL) were first extracted by column extraction and then purified with column chromatorgraphies. Final extracts (20μL) were introduced to the PTV injector under the solvent vent mode and detected by GC-HRMS (SIM mode). PTV parameters were observed by changing one factor at a time (practical conditions: vent flow: 50mLmin(-1), vent pressure: 0kPa and vent time: 0.1min), recoveries of most PCB congeners ranged from 55.1% to 108%, and method detection limits were in the range of 0.11-1.63pgg(-1). Copyright © 2013 Elsevier B.V. All rights reserved.

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

PubMed Central

Sun, Xiaocun; Flatland, Bente

2016-01-01

Background Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Methods Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV) of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia). Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Results Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature), while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice. Discussion Canine EDTA whole blood samples cool most rapidly and to a greater degree when placed in an ice-water bath rather than in ice. Samples stored on ice water can rapidly drop below normal refrigeration temperatures; this should be taken into consideration when using this cooling modality. PMID:27917319

Biomonitoring of Toxic Effects of Pesticides in Occupationally Exposed Individuals.

PubMed

Arshad, Muhammad; Siddiqa, Maryam; Rashid, Saddaf; Hashmi, Imran; Awan, Muhammad Ali; Ali, Muhammad Arif

2016-06-01

Workers in pesticide manufacturing industries are constantly exposed to pesticides. Genetic biomonitoring provides an early identification of potential cancer and genetic diseases in exposed populations. The objectives of this biomonitoring study were to assess DNA damage through comet assay in blood samples collected from industry workers and compare these results with those of classical analytical techniques used for complete blood count analysis. Samples from controls (n = 20) and exposed workers (n = 38) from an industrial area in Multan, Pakistan, were subjected to various tests. Malathion residues in blood samples were measured by gas chromatography. The exposed workers who were employed in the pesticide manufacturing industry for a longer period (i.e., 13-25 years) had significantly higher DNA tail length (7.04 μm) than the controls (0.94 μm). Workers in the exposed group also had higher white blood cell and red blood cell counts, and lower levels of mean corpuscular hemoglobin (MCH), MCH concentration, and mean corpuscular volume in comparison with normal levels for these parameters. Malathion was not detected in the control group. However, in the exposed group, 72% of whole blood samples had malathion with a mean value of 0.14 mg/L (range 0.01-0.31 mg/L). We found a strong correlation (R (2) = 0.91) between DNA damage in terms of tail length and malathion concentration in blood. Intensive efforts and trainings are thus required to build awareness about safety practices and to change industrial workers' attitude to prevent harmful environmental and anthropogenic effects.

Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo

NASA Astrophysics Data System (ADS)

Zettergren, Eric; Swamy, Tushar; Runnels, Judith; Lin, Charles P.; Niedre, Mark

2012-07-01

Sensing and enumeration of specific types of circulating cells in small animals is an important problem in many areas of biomedical research. Microscopy-based fluorescence in vivo flow cytometry methods have been developed previously, but these are typically limited to sampling of very small blood volumes, so that very rare circulating cells may escape detection. Recently, we described the development of a ‘diffuse fluorescence flow cytometer’ (DFFC) that allows sampling of much larger blood vessels and therefore circulating blood volumes in the hindlimb, forelimb or tail of a mouse. In this work, we extend this concept by developing and validating a method to tomographically localize circulating fluorescently labeled cells in the cross section of a tissue simulating optical flow phantom and mouse limb. This was achieved using two modulated light sources and an array of six fiber-coupled detectors that allowed rapid, high-sensitivity acquisition of full tomographic data sets at 10 Hz. These were reconstructed into two-dimensional cross-sectional images using Monte Carlo models of light propagation and the randomized algebraic reconstruction technique. We were able to obtain continuous images of moving cells in the sample cross section with 0.5 mm accuracy or better. We first demonstrated this concept in limb-mimicking optical flow photons with up to four flow channels, and then in the tails of mice with fluorescently labeled multiple myeloma cells. This approach increases the overall diagnostic utility of our DFFC instrument.

Blood volume regulating hormones, fluid and electrolyte modifications during 21 and 198-day space flights (Altair-MIR 1993)

NASA Astrophysics Data System (ADS)

Vorobiev, D.; Maillet, A.; Fortrat, J. O.; Pastushkova, L.; Allevard, A. M.; Sigaudo, D.; Cartier, R.; Patricot, M.; Andre-Deshays, C.; Kotovskaya, A.; Grigoriev, A.; Gharib, C.; Gauquelin, G.

During the Altair MIR' 93 mission we studied several parameters involved in blood volume regulation. The experiment was done on two cosmonauts before (B-60, B-30), during (D6, D12, D18 for French and D7, D12, D17 for Russian) and after the flight (R+1, R+3 and R+7). Space flight durations were different for two cosmonauts: for the Russian the flight duration was 198 days and for the French 21 days. On board the MIR station only urinary (volume and electrolytes, atrial natriuretic peptide (ANP), cyclic guanosine monophosphate (cGMP) and catecholamines) and salivary (cGMP and cortisol) samples were collected, centrifuged and stored in freezer. Lithium was used as a tracer to know exactly the 24 h urine output (CNES urine collection Kit). Before and after flight, blood was drawn with an epicite needle and vacutainer system for hormonal assays (renin, antidiuretic hormone, cGMP, ANP and aldosterone) in two positions: after 30 min rest in upright seated position and after 90 min of supine position. Salivary samples were collected simultaneously. During flight a decrease of diuresis and ANP and an increase of osmolality were found. No modifications of hematocrit, but an increase of salivary cGMP and cortisol were also observed. The decrease of urinary ANP is in favor of hypovolemia as described in previous flights. The postflight examinations revealed changes in fluid-electrolyte metabolism which indicate a hypohydration status and a stimulation of hormonal system responsible for water and electrolyte retention in order to readapt to the normal gravity.

A Portable Immunoassay Platform for Multiplexed Detection of Biotoxins in Clinical and Environmental Samples

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

Koh, Chung-Yan; Piccini, Matthew Ernest; Schaff, Ulrich Y.

Multiple cases of attempted bioterrorism events using biotoxins have highlighted the urgent need for tools capable of rapid screening of suspect samples in the field (e.g., mailroom and public events). We present a portable microfluidic device capable of analyzing environmental (e.g., white powder), food (e.g., milk) and clinical (e.g., blood) samples for multiplexed detection of biotoxins. The device is rapid (<15-30 min sample-to-answer), sensitive (< 0.08 pg/mL detection limit for botulinum toxin), multiplexed (up to 64 parallel assays) and capable of analyzing small volume samples (< 20 μL total sample input). The immunoassay approach (SpinDx) is based on binding ofmore » toxins in a sample to antibody-laden capture particles followed by sedimentation of particles through a density-media in a microfluidic disk and quantification using a laser-induced fluorescence detector. A direct, blinded comparison with a gold standard ELISA revealed a 5-fold more sensitive detection limit for botulinum toxin while requiring 250-fold less sample volume and a 30 minute assay time with a near unity correlation. A key advantage of the technique is its compatibility with a variety of sample matrices with no additional sample preparation required. Ultrasensitive quantification has been demonstrated from direct analysis of multiple clinical, environmental and food samples, including white powder, whole blood, saliva, salad dressing, whole milk, peanut butter, half and half, honey, and canned meat. We believe that this device can met an urgent need in screening both potentially exposed people as well as suspicious samples in mail-rooms, airports, public sporting venues and emergency rooms. The general-purpose immunodiagnostics device can also find applications in screening of infectious and systemic diseases or serve as a lab device for conducting rapid immunoassays.« less

Regional Blood Volume and Peripheral Blood Flow in the Postural Tachycardia Syndrome

PubMed Central

Stewart, Julian M.; Montgomery, Leslie D.

2015-01-01

Variants of postural tachycardia syndrome (POTS) are associated with increased (“high flow” POTS, HFP), decreased (“low flow POTS”, LFP) and normal (“normal flow POTS”, NFP) blood flow measured in the lower extremities while supine. We propose that postural tachycardia is related to thoracic hypovolemia during orthostasis but that the patterns of peripheral blood flow relate to different mechanisms for thoracic hypovolemia. We studied 37 POTS patients aged 14-21 years: 14 LFP, 15 NFP and 8 HFP patients and 12 healthy control subjects. Peripheral blood flow was measured supine by venous occlusion strain gauge plethysmography of the forearm and calf in order to subgroup patients. Using indocyanine green techniques we showed decreased cardiac index (CI) and increased total peripheral resistance (TPR) in LFP, increased CI and decreased TPR in HFP, and unchanged CI and TPR in NFP while supine compared to control subjects. Blood volume tended to be decreased in LFP compared to control subjects. We used impedance plethysmography to assess regional blood volume redistribution during upright tilt. Thoracic blood volume decreased while splanchnic, pelvic and leg blood volumes increased for all subjects during orthostasis, but were markedly lower than control for all POTS groups. Splanchnic volume was increased in NFP and LFP. Pelvic blood volume was increased in HFP only. Calf volume was increased above control in HFP and LFP. The results support the hypothesis of [at least] three pathophysiologic variants of POTS distinguished by peripheral blood flow related to characteristic changes in regional circulations. The data demonstrate enhanced thoracic hypovolemia during upright tilt and confirm that POTS is related to inadequate cardiac venous return during orthostasis. PMID:15117717

Red Blood Cell Volume, Plasma Volume and Total Blood Volume in Healthy Elderly Men and Women Aged 64 to 100

DTIC Science & Technology

1992-05-06

Robert Valeri, Linda E. Pivacek, Hiliary Siebens, and Mark D. Altschule ». PERFORMING ORGANIZATION NAME AND AOORESS Naval Blood Research Laboratory...Gibson JG, Peacock WC, Seligman AM, Sack T: Circulating red cell volume measured simultaneously by the radioactive iron and dye methods. J Clin

Ad libitum fluid intake and plasma responses after pickle juice, hypertonic saline, or deionized water ingestion.

PubMed

Allen, Scott; Miller, Kevin C; Albrecht, Jay; Garden-Robinson, Julie; Blodgett-Salafia, Elizabeth

2013-01-01

Adding sodium (Na(+)) to drinks improves rehydration and ad libitum fluid consumption. Clinicians (∼25%) use pickle juice (PJ) to treat cramping. Scientists warn against PJ ingestion, fearing it will cause rapid plasma volume restoration and thereby decrease thirst and delay rehydration. Advice about drinking PJ has been developed but never tested. To determine if drinking small volumes of PJ, hypertonic saline (HS), or deionized water (DIW) affects ad libitum DIW ingestion, plasma variables, or perceptual indicators. Crossover study. Laboratory. Fifteen, euhydrated (urine specific gravity ≤ 1.01) men (age = 22 ± 2 years, height = 178 ± 6 cm, mass = 82.9 ± 8.4 kg). Participants completed 3 testing days (≥ 72 hours between days). After a 30-minute rest, a blood sample was collected. Participants completed 60 minutes of hard exercise (temperature = 36 ± 2°C, relative humidity = 16 ± 1%). Postexercise, they rested for 30 minutes; had a blood sample collected; rated thirst, fullness, and nausea; and ingested 83 ± 8 mL of PJ, HS, or DIW. They rated drink palatability (100-mm visual analog scale) and were allowed to drink DIW ad libitum for 60 minutes. Blood samples and thirst, fullness, and nausea ratings (100-mm visual analog scales) were collected at 15, 30, 45, and 60 minutes posttreatment drink ingestion. Ad libitum DIW volume, percentage change in plasma volume, plasma osmolality (OSMp,) plasma sodium concentration ([Na(+)]p), and thirst, fullness, nausea, and palatability ratings. Participants consumed more DIW ad libitum after HS (708.03 ± 371.03 mL) than after DIW (532.99 ± 337.14 mL, P < .05). Ad libitum DIW ingested after PJ (700.35 ± 366.15 mL) was similar to that after HS and DIW (P > .05). Plasma sodium concentration, OSMp, percentage change in plasma volume, thirst, fullness, and nausea did not differ among treatment drinks over time (P > .05). Deionized water (73 ± 14 mm) was more palatable than HS (17 ± 13 mm) or PJ (26 ± 16 mm, P < .05). The rationale behind advice about drinking PJ is questionable. Participants drank more, not less, after PJ ingestion, and plasma variables and perceptual indicators were similar after PJ and DIW ingestion. Pickle juice did not inhibit short-term rehydration.

Evaluation of Trapper-Collected Nobuto Filter-Paper Blood Samples for Distemper and Parvovirus Antibody Detection in Coyotes (Canis latrans) and Raccoons (Procyon lotor).

PubMed

Kamps, Amanda J; Dubay, Shelli A; Langenberg, Julie; Maes, Roger K

2015-07-01

Blood samples are often collected from free-ranging wildlife for antibody detection. However, filter-paper (FP) strips are more cost efficient and easy to collect and store. We evaluated trapper-collected FP strips and body-cavity blood for canine distemper (CDV) and parvovirus (CPV-2) antibody detection in raccoons (Procyon lotor) and coyotes (Canis latrans). From 2008 to 2010, licensed trappers near Madison and Milwaukee, Wisconsin, US collected paired samples from harvested animals. Canine distemper antibodies were detected using virus neutralization and parvovirus antibodies were detected using hemagglutination inhibition. Titers ≥ 1:32 for CDV and ≥ 1:25 for CPV-2 were considered evidence of exposure. Using Cohen's kappa test of agreement, FP strip titers agreed with sera for CDV in coyotes (n = 28, K = 0.772) and raccoons (n = 29, K = 0.858) and for CPV-2 in coyotes (n = 40, K = 0.775) and raccoons (n = 70, K = 0.646). However, raccoons determined to be exposed to CPV-2 from sera were unexposed by FP strips in 35% of the samples. Titer results may be affected by quality and volume of blood samples, interval between collection and processing, small sample sizes, and diagnostic testing procedures. Filter-paper strips can be useful for detecting CDV and CPV-2 exposure in coyotes and raccoons with correct field sample collection and appropriate diagnostic testing procedures.

Drop-to-drop solvent microextraction coupled with gas chromatography/mass spectrometry for rapid determination of trimeprazine in urine and blood of rats: application to pharmacokinetic studies.

PubMed

Agrawal, Kavita; Wu, Hui-Fen

2007-01-01

A simple and rapid method based on drop-to-drop solvent microextraction (DDSME) coupled with gas chromatography/mass spectrometry (GC/MS) has been successfully applied for the pharmacokinetic studies of trimeprazine in 8 microL of urine and blood samples of rats. Several factors that influenced the extraction efficiency of DDSME, such as selection of organic solvent, extraction time, exposure volume of organic phase, addition of salt and pH, were optimized. Linearity was obtained over the concentration ranges of 0.2-10, 0.25-7.0 and 0.5-6.0 microg/mL with correlation coefficients of 0.998, 0.996 and 0.993 in deionized water, urine and blood samples of rats, respectively. The limits of detection (LODs) of trimeprazine were 0.05, 0.06 and 0.1 microg/mL in deionized water, urine and blood samples. The concentrations of trimeprazine obtained in urine and blood samples of rats were 0.21-1.25 and 2.72-0.22 microg/mL, respectively, after a single intravenous administration of this drug. The enrichment factors and LOD values obtained by DDSME coupled to GC/MS were compared with those of hollow fiber liquid-phase microextraction (HF-LPME) combined with GC/MS. We believe that this novel approach can be very useful in clinical application since only one microdrop of biological samples was required to perform the pharmacokinetic studies from rats, so the sample pretreatments for animal experiments can be very easy too. Copyright (c) 2007 John Wiley & Sons, Ltd.

Multiple-wavelength spectroscopic quantitation of light-absorbing species in scattering media

DOEpatents

Nathel, Howard; Cartland, Harry E.; Colston, Jr., Billy W.; Everett, Matthew J.; Roe, Jeffery N.

2000-01-01

An oxygen concentration measurement system for blood hemoglobin comprises a multiple-wavelength low-coherence optical light source that is coupled by single mode fibers through a splitter and combiner and focused on both a target tissue sample and a reference mirror. Reflections from both the reference mirror and from the depths of the target tissue sample are carried back and mixed to produce interference fringes in the splitter and combiner. The reference mirror is set such that the distance traversed in the reference path is the same as the distance traversed into and back from the target tissue sample at some depth in the sample that will provide light attenuation information that is dependent on the oxygen in blood hemoglobin in the target tissue sample. Two wavelengths of light are used to obtain concentrations. The method can be used to measure total hemoglobin concentration [Hb.sub.deoxy +Hb.sub.oxy ] or total blood volume in tissue and in conjunction with oxygen saturation measurements from pulse oximetry can be used to absolutely quantify oxyhemoglobin [HbO.sub.2 ] in tissue. The apparatus and method provide a general means for absolute quantitation of an absorber dispersed in a highly scattering medium.

Using Hematology Data from Malaria Vaccine Research Trials in Humans and Rhesus Macaques (Macaca mulatta) To Guide Volume Limits for Blood Withdrawal.

PubMed

Hegge, Sara R; Hickey, Bradley W; Mcgrath, Shannon M; Stewart, V Ann

2016-12-01

Guidelines on safe volume limits for blood collection from research participants in both humans and laboratory animals vary widely between institutions. The main adverse event that may be encountered in large blood volume withdrawal is iron-deficiency anemia. Monitoring various parameters in a standard blood panel may help to prevent this outcome. To this end, we analyzed the Hgb and MCV values from 43 humans and 46 macaques in malaria vaccine research trials. Although the percentage of blood volume removed was greater for macaques than humans, macaques demonstrated an overall increase of MCV over time, indicating the ability to respond appropriately to frequent volume withdrawals. In contrast, humans showed a consistent declining trend in MCV. These declines in human MCV and Hgb were significant from the beginning to end of the study despite withdrawals that were smaller than recommended volume limits. Limiting the volume withdrawn to no more than 12.5% seemed to be sufficient for macaques, and at 14% or more individual animals tended to fail to respond appropriately to large-volume blood loss, as demonstrated by a decrease in MCV. The overall positive erythropoietic response seen in macaques was likely due to the controlled, iron-fortified diet they received. The lack of erythropoietic response in the human subjects may warrant iron supplementation or reconsideration of current blood volume withdrawal guidelines.

Distribution of extravascular fluid volumes in isolated perfused lungs measured with H215O.

PubMed Central

Jones, T; Jones, H A; Rhodes, C G; Buckingham, P D; Hughes, J M

1976-01-01

The distributions per unit volume of extravascular water (EVLW), blood volume, and blood flow were measured in isolated perfused vertical dog lungs. A steady-state tracer technique was employed using oxygen-15, carbon-11, and nitrogen-13 isotopes and external scintillation counting of the 511-KeV annihilation radiation common to all three radionuclides. EVLW, and blood volume and flow increased from apex to base in all preparations, but the gradient of increasing flow exceeded that for blood and EVLW volumes. The regional distributions of EVLW and blood volume were almost identical. With increasing edema, lower-zone EVLW increased slightly relative to that in the upper zone. There was no change in the distribution of blood volume or flow until gross edema (100% wt gain) occurred when lower zone values were reduced. In four lungs the distribution of EVLW was compared with wet-to-dry ratios from lung biopsies taken immediately afterwards. Whereas the isotopically measured EVLW increased from apex to base, the wet-to-dry weight ratios remained essentially uniform. We concluded that isotopic methods measure only an "exchangeable" water pool whose volume is dependent on regional blood flow and capillary recruitment. Second, the isolated perfused lung can accommodate up to 60% wt gain without much change in the regional distribution of EVLW, volume, or flow. PMID:765354

Optimised cord blood sample selection for small‑scale CD34+ cell immunomagnetic isolation.

PubMed

Perdomo-Arciniegas, Ana-María; Vernot, Jean-Paul

2012-03-01

Haematopoietic stem cells (HSCs) are defined as multipotential cells, capable of self-renewal and reconstituting in vivo the haematopoietic compartment. The CD34 antigen is considered an important HSCs marker in humans. Immunomagnetic isolation, by targeting CD34 antigen, is widely used for human HSC separation. This method allows the enrichment of human HSCs that are present at low frequencies in umbilical cord blood (CB). Immunomagnetic CD34+-cell isolation reproducibility, regarding cell yield and purity, is affected by the CD34+ cell frequency and total cell numbers present in a given sample; CB HSC purification may thus yield variable results, which also depend on the volume and density fractionation-derived cell loss of a CB sample. The uncertainty of such an outcome and associated technical costs call for a cost-effective sample screening strategy. A correlation analysis using clinical and laboratory data from 59 CB samples was performed to establish predictive variables for CD34+-immunomagnetic HSCs isolation. This study described the positive association of CD34+-cell isolation with white and red cell numbers present after cell fractionation. Furthermore, purity has been correlated with lymphocyte percentages. Predictive variable cut-off values, which are particularly useful in situations involving low CB volumes being collected (such as prevalent late umbilical cord clamping clinical practice), were proposed for HSC isolation sampling. Using the simple and cost-effective CB sample screening criteria described here would lead to avoiding costly inefficient sample purification, thereby ensuring that pure CD34+ cells are obtained in the desired numbers following CD34 immunomagnetic isolation.

High quality cord blood banking is feasible with delayed clamping practices. The eight-year experience and current status of the national Swedish Cord Blood Bank.

PubMed

Frändberg, Sofia; Waldner, Berit; Konar, Jan; Rydberg, Lennart; Fasth, Anders; Holgersson, Jan

2016-09-01

The National Swedish Cord Blood Bank (NS-CBB) is altruistic and publicly funded. Herein we describe the status of the bank and the impact of delayed versus early clamping on cell number and volume. Cord Blood Units (CBUs) were collected at two University Hospitals in Sweden. Collected volume and nucleated cell content (TNC) were investigated in 146 consecutive Cord Blood (CB) collections sampled during the first quarter of 2012 and in 162 consecutive CB collections done in the first quarter of 2013, before and after clamping practices were changed from immediate to late (60 s) clamping. NS-CBB now holds close to 5000 units whereof 30 % are from non-Caucasian or mixed origins. Delayed clamping had no major effect on collection efficiency. The volume collected was slightly reduced (mean difference, 8.1 ml; 95 % CI, 1.3-15.0 ml; p = 0.02), while cell recovery was not (p = 0.1). The proportion of CBUs that met initial total TNC banking criteria was 60 % using a TNC threshold of 12.5 × 10(8), and 47 % using a threshold of 15 × 10(8) for the early clamping group and 52 and 37 % in the late clamping group. Following implementation of delayed clamping practices at NS-CBB; close to 40 % of the collections in the late clamping group still met the high TNC banking threshold and were eligible for banking, implicating that that cord blood banking is feasible with delayed clamping practices.

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.

Detection of bacteria in platelet concentrates prepared from spiked single donations using cultural and molecular genetic methods.

PubMed

Störmer, M; Cassens, U; Kleesiek, K; Dreier, J

2007-02-01

Bacteria show differences in their growth kinetics depending on the type of blood component. On to storage at 22 degrees C, platelet concentrates (PCs) seem to be more prone to bacterial multiplication than red cell concentrates. Knowledge of the potential for bacterial proliferation in blood components, which are stored at a range of temperatures, is essential before considering implementation of a detection strategy. The efficacy of bacterial detection was determined, using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), following bacterial growth in blood components obtained from a deliberately contaminated whole-blood (WB) unit. Cultivation was used as the reference method. WB was spiked with 2 colony-forming units mL(-1)Staphylococcus epidermidis or Klebsiella pneumoniae, kept for 15 h at room temperature and component preparation was processed. Samples were drawn, at intervals throughout the whole separation process, from each blood component. Nucleic acids were extracted using an automated high-volume extraction method. The 15-h storage revealed an insignificant increase in bacterial titre. No bacterial growth was detected in red blood cell or plasma units. K. pneumoniae showed rapid growth in the pooled PC and could be detected immediately after preparation using RT-PCR. S. epidermidis grew slowly and was detected 24 h after separation. These experiments show that sampling is indicative at 24 h after preparation of PCs at the earliest to minimize the sampling error.

The influence of moderate-intensity physical effort on peripheral blood in adults with Down syndrome - a pilot study.

PubMed

Aleksander-Szymanowicz, P; Marchewka, A; Dabrowski, Z; Teleglow, A; Bac, A; Glodzik, J

2014-10-01

The aim of this study was to evaluate the influence of a six-week aerobic training on peripheral blood in adults with Down syndrome. Fifteen men with Down syndrome (average age 22.4 years ± 0.91) with moderate or severe intellectual disability took part in the study. Patients underwent a training program three times a week for six weeks. Venous blood samples of 10 ml were collected from every examined patient, 24 hours before and after the exercise. The blood samples were submitted to hematological examination (hematocrit, fibrinogen, plasma viscosity, red blood cell (RBC) number, RBC indicators: mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH)), reduced glutathione (GSH) level and number of macrocytes, polikilocytrometric examination of RBC and rheological blood examination (elongation index (EI), aggregation index (AI), syllectogram amplitude (AMP), aggregation half time (t1/2)) was made by LORCA. Amoderate six-week physical training performed on a cycloergometer resulted in a statistically significant decrease in the MCV value, hematocrit and plasma viscosity. The six-week cycloergometer training caused a statistically significant increase in the GSH level and erythrocyte pliability at a shear stress of 0.58 Pa.

Lung function, transfusion, pulmonary capillary blood volume and sickle cell disease.

PubMed

Lunt, Alan; McGhee, Emily; Robinson, Polly; Rees, David; Height, Susan; Greenough, Anne

2016-02-01

Lung function abnormalities occur in children with sickle cell disease (SCD) and may be associated with elevated pulmonary blood volume. To investigate that association, we determined whether blood transfusion in SCD children acutely increased pulmonary capillary blood volume (PCBV) and increased respiratory system resistance (Rrs5). Measurements of Rrs5 and spirometry were made before and after blood transfusion in 18 children, median age 14.2 (6.6-18.5) years. Diffusing capacity for carbon monoxide and nitric oxide were assessed to calculate the PCBV. Post transfusion, the median Rrs5 had increased from 127.4 to 141.3% predicted (p<0.0001) and pulmonary capillary blood volume from 39.7 to 64.1 ml/m2 (p<0.0001); forced expiratory volume in one second (p=0.0056) and vital capacity (p=0.0008) decreased. The increase in Rrs5 correlated with the increase in PCBV (r=0.50, p=0.0493). Increased pulmonary capillary blood volume may at least partially explain the lung function abnormalities in SCD children. Copyright © 2015 Elsevier B.V. All rights reserved.

Effects of nifedipine and captopril on vascular capacitance of ganglion-blocked anesthetized dogs.

PubMed

Ogilvie, R I; Zborowska-Sluis, D

1990-03-01

The hemodynamic effects of nifedipine and captopril at doses producing similar reductions in arterial pressure were studied in pentobarbital-anesthetized ventilated dogs after splenectomy during ganglion blockade with hexamethonium. Mean circulatory filling pressure (Pmcf) was determined during transient circulatory arrest induced by acetylcholine at baseline circulating blood volumes and after increases of 5 and 10 mL/kg. Central blood volumes (pulmonary artery to aortic root) were determined from transit times, and separately determined cardiac outputs (right atrium to pulmonary artery) were estimated by thermodilution. Nifedipine (n = 5) increased Pmcf at all circulating blood volumes and reduced total vascular capacitance without a change in total vascular compliance. Central blood volume, right atrial pressure, and cardiac output were increased with induced increases in circulating blood volume. In contrast, captopril (n = 5) did not alter total vascular capacitance, central blood volume, right atrial pressure, or cardiac output at baseline or with increased circulating volume. Thus, at doses producing similar reductions in arterial pressure, nifedipine but not captopril increased venous return and cardiac output in ganglion-blocked dogs.

Capillary blood islet autoantibody screening for identifying pre-type 1 diabetes in the general population: design and initial results of the Fr1da study

PubMed Central

Raab, Jennifer; Haupt, Florian; Scholz, Marlon; Matzke, Claudia; Warncke, Katharina; Lange, Karin; Assfalg, Robin; Weininger, Katharina; Wittich, Susanne; Löbner, Stephanie; Beyerlein, Andreas; Nennstiel-Ratzel, Uta; Lang, Martin; Laub, Otto; Dunstheimer, Desiree; Bonifacio, Ezio; Achenbach, Peter; Winkler, Christiane; Ziegler, Anette-G

2016-01-01

Introduction Type 1 diabetes can be diagnosed at an early presymptomatic stage by the detection of islet autoantibodies. The Fr1da study aims to assess whether early staging of type 1 diabetes (1) is feasible at a population-based level, (2) prevents severe metabolic decompensation observed at the clinical manifestation of type 1 diabetes and (3) reduces psychological distress through preventive teaching and care. Methods and analysis Children aged 2–5 years in Bavaria, Germany, will be tested for the presence of multiple islet autoantibodies. Between February 2015 and December 2016, 100 000 children will be screened by primary care paediatricians. Islet autoantibodies are measured in capillary blood samples using a multiplex three-screen ELISA. Samples with ELISA results >97.5th centile are retested using reference radiobinding assays. A venous blood sample is also obtained to confirm the autoantibody status of children with at least two autoantibodies. Children with confirmed multiple islet autoantibodies are diagnosed with pre-type 1 diabetes. These children and their parents are invited to participate in an education and counselling programme at a local diabetes centre. Depression and anxiety, and burden of early diagnosis are also assessed. Results Of the 1027 Bavarian paediatricians, 39.3% are participating in the study. Overall, 26 760 children have been screened between February 2015 and November 2015. Capillary blood collection was sufficient in volume for islet autoantibody detection in 99.46% of the children. The remaining 0.54% had insufficient blood volume collected. Of the 26 760 capillary samples tested, 0.39% were positive for at least two islet autoantibodies. Discussion Staging for early type 1 diabetes within a public health setting appears to be feasible. The study may set new standards for the early diagnosis of type 1 diabetes and education. Ethics dissemination The study was approved by the ethics committee of Technische Universität München (Nr. 70/14). PMID:27194320

Capillary blood islet autoantibody screening for identifying pre-type 1 diabetes in the general population: design and initial results of the Fr1da study.

PubMed

Raab, Jennifer; Haupt, Florian; Scholz, Marlon; Matzke, Claudia; Warncke, Katharina; Lange, Karin; Assfalg, Robin; Weininger, Katharina; Wittich, Susanne; Löbner, Stephanie; Beyerlein, Andreas; Nennstiel-Ratzel, Uta; Lang, Martin; Laub, Otto; Dunstheimer, Desiree; Bonifacio, Ezio; Achenbach, Peter; Winkler, Christiane; Ziegler, Anette-G

2016-05-18

Type 1 diabetes can be diagnosed at an early presymptomatic stage by the detection of islet autoantibodies. The Fr1da study aims to assess whether early staging of type 1 diabetes (1) is feasible at a population-based level, (2) prevents severe metabolic decompensation observed at the clinical manifestation of type 1 diabetes and (3) reduces psychological distress through preventive teaching and care. Children aged 2-5 years in Bavaria, Germany, will be tested for the presence of multiple islet autoantibodies. Between February 2015 and December 2016, 100 000 children will be screened by primary care paediatricians. Islet autoantibodies are measured in capillary blood samples using a multiplex three-screen ELISA. Samples with ELISA results >97.5th centile are retested using reference radiobinding assays. A venous blood sample is also obtained to confirm the autoantibody status of children with at least two autoantibodies. Children with confirmed multiple islet autoantibodies are diagnosed with pre-type 1 diabetes. These children and their parents are invited to participate in an education and counselling programme at a local diabetes centre. Depression and anxiety, and burden of early diagnosis are also assessed. Of the 1027 Bavarian paediatricians, 39.3% are participating in the study. Overall, 26 760 children have been screened between February 2015 and November 2015. Capillary blood collection was sufficient in volume for islet autoantibody detection in 99.46% of the children. The remaining 0.54% had insufficient blood volume collected. Of the 26 760 capillary samples tested, 0.39% were positive for at least two islet autoantibodies. Staging for early type 1 diabetes within a public health setting appears to be feasible. The study may set new standards for the early diagnosis of type 1 diabetes and education. The study was approved by the ethics committee of Technische Universität München (Nr. 70/14). Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

Comparison of whole body and tissue blood volumes in rainbow trout (Salmo gairdneri) with 125I bovine serum albumin and 51Cr-erythrocyte tracers

USGS Publications Warehouse

Gingerich, W.H.; Pityer, R.A.

1989-01-01

Total, packed cell and, plasma volume estimates were made for the whole body and selected tissues of rainbow trout by the simultaneous injection of radiolabelled trout erythrocyte (51Cr-RBC) and radioiodinated bovine serum albumin (125I-BSA) tracers. Blood volumes were estimated with both markers separately by the tracer-hematocrit method and as the combination of the 51Cr-RBC packed cell and 125I-BSA plasma volumes. Mean whole body blood volume was significantly less when calculated from the 51Cr-RBC tracer data (3.52±0.78 ml/100 g; ±SD) than when calculated with the 125I-BSA tracer (5.06±0.86 ml/100 g) or as the sum of the two volumes combined (4.49±0.60 ml/100 g). The whole body hematocrit (28±5%), estimated as the quotient of the 51Cr-RBC volume divided by the sum of the 125I-BSA and the 51Cr-RBC volumes, also was significantly less than the dorsal aortic microhematocrit (36±4%). Estimates of total blood volumes in most tissues were significantly smaller when calculated from the51Cr-RBC data than when calculated by the other two methods. Tissue blood volumes were greatest in highly vascularized and well perfused tissues and least in poorly vascularized tissues. The relative degree of vascularization among tissues generally remained the same regardless of whether the red cell or the plasma tracer was used to calculated blood volume. It is not clear whether the expanded plasma volume is the result of the distribution of erythrocyte-poor blood into the secondary circulation or the result of extravascular exchange of plasma proteins.

Life-course blood pressure in relation to brain volumes

PubMed Central

Power, Melinda C.; Schneider, Andrea L. C.; Wruck, Lisa; Griswold, Michael; Coker, Laura H.; Alonso, Alvaro; Jack, Clifford R.; Knopman, David; Mosley, Thomas H.; Gottesman, Rebecca F

2016-01-01

INTRODUCTION The impact of blood pressure on brain volumes may be time- or pattern-dependent. METHODS In 1678 participants from the Atherosclerosis Risk in Communities Neurocognitive Study, we quantified the association between measures and patterns of blood pressure over three time points (~24 or ~15 years prior and concurrent with neuroimaging) with late life brain volumes. RESULTS Higher diastolic blood pressure ~24 years prior, higher systolic and pulse pressure ~15 years prior, and consistently elevated or rising systolic blood pressure from ~15 years prior to concurrent with neuroimaging, but not blood pressures measured concurrent with neuroimaging, were associated with smaller volumes. The pattern of hypertension ~15 years prior and hypotension concurrent with neuroimaging was associated with smaller volumes in regions preferentially affected by Alzheimer’s disease (e.g., hippocampus: −0.27 standard units, 95%CI:−0.51,−0.03). DISCUSSION Hypertension 15 to 24 years prior is relevant to current brain volumes. Hypertension followed by hypotension appears particularly detrimental. PMID:27139841

Effects of growth, diving history, and high altitude on blood oxygen capacity in harbor seals

NASA Technical Reports Server (NTRS)

Kodama, A. M.; Elsner, R.; Pace, N.

1977-01-01

Blood volume and body composition for diving and nondiving harbor seals were measured at six-week intervals during a 10-month period of captitivity. Whole body hematocrit, red cell volume per kg of lean body mass, and total circulating hemoglobin per kg lean body mass were significantly higher in the diving group, but relatively large blood volumes expressed in terms of body weight (11-12%) were found in both groups. A pair of harbor seals exposed to high altitude for about three months registered significant increases in red cell volume, blood hemoglobin levels, and blood volume expressed in terms of body weight; results of alveolar gas analyses indicate that hyperventilation also occurred. These typical mammalian responses to hypoxia suggest that the harbor seal's large blood volume and high hemoglobin content are an expression of phylogenetic control, and that in spite of its adaptability to apnea during its diving life, the animal cannot be considered preacclimatized to high altitude.

Brain Structure and Function Associated with a History of Sport Concussion: A Multi-Modal Magnetic Resonance Imaging Study.

PubMed

Churchill, Nathan; Hutchison, Michael; Richards, Doug; Leung, General; Graham, Simon; Schweizer, Tom A

2017-02-15

There is growing concern about the potential long-term consequences of sport concussion for young, currently active athletes. However, there remains limited information about brain abnormalities associated with a history of concussion and how they relate to clinical factors. In this study, advanced MRI was used to comprehensively describe abnormalities in brain structure and function associated with a history of sport concussion. Forty-three athletes (21 male, 22 female) were recruited from interuniversity teams at the beginning of the season, including 21 with a history of concussion and 22 without prior concussion; both groups also contained a balanced sample of contact and noncontact sports. Multi-modal MRI was used to evaluate abnormalities in brain structure and function. Athletes with a history of concussion showed frontal decreases in brain volume and blood flow. However, they also demonstrated increased posterior cortical volume and elevated markers of white matter microstructure. A greater number of prior concussions was associated with more extensive decreases in cerebral blood flow and insular volume, whereas recovery time from most recent concussion was correlated with reduced frontotemporal volume. White matter showed limited correlations with clinical factors, predominantly in the anterior corona radiata. This study provides the first evidence of the long-term effects of concussion on gray matter volume, blood flow, and white matter microstructure within a single athlete cohort. This was examined for a mixture of male and female athletes in both contact and noncontact sports, demonstrating the relevance of these findings for the overall sporting community.

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.

Successive changes of hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta).

PubMed

Kakizoe, Yuka; Sakaoka, Ken; Kakizoe, Futoshi; Yoshii, Makoto; Nakamura, Hitoshi; Kanou, Yoshihiko; Uchida, Itaru

2007-03-01

Hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta) from the ages of 1 mo to 3 yr were obtained to establish baseline values. Five clinically normal loggerhead turtles were selected from the same clutch and raised in an indoor artificial nesting beach. Blood samples were successively collected and examined for various blood characteristics for a maximum total of 15 times. Hematologic characteristics, including packed cell volume, white blood cell counts, and white blood cell differentials; and plasma chemistry values, including total bilirubin, total protein, albumin, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, gamma-glutamic transpeptidase, creatinine, blood urea nitrogen, uric acid, alkaline phosphatase, amylase, triglyceride, total cholesterol, ionized sodium, ionized potassium and ionized chlorine, were measured. These results were used to establish a hematology and blood chemistry baseline for captive juvenile loggerhead turtles and will aid in their medical management.

C-arm flat detector computed tomography parenchymal blood volume imaging: the nature of parenchymal blood volume parameter and the feasibility of parenchymal blood volume imaging in aneurysmal subarachnoid haemorrhage patients.

PubMed

Kamran, Mudassar; Byrne, James V

2015-09-01

C-arm flat detector computed tomography (FDCT) parenchymal blood volume (PBV) measurements allow assessment of cerebral haemodynamics in the neurointerventional suite. This paper explores the feasibility of C-arm computed tomography (CT) PBV imaging and the relationship between the C-arm CT PBV and the MR-PWI-derived cerebral blood volume (CBV) and cerebral blood flow (CBF) parameters in aneurysmal subarachnoid haemorrhage (SAH) patients developing delayed cerebral ischemia (DCI). Twenty-six patients with DCI following aneurysmal SAH underwent a research C-arm CT PBV scan using a biplane angiography system and contemporaneous MR-PWI scan as part of a prospective study. Quantitative whole-brain atlas-based volume-of-interest analysis in conjunction with Pearson correlation and Bland-Altman tests was performed to explore the agreement between C-arm CT PBV and MR-derived CBV and CBF measurements. All patients received medical management, while eight patients (31%) underwent selective intra-arterial chemical angioplasty. Colour-coded C-arm CT PBV maps were 91% sensitive and 100% specific in detecting the perfusion abnormalities. C-arm CT rPBV demonstrated good agreement and strong correlation with both MR-rCBV and MR-rCBF measurements; the agreement and correlation were stronger for MR-rCBF relative to MR-rCBV and improved for C-arm CT PBV versus the geometric mean of MR-rCBV and MR-rCBF. Analysis of weighted means showed that the C-arm CT PBV has a preferential blood flow weighting (≈ 60% blood flow and ≈ 40% blood volume weighting). C-arm CT PBV imaging is feasible in DCI following aneurysmal SAH. PBV is a composite perfusion parameter incorporating both blood flow and blood volume weightings. That PBV has preferential (≈ 60%) blood flow weighting is an important finding, which is of clinical significance when interpreting the C-arm CT PBV maps, particularly in the setting of acute brain ischemia.

Hematology and erythrocyte osmotic fragility of the Franquet's fruit bat (Epomops franqueti).

PubMed

Ekeolu, Oyetunde Kazeem; Adebiyi, Olamide Elizabeth

2018-03-15

Hematological parameters are vital diagnostic tools for understanding health dynamics of humans and animals. Franquet's fruit bat (Epomops franqueti) is host to several parasites such as protozoa, bacteria, viruses and mites. Yet, studies exploring the values of its blood components with interest for research or food purposes are scarce. Thus, this study was carried out to investigate the hematological values of the adult E. franqueti. Seventeen (nine female and eight male) apparently healthy adult E. franqueti were captured from their roosting colony. Blood samples were collected for determination of erythrocyte indices [red blood cell count (RBC), packed cell volume (PCV), hemoglobin (Hb) concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC)] and leukocyte indices [total white blood cell counts (WBC), lymphocytes, eosinophil, monocytes, neutrophil count and erythrocytes osmotic fragility]. There were no significant (p≥0.05) sex-related differences in RBC, PCV, Hb concentration, MCV, MCH, MCHC and total and differential WBC of E. franqueti. Erythrocyte osmotic fragility was significantly higher in female than in male E. franqueti at 0.1% NaCl. These considerations are critical in establishing reference ranges of blood parameters for E. franqueti and may provide insight to why they serve as reservoir hosts for several microorganisms.

Blood venous sample collection: Recommendations overview and a checklist to improve quality.

PubMed

Giavarina, Davide; Lippi, Giuseppe

2017-07-01

The extra-analytical phases of the total testing process have substantial impact on managed care, as well as an inherent high risk of vulnerability to errors which is often greater than that of the analytical phase. The collection of biological samples is a crucial preanalytical activity. Problems or errors occurring shortly before, or soon after, this preanalytical step may impair sample quality and characteristics, or else modify the final results of testing. The standardization of fasting requirements, rest, patient position and psychological state of the patient are therefore crucial for mitigating the impact of preanalytical variability. Moreover, the quality of materials used for collecting specimens, along with their compatibility, can guarantee sample quality and persistence of chemical and physical characteristics of the analytes over time, so safeguarding the reliability of testing. Appropriate techniques and sampling procedures are effective to prevent problems such as hemolysis, undue clotting in the blood tube, draw of insufficient sample volume and modification of analyte concentration. An accurate identification of both patient and blood samples is a key priority as for other healthcare activities. Good laboratory practice and appropriate training of operators, by specifically targeting collection of biological samples, blood in particular, may greatly improve this issue, thus lowering the risk of errors and their adverse clinical consequences. The implementation of a simple and rapid check-list, including verification of blood collection devices, patient preparation and sampling techniques, was found to be effective for enhancing sample quality and reducing some preanalytical errors associated with these procedures. The use of this tool, along with implementation of objective and standardized systems for detecting non-conformities related to unsuitable samples, can be helpful for standardizing preanalytical activities and improving the quality of laboratory diagnostics, ultimately helping to reaffirm a "preanalytical" culture founded on knowledge and real risk perception. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Transcutaneous blood glucose monitoring system based on an ISFET glucose sensor and studies on diabetic patients.

PubMed

Ito, N; Saito, A; Kayashima, S; Kimura, J; Kuriyama, T; Nagata, N; Arai, T; Kikuchi, M

1995-01-01

A transcutaneous blood glucose monitoring system consists of an ion-sensitive field-effect transistor (ISFET) glucose sensor unit and a suction effusion fluid (SEF) collecting unit. The SEF is directly collected by a weak suction (400 mmHg absolute pressure) through the skin from which the corneum layer of the epidermis has been previously removed. An ISFET glucose sensor unit is able to measure glucose concentrations in a microliter order sampling volume. The system was applied to three diabetic patients during a 75 g oral glucose tolerance test for monitoring blood glucose levels. During the experiments, glucose changes in the SEF followed actual blood glucose levels with 10 min delays. Results suggest the feasibility of utilizing quasi-continuous, transcutaneous blood glucose monitoring for individual patients with various diabetic histories or diabetic complications.

Multiplex Profiling Identifies Distinct Local and Systemic Alterations during Intraperitoneal Chemotherapy for Ovarian Cancer: an NRG Oncology/Gynecologic Oncology Group Study

PubMed Central

Grabosch, Shannon; Tseng, George; Edwards, Robert P; Lankes, Heather A.; Moore, Kathleen; Odunsi, Kunle; Vlad, Anda; Ma, Tianzhou; Strange, Mary; Brozick, Joan; Lugade, Amit; Omilian, Angela; Bshara, Wiam; Stuckey, Ashley R.; Walker, Joan L.; Birrer, Michael

2017-01-01

OBJECTIVES Ovarian cancer leads to abdominal carcinomatosis and late stage (III/IV) diagnosis in 75% of patients. Three randomized phase III trials have demonstrated that intraperitoneal (IP) chemotherapy improves outcomes in epithelial ovarian cancer. While IP treatment is validated by clinical trials, there is a poor understanding of the mechanism(s) leading to the survival advantage other than the increased concentration of cytotoxic drugs within the tumor microenvironment. A better understanding of this process through analysis of dynamic biomarkers should promote novel approaches that may enhance tumor clearance. We propose this pilot study to confirm the feasibility of collecting serial peritoneal samples from implanted catheters in women receiving IP chemotherapy. We believe these specimens may be used for multiplex analysis to reveal unique biomarker fluctuations when compared to peripheral blood. METHODS From 13 women participating on GOG 252, 30 whole blood, 12 peritoneal fluid (PF), and 20 peritoneal wash (PW) with 30 mL saline were obtained. Samples were requested prior to the first three chemotherapy cycles. Samples were assessed for volume, cell populations, protein, RNA, and miRNA content changes. RESULTS Median volume for PF was 1.6 mL and 3.1 mL for PW. PW is a dilution of PF capable of capturing measurable biomarkers. Peritoneal aspirates contain a unique profile of biomarkers distinct from blood. miRNA undergo earlier alteration with chemotherapy than genes. Flow cytometry does not adequately capture biomarker fluctuations. CONCLUSIONS As a proof of principle study, this trial provides evidence that sampling the peritoneal cavity can be adapted for biomarker analysis. PMID:28483269

Coinfusion of dextrose-containing fluids and red blood cells does not adversely affect in vitro red blood cell quality.

PubMed

Keir, Amy K; Hansen, Adele L; Callum, Jeannie; Jankov, Robert P; Acker, Jason P

2014-08-01

Transfusion guidelines advise against coinfusing red blood cells (RBCs) with solutions other than 0.9% saline. We evaluated the impact of coinfusion with dextrose-containing fluids (DW) on markers of RBC quality. A pool-and-split design was used to allow conditions to be tested on each pool within 2 hours of irradiation. Three pools at each storage age (5, 14, and 21 days) were created for each phase. In Phase 1, samples were infused through a neonatal transfusion apparatus alone or with treatment solutions: D5W, D10W, D5W/0.2% saline, and 0.9% saline. In Phase 2, samples were incubated alone or in a 1:1 ratio with treatment solutions and tested after 5, 30, and 180 minutes. Hemolysis, supernatant potassium, RBC indices, morphology, and deformability were measured on all samples. In Phase 1, RBCs transfused alone through the apparatus had higher (p<0.01) hematocrit, total hemoglobin, and supernatant potassium compared to all other groups. No statistical differences were identified between groups for other measured variables. In Phase 2, mean corpuscular volume of all samples containing DW increased with incubation length and were higher (p<0.01) than RBCs incubated alone or with 0.9% saline after 30 and 180 minutes. RBCs incubated with D5W and D5W/0.2% saline had greater (p<0.05) hemolysis than RBCs alone after 180 minutes. In vitro characteristics of RBCs coinfused with 0.9% saline or D10W were not adversely impacted. When developing clinical studies in neonates, we recommend use of D10W and a transfusion apparatus that minimizes the contact volume of the coinfusate with the RBC. © 2014 AABB.

Pharmacokinetic-pharmacodynamic study of subcutaneous injection of depot nandrolone decanoate using dried blood spots sampling coupled with ultrapressure liquid chromatography tandem mass spectrometry assays.

PubMed

Singh, Gurmeet K S; Turner, Leo; Desai, Reena; Jimenez, Mark; Handelsman, David J

2014-07-01

Testosterone (T) and nandrolone (N) esters require deep im injections by medical personnel but these often deposit injectate into sc fat so that more convenient sc self-administration may be feasible. To investigate the feasibility and pharmacology of sc injection of N decanoate in healthy men using dried blood spot (DBS) for frequent blood sampling without clinic visits. Healthy male volunteers received 100 mg N decanoate by a single sc injection. Finger-prick capillary blood was spotted onto filter paper before injection daily at home for 21 d and stored at room temperature. Venous whole blood was also spotted onto filter paper before and weekly for 3 wk after injection. DBS were extracted for assay of N and T by liquid chromatography tandem mass spectrometry in a single batch with serum concentrations estimated with adjustment for capillary blood sample volume and hematocrit to define peak (N) or nadir (T) time and concentration from individual daily measurements. Daily serum N peaked 2.50 ± 0.25 (SEM) ng/mL at a median (range) of 6 (4-13) days causing a reduction in serum T from 3.50 ± 0.57 ng/mL at baseline to a nadir of 0.38 ± 0.13 (SEM) ng/mL (89 ± 3% suppression) at a median (range) of 8 (5-16) days. Simultaneously sampled capillary, venous whole blood, and serum gave almost identical results for serum T and N. Finger-pricks and sc injections were well tolerated. This study demonstrates that A) DBS sampling with liquid chromatography mass spectrometry steroid analysis achieves frequent time sampling in the community without requiring clinic visits, venesection, or frozen serum storage, and B) androgen esters in an oil vehicle can be delivered effectively by sc injection, thus avoiding the need for medically supervised deep-im injections.

Stable-Isotope Dilution HPLC-Electrospray Ionization Tandem Mass Spectrometry Method for Quantifying Hydroxyurea in Dried Blood Samples.

PubMed

Marahatta, Anu; Megaraj, Vandana; McGann, Patrick T; Ware, Russell E; Setchell, Kenneth D R

2016-12-01

Sickle cell anemia (SCA) is a life-threatening blood disorder characterized by the presence of sickle-shaped erythrocytes. Hydroxyurea is currently the only US Food and Drug Administration-approved treatment and there is a need for a convenient method to monitor compliance and hydroxyurea concentrations, especially in pediatric SCA patients. We describe a novel approach to the determination of hydroxyurea concentrations in dried whole blood collected on DMPK-C cards or volumetric absorptive microsampling (VAMS) devices. Hydroxyurea was quantified by electrospray ionization LC-MS/MS using [ 13 C 15 N 2 ]hydroxyurea as the internal standard. Calibrators were prepared in whole blood applied to DMPK-C cards or VAMS devices. Calibration curves for blood hydroxyurea measured from DMPK-C cards and VAMS devices were linear over the range 0.5-60 μg/mL. Interassay and intraassay CVs were <15% for blood collected by both methods, and the limit of detection was 5 ng/mL. Whole blood hydroxyurea was stable for up to 60 days on DMPK-C cards and VAMS devices when frozen at -20 °C or -80 °C. Whole blood hydroxyurea concentrations in samples collected on DMPK-C cards or VAMS devices from SCA patients were in close agreement. This tandem mass spectrometry method permits measurement of hydroxyurea concentrations in small volumes of dried blood applied to either DMPK-C cards or VAMS devices with comparable performance. This method for measuring hydroxyurea from dried blood permits the evaluation of therapeutic drug monitoring, individual pharmacokinetics, and medication adherence using heel/finger-prick samples from pediatric patients with SCA treated with hydroxyurea. © 2016 American Association for Clinical Chemistry.

Automated processing of first-pass radioisotope ventriculography data to determine essential central circulation parameters

NASA Astrophysics Data System (ADS)

Krotov, Aleksei; Pankin, Victor

2017-09-01

The assessment of central circulation (including heart function) parameters is vital in the preventive diagnostics of inherent and acquired heart failures and during polychemotherapy. The protocols currently applied in Russia do not fully utilize the first-pass assessment (FPRNA) and that results in poor data formalization, while the FPRNA is the one of the fastest, affordable and compact methods among other radioisotope diagnostics protocols. A non-imaging algorithm basing on existing protocols has been designed to use the readings of an additional detector above vena subclavia to determine the total blood volume (TBV), not requiring blood sampling in contrast to current protocols. An automated processing of precordial detector readings is presented, in order to determine the heart strike volume (SV). Two techniques to estimate the ejection fraction (EF) of the heart are discussed.

Detection of low-volume blood loss: compensatory reserve versus traditional vital signs.

PubMed

Stewart, Camille L; Mulligan, Jane; Grudic, Greg Z; Convertino, Victor A; Moulton, Steven L

2014-12-01

Humans are able to compensate for low-volume blood loss with minimal change in traditional vital signs. We hypothesized that a novel algorithm, which analyzes photoplethysmogram (PPG) wave forms to continuously estimate compensatory reserve would provide greater sensitivity and specificity to detect low-volume blood loss compared with traditional vital signs. The compensatory reserve index (CRI) is a measure of the reserve remaining to compensate for reduced central blood volume, where a CRI of 1 represents supine normovolemia and 0 represents the circulating blood volume at which hemodynamic decompensation occurs; values between 1 and 0 indicate the proportion of reserve remaining. Subjects underwent voluntary donation of 1 U (approximately 450 mL) of blood. Demographic and continuous noninvasive vital sign wave form data were collected, including PPG, heart rate, systolic blood pressure, cardiac output, and stroke volume. PPG wave forms were later processed by the algorithm to estimate CRI values. Data were collected from 244 healthy subjects (79 males and 165 females), with a mean (SD) age of 40.1 (14.2) years and mean (SD) body mass index of 25.6 (4.7). After blood donation, CRI significantly decreased in 92% (α = 0.05; 95% confidence interval [CI], 88-95%) of the subjects. With the use of a threshold decrease in CRI of 0.05 or greater for the detection of 1 U of blood loss, the receiver operating characteristic area under the curve was 0.90, with a sensitivity of 0.84 and specificity of 0.86. In comparison, systolic blood pressure (52%; 95% CI, 45-59%), heart rate (65%; 95% CI, 58-72%), cardiac output (47%; 95% CI, 40-54%), and stroke volume (74%; 95% CI, 67-80%) changed in fewer subjects, had significantly lower receiver operating characteristic area under the curve values, and significantly lower specificities for detecting the same volume of blood loss. Consistent with our hypothesis, CRI detected low-volume blood loss with significantly greater specificity than other traditional physiologic measures. These findings warrant further evaluation of the CRI algorithm in actual trauma settings. Diagnostic study, level II.

Estimates of plasma, packed cell and total blood volume in tissues of the rainbow trout (Salmo gairdneri)

USGS Publications Warehouse

Gingerich, W.H.; Pityer, R.A.; Rach, J.J.

1987-01-01

1. Total blood volume and relative blood volumes in selected tissues were determined in non-anesthetized, confined rainbow trout by using 51Cr-labelled trout erythrocytes as a vascular space marker.2. Mean total blood volume was estimated to be 4.09 ± 0.55 ml/100 g, or about 75% of that estimated with the commonly used plasma space marker Evans blue dye.3. Relative tissue blood volumes were greatest in highly perfused tissues such as kidney, gills, brain and liver and least in mosaic muscle.4. Estimates of tissue vascular spaces, made using radiolabelled erythrocytes, were only 25–50% of those based on plasma space markers.5. The consistently smaller vascular volumes obtained with labelled erythrocytes could be explained by assuming that commonly used plasma space markers diffuse from the vascular compartment.

Effects of etravirine on the pharmacokinetics and pharmacodynamics of warfarin in rats

PubMed Central

John, J; John, M; Wu, L; Hsiao, C; Abobo, CV; Liang, D

2013-01-01

Background and Purpose Warfarin is often used with etravirine (ETV) to prevent HIV-related thromboembolic events. As both warfarin and ETV bind to plasma proteins and are metabolized by hepatic cytochrome P450s, they are likely to interact. Hence, we evaluated the effect of ETV on the pharmacokinetics and blood clotting time of racemic warfarin in rats. Experimental Approach Two groups of male Sprague-Dawley rats, in which the jugular vein had been cannulated, were studied. The control group (n = 10) received 1 mg·kg−1 racemic warfarin i.v., and the test group (n = 13) 1 mg·kg−1 of racemic warfarin followed by 25 mg·kg−1 ETV i.v. Serial blood samples were collected for up to 144 h and the blood clotting time (calculated as international normalized ratio [INR]) measured in blood plasma at each sample point. Plasma concentrations of R-warfarin, S-warfarin, R-7-hydroxywarfarin and S-7-hydroxywarfarin were measured by a LC/MS/MS method using a chiral lux cellulose-1 column. Pharmacokinetic parameters were analysed using non-compartmental methods. Key Results ETV significantly increased, by threefold, the systemic clearance and volume of distribution of S-warfarin, but not those of R-warfarin. ETV decreased the total AUC of warfarin, but had no effect on its elimination half-life. ETV also increased the systemic clearance of both R-7-hydroxywarfarin and S-7-hydroxywarfarin but only increased the volume of distribution of R-7-hydroxywarfarin. Interestingly, the effect of warfarin on blood clotting time (INR) was significantly increased in the presence of etravirine. Conclusion and Implications Our data suggest that etravirine may potentiate the anticoagulant effect of warfarin and this could have clinical significance. PMID:23215758

Rapid Point of Care Analyzer for the Measurement of Cyanide in Blood

PubMed Central

Ma, Jian; Ohira, Shin-Ichi; Mishra, Santosh K.; Puanngam, Mahitti; Dasgupta, Purnendu K.; Mahon, Sari B.; Brenner, Matthew; Blackledge, William; Boss, Gerry R.

2011-01-01

A simple, sensitive optical analyzer for the rapid determination of cyanide in blood in point of care applications is described. HCN is liberated by the addition of 20% H3PO4 and is absorbed by a paper filter impregnated with borate-buffered (pH 9.0) hydroxoaquocobinamide Hereinafter called cobinamide). Cobinamide on the filter changes color from orange (λmax = 510 nm) to violet (λmax = 583 nm) upon reaction with cyanide. This color change is monitored in the transmission mode by a light emitting diode (LED) with a 583 nm emission maximum and a photodiode detector. The observed rate of color change increases 10x when the cobinamide solution for filter impregnation is prepared in borate-buffer rather than in water. The use of a second LED emitting at 653 nm and alternate pulsing of the LEDs improve the limit of detection by 4x to ~ 0.5 μM for a 1 mL blood sample. Blood cyanide levels of imminent concern (≥ 10 μM) can be accurately measured in ~ 2 min. The response is proportional to the mass of cyanide in the sample – smaller sample volumes can be successfully used with proportionate change in the concentration LODs. Bubbling air through the blood-acid mixture was found effective for mixing of the acid with the sample and the liberation of HCN. A small amount of ethanol added to the top of the blood was found to be the most effective means to prevent frothing during aeration. The relative standard deviation (RSD) for repetitive determination of blood samples containing 9 μM CN was 1.09% (n=5). The technique was compared blind with a standard microdiffusion-spectrophotometric method used for the determination of cyanide in rabbit blood. The results showed good correlation (slope 1.05, r2 0.9257); independent calibration standards were used. PMID:21553921

Cell Death, Inflammation, Tumor Burden, and Proliferation Blood Biomarkers Predict Lung Cancer Radiotherapy Response and Correlate With Tumor Volume and Proliferation Imaging.

PubMed

Salem, Ahmed; Mistry, Hitesh; Backen, Alison; Hodgson, Clare; Koh, Pek; Dean, Emma; Priest, Lynsey; Haslett, Kate; Trigonis, Ioannis; Jackson, Alan; Asselin, Marie-Claude; Dive, Caroline; Renehan, Andrew; Faivre-Finn, Corinne; Blackhall, Fiona

2018-05-01

There is an unmet need to develop noninvasive biomarkers to stratify patients in drug-radiotherapy trials. In this pilot study we investigated lung cancer radiotherapy response and toxicity blood biomarkers and correlated findings with tumor volume and proliferation imaging. Blood samples were collected before and during (day 21) radiotherapy. Twenty-six cell-death, hypoxia, angiogenesis, inflammation, proliferation, invasion, and tumor-burden biomarkers were evaluated. Clinical and laboratory data were collected. Univariate analysis was performed on small-cell and non-small-cell lung cancer (NSCLC) whereas multivariate analysis focused on NSCLC. Blood samples from 78 patients were analyzed. Sixty-one (78.2%) harbored NSCLC, 48 (61.5%) received sequential chemoradiotherapy. Of tested baseline biomarkers, undetectable interleukin (IL)-1b (hazard ratio [HR], 4.02; 95% confidence interval [CI], 2.04-7.93; P < .001) was the only significant survival covariate. Of routinely collected laboratory tests, high baseline neutrophil count was a significant survival covariate (HR, 1.07; 95% CI, 1.02-1.11; P = .017). Baseline IL-1b and neutrophil count were prognostic for survival in a multivariate model. The addition of day-21 cytokeratin-19 antigen modestly improved this model's survival prediction (concordance probability, 0.75-0.78). Chemotherapy (P < .001) and baseline keratinocyte growth factor (P = .019) predicted acute esophagitis, but only chemotherapy remained significant after Bonferroni correction. Baseline angioprotein-1 and hepatocyte growth factor showed a direct correlation with tumor volume whereas changes in vascular cell adhesion molecule 1 showed significant correlations with 18F-fluorothymidine (FLT) positron emission tomography (PET). Select biomarkers are prognostic after radiotherapy in this lung cancer series. The correlation between circulating biomarkers and 18F-FLT PET is shown, to our knowledge for the first time, highlighting their potential role as imaging surrogates. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Urtica Dioica Distillate Regenerates Pancreatic Beta Cells in Streptozotocin-Induced Diabetic Rats.

PubMed

Gohari, Ali; Noorafshan, Ali; Akmali, Masoumeh; Zamani-Garmsiri, Fahimeh; Seghatoleslam, Atefeh

2018-03-01

Urtica dioica is known as an anti-hyperglycemic plant. Urtica dioica distillate (UD) is a traditional Iranian drink, locally known as "aragh gazaneh". In spite of its widespread consumption in Iran, according to traditional Iranian medicine, there is no scientific report on the usefulness of UD for diabetic patients. This survey was designed to evaluate its protective effects for the recovery from diabetes by determining the serum insulin, blood glucose, volume of pancreatic islets, and the number and volume of β-cells in diabetic rats. A total of 48 Sprague-Dawley male rats (200-250 g) were randomly distributed into 6 groups (n=8), including non-diabetic plus distilled water (DW), non-diabetic plus UD, diabetic plus DW, diabetic plus UD, diabetic plus insulin, and diabetic plus glibenclamide. DW, UD, and glibenclamide were administered via intragastric gavage and insulin was injected subcutaneously. After four weeks of experiments, blood samples were collected for serum insulin and blood glucose assay. Pancreas was also evaluated using stereological method. The SPSS software was used for statistical analysis. Kruskal-Wallis, repeated measurements, and Mann-Whitney U test were applied for comparisons between the groups. The treatment of diabetic rats with UD reduced the blood glucose dramatically (P<0.001) and increased serum insulin levels significantly (P=0.03) in comparison to the diabetic plus DW rats. Treatment with UD did not affect the mean β-cell volumes in the diabetic rats when compared to the diabetic plus DW rats, but the islet volumes and β-cell numbers were significantly recovered. UD treatment in diabetic rats improves hyperglycemia by partially restoring plasma insulin levels. The data suggest that UD prevents islet atrophy and/or regenerate pancreatic β-cells.

A step towards removing plasma volume variance from the Athlete's Biological Passport: The use of biomarkers to describe vascular volumes from a simple blood test.

PubMed

Lobigs, Louisa M; Sottas, Pierre-Edouard; Bourdon, Pitre C; Nikolovski, Zoran; El-Gingo, Mohamed; Varamenti, Evdokia; Peeling, Peter; Dawson, Brian; Schumacher, Yorck O

2018-02-01

The haematological module of the Athlete's Biological Passport (ABP) has significantly impacted the prevalence of blood manipulations in elite sports. However, the ABP relies on a number of concentration-based markers of erythropoiesis, such as haemoglobin concentration ([Hb]), which are influenced by shifts in plasma volume (PV). Fluctuations in PV contribute to the majority of biological variance associated with volumetric ABP markers. Our laboratory recently identified a panel of common chemistry markers (from a simple blood test) capable of describing ca 67% of PV variance, presenting an applicable method to account for volume shifts within anti-doping practices. Here, this novel PV marker was included into the ABP adaptive model. Over a six-month period (one test per month), 33 healthy, active males provided blood samples and performed the CO-rebreathing method to record PV (control). In the final month participants performed a single maximal exercise effort to promote a PV shift (mean PV decrease -17%, 95% CI -9.75 to -18.13%). Applying the ABP adaptive model, individualized reference limits for [Hb] and the OFF-score were created, with and without the PV correction. With the PV correction, an average of 66% of [Hb] within-subject variance is explained, narrowing the predicted reference limits, and reducing the number of atypical ABP findings post-exercise. Despite an increase in sensitivity there was no observed loss of specificity with the addition of the PV correction. The novel PV marker presented here has the potential to improve the ABP's rate of correct doping detection by removing the confounding effects of PV variance. Copyright © 2017 John Wiley & Sons, Ltd.

Urtica Dioica Distillate Regenerates Pancreatic Beta Cells in Streptozotocin-Induced Diabetic Rats

PubMed Central

Gohari, Ali; Noorafshan, Ali; Akmali, Masoumeh; Zamani-Garmsiri, Fahimeh; Seghatoleslam, Atefeh

2018-01-01

Background Urtica dioica is known as an anti-hyperglycemic plant. Urtica dioica distillate (UD) is a traditional Iranian drink, locally known as “aragh gazaneh”. In spite of its widespread consumption in Iran, according to traditional Iranian medicine, there is no scientific report on the usefulness of UD for diabetic patients. This survey was designed to evaluate its protective effects for the recovery from diabetes by determining the serum insulin, blood glucose, volume of pancreatic islets, and the number and volume of β-cells in diabetic rats. Methods A total of 48 Sprague-Dawley male rats (200-250 g) were randomly distributed into 6 groups (n=8), including non-diabetic plus distilled water (DW), non-diabetic plus UD, diabetic plus DW, diabetic plus UD, diabetic plus insulin, and diabetic plus glibenclamide. DW, UD, and glibenclamide were administered via intragastric gavage and insulin was injected subcutaneously. After four weeks of experiments, blood samples were collected for serum insulin and blood glucose assay. Pancreas was also evaluated using stereological method. The SPSS software was used for statistical analysis. Kruskal-Wallis, repeated measurements, and Mann-Whitney U test were applied for comparisons between the groups. Results The treatment of diabetic rats with UD reduced the blood glucose dramatically (P<0.001) and increased serum insulin levels significantly (P=0.03) in comparison to the diabetic plus DW rats. Treatment with UD did not affect the mean β-cell volumes in the diabetic rats when compared to the diabetic plus DW rats, but the islet volumes and β-cell numbers were significantly recovered. Conclusion UD treatment in diabetic rats improves hyperglycemia by partially restoring plasma insulin levels. The data suggest that UD prevents islet atrophy and/or regenerate pancreatic β-cells. PMID:29749986

Further evaluation of the NWF filter for the purification of Plasmodium vivax-infected erythrocytes.

PubMed

Li, Jiangyan; Tao, Zhiyong; Li, Qian; Brashear, Awtum; Wang, Ying; Xia, Hui; Fang, Qiang; Cui, Liwang

2017-05-17

Isolation of Plasmodium-infected red blood cells (iRBCs) from clinical blood samples is often required for experiments, such as ex vivo drug assays, in vitro invasion assays and genome sequencing. Current methods for removing white blood cells (WBCs) from malaria-infected blood are time-consuming or costly. A prototype non-woven fabric (NWF) filter was developed for the purification of iRBCs, which showed great efficiency for removing WBCs in a pilot study. Previous work was performed with prototype filters optimized for processing 5-10 mL of blood. With the commercialization of the filters, this study aims to evaluate the efficiency and suitability of the commercial NWF filter for the purification of Plasmodium vivax-infected RBCs in smaller volumes of blood and to compare its performance with that of Plasmodipur ® filters. Forty-three clinical P. vivax blood samples taken from symptomatic patients attending malaria clinics at the China-Myanmar border were processed using the NWF filters in a nearby field laboratory. The numbers of WBCs and iRBCs and morphology of P. vivax parasites in the blood samples before and after NWF filtration were compared. The viability of P. vivax parasites after filtration from 27 blood samples was examined by in vitro short-term culture. In addition, the effectiveness of the NWF filter for removing WBCs was compared with that of the Plasmodipur ® filter in six P. vivax blood samples. Filtration of 1-2 mL of P. vivax-infected blood with the NWF filter removed 99.68% WBCs. The densities of total iRBCs, ring and trophozoite stages before and after filtration were not significantly different (P > 0.05). However, the recovery rates of schizont- and gametocyte-infected RBCs, which were minor parasite stages in the clinical samples, were relatively low. After filtration, the P. vivax parasites did not show apparent morphological changes. Culture of 27 P. vivax-infected blood samples after filtration showed that parasites successfully matured into the schizont stage. The WBC removal rates and iRBC recovery rates were not significantly different between the NWF and Plasmodipur ® filters (P > 0.05). When tested with 1-2 mL of P. vivax-infected blood, the NWF filter could effectively remove WBCs and the recovery rates for ring- and trophozoite-iRBCs were high. P. vivax parasites after filtration could be successfully cultured in vitro to reach maturity. The performance of the NWF and Plasmodipur ® filters for removing WBCs and recovering iRBCs was comparable.

Low volume tubes are not effective to reduce the rate of hemolyzed specimens from the emergency department.

PubMed

Lippi, Giuseppe; Bonelli, Patrizia; Graiani, Virna; Caleffi, Catia; Cervellin, Gianfranco

2014-02-01

Spurious hemolysis is the leading source of nonconformities that can be recorded in diagnostic samples, especially those collected in the emergency department (ED). The aim of this study was to assess whether the shift from regular to low volume blood collection tubes may reduce the rate of hemolysis in a large urban ED, where approximately 80% of blood collections are performed through catheters. In a former 5-month period, blood collection in the ED was performed using 5.0mL (13×100mm) plastic serum tubes, which were then completely replaced with 3.5mL (13×75mm) plastic serum tubes for another period of 5months. The rate of hemolyzed specimens (i.e., those containing a cell-free hemoglobin ≥0.5gL) collected in the two periods was compared by Fisher exact test. The rate of hemolyzed specimens received from the ED increased from 3.5% using 5.0mL plastic serum tubes to 5.2% after introduction of 3.5mL plastic serum tubes (p<0.001). The use of low volume tubes was not effective to decrease the hemolysis rate in a large urban ED. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Cardiovascular regulatory response to lower body negative pressure following blood volume loss

NASA Technical Reports Server (NTRS)

Shimizu, M.; Ghista, D. N.; Sandler, H.

1979-01-01

An attempt is made to explain the cardiovascular regulatory responses to lower body negative pressure (LBNP) stress, both in the absence of and following blood or plasma volume loss, the latter being factors regularly observed with short- or long-term recumbency or weightlessness and associated with resulting cardiovascular deconditioning. Analytical expressions are derived for the responses of mean venous pressure and blood volume pooled in the lower body due to LBNP. An analysis is presented for determining the HR change due to LBNP stress following blood volume loss. It is concluded that the reduced orthostatic tolerance following long-term space flight or recumbency can be mainly attributed to blood volume loss, and that the associated cardiovascular responses characterizing this orthostatic intolerance is elicited by the associated central venous pressure response.

Microfluidic Blood Cell Preparation: Now and Beyond

PubMed Central

Yu, Zeta Tak For; Yong, Koh Meng Aw; Fu, Jianping

2014-01-01

Blood plays an important role in homeostatic regulation with each of its cellular components having important therapeutic and diagnostic uses. Therefore, separation and sorting of blood cells has been of a great interest to clinicians and researchers. However, while conventional methods of processing blood have been successful in generating relatively pure fractions, they are time consuming, labor intensive, and are not optimal for processing small volume blood samples. In recent years, microfluidics has garnered great interest from clinicians and researchers as a powerful technology for separating blood into different cell fractions. As microfluidics involves fluid manipulation at the microscale level, it has the potential for achieving high-resolution separation and sorting of blood cells down to a single-cell level, with an added benefit of integrating physical and biological methods for blood cell separation and analysis on the same single chip platform. This paper will first review the conventional methods of processing and sorting blood cells, followed by a discussion on how microfluidics is emerging as an efficient tool to rapidly change the field of blood cell sorting for blood-based therapeutic and diagnostic applications. PMID:24515899

Field performance of a low-cost and fully-automated blood counting system operated by trained and untrained users (Conference Presentation)

NASA Astrophysics Data System (ADS)

Xie, Dengling; Xie, Yanjun; Liu, Peng; Tong, Lieshu; Chu, Kaiqin; Smith, Zachary J.

2017-02-01

Current flow-based blood counting devices require expensive and centralized medical infrastructure and are not appropriate for field use. In this paper we report a method to count red blood cells, white blood cells as well as platelets through a low-cost and fully-automated blood counting system. The approach consists of using a compact, custom-built microscope with large field-of-view to record bright-field and fluorescence images of samples that are diluted with a single, stable reagent mixture and counted using automatic algorithms. Sample collection is performed manually using a spring loaded lancet, and volume-metering capillary tubes. The capillaries are then dropped into a tube of pre-measured reagents and gently shaken for 10-30 seconds. The sample is loaded into a measurement chamber and placed on a custom 3D printed platform. Sample translation and focusing is fully automated, and a user has only to press a button for the measurement and analysis to commence. Cost of the system is minimized through the use of custom-designed motorized components. We performed a series of comparative experiments by trained and untrained users on blood from adults and children. We compare the performance of our system, as operated by trained and untrained users, to the clinical gold standard using a Bland-Altman analysis, demonstrating good agreement of our system to the clinical standard. The system's low cost, complete automation, and good field performance indicate that it can be successfully translated for use in low-resource settings where central hematology laboratories are not accessible.

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

NASA Astrophysics Data System (ADS)

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

2015-03-01

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

Measuring the level of agreement in hematologic and biochemical values between blood sampling sites in leatherback sea turtles (Dermochelys coriacea).

PubMed

Stewart, Kimberly; Mitchell, Mark A; Norton, Terry; Krecek, Rosina C

2012-12-01

Conservation programs to protect endangered sea turtles are being instituted worldwide. A common practice in these programs is to collect blood to evaluate the health of the turtles. Several different venipuncture sites are used to collect blood from sea turtles for hematologic and biochemistry tests, depending on the species. To date, it is unknown what affect venipuncture site may have on sample results. The purpose of this study was to measure the level of agreement between hematologic and biochemistry values collected from the dorsal cervical sinus and the interdigital vein of leatherback (Dermochelys coriacea) sea turtles. Paired heparinized blood samples were obtained from the dorsal cervical sinus and the interdigital vein of 12 adult female nesting leatherback sea turtles on Keys Beach, St. Kitts, West Indies. Even though the sample population was small, the data for each chemistry were normally distributed, except for creatine kinase (CK). There was no significant difference when comparing biochemistry or hematologic values by venipuncture site, except for CK (P = 0.02). The level of agreement between sampling sites was considered good for albumin, calcium, globulin, glucose, packed cell volume, phosphorus, potassium, sodium, total protein, total solids, uric acid, white blood cell count, and all of the individual white cell types, while the level of agreement for aspartate aminotransferase and CK were considered poor. This information, coupled with the fact that the interdigital vein affords a less-invasive procedure, demonstrates that the interdigital vein is an appropriate location to use when establishing a hematologic and biochemical profile for leatherback sea turtles.

Detection of Theileria annulata in blood samples of carrier cattle by PCR.

PubMed Central

d'Oliveira, C; van der Weide, M; Habela, M A; Jacquiet, P; Jongejan, F

1995-01-01

We report the detection of Theileria annulata, the causative agent of tropical theileriosis, by PCR in blood samples obtained from carrier cattle. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata. A 721-bp fragment was amplified from blood samples taken monthly from calves experimentally infected with one of four different stocks of T. annulata originating in either Mauritania, Portugal, Spain, or Turkey. At the end of the experiment, five animals carried the infection for 12 months and two animals remained infected for 15 months. DNAs from six other Theileria species, T. parva, T. mutans, T. sergenti, T. buffeli, T. velifera, and T. taurotragi, were not amplified. Moreover, DNAs from four other hemoparasites (Anaplasma centrale, Anaplasma marginale, Babesia bovis, and Babesia bigemina) were also not amplified. As a control, primers derived from the small subunit rRNA gene of Theileria spp. amplified a 1.1-kb DNA fragment from all Theileria species examined but not from the other four hemoparasites. As few as two to three parasites per microliter of infected blood in a 50-microliters sample volume were detected by Southern or microplate hybridization with a T. annulata-specific cDNA probe. In addition, 92 field samples obtained from cattle in Spain were tested; 22% were positive in blood smears, 40% were positive by immunofluorescent antibody test, and 75% were positive for T. annulata by PCR. The method provides a useful diagnostic tool for detecting T. annulata carrier cattle. PMID:8567902

Noninvasive, low-noise, fast imaging of blood volume and deoxygenation changes in muscles using light-emitting diode continuous-wave imager

NASA Astrophysics Data System (ADS)

Lin, Yuanqing; Lech, Gwen; Nioka, Shoko; Intes, Xavier; Chance, Britton

2002-08-01

This article focuses on optimizing the signal to noise ratio (SNR) of a three-wavelength light-emitting diode (LED) near-infrared continuous-wave (cw) imager and its application to in vivo muscle metabolism measurement. The shot-noise limited SNR is derived and calculated to be 2 x104 for the physiological blood concentrations of muscle. Aiming at shot-noise limited SNR performance and fast imaging, we utilize sample and hold circuits to reduce high-frequency noise. These circuits have also been designed to be parallel integrating, through which SNR of 2 x103 and 2 Hz imaging acquisition rate have been achieved when the probe is placed on a muscle model. The noise corresponds to 2 x10-4 optical density error, which suggests an in vitro resolution of 15. 4 nM blood volume and 46.8 nM deoxygenation changes. A 48 dB digital gain control circuit with 256 steps is employed to enlarge the dynamic range of the imager. We utilize cuff ischemia as a living model demonstration and its results are reported. The instrument is applied during exercise to measure the changes of blood volume and deoxygenation, which provides important information about muscle metabolism. We find that the primary source of noise encountered during exercise experiment is from the random motion of muscle. The results demonstrate that the LED cw imager is ideal for the noninvasive study of muscle metabolism.

Effects of passive heating on central blood volume and ventricular dimensions in humans

PubMed Central

Crandall, C G; Wilson, T E; Marving, J; Vogelsang, T W; Kjaer, A; Hesse, B; Secher, N H

2008-01-01

Mixed findings regarding the effects of whole-body heat stress on central blood volume have been reported. This study evaluated the hypothesis that heat stress reduces central blood volume and alters blood volume distribution. Ten healthy experimental and seven healthy time control (i.e. non-heat stressed) subjects participated in this protocol. Changes in regional blood volume during heat stress and time control were estimated using technetium-99m labelled autologous red blood cells and gamma camera imaging. Whole-body heating increased internal temperature (≥ 1.0°C), cutaneous vascular conductance (approximately fivefold), and heart rate (52 ± 2 to 93 ± 4 beats min−1), while reducing central venous pressure (5.5 ± 07 to 0.2 ± 0.6 mmHg) accompanied by minor decreases in mean arterial pressure (all P < 0.05). The heat stress reduced the blood volume of the heart (18 ± 2%), heart plus central vasculature (17 ± 2%), thorax (14 ± 2%), inferior vena cava (23 ± 2%) and liver (23 ± 2%) (all P≤ 0.005 relative to time control subjects). Radionuclide multiple-gated acquisition assessment revealed that heat stress did not significantly change left ventricular end-diastolic volume, while ventricular end-systolic volume was reduced by 24 ± 6% of pre-heat stress levels (P < 0.001 relative to time control subjects). Thus, heat stress increased left ventricular ejection fraction from 60 ± 1% to 68 ± 2% (P= 0.02). We conclude that heat stress shifts blood volume from thoracic and splanchnic regions presumably to aid in heat dissipation, while simultaneously increasing heart rate and ejection fraction. PMID:17962331

Anesthesia and blood sampling of wild big brown bats (eptesicus fuscus) with an assessment of impacts on survival.

PubMed

Wimsatt, Jeffrey; O'Shea, Thomas J; Ellison, Laura E; Pearce, Roger D; Price, Valerie R

2005-01-01

We anesthetized and blood sampled wild big brown bats (Eptesicus fuscus) in Fort Collins, Colorado (USA) in 2001 and 2002 and assessed effects on survival. Inhalant anesthesia was delivered into a specially designed restraint and inhalation capsule that minimized handling and bite exposures. Bats were immobilized an average of 9.1+/-5.1 (SD) min (range 1-71, n=876); blood sample volumes averaged 58+/-12 microl (range 13-126, n=718). We randomly selected control (subject to multiple procedures before release) and treatment (control procedures plus inhalant anesthesia and 1% of body weight blood sampling) groups in 2002 to assess treatment effects on daily survival over a 14-day period for adult female and volant juvenile bats captured at maternity roosts in buildings. We monitored survival after release using passive integrated transponder tag detection hoops placed at openings to selected roosts. Annual return rates of bats sampled in 2001 were used to assess long-term outcomes. Comparison of 14-day maximum-likelihood daily survival estimates from control (86 adult females, 92 volant juveniles) and treated bats (187 adult females, 87 volant juveniles) indicated no adverse effect from anesthesia and blood sampling (juveniles: chi2=22.22, df=27, P>0.05; adults: chi2=9.72, df=18, P>0.05). One-year return rates were similar among adult female controls (81%, n=72, 95% confidence interval [CI]=70-91%), females treated once (82%, n=276, 95% CI=81-84%), and females treated twice (84%, n=50, 95% CI=74-94%). Lack of an effect was also noted in 1-yr return rates of juvenile female controls (55%, n=29, 95% CI=37-73%), juveniles treated once (66%, n=113, 95% CI=58-75%), and juveniles treated twice (71%, n=17, 95% CI=49-92%). These data suggest that anesthesia and blood sampling for health monitoring did not measurably affect survival of adult female and volant juvenile big brown bats.

Poor sleep quality is associated with increased cortical atrophy in community-dwelling adults.

PubMed

Sexton, Claire E; Storsve, Andreas B; Walhovd, Kristine B; Johansen-Berg, Heidi; Fjell, Anders M

2014-09-09

To examine the relationship between sleep quality and cortical and hippocampal volume and atrophy within a community-based sample, explore the influence of age on results, and assess the possible confounding effects of physical activity levels, body mass index (BMI), and blood pressure. In 147 community-dwelling adults (92 female; age 53.9 ± 15.5 years), sleep quality was measured using the Pittsburgh Sleep Quality Index and correlated with cross-sectional measures of volume and longitudinal measures of atrophy derived from MRI scans separated by an average of 3.5 years. Exploratory post hoc analysis compared correlations between different age groups and included physical activity, BMI, and blood pressure as additional covariates. Poor sleep quality was associated with reduced volume within the right superior frontal cortex in cross-sectional analyses, and an increased rate of atrophy within widespread frontal, temporal, and parietal regions in longitudinal analyses. Results were largely driven by correlations within adults over the age of 60, and could not be explained by variation in physical activity, BMI, or blood pressure. Sleep quality was not associated with hippocampal volume or atrophy. We found that longitudinal measures of cortical atrophy were widely correlated with sleep quality. Poor sleep quality may be a cause or a consequence of brain atrophy, and future studies examining the effect of interventions that improve sleep quality on rates of atrophy may hold key insights into the direction of this relationship. © 2014 American Academy of Neurology.

Impact of Infarct Size on Blood Pressure in Young Patients with Acute Stroke.

PubMed

Bonardo, Pablo; Pantiú, Fátima; Ferraro, Martín; Chertcoff, Anibal; Bandeo, Lucrecia; Cejas, Luciana León; Pacha, Sol; Roca, Claudia Uribe; Rugilo, Carlos; Pardal, Manuel Maria Fernández; Reisin, Ricardo

2018-06-01

Hypertension can be found in up to 80% of patients with acute stroke. Many factors have been related to this phenomenon such as age, history of hypertension, and stroke severity. The aim of our study was to determine the relationship between infarct volume and blood pressure, at admission, in young patients with acute ischemic stroke. Patients younger than 55 years old admitted within 24 hours of ischemic stroke were included. Socio-demographic variables, systolic blood pressure, diastolic blood pressure, and infarct volume at admission were assessed. Statistical analysis: mean and SEM for quantitative variables, percentages for qualitative, and Spearman correlations ( p value < 0.05 was considered statistically significant). Twenty-two patients (12 men), mean age: 44.64 ± 1.62 years. The most frequent vascular risk factors were: hypertension, smoking, and overweight (40.9%). Mean systolic and diastolic blood pressure on admission were: 143.27 ± 6.57 mmHg and 85.14 ± 3.62 mmHg, respectively. Infarct volume: 11.55 ± 4.74 ml. Spearman correlations: systolic blood pressure and infarct volume: p = 0.15 r : -0.317; diastolic blood pressure and infarct volume: p = 0.738 r: -0.76. In our series of young patients with acute ischemic stroke, large infarct volume was not associated with high blood pressure at admission.

Pediatric blood volumes: a one-page reference guide.

PubMed

Smiley, J; Reitan, J

1998-10-01

At our institution, a multidisciplinary team met to work out a blood volume policy for our pediatric patients' laboratory testing. Because we are a cancer center, many of our patients are on protocols and/or are in the hospital for an extended period of time. These factors result in multiple blood draws. It is important to manage the volumes used so that we do not compromise the hematological status of our pediatric patients. The concerns of nurses and laboratory technologists were discussed and a three-tiered system was designed consisting of adult volumes, volumes for < 20-kg patients, and volumes for < 10-kg patients. Each tier consists of less blood than the one above it but the cost for such a procedure is increased. Time to conduct the draw, the cost of equipment to do the draw, and inflexibility to verify or add to the original order without resticking the patient increase at each tier. It is imperative to the overall quality of care for all patients that discretion is used when following these guidelines. When it is medically prudent to restrict the blood volumes taken from any patient, it should be done. However, when it is not medically necessary, the increased costs and potential decrease in the quality of laboratory service outweighs the desire to use smaller blood volumes.

Analytical Comparison of In Vitro-Spiked Human Serum and Plasma for PCR-Based Detection of Aspergillus fumigatus DNA: a Study by the European Aspergillus PCR Initiative

PubMed Central

Mengoli, Carlo; Springer, Jan; Bretagne, Stéphane; Cuenca-Estrella, Manuel; Klingspor, Lena; Lagrou, Katrien; Melchers, Willem J. G.; Morton, C. Oliver; Barnes, Rosemary A.; Donnelly, J. Peter; White, P. Lewis

2015-01-01

The use of serum or plasma for Aspergillus PCR testing facilitates automated and standardized technology. Recommendations for serum testing are available, and while serum and plasma are regularly considered interchangeable for use in fungal diagnostics, differences in galactomannan enzyme immunoassay (GM-EIA) performance have been reported and are attributed to clot formation. Therefore, it is important to assess plasma PCR testing to determine if previous recommendations for serum are applicable and also to compare analytical performance with that of serum PCR. Molecular methods testing serum and plasma were compared through multicenter distribution of quality control panels, with additional studies to investigate the effect of clot formation and blood fractionation on DNA availability. Analytical sensitivity and time to positivity (TTP) were compared, and a regression analysis was performed to identify variables that enhanced plasma PCR performance. When testing plasma, sample volume, preextraction-to-postextraction volume ratio, PCR volume, duplicate testing, and the use of an internal control for PCR were positively associated with performance. When whole-blood samples were spiked and then fractionated, the analytical sensitivity and TTP were superior when testing plasma. Centrifugation had no effect on DNA availability, whereas the presence of clot material significantly lowered the concentration (P = 0.028). Technically, there are no major differences in the molecular processing of serum and plasma, but the formation of clot material potentially reduces available DNA in serum. During disease, Aspergillus DNA burdens in blood are often at the limits of PCR performance. Using plasma might improve performance while maintaining the methodological simplicity of serum testing. PMID:26085614

[Evaluation of immunoglobulin G concentration in colostrum of mares by ELISA, refractometry and colostrometry].

PubMed

Venner, Monica; Markus, R G; Strutzberg-Minder, K; Nogai, K; Beyerbach, M; Klug, E

2008-01-01

In 360 samples of colostrum and 36 samples of blood of warmblood mares, the concentration of immunoglobulin G (IgG) was evaluated in the post partal period with an ELISA and the results were compared to values obtained with 2 field methods--refractometry and colostrometry. A significant correlation (p < 0.0001) was determined between ELISA and colostrometry (r = +0.88) and between ELISA and refractometry (r = +0.93). So both field-methods seem suitable for evaluation of the colostral IgG-concentration in mares. Further the kinetic of the IgG concentration in colostrum, the volume of colostrum and the total amount of IgG was measured in the 12 hours post partum (p.p.) in each half udder of 36 mares of different parity. Immediately p.p. primiparous mares have a greater mean concentration of IgG (68 mg/ml) than multiparous mares (51 mg/ml). However, multiparous mares have a mean colostral volume of 1020 ml whereas, in primiparous mares, a mean volume of 527 ml was determined within the first three hours p.p. As a result of this the total amount of IgG was lower in primiparous (31.5 g) than in multiparous mares (48.5 g). A significant decrease of IgG concentration was measured in multiparous mares in the 1.5 hours following partum versus 3 hours in primiparous mares. The mean IgG concentration in the blood serum of the 36 mares immediately p.p. was 13.4 +/- 3.6 mg/ml. No significant correlation was observed between values of IgG concentration in the blood and in the colostrum of the mares.

Analytical Comparison of In Vitro-Spiked Human Serum and Plasma for PCR-Based Detection of Aspergillus fumigatus DNA: a Study by the European Aspergillus PCR Initiative.

PubMed

Loeffler, Juergen; Mengoli, Carlo; Springer, Jan; Bretagne, Stéphane; Cuenca-Estrella, Manuel; Klingspor, Lena; Lagrou, Katrien; Melchers, Willem J G; Morton, C Oliver; Barnes, Rosemary A; Donnelly, J Peter; White, P Lewis

2015-09-01

The use of serum or plasma for Aspergillus PCR testing facilitates automated and standardized technology. Recommendations for serum testing are available, and while serum and plasma are regularly considered interchangeable for use in fungal diagnostics, differences in galactomannan enzyme immunoassay (GM-EIA) performance have been reported and are attributed to clot formation. Therefore, it is important to assess plasma PCR testing to determine if previous recommendations for serum are applicable and also to compare analytical performance with that of serum PCR. Molecular methods testing serum and plasma were compared through multicenter distribution of quality control panels, with additional studies to investigate the effect of clot formation and blood fractionation on DNA availability. Analytical sensitivity and time to positivity (TTP) were compared, and a regression analysis was performed to identify variables that enhanced plasma PCR performance. When testing plasma, sample volume, preextraction-to-postextraction volume ratio, PCR volume, duplicate testing, and the use of an internal control for PCR were positively associated with performance. When whole-blood samples were spiked and then fractionated, the analytical sensitivity and TTP were superior when testing plasma. Centrifugation had no effect on DNA availability, whereas the presence of clot material significantly lowered the concentration (P = 0.028). Technically, there are no major differences in the molecular processing of serum and plasma, but the formation of clot material potentially reduces available DNA in serum. During disease, Aspergillus DNA burdens in blood are often at the limits of PCR performance. Using plasma might improve performance while maintaining the methodological simplicity of serum testing. Copyright © 2015 Loeffler et al.

Dissociation of local and global skeletal muscle oxygen transport metrics in type 2 diabetes.

PubMed

Mason McClatchey, P; Bauer, Timothy A; Regensteiner, Judith G; Schauer, Irene E; Huebschmann, Amy G; Reusch, Jane E B

2017-08-01

Exercise capacity is impaired in type 2 diabetes, and this impairment predicts excess morbidity and mortality. This defect appears to involve excess skeletal muscle deoxygenation, but the underlying mechanisms remain unclear. We hypothesized that reduced blood flow, reduced local recruitment of blood volume/hematocrit, or both contribute to excess skeletal muscle deoxygenation in type 2 diabetes. In patients with (n=23) and without (n=18) type 2 diabetes, we recorded maximal reactive hyperemic leg blood flow, peak oxygen utilization during cycling ergometer exercise (VO 2peak ), and near-infrared spectroscopy-derived measures of exercise-induced changes in skeletal muscle oxygenation and blood volume/hematocrit. We observed a significant increase (p<0.05) in skeletal muscle deoxygenation in type 2 diabetes despite similar blood flow and recruitment of local blood volume/hematocrit. Within the control group skeletal muscle deoxygenation, local recruitment of microvascular blood volume/hematocrit, blood flow, and VO 2peak are all mutually correlated. None of these correlations were preserved in type 2 diabetes. These results suggest that in type 2 diabetes 1) skeletal muscle oxygenation is impaired, 2) this impairment may occur independently of bulk blood flow or local recruitment of blood volume/hematocrit, and 3) local and global metrics of oxygen transport are dissociated. Copyright © 2017 Elsevier Inc. All rights reserved.

Method of freezing living cells and tissues with improved subsequent survival

DOEpatents

Senkan, Selim M.; Hirsch, Gerald P.

1980-01-01

This invention relates to an improved method for freezing red blood cells, ther living cells, or tissues with improved subsequent survival, wherein constant-volume freezing is utilized that results in significantly improved survival compared with constant-pressure freezing; optimization is attainable through the use of different vessel geometries, cooling baths and warming baths, and sample concentrations.

Combining image-derived and venous input functions enables quantification of serotonin-1A receptors with [carbonyl-11C]WAY-100635 independent of arterial sampling.

PubMed

Hahn, Andreas; Nics, Lukas; Baldinger, Pia; Ungersböck, Johanna; Dolliner, Peter; Frey, Richard; Birkfellner, Wolfgang; Mitterhauser, Markus; Wadsak, Wolfgang; Karanikas, Georgios; Kasper, Siegfried; Lanzenberger, Rupert

2012-08-01

image- derived input functions (IDIFs) represent a promising technique for a simpler and less invasive quantification of PET studies as compared to arterial cannulation. However, a number of limitations complicate the routine use of IDIFs in clinical research protocols and the full substitution of manual arterial samples by venous ones has hardly been evaluated. This study aims for a direct validation of IDIFs and venous data for the quantification of serotonin-1A receptor binding (5-HT(1A)) with [carbonyl-(11)C]WAY-100635 before and after hormone treatment. Fifteen PET measurements with arterial and venous blood sampling were obtained from 10 healthy women, 8 scans before and 7 after eight weeks of hormone replacement therapy. Image-derived input functions were derived automatically from cerebral blood vessels, corrected for partial volume effects and combined with venous manual samples from 10 min onward (IDIF+VIF). Corrections for plasma/whole-blood ratio and metabolites were done separately with arterial and venous samples. 5-HT(1A) receptor quantification was achieved with arterial input functions (AIF) and IDIF+VIF using a two-tissue compartment model. Comparison between arterial and venous manual blood samples yielded excellent reproducibility. Variability (VAR) was less than 10% for whole-blood activity (p>0.4) and below 2% for plasma to whole-blood ratios (p>0.4). Variability was slightly higher for parent fractions (VARmax=24% at 5 min, p<0.05 and VAR<13% after 20 min, p>0.1) but still within previously reported values. IDIFs after partial volume correction had peak values comparable to AIFs (mean difference Δ=-7.6 ± 16.9 kBq/ml, p>0.1), whereas AIFs exhibited a delay (Δ=4 ± 6.4s, p<0.05) and higher peak width (Δ=15.9 ± 5.2s, p<0.001). Linear regression analysis showed strong agreement for 5-HT(1A) binding as obtained with AIF and IDIF+VIF at baseline (R(2)=0.95), after treatment (R(2)=0.93) and when pooling all scans (R(2)=0.93), with slopes and intercepts in the range of 0.97 to 1.07 and -0.05 to 0.16, respectively. In addition to the region of interest analysis, the approach yielded virtually identical results for voxel-wise quantification as compared to the AIF. Despite the fast metabolism of the radioligand, manual arterial blood samples can be substituted by venous ones for parent fractions and plasma to whole-blood ratios. Moreover, the combination of image-derived and venous input functions provides a reliable quantification of 5-HT(1A) receptors. This holds true for 5-HT(1A) binding estimates before and after treatment for both regions of interest-based and voxel-wise modeling. Taken together, the approach provides less invasive receptor quantification by full independence of arterial cannulation. This offers great potential for the routine use in clinical research protocols and encourages further investigation for other radioligands with different kinetic characteristics. Copyright © 2012 Elsevier Inc. All rights reserved.

Evaluation of liver functional reserve by combining D-sorbitol clearance rate and CT measured liver volume

PubMed Central

Li, Yi-Ming; Lv, Fan; Xu, Xin; Ji, Hong; Gao, Wen-Tao; Lei, Tuan-Jie; Ren, Gui-Bing; Bai, Zhi-Lan; Li, Qiang

2003-01-01

AIM: Our research attempted to evaluate the overall functional reserve of cirrhotic liver by combination of hepatic functional blood flow, liver volume, and Child-Pugh’s classification, and to discuss its value of clinical application. METHODS: Ninety two patients with portal hypertension due to hepatic cirrhosis were investigated. All had a history of haematemesis and hematochezia, esophageal and gastric fundus varices, splenomegaly and hypersplenia. A 2-year follow-up was routinely performed and no one was lost. Twenty two healthy volunteers were used as control group. Blood and urine samples were collected 4 times before and after intravenous D-sorbitol infusion. The hepatic clearance (CLH) of D-sorbitol was then calculated according to enzymatic spectrophotometric method while the total blood flow (QTOTAL) and intrahepatic shunt (RINS) were detected by multicolor Doppler ultrasound, and the liver volume was measured by spiral CT. Data were estimated by t-test, variance calculation and chi-squared test. The relationships between all these parameters and different groups were investigated according to Child-Pugh classification and postoperative complications respectively. RESULTS: Steady blood concentration was achieved 120 mins after D-sorbitol intravenous infusion, which was (0.358 ± 0.064) mmol·L-1 in cirrhotic group and (0.189 ± 0.05) mmol·L-1 in control group (P < 0.01). CLH = (812.7 ± 112.4) mL·min-1, QTOTAL = (1280.6 ± 131.4) mL·min-1, and RINS = (36.54 ± 10.65)% in cirrhotic group and CLH = (1248.3 ± 210.5) mL·min-1, QTOTAL = (1362.4 ± 126.9) mL·min-1, and RINS = (8.37 ± 3.32)% in control group (P < 0.01). The liver volume of cirrhotic group was 1057 ± 249 cm3, 851 ± 148 cm3 and 663 ± 77 cm3 in Child A, B and C group respectively with significant difference (P < 0.001). The average volume of cirrhotic liver in Child B, C group was significantly reduced in comparison with that in control group (P < 0.001). The patient, whose liver volume decreased by 40% with the CLH below 600 mL·min-1, would have a higher incidence of postoperative complications. There was no strict correspondent relationship between CLH, liver volume and Child-Pugh’s classification. CONCLUSION: The hepatic clearance of D-sorbitol, CT measured liver volume can be reliably used for the evaluation of hepatic functional blood flow and liver metabolic volume. Combined with the Child-Pugh’s classification, it could be very useful for further understanding the liver functional reserve, therefore help determine reasonable therapeutic plan, choose surgical procedures and operating time. PMID:12970913

Altered baseline blood volume and the norepinephrine response to stress in humans

NASA Technical Reports Server (NTRS)

Vernikos, J.; Convertino, V. A.

1992-01-01

A hypothesis is proposed that a primary physiological purpose of the neural and endocrine response to stressors is the preservation of the blood volume/blood pressure relationship. Changes in blood volume caused by an adaptation to the environmental challenge serve to modulate the neural and endocrine responsiveness to stress. Relationships between changes in vascular volume, vasoconstriction, and norepinephrine (NE) responses during acute and chronic exposure to various stressors are examined. It is noted that the hypothesis is based on numerous observations rather than definitive cause-effect experiments and further investigation is required to prove it.

Dehydration and fluid volume kinetics before major open abdominal surgery.

PubMed

Hahn, R G; Bahlmann, H; Nilsson, L

2014-11-01

Assessment of dehydration in the preoperative setting is of potential clinical value. The present study uses urine analysis and plasma volume kinetics, which have both been validated against induced changes in body water in volunteers, to study the incidence and severity of dehydration before open abdominal surgery begins. Thirty patients (mean age 64 years) had their urine analysed before major elective open abdominal surgery for colour, specific weight, osmolality and creatinine. The results were scored and the mean taken to represent a 'dehydration index'. Thereafter, the patients received an infusion of 5 ml/kg of Ringer's acetate intravenously for over 15 min. Blood was sampled for 70 min and the blood haemoglobin concentration used to estimate the plasma volume kinetics. Distribution of fluid occurred more slowly (P < 0.01) and the elimination half-life was twice as long (median 40 min, not significant) in the 11 patients (37%) diagnosed to be moderately dehydrated as compared with euhydrated patients. The dehydration index indicated that the fluid deficit in these patients corresponded to 2.5% of the body weight, whereas the deficit in the others was 1%. In contrast, the 11 patients who later developed postoperative nausea and vomiting had a very short elimination half-life, only 9 min (median, P < 0.01). These patients were usually euhydrated but had microalbuminuria (P < 0.03) and higher natriuresis (P < 0.01). The degree of dehydration before major surgery was modest as evidenced both by urine sampling and volume kinetic analysis. © 2014 The Acta Anaesthesiologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Direct detection of cancer biomarkers in blood using a "place n play" modular polydimethylsiloxane pump.

PubMed

Zhang, Honglian; Li, Gang; Liao, Lingying; Mao, Hongju; Jin, Qinghui; Zhao, Jianlong

2013-01-01

Cancer biomarkers have significant potential as reliable tools for the early detection of the disease and for monitoring its recurrence. However, most current methods for biomarker detection have technical difficulties (such as sample preparation and specific detector requirements) which limit their application in point of care diagnostics. We developed an extremely simple, power-free microfluidic system for direct detection of cancer biomarkers in microliter volumes of whole blood. CEA and CYFRA21-1 were chosen as model cancer biomarkers. The system automatically extracted blood plasma from less than 3 μl of whole blood and performed a multiplex sample-to-answer assay (nano-ELISA (enzyme-linked immunosorbent assay) technique) without the use of external power or extra components. By taking advantage of the nano-ELISA technique, this microfluidic system detected CEA at a concentration of 50 pg/ml and CYFRA21-1 at a concentration of 60 pg/ml within 60 min. The combination of PnP polydimethylsiloxane (PDMS) pump and nano-ELISA technique in a single microchip system shows great promise for the detection of cancer biomarkers in a drop of blood.

The effect of chronic erythrocytic polycythemia and high altitude upon plasma and blood volumes.

NASA Technical Reports Server (NTRS)

Burton, R. R.; Smith, A. H.

1972-01-01

Comparison of two kinds of physiological chronic erythrocytic polycythemias in order to differentiate the specific effect of erythrocytic polycythemia from the general effects of high altitude upon the plasma volume. The two kinds were produced hormonally in female chickens, at sea level, or by protracted high-altitude exposures. It appears that the vascular system of the body may account for an increase in red blood cell mass either by reduction in plasma volume, or by no change in plasma volume, resulting in differential changes in total blood volumes.

How large is the typical subarachnoid hemorrhage? A review of current neurosurgical knowledge.

PubMed

Whitmore, Robert G; Grant, Ryan A; LeRoux, Peter; El-Falaki, Omar; Stein, Sherman C

2012-01-01

Despite the morbidity and mortality of subarachnoid hemorrhage (SAH), the average volume of a typical hemorrhage is not well defined. Animal models of SAH often do not accurately mimic the human disease process. The purpose of this study is to estimate the average SAH volume, allowing standardization of animal models of the disease. We performed a MEDLINE search of SAH volume and erythrocyte counts in human cerebrospinal fluid as well as for volumes of blood used in animal injection models of SAH, from 1956 to 2010. We polled members of the American Association of Neurological Surgeons (AANS) for estimates of typical SAH volume. Using quantitative data from the literature, we calculated the total volume of SAH as equal to the volume of blood clotted in basal cisterns plus the volume of dispersed blood in cerebrospinal fluid. The results of the AANS poll confirmed our estimates. The human literature yielded 322 publications and animal literature, 237 studies. Four quantitative human studies reported blood clot volumes ranging from 0.2 to 170 mL, with a mean of ∼20 mL. There was only one quantitative study reporting cerebrospinal fluid red blood cell counts from serial lumbar puncture after SAH. Dispersed blood volume ranged from 2.9 to 45.9 mL, and we used the mean of 15 mL for our calculation. Therefore, total volume of SAH equals 35 mL. The AANS poll yielded 176 responses, ranging from 2 to 350 mL, with a mean of 33.9 ± 4.4 mL. Based on our estimate of total SAH volume of 35 mL, animal injection models may now become standardized for more accurate portrayal of the human disease process. Copyright © 2012 Elsevier Inc. All rights reserved.

Isolation of Circulating Plasma Cells in Multiple Myeloma Using CD138 Antibody-Based Capture in a Microfluidic Device

NASA Astrophysics Data System (ADS)

Qasaimeh, Mohammad A.; Wu, Yichao C.; Bose, Suman; Menachery, Anoop; Talluri, Srikanth; Gonzalez, Gabriel; Fulciniti, Mariateresa; Karp, Jeffrey M.; Prabhala, Rao H.; Karnik, Rohit

2017-04-01

The necessity for bone marrow aspiration and the lack of highly sensitive assays to detect residual disease present challenges for effective management of multiple myeloma (MM), a plasma cell cancer. We show that a microfluidic cell capture based on CD138 antigen, which is highly expressed on plasma cells, permits quantitation of rare circulating plasma cells (CPCs) in blood and subsequent fluorescence-based assays. The microfluidic device is based on a herringbone channel design, and exhibits an estimated cell capture efficiency of ~40-70%, permitting detection of <10 CPCs/mL using 1-mL sample volumes, which is difficult using existing techniques. In bone marrow samples, the microfluidic-based plasma cell counts exhibited excellent correlation with flow cytometry analysis. In peripheral blood samples, the device detected a baseline of 2-5 CD138+ cells/mL in healthy donor blood, with significantly higher numbers in blood samples of MM patients in remission (20-24 CD138+ cells/mL), and yet higher numbers in MM patients exhibiting disease (45-184 CD138+ cells/mL). Analysis of CPCs isolated using the device was consistent with serum immunoglobulin assays that are commonly used in MM diagnostics. These results indicate the potential of CD138-based microfluidic CPC capture as a useful ‘liquid biopsy’ that may complement or partially replace bone marrow aspiration.

COMPARISON OF PRE- AND POSTQUARANTINE BLOOD CHEMISTRY AND HEMATOLOGY VALUES FROM WILD-CAUGHT COWNOSE RAYS (RHINOPTERA BONASUS).

PubMed

Cusack, Lara; Field, Cara L; Hoopes, Lisa; McDermott, Alexa; Clauss, Tonya

2016-06-01

Though one of the most widely kept elasmobranchs in human care, the cownose ray (CNR; Rhinoptera bonasus ), remains a species with minimal published information on hematologic reference intervals. As part of a larger study investigating the health and nutrition of the CNR, this study established a preliminary data set of plasma chemistry and hematology values specific to animals recently caught from the wild and compared this data set (intake sample) to values obtained following a period of quarantine (27-40 days) in an aquarium (exit sample). Blood samples were collected from 47 wild female (n = 46) and male (n = 1) CNR caught in pound nets off the coast of North Carolina and South Carolina. Differences between intake and exit values were analyzed. Due to the preponderance of female animals, data were not analyzed for sex differences. Plasma biochemical profiles were performed and analyzed. A select number of complete blood cell counts were performed (n = 24 from 12 animals). Statistically significant differences (P < 0.05) specific to time of sampling were determined for packed cell volume, total solids, blood urea nitrogen, sodium, chloride, potassium, phosphorus, cholesterol, glucose, and aspartate aminotransferase. Values reported are a significant expansion on the existing limited data for CNRs and will serve as a reference for health assessment of individuals both in the wild and in exhibit populations.

A Single Session of Low-Volume High-Intensity Interval Exercise Reduces Ambulatory Blood Pressure in Normotensive Men.

PubMed

Dantas, Teresa C B; Farias Junior, Luiz F; Frazão, Danniel T; Silva, Paulo H M; Sousa Junior, Altieres E; Costa, Ingrid B B; Ritti-Dias, Raphael M; Forjaz, Cláudia L M; Duhamel, Todd A; Costa, Eduardo C

2017-08-01

Dantas, TCB, Farias Junior, LF, Frazão, DT, Silva, PHM, Sousa Junior, AE, Costa, IBB, Ritti-Dias, RM, Forjaz, CLM, Duhamel, TA, and Costa, EC. A single session of low-volume high-intensity interval exercise reduces ambulatory blood pressure in normotensive men. J Strength Cond Res 31(8): 2263-2269, 2017-The magnitude and duration of postexercise hypotension (PEH) may provide valuable information on the efficacy of an exercise approach to blood pressure (BP) control. We investigated the acute effect of a time-efficient high-intensity interval exercise (HIIE) on ambulatory BP. Twenty-one normotensive men (23.6 ± 3.6 years) completed 2 experimental sessions in a randomized order: (a) control (no exercise) and (b) low-volume HIIE: 10 × 1 minute at 100% of maximal treadmill velocity interspersed with 1 minute of recovery. After each experimental session, an ambulatory BP monitoring was initiated. Paired sample t-test was used to compare BP averages for awake, asleep, and 20-hour periods between the control and the low-volume HIIE sessions. A 2-way repeated measures analysis of variance was used to analyze hourly BP after both experimental sessions. Blood pressure averages during the awake (systolic: 118 ± 6 vs. 122 ± 6 mm Hg; diastolic: 65 ± 7 vs. 67 ± 7 mm Hg) and 20-hour (systolic: 115 ± 7 vs. 118 ± 6 mm Hg; diastolic: 62 ± 7 vs. 64 ± 7 mm Hg) periods were lower after the low-volume HIIE compared with the control (p ≤ 0.05). Systolic and diastolic PEH presented medium (Cohen's d = 0.50-0.67) and small (Cohen's d = 0.29) effect sizes, respectively. Systolic PEH occurred in a greater magnitude during the first 5 hours (3-5 mm Hg). No changes were found in asleep BP (p > 0.05). In conclusion, a single session of low-volume HIIE reduced ambulatory BP in normotensive men. The PEH occurred mainly in systolic BP during the first 5 hours postexercise.

Sports anaemia and anthropometric evaluation of footballers at Kwame Nkrumah University of Science and Technology (KNUST)

PubMed Central

Opoku-Okrah, Clement; Sam, Daniel Kwasi; Nkum, Bernard; Dogbe, Elliot Eli; Antwi-Boateng, Lilian; Sackey, Benedict; Gyamfi, Daniel; Danquah, Kwabena Owusu

2016-01-01

Introduction Sports anaemia is a physiological activity that occurs amongst footballers and may be due to poor diet, over-training, as well as an increase in plasma volume in endurance training activities. High plasma volume leads to changes in haematological parameters that may impact on endurance of footballers. The objective of the study was to determine the correlation between haematological and an-thropometric indices and their role in sports anaemia in a tropical setting. Methods Venous blood was taken into EDTA for 12 soccer players of KNUST soccer team before training and after training for the first (W1) and fifth (W5) weeks of training sessions. Complete blood count analysis was done for each blood sample and anthropometric parameters such as height, weight, body mass index, body fat percent and lean body mass were also measured. Cross-tabulations with mean and standard deviation or median and range were computed. Paired t-test & and Mann-Whitney test for parametric and non-parametric data computations were carried out and a p-value ≤ 0.05 was taken to rep-resent significant difference between data groups. Results There was significant reduction in haemoglobin (p = 0.003), haematocrit (p = 0.002), mean cell volume (MCV) (p = 0.034) and red blood cell (RBC) count (p = 0.011) as a result of a significant expansion of plasma volume (p= 0.006). Neutrophil, lymphocyte and eosinophil counts were reduced significantly (p= 0.043, 0.001 and 0.007, respectively) after the training at W5. Lean body mass (LBM) inversely correlated with haemoglobin (r = -0.787, p = 0.002) and haematocrit (r = -0.588, p = 0.044). Body fat percentage (BFP) also negatively correlated with lymphocyte count (r = -0.700, p = 0.011). Furthermore, there was a positive correlation between body mass index (BMI) and plasma volume change after the training programme (r = 0.689, p = 0.013). Conclusion The results suggest that sports anaemia was induced by an increase in plasma volume that resulted in changes in haematological parameters. PMID:27583089

Sports anaemia and anthropometric evaluation of footballers at Kwame Nkrumah University of Science and Technology (KNUST).

PubMed

Opoku-Okrah, Clement; Sam, Daniel Kwasi; Nkum, Bernard; Dogbe, Elliot Eli; Antwi-Boateng, Lilian; Sackey, Benedict; Gyamfi, Daniel; Danquah, Kwabena Owusu

2016-01-01

Sports anaemia is a physiological activity that occurs amongst footballers and may be due to poor diet, over-training, as well as an increase in plasma volume in endurance training activities. High plasma volume leads to changes in haematological parameters that may impact on endurance of footballers. The objective of the study was to determine the correlation between haematological and an-thropometric indices and their role in sports anaemia in a tropical setting. Venous blood was taken into EDTA for 12 soccer players of KNUST soccer team before training and after training for the first (W1) and fifth (W5) weeks of training sessions. Complete blood count analysis was done for each blood sample and anthropometric parameters such as height, weight, body mass index, body fat percent and lean body mass were also measured. Cross-tabulations with mean and standard deviation or median and range were computed. Paired t-test & and Mann-Whitney test for parametric and non-parametric data computations were carried out and a p-value ≤ 0.05 was taken to rep-resent significant difference between data groups. There was significant reduction in haemoglobin (p = 0.003), haematocrit (p = 0.002), mean cell volume (MCV) (p = 0.034) and red blood cell (RBC) count (p = 0.011) as a result of a significant expansion of plasma volume (p= 0.006). Neutrophil, lymphocyte and eosinophil counts were reduced significantly (p= 0.043, 0.001 and 0.007, respectively) after the training at W5. Lean body mass (LBM) inversely correlated with haemoglobin (r = -0.787, p = 0.002) and haematocrit (r = -0.588, p = 0.044). Body fat percentage (BFP) also negatively correlated with lymphocyte count (r = -0.700, p = 0.011). Furthermore, there was a positive correlation between body mass index (BMI) and plasma volume change after the training programme (r = 0.689, p = 0.013). The results suggest that sports anaemia was induced by an increase in plasma volume that resulted in changes in haematological parameters.

Increased hippocampal blood volume and normal blood flow in schizophrenia

PubMed Central

Talati, Pratik; Rane, Swati; Skinner, Jack; Gore, John; Heckers, Stephan

2015-01-01

Neuroimaging studies have provided compelling evidence for abnormal hippocampal activity in schizophrenia. Most studies made inferences about baseline hippocampal activity using a single hemodynamic parameter (e.g., blood volume or blood flow). Here we studied several hemodynamic measures in the same cohort to test the hypothesis of increased hippocampal activity in schizophrenia. We used dynamic susceptibility contrast- (DSC-) magnetic resonance imaging to assess blood volume, blood flow, and mean transit time in the hippocampus of 15 patients with chronic schizophrenia and 15 healthy controls. Left and right hippocampal measurements were combined for absolute measures of cerebral blood volume (CBV), blood flow (CBF), and mean transit time (MTT). We found significantly increased hippocampal CBV, but normal CBF and MTT, in schizophrenia. The uncoupling of CBV and CBF could be due to several factors, including antipsychotic medication, loss of cerebral perfusion pressure, or angiogenesis. Further studies need to incorporate several complementary imaging modalities to better characterize hippocampal dysfunction in schizophrenia. PMID:25896442

Cardiac-gated, phase contrast magnetic resonance angiography is a reliable and reproducible technique for quantifying blood flow in canine major cranial abdominal vessels.

PubMed

Del Chicca, Francesca; Schwarz, Andrea; Grest, Paula; Willmitzer, Florian; Dennler, Matthias; Kircher, Patrick R

2018-04-17

Blood flow changes in cranial abdominal vessels are important contributing factors for canine hepatic disease. This prospective, experimental, pilot study aimed to evaluate cardiac-gated, phase contrast magnetic resonance angiography (PCMRA) as a method for characterizing blood flow in canine major cranial abdominal vessels. Eleven, healthy, adult beagle dogs were sampled. Cardiac-gated, phase contrast magnetic resonance angiography of the cranial abdomen was performed in each dog and blood flow was independently measured in each of the major cranial abdominal vessels by three observers, with two observers recording blood flow values once and one observer recording blood flow values three times. Each dog then underwent ultrasonographic examination of the liver with fine needle aspirations and biopsies submitted to cytologic and histologic examination. The mean absolute stroke volume and velocity were respectively 9.6 ± 1.9 ml and -11.1 ± 1.1 cm/s for the cranial abdominal aorta, 2.1 ± 0.6 ml and -6.6 ± 1.9 cm/s for the celiac artery, and 2.3 ± 1.0 ml and -7.9 ± 3.1 cm/s for the cranial mesenteric artery. The mean absolute stroke volume and velocity were respectively 6.7 ± 1.3 ml and 3.9 ± 0.9 cm/s for the caudal vena cava and 2.6 ± 0.9 ml and 3.2 ± 1.2 cm/s for the portal vein. Intraobserver reliability was excellent (intraclass correlation coefficient > 0.9). Interobserver reproducibility was also excellent (intraclass correlation coefficient 0.89-0.99). Results of liver ultrasonography, cytology, and histopathology were unremarkable. Findings indicated that cardiac-gated, phase contrast magnetic resonance angiography is a feasible technique for quantifying blood blow in canine major cranial abdominal vessels. Blood flow values from this sample of healthy beagles can be used as background for future studies on canine hepatic disease. © 2018 American College of Veterinary Radiology.

Prenatal lead exposure and childhood blood pressure and kidney function.

PubMed

Skröder, Helena; Hawkesworth, Sophie; Moore, Sophie E; Wagatsuma, Yukiko; Kippler, Maria; Vahter, Marie

2016-11-01

Exposure to lead, a common environmental pollutant, is known to cause cardiovascular and nephrotoxic effects in adults. Potential effects of early-life lead exposure on these functions are, however, less well characterized. To assess blood pressure and kidney function in preschool-aged children in relation to prenatal lead exposure. This prospective study in rural Bangladesh measured children's systolic and diastolic blood pressure in triplicate at the follow-up at 4.5±0.11 years. Their kidney function was assessed by the estimated glomerular filtration rate (eGFR), calculated based on serum cystatin C concentrations, and by kidney volume, measured by sonography. Exposure to lead was assessed by concentrations in the mothers' blood (erythrocyte fraction; Ery-Pb) in gestational weeks (GW) 14 and 30, the effects of which were evaluated separately in multivariable-adjusted linear regression analyses. We found no associations between maternal exposure to lead [n~1500 for GW14 and 700 for GW30] and children's blood pressure or eGFR. However, we found an inverse association between late gestation lead and kidney volume, although the sample size was limited (n=117), but not with early gestation lead (n=573). An increase of 85µg/kg in Ery-Pb (median concentration at GW30) was associated with a 6.0cm 3 /m 2 decrease in kidney volume (=0.4SD; p=0.041). After stratifying on gender, there seemed to be a somewhat stronger association in girls. Prenatal lead exposure may cause long-lasting effects on the kidney. This warrants follow-up studies in older children, as well as additional studies in other populations. Copyright © 2016. Published by Elsevier Inc.

Pulmonary Edema and Plasma Volume Changes in Dysbarism. M.S. Thesis - Texas Univ.

NASA Technical Reports Server (NTRS)

Joki, J. A.

1972-01-01

Two groups of anesthetized, fasted pigs were utilized. One group of 13 animals (8.5 to 16.6 kilograms) was exposed to a high-pressure environment, and the other group of eight animals (6.9 to 20.0 kilograms) constituted the control group. The experimental group was subjected to an atmosphere of 90 percent nitrogen and 10 percent oxygen at a pressure of 50 psig for 30 minutes and then decompressed at a rate 10 psi/min. Plasma volumes, using both iodine-125-tagged-albumin and chromium-51-tagged-cell dilution techniques, were measured before, immediately after, and at 30 and 60 minutes after decompression. Aortic and right-ventricular systolic pressures were also recorded. At 60 minutes after decompression, blood samples were taken, the animals were sacrificed, and the water content of the lungs, kidneys, livers, and spleens was estimated by measuring tissue wet weight and dry weight. Protein extravasation and tissue blood volumes were determined by measuring the iodine-125-tagged-albumin and chromium-51-tagged-cell spaces in homo-genates of the organs under investigation.

Accurate measurement of volume and shape of resting and activated blood platelets from light scattering.

PubMed

Moskalensky, Alexander E; Yurkin, Maxim A; Konokhova, Anastasiya I; Strokotov, Dmitry I; Nekrasov, Vyacheslav M; Chernyshev, Andrei V; Tsvetovskaya, Galina A; Chikova, Elena D; Maltsev, Valeri P

2013-01-01

We introduce a novel approach for determination of volume and shape of individual blood platelets modeled as an oblate spheroid from angle-resolved light scattering with flow-cytometric technique. The light-scattering profiles (LSPs) of individual platelets were measured with the scanning flow cytometer and the platelet characteristics were determined from the solution of the inverse light-scattering problem using the precomputed database of theoretical LSPs. We revealed a phenomenon of parameter compensation, which is partly explained in the framework of anomalous diffraction approximation. To overcome this problem, additional a priori information on the platelet refractive index was used. It allowed us to determine the size of each platelet with subdiffraction precision and independent of the particular value of the platelet aspect ratio. The shape (spheroidal aspect ratio) distributions of platelets showed substantial differences between native and activated by 10 μM adenosine diphosphate samples. We expect that the new approach may find use in hematological analyzers for accurate measurement of platelet volume distribution and for determination of the platelet activation efficiency.

In vitro platelet quality in storage containers used for pediatric transfusions.

PubMed

Weiss, Sandra; Scammell, Kenneth; Levin, Elena; Culibrk, Brankica; Zolfaghari, Sima; Gyöngyössy-Issa, Maria I C; Acker, Jason P

2012-08-01

The in vitro quality of small-volume platelet (PLT) aliquots for pediatric transfusions was assessed to determine the best practice approach. Small volumes (50 mL) of single apheresis PLT components (APCs), collected on either CaridianBCT Trima or Haemonetics MCS+ instruments, were aliquoted on Days 2, 3, 4, and 5 postcollection into Fenwal PL1240 or 4R2014 bags or 60-mL polypropylene syringes. Samples were tested for in vitro quality at their recommended expiry times (4 hr for 4R2014 bags and syringes or Day 5 for PL1240 bags). Assays included pH, CD62P expression, and metabolic measures. CD62P expression increased throughout storage in all containers. Among the small-volume containers, pH, pCO(2) , lactate, and bicarbonate varied considerably. Regardless of the day of aliquoting, pCO(2) was significantly higher and pO(2) was significantly lower in gas-impermeable syringes than other containers. No bacterial growth was detected in any sample. The quality of APCs aliquoted into small-volume containers meets regulatory requirements and is generally equivalent to that of full-volume APCs at expiry. © 2012 American Association of Blood Banks.

Exsanguinated blood volume estimation using fractal analysis of digital images.

PubMed

Sant, Sonia P; Fairgrieve, Scott I

2012-05-01

The estimation of bloodstain volume using fractal analysis of digital images of passive blood stains is presented. Binary digital photos of bloodstains of known volumes (ranging from 1 to 7 mL), dispersed in a defined area, were subjected to image analysis using FracLac V. 2.0 for ImageJ. The box-counting method was used to generate a fractal dimension for each trial. A positive correlation between the generated fractal number and the volume of blood was found (R(2) = 0.99). Regression equations were produced to estimate the volume of blood in blind trials. An error rate ranging from 78% for 1 mL to 7% for 6 mL demonstrated that as the volume increases so does the accuracy of the volume estimation. This method used in the preliminary study proved that bloodstain patterns may be deconstructed into mathematical parameters, thus removing the subjective element inherent in other methods of volume estimation. © 2012 American Academy of Forensic Sciences.

Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.

PubMed

Khoo, Bee Luan; Warkiani, Majid Ebrahimi; Tan, Daniel Shao-Weng; Bhagat, Ali Asgar S; Irwin, Darryl; Lau, Dawn Pingxi; Lim, Alvin S T; Lim, Kiat Hon; Krisna, Sai Sakktee; Lim, Wan-Teck; Yap, Yoon Sim; Lee, Soo Chin; Soo, Ross A; Han, Jongyoon; Lim, Chwee Teck

2014-01-01

Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation. Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12-1275 CTCs/ml; Lung cancer samples: 10-1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples. We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.

Effects of regional hemoconcentration during LBNP on plasma volume determinations. [Lower Body Negative Pressure

NASA Technical Reports Server (NTRS)

Loeppky, J. A.; Kobayashi, Y.; Venters, M. D.; Luft, U. C.

1979-01-01

Blood samples were obtained from forearm vein or artery with indwelling cannula (1) before, (2) during the last min, and (3) about 2 min after lower body negative pressure (LBNP) in 16 experiments to determine whether plasma volume (PV) estimates were affected by regional hemoconcentration in the lower body. Total hemoglobin (THb) was estimated with the CO method prior to LBNP. Hemoglobin (Hb) and hematocrit (Hct) values from (2) gave only a 3% (87 ml) loss in PV due to LBNP, assuming no change in THb. However, Hb and Hct values from (3) showed an 11% loss in PV (313 ml). This 72% underestimation of PV loss with (2) must have resulted from the sequestration of blood and subsequent hemoconcentration in the lower body during LBNP. The effects of LBNP on PV should be estimated 1-3 min after exposure, after mixing but before extravascular fluid returns to the circulation.

Microchip Module for Blood Sample Preparation and Nucleic Acid Amplification Reactions

PubMed Central

Yuen, Po Ki; Kricka, Larry J.; Fortina, Paolo; Panaro, Nicholas J.; Sakazume, Taku; Wilding, Peter

2001-01-01

A computer numerical control-machined plexiglas-based microchip module was designed and constructed for the integration of blood sample preparation and nucleic acid amplification reactions. The microchip module is comprised of a custom-made heater-cooler for thermal cycling, a series of 254 μm × 254 μm microchannels for transporting human whole blood and reagents in and out of an 8–9 μL dual-purpose (cell isolation and PCR) glass-silicon microchip. White blood cells were first isolated from a small volume of human whole blood (<3 μL) in an integrated cell isolation–PCR microchip containing a series of 3.5-μm feature-sized “weir-type” filters, formed by an etched silicon dam spanning the flow chamber. A genomic target, a region in the human coagulation Factor V gene (226-bp), was subsequently directly amplified by microchip-based PCR on DNA released from white blood cells isolated on the filter section of the microchip mounted onto the microchip module. The microchip module provides a convenient means to simplify nucleic acid analyses by integrating two key steps in genetic testing procedures, cell isolation and PCR and promises to be adaptable for additional types of integrated assays. PMID:11230164

On the Mechanism of Human Red Blood Cell Longevity: Roles of Calcium, the Sodium Pump, PIEZO1, and Gardos Channels.

PubMed

Lew, Virgilio L; Tiffert, Teresa

2017-01-01

In a healthy adult, the transport of O 2 and CO 2 between lungs and tissues is performed by about 2 · 10 13 red blood cells, of which around 1.7 · 10 11 are renewed every day, a turnover resulting from an average circulatory lifespan of about 120 days. Cellular lifespan is the result of an evolutionary balance between the energy costs of maintaining cells in a fit functional state versus cell renewal. In this Review we examine how the set of passive and active membrane transporters of the mature red blood cells interact to maximize their circulatory longevity thus minimizing costs on expensive cell turnover. Red blood cell deformability is critical for optimal rheology and gas exchange functionality during capillary flow, best fulfilled when the volume of each human red blood cell is kept at a fraction of about 0.55-0.60 of the maximal spherical volume allowed by its membrane area, the optimal-volume-ratio range. The extent to which red blood cell volumes can be preserved within or near these narrow optimal-volume-ratio margins determines the potential for circulatory longevity. We show that the low cation permeability of red blood cells allows volume stability to be achieved with extraordinary cost-efficiency, favouring cell longevity over cell turnover. We suggest a mechanism by which the interplay of a declining sodium pump and two passive membrane transporters, the mechanosensitive PIEZO1 channel, a candidate mediator of P sickle in sickle cells, and the Ca 2+ -sensitive, K + -selective Gardos channel, can implement red blood cell volume stability around the optimal-volume-ratio range, as required for extended circulatory longevity.

On the Mechanism of Human Red Blood Cell Longevity: Roles of Calcium, the Sodium Pump, PIEZO1, and Gardos Channels

PubMed Central

Lew, Virgilio L.; Tiffert, Teresa

2017-01-01

In a healthy adult, the transport of O2 and CO2 between lungs and tissues is performed by about 2 · 1013 red blood cells, of which around 1.7 · 1011 are renewed every day, a turnover resulting from an average circulatory lifespan of about 120 days. Cellular lifespan is the result of an evolutionary balance between the energy costs of maintaining cells in a fit functional state versus cell renewal. In this Review we examine how the set of passive and active membrane transporters of the mature red blood cells interact to maximize their circulatory longevity thus minimizing costs on expensive cell turnover. Red blood cell deformability is critical for optimal rheology and gas exchange functionality during capillary flow, best fulfilled when the volume of each human red blood cell is kept at a fraction of about 0.55–0.60 of the maximal spherical volume allowed by its membrane area, the optimal-volume-ratio range. The extent to which red blood cell volumes can be preserved within or near these narrow optimal-volume-ratio margins determines the potential for circulatory longevity. We show that the low cation permeability of red blood cells allows volume stability to be achieved with extraordinary cost-efficiency, favouring cell longevity over cell turnover. We suggest a mechanism by which the interplay of a declining sodium pump and two passive membrane transporters, the mechanosensitive PIEZO1 channel, a candidate mediator of Psickle in sickle cells, and the Ca2+-sensitive, K+-selective Gardos channel, can implement red blood cell volume stability around the optimal-volume-ratio range, as required for extended circulatory longevity. PMID:29311949

The Effect of Pressure-Controlled Ventilation and Volume-Controlled Ventilation in Prone Position on Pulmonary Mechanics and Inflammatory Markers.

PubMed

Şenay, Hasan; Sıvacı, Remziye; Kokulu, Serdar; Koca, Buğra; Bakı, Elif Doğan; Ela, Yüksel

2016-08-01

The aim of this present study is to compare the effect of pressure-controlled ventilation and volume-controlled ventilation on pulmonary mechanics and inflammatory markers in prone position. The study included 41 patients undergoing to vertebrae surgery. The patients were randomized into two groups: Group 1 received volume-controlled ventilation, while group 2 received pressure-controlled ventilation. The demographic data, pulmonary mechanics, the inflammatory marker levels just after the induction of anesthetics, at the 6th and 12th hours, and gas analysis from arterial blood samples taken at the beginning and the 30th minute were recorded. The inflammatory marker levels increased in both groups, without any significant difference among groups. Peak inspiratory pressure level was higher in the volume-controlled ventilation group. This study revealed that there is no difference regarding inflammatory marker levels between volume- and pressure-controlled ventilation.

Chemistry Testing on Plasma Versus Serum Samples in Dialysis Patients: Clinical and Quality Improvement Implications.

PubMed

Carey, Roger Neill; Jani, Chinu; Johnson, Curtis; Pearce, Jim; Hui-Ng, Patricia; Lacson, Eduardo

2016-09-07

Plasma samples collected in tubes containing separator gels have replaced serum samples for most chemistry tests in many hospital and commercial laboratories. Use of plasma samples for blood tests in the dialysis population eliminates delays in sample processing while waiting for clotting to complete, laboratory technical issues associated with fibrin formation, repeat sample collection, and patient care issues caused by delay of results because of incompletely clotted specimens. Additionally, a larger volume of plasma is produced than serum for the same amount of blood collected. Plasma samples are also acceptable for most chemical tests involved in the care of patients with ESRD. This information becomes very important when United States regulatory requirements for ESRD inadvertently limit the type of sample that can be used for government reporting, quality assessment, and value-based payment initiatives. In this narrative, we summarize the renal community experience and how the subsequent resolution of the acceptability of phosphorus levels measured from serum and plasma samples may have significant implications in the country's continued development of a value-based Medicare ESRD Quality Incentive Program. Copyright © 2016 by the American Society of Nephrology.

Prevalence of Candidatus Mycoplasma haemolamae, bovine viral diarrhoea virus, and gastrointestinal parasitism in a sample of adult New Zealand alpaca (Vicugna pacos).

PubMed

Dittmer, K E; Hinkson, J A; Dwyer, C; Adlington, B; van Andel, M

2018-01-01

To determine the prevalence of infection with Candidatus Mycoplasma haemolamae (Mhl), antibodies to bovine viral diarrhoea virus (BVDV), and BVDV antigen, and the prevalence of animals with elevated faecal nematode egg counts (FEC) in a sample of adult New Zealand alpaca (Vicugna pacos). Blood samples were obtained from 175 alpaca, collected from 15 farms around New Zealand, and from 31 samples sent to a diagnostic laboratory for routine haematology. Blood smears (n=170) were examined microscopically for the presence of haemoplasma, and DNA was extracted from whole blood (n=206) for real-time PCR testing for Mhl. Packed cell volume (PCV) was determined for 193 samples. Serum samples (n=195) were tested for BVDV antibody using ELISA, and for BVDV antigen using a real-time PCR assay. Faecal samples were collected from 143 animals; FEC were measured, and samples pooled for larval culture. No haemoplasma organisms were present on blood smear examination. Of the 206 blood samples, two (from the same farm) were positive for Mhl by real-time PCR testing, giving a prevalence of infection with Mhl of 0.97%. Of the 195 serum samples tested, four (2.1%) were positive for antibodies to BVDV; animals with BVDV antibodies were from 3/15 (20%) farms, none of which farmed cattle. None of the serum samples were positive by PCR for BVDV antigen. The median FEC was 50 epg (min 0, max 4,700), with 55/143 (38.5%) samples having 0 epg, and 33/143 (23.1%) having ≥250 epg. Haemonchus spp. were the most common nematodes present in faecal larval cultures from the North Island. Log 10 FEC was negatively associated with PCV (p=0.02), and was higher in males than females (p<0.001), and in animals that were positive compared with negative for Mhl (p=0.022). The number of alpaca infected with Mhl was low, as was the seroprevalence of BVDV. Gastrointestinal parasitism was, however, a common finding in this sample of New Zealand alpaca.

Selling blood and gametes during tough economic times: insights from Google search.

PubMed

Wu, Jonathan A; Ngo, Tin C; Rothman, Cappy; Breyer, Benjamin N; Eisenberg, Michael L

2015-10-01

To use Google Insights search volume and publicly available economic indicators to test the hypothesis that sperm, egg, and blood donations increase during economic downturns and to demonstrate the feasibility of using Google search volume data to predict national trends in actual sperm, egg, and blood donations rates. Cross-correlation statistical analysis comparing Google search data for terms relating to blood, egg, and sperm donations with various economic indicators including the S&P 500 closing values, gross domestic product (GDP), the U.S. Index of Leading Indicators (U.S. Leading Index), gross savings rate, mortgage interest rates, unemployment rate, and consumer price index (CPI) from 2004-2011. A secondary analysis determined the Pearson correlation coefficient between Google search data with actual sperm, egg, and blood donation volume in the U.S. as measured by California Cryobank, the National Assisted Reproductive Technology Surveillance System, and the National Blood Collection and Utilization Survey, respectively. Significance of cross-correlation and Pearson correlation analysis as indicated by p value. There were several highly significant cross-correlation relationships between search volume and various economic indicators. Correlation between Google search volume for the term 'sperm donation,' 'egg donation,' and 'blood donation' with actual number of sperm, egg and blood donations in the United States demonstrated Pearson correlation coefficients of 0.2 (p > 0.10), -0.1 (p > 0.10), and 0.07 (p > 0.10), respectively. Temporal analysis showed an improved correlation coefficient of 0.9 (p < 0.05) for blood donation when shifted 12 months later relative to Google search volume. Google search volume data for search terms relating to sperm, egg, and blood donation increase during economic downturns. This finding suggests gamete and bodily fluid donations are influenced by market forces like other commodities. Google search may be useful for predicting blood donation trends but is more limited in predicting actual semen and oocyte donation patterns.

Blood transfusion in pediatric cardiac surgery.

PubMed

Durandy, Yves

2010-11-01

The aim of the study is to measure the volume of homologous blood needed for one pediatric patient during his hospital stay. Over a 4-month period, all the patients operated upon with a blood prime or requiring blood transfusion during their hospital stay were included in this study.The cardiopulmonary bypass protocol associates a miniaturized bypass circuit, vacuum-assisted venous drainage, and microplegia. The volume of each blood product opened is known and the volume of blood product remaining, following the last transfusion, is measured. Data collected areas follows: patient weight; hemoglobin level before surgery,during bypass, and in intensive care after the last transfusion;time to extubation; and degree of inotropic support.Forty-six patients weighing 5.1 1.5 kg were included in this study. Cardiopulmonary bypass priming volume was 100 mL for patients up to 3.5 kg, 120 mL for patients between 3.6 and 7.5 kg, and 160 mL for patients between 7.6 and 8.6 kg. The volume of blood transfusion was 271 112 mL, hemoglobin level before surgery was 10.3 1.7 g/dL, hemoglobin level during surgery was 11.0 1.5 g/dL, and hemoglobin level after the last transfusion was 12.3 2.4 g/dL. Time to extubation was 12 3.3 h, and inotropic support was enoximone in 37 patients,whereas 6 patients needed enoximone and epinephrine.No patient needed reexploration for bleeding and one patient received a platelet transfusion.The mean blood transfusion volume was equivalent to 60% of the patient’s total blood volume (estimated to be 80 mL/kg).

Blood Infusion and the Risk of Haemorrhage in Patients Undergoing Cardiac Surgery with Extracorporeal Circulation.

PubMed

Luque-Oliveros, Manuel; Garcia-Carpintero, Maria Angeles; Cauli, Omar

2017-01-01

Patients undergoing cardiac surgery with extracorporeal circulation (ECC) frequently present haemorrhages as a complication associated with high morbidity and mortality. One of the factors that influences this risk is the volume of blood infused during surgery. The objective of this study was to determine the optimal volume of autologous blood that can be processed during cardiac surgery with ECC. We also determined the number of salvaged red blood cells to be reinfused into the patient in order to minimize the risk of haemorrhage in the postoperative period. This was an observational retrospective cross-sectional study performed in 162 ECC cardiac surgery patients. Data regarding the sociodemographic profiles of the patients, their pathologies and surgical treatments, and the blood volume recovered, processed, and reinfused after cell salvage were collected. We also evaluated the occurrence of postoperative haemorrhage. The volume of blood infused after cell salvage had a statistically significant effect (p < 0.01) on the risk of post-operative haemorrhage; the receiver operating characteristic sensitivity was 0.813 and the optimal blood volume cut-off was 1800 ml. The best clinical outcome (16.7% of patients presenting haemorrhages) was in patients that had received less than 1800 ml of recovered and processed autologous blood, which represented a volume of up to 580 ml reinfused red blood cells. The optimum thresholds for autologous processed blood and red blood cells reinfused into the patient were 1800 and 580 ml, respectively. Increasing these thresholds augmented the risk of haemorrhage as an immediate postoperative period complication. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

A simple microviscometric approach based on Brownian motion tracking.

PubMed

Hnyluchová, Zuzana; Bjalončíková, Petra; Karas, Pavel; Mravec, Filip; Halasová, Tereza; Pekař, Miloslav; Kubala, Lukáš; Víteček, Jan

2015-02-01

Viscosity-an integral property of a liquid-is traditionally determined by mechanical instruments. The most pronounced disadvantage of such an approach is the requirement of a large sample volume, which poses a serious obstacle, particularly in biology and biophysics when working with limited samples. Scaling down the required volume by means of microviscometry based on tracking the Brownian motion of particles can provide a reasonable alternative. In this paper, we report a simple microviscometric approach which can be conducted with common laboratory equipment. The core of this approach consists in a freely available standalone script to process particle trajectory data based on a Newtonian model. In our study, this setup allowed the sample to be scaled down to 10 μl. The utility of the approach was demonstrated using model solutions of glycerine, hyaluronate, and mouse blood plasma. Therefore, this microviscometric approach based on a newly developed freely available script can be suggested for determination of the viscosity of small biological samples (e.g., body fluids).

Self-driven filter-based blood plasma separator microfluidic chip for point-of-care testing.

PubMed

Madadi, Hojjat; Casals-Terré, Jasmina; Mohammadi, Mahdi

2015-05-22

There is currently a growing need for lab-on-a-chip devices for use in clinical analysis and diagnostics, especially in the area of patient care. The first step in most blood assays is plasma extraction from whole blood. This paper presents a novel, self-driven blood plasma separation microfluidic chip, which can extract more than 0.1 μl plasma from a single droplet of undiluted fresh human blood (~5 μl). This volume of blood plasma is extracted from whole blood with high purity (more than 98%) in a reasonable time frame (3 to 5 min), and without the need for any external force. This would be the first step towards the realization of a single-use, self-blood test that does not require any external force or power source to deliver and analyze a fresh whole-blood sample, in contrast to the existing time-consuming conventional blood analysis. The prototypes are manufactured in polydimethylsiloxane that has been modified with a strong nonionic surfactant (Silwet L-77) to achieve hydrophilic behavior. The main advantage of this microfluidic chip design is the clogging delay in the filtration area, which results in an increased amount of extracted plasma (0.1 μl). Moreover, the plasma can be collected in one or more 10 μm-deep channels to facilitate the detection and readout of multiple blood assays. This high volume of extracted plasma is achieved thanks to a novel design that combines maximum pumping efficiency without disturbing the red blood cells' trajectory through the use of different hydrodynamic principles, such as a constriction effect and a symmetrical filtration mode. To demonstrate the microfluidic chip's functionality, we designed and fabricated a novel hybrid microdevice that exhibits the benefits of both microfluidics and lateral flow immunochromatographic tests. The performance of the presented hybrid microdevice is validated using rapid detection of thyroid stimulating hormone within a single droplet of whole blood.

Development, validation and clinical application of a LC-MS/MS method for the simultaneous quantification of hydroxychloroquine and its active metabolites in human whole blood.

PubMed

Soichot, Marion; Mégarbane, Bruno; Houzé, Pascal; Chevillard, Lucie; Fonsart, Julien; Baud, Frédéric J; Laprévote, Olivier; Bourgogne, Emmanuel

2014-11-01

A rapid, sensitive and specific method using liquid chromatography coupled to tandem mass spectrometry was developed for the simultaneous quantification of hydroxychloroquine (HCQ) and its three major metabolites in human whole blood. The assay, using a sample volume of 100μL, was linear in a dynamic 25-2000ng/mL range (R(2)>0.99) for all four compounds and suitable for the determination of elevated HCQ concentrations up to 20,000ng/mL, after appropriate sample dilution. Inter- and intra-assay precisions were <18.2% and accuracies were between 84% and 113% for any analyte. No matrix effects were observed. The assay was successfully applied to a blood sample obtained from one poisoned patient following a massive HCQ self-ingestion resulting in an estimated concentration of 19,500ng/mL on hospital admission. In this patient, HCQ metabolites were identified and quantified at 1123, 465 and 91ng/mL for monodesethylhydroxychloroquine, desethylchloroquine and bisdesethylchloroquine, respectively. Further investigations are still required to assess the usefulness of the simultaneous measurement of blood concentrations of HCQ and its three active metabolites for monitoring HCQ treatment and managing HCQ poisoning. Copyright © 2014 Elsevier B.V. All rights reserved.

A Volume-Fraction Based Two-Phase Constitutive Model for Blood

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

Zhao, Rui; Massoudi, Mehrdad; Hund, S.J.

2008-06-01

Mechanically-induced blood trauma such as hemolysis and thrombosis often occurs at microscopic channels, steps and crevices within cardiovascular devices. A predictive mathematical model based on a broad understanding of hemodynamics at micro scale is needed to mitigate these effects, and is the motivation of this research project. Platelet transport and surface deposition is important in thrombosis. Microfluidic experiments have previously revealed a significant impact of red blood cell (RBC)-plasma phase separation on platelet transport [5], whereby platelet localized concentration can be enhanced due to a non-uniform distribution of RBCs of blood flow in a capillary tube and sudden expansion. However,more » current platelet deposition models either totally ignored RBCs in the fluid by assuming a zero sample hematocrit or treated them as being evenly distributed. As a result, those models often underestimated platelet advection and deposition to certain areas [2]. The current study aims to develop a two-phase blood constitutive model that can predict phase separation in a RBC-plasma mixture at the micro scale. The model is based on a sophisticated theory known as theory of interacting continua, i.e., mixture theory. The volume fraction is treated as a field variable in this model, which allows the prediction of concentration as well as velocity profiles of both RBC and plasma phases. The results will be used as the input of successive platelet deposition models.« less

Evaluation of the agreement among three handheld blood glucose meters and a laboratory blood analyzer for measurement of blood glucose concentration in Hispaniolan Amazon parrots (Amazona ventralis).

PubMed

Acierno, Mark J; Mitchell, Mark A; Schuster, Patricia J; Freeman, Diana; Sanchez-Migallon Guzman, David; Tully, Thomas N

2009-02-01

To determine the degree of agreement between 3 commercially available point-of-care blood glucose meters and a laboratory analyzer for measurement of blood glucose concentrations in Hispaniolan Amazon parrots (Amazona ventralis). 20 healthy adult Hispaniolan Amazon parrots. A 26-gauge needle and 3-mL syringe were used to obtain a blood sample (approx 0.5 mL) from a jugular vein of each parrot. Small volumes of blood (0.6 to 1.5 microL) were used to operate each of the blood glucose meters, and the remainder was placed into lithium heparin microtubes and centrifuged. Plasma was harvested and frozen at -30 degrees C. Within 5 days after collection, plasma samples were thawed and plasma glucose concentrations were measured by means of the laboratory analyzer. Agreement between pairs of blood glucose meters and between each blood glucose meter and the laboratory analyzer was evaluated by means of the Bland-Altman method, and limits of agreement (LOA) were calculated. None of the results of the 3 blood glucose meters agreed with results of the laboratory analyzer. Each point-of-care blood glucose meter underestimated the blood glucose concentration, and the degree of negative bias was not consistent (meter A bias, -94.9 mg/dL [LOA, -148.0 to -41.7 mg/dL]; meter B bias, -52 mg/dL [LOA, -107.5 to 3.5 mg/dL]; and meter C bias, -78.9 mg/dL [LOA, -137.2 to -20.6 mg/dL]). On the basis of these results, use of handheld blood glucose meters in the diagnosis or treatment of Hispaniolan Amazon parrots and other psittacines cannot be recommended.

Pinched flow coupled shear-modulated inertial microfluidics for high-throughput rare blood cell separation.

PubMed

Bhagat, Ali Asgar S; Hou, Han Wei; Li, Leon D; Lim, Chwee Teck; Han, Jongyoon

2011-06-07

Blood is a highly complex bio-fluid with cellular components making up >40% of the total volume, thus making its analysis challenging and time-consuming. In this work, we introduce a high-throughput size-based separation method for processing diluted blood using inertial microfluidics. The technique takes advantage of the preferential cell focusing in high aspect-ratio microchannels coupled with pinched flow dynamics for isolating low abundance cells from blood. As an application of the developed technique, we demonstrate the isolation of cancer cells (circulating tumor cells (CTCs)) spiked in blood by exploiting the difference in size between CTCs and hematologic cells. The microchannel dimensions and processing parameters were optimized to enable high throughput and high resolution separation, comparable to existing CTC isolation technologies. Results from experiments conducted with MCF-7 cells spiked into whole blood indicate >80% cell recovery with an impressive 3.25 × 10(5) fold enrichment over red blood cells (RBCs) and 1.2 × 10(4) fold enrichment over peripheral blood leukocytes (PBL). In spite of a 20× sample dilution, the fast operating flow rate allows the processing of ∼10(8) cells min(-1) through a single microfluidic device. The device design can be easily customized for isolating other rare cells from blood including peripheral blood leukocytes and fetal nucleated red blood cells by simply varying the 'pinching' width. The advantage of simple label-free separation, combined with the ability to retrieve viable cells post enrichment and minimal sample pre-processing presents numerous applications for use in clinical diagnosis and conducting fundamental studies.

Measuring the level of agreement between a veterinary and a human point-of-care glucometer and a laboratory blood analyzer in Hispaniolan Amazon parrots (Amazona ventralis).

PubMed

Acierno, Mark J; Schnellbacher, Rodney; Tully, Thomas N

2012-12-01

Although abnormalities in blood glucose concentrations in avian species are not as common as they are in mammals, the inability to provide point-of-care glucose measurement likely results in underreporting and missed treatment opportunities. A veterinary glucometer that uses different optimization codes for specific groups of animals has been produced. To obtain data for a psittacine bird-specific optimization code, as well as to calculate agreement between the veterinary glucometer, a standard human glucometer, and a laboratory analyzer, blood samples were obtained from 25 Hispaniolan Amazon parrots (Amazona ventralis) in a 2-phase study. In the initial phase, blood samples were obtained from 20 parrots twice at a 2-week interval. For each sample, the packed cell volume was determined, and the blood glucose concentration was measured by the veterinary glucometer. The rest of each sample was placed into a lithium heparin microtainer tube and centrifuged, and plasma was removed and frozen at -30 degrees C. Within 5 days, tubes were thawed, and blood glucose concentrations were measured with a laboratory analyzer. The data from both procedures were used to develop a psittacine bird-specific code. For the second phase of the study, the same procedure was repeated twice at a 2-week interval in 25 birds to determine agreement between the veterinary glucometer, a standard human glucometer, and a laboratory analyzer. Neither glucometer was in good agreement with the laboratory analyzer (veterinary glucometer bias, 9.0; level of agreement, -38.1 to 56.2; standard glucometer bias, 69.4; level of agreement -17.8 to 156.7). Based on these results, the use of handheld glucometers in the diagnostic testing of Hispaniolan Amazon parrots and other psittacine birds cannot be recommended.

Low vacuum and discard tubes reduce hemolysis in samples drawn from intravenous catheters.

PubMed

Heiligers-Duckers, Connie; Peters, Nathalie A L R; van Dijck, Jose J P; Hoeijmakers, Jan M J; Janssen, Marcel J W

2013-08-01

In-vitro hemolysis is a great challenge to emergency departments where blood is drawn from intravenous catheters (IVCs). Although high quality samples can be obtained by straight needle venipuncture, IVCs are preferred for various reasons. The aim of this study was to identify blood collection practices that reduce hemolysis while using IVC. The study was conducted at an emergency department where blood is drawn in ≥ 90% of patients from IVC. Hemolysis, measured spectrophotometrically, was compared between syringe and vacuum tubes. The following practices were tested in combination with vacuum collection; a Luer-slip adapter, a Luer-lock adapter, discard tubes and low vacuum tubes. Each intervention lasted 1 week and retrieved 154 to 297 samples. As reference, hemolysis was also measured in vacuum tubes retrieved from departments where only straight needle venipuncture is performed. Vacuum collection led to more hemolytic samples compared with syringe tubes (24% versus 16% respectively, p=0.008). No difference in hemolysis was observed between the Luer-slip and the Luer-lock adapter. The use of discard (17% hemolytic, p=0.045) and low vacuum tubes (12% hemolytic, p<0.001) substantially decreased hemolysis. None of the interventions reduced the hemolysis rate to the level observed when drawing blood by straight needle venipuncture (3%, p<0.02). In summary, both discard and low vacuum tubes reduce hemolysis while drawing blood from IVC. Of these practices the use of a low vacuum tube is preferred considering the less volume of blood and the amount of tubes drawn. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Dried Blood Spot Methodology in Combination With Liquid Chromatography/Tandem Mass Spectrometry Facilitates the Monitoring of Teriflunomide

PubMed Central

Lunven, Catherine; Turpault, Sandrine; Beyer, Yann-Joel; O'Brien, Amy; Delfolie, Astrid; Boyanova, Neli; Sanderink, Ger-Jan; Baldinetti, Francesca

2016-01-01

Background: Teriflunomide, a once-daily oral immunomodulator approved for treatment of relapsing-remitting multiple sclerosis, is eliminated slowly from plasma. If necessary to rapidly lower plasma concentrations of teriflunomide, an accelerated elimination procedure using cholestyramine or activated charcoal may be used. The current bioanalytical assay for determination of plasma teriflunomide concentration requires laboratory facilities for blood centrifugation and plasma storage. An alternative method, with potential for greater convenience, is dried blood spot (DBS) methodology. Analytical and clinical validations are required to switch from plasma to DBS (finger-prick sampling) methodology. Methods: Using blood samples from healthy subjects, an LC-MS/MS assay method for quantification of teriflunomide in DBS over a range of 0.01–10 mcg/mL was developed and validated for specificity, selectivity, accuracy, precision, reproducibility, and stability. Results were compared with those from the current plasma assay for determination of plasma teriflunomide concentration. Results: Method was specific and selective relative to endogenous compounds, with process efficiency ∼88%, and no matrix effect. Inaccuracy and imprecision for intraday and interday analyses were <15% at all concentrations tested. Quantification of teriflunomide in DBS assay was not affected by blood deposit volume and punch position within spot, and hematocrit level had a limited but acceptable effect on measurement accuracy. Teriflunomide was stable for at least 4 months at room temperature, and for at least 24 hours at 37°C with and without 95% relative humidity, to cover sampling, drying, and shipment conditions in the field. The correlation between DBS and plasma concentrations (R2 = 0.97), with an average blood to plasma ratio of 0.59, was concentration independent and constant over time. Conclusions: DBS sampling is a simple and practical method for monitoring teriflunomide concentrations. PMID:27015245

The Blood Volume of the Guinea Pig: Effects of Epinephrine and Isoproterenol upon the Red Cell and Plasma Volumes, Heart Rate, and Mean Arterial Pressure,

DTIC Science & Technology

1987-09-01

capillaries (4), blood volumes calculated from plasma volume measures must correct for label that has left the system between the time of the injected dose...Splenic sequestration and contraction are mediated by the autonomic nervous system and blood-borne agents (10). Sympathetic nerve fibers from the truncus...sympathlcus and parasympathetic neurons of the nervus vagus (cranial nerve X) innervate the celiac plexus (8, 11). A subdivision of the celiac plexus

Continuous monitoring of blood volume changes in humans

NASA Technical Reports Server (NTRS)

Hinghofer-Szalkay, H.; Greenleaf, J. E.

1987-01-01

Use of on-line high-precision mass densitometry for the continuous monitoring of blood volume changes in humans was demonstrated by recording short-term blood volume alterations produced by changes in body position. The mass density of antecubital venous blood was measured continuously for 80 min per session with 0.1 g/l precision at a flow rate of 1.5 ml/min. Additional discrete plasma density and hematocrit measurements gave linear relations between all possible combinations of blood density, plasma density, and hematocrit. Transient filtration phenomena were revealed that are not amenable to discontinuous measurements.

Optimization of Evans blue quantitation in limited rat tissue samples

PubMed Central

Wang, Hwai-Lee; Lai, Ted Weita

2014-01-01

Evans blue dye (EBD) is an inert tracer that measures plasma volume in human subjects and vascular permeability in animal models. Quantitation of EBD can be difficult when dye concentration in the sample is limited, such as when extravasated dye is measured in the blood-brain barrier (BBB) intact brain. The procedure described here used a very small volume (30 µl) per sample replicate, which enabled high-throughput measurements of the EBD concentration based on a standard 96-well plate reader. First, ethanol ensured a consistent optic path length in each well and substantially enhanced the sensitivity of EBD fluorescence spectroscopy. Second, trichloroacetic acid (TCA) removed false-positive EBD measurements as a result of biological solutes and partially extracted EBD into the supernatant. Moreover, a 1:2 volume ratio of 50% TCA ([TCA final] = 33.3%) optimally extracted EBD from the rat plasma protein-EBD complex in vitro and in vivo, and 1:2 and 1:3 weight-volume ratios of 50% TCA optimally extracted extravasated EBD from the rat brain and liver, respectively, in vivo. This procedure is particularly useful in the detection of EBD extravasation into the BBB-intact brain, but it can also be applied to detect dye extravasation into tissues where vascular permeability is less limiting. PMID:25300427

Optimization of Evans blue quantitation in limited rat tissue samples

NASA Astrophysics Data System (ADS)

Wang, Hwai-Lee; Lai, Ted Weita

2014-10-01

Evans blue dye (EBD) is an inert tracer that measures plasma volume in human subjects and vascular permeability in animal models. Quantitation of EBD can be difficult when dye concentration in the sample is limited, such as when extravasated dye is measured in the blood-brain barrier (BBB) intact brain. The procedure described here used a very small volume (30 µl) per sample replicate, which enabled high-throughput measurements of the EBD concentration based on a standard 96-well plate reader. First, ethanol ensured a consistent optic path length in each well and substantially enhanced the sensitivity of EBD fluorescence spectroscopy. Second, trichloroacetic acid (TCA) removed false-positive EBD measurements as a result of biological solutes and partially extracted EBD into the supernatant. Moreover, a 1:2 volume ratio of 50% TCA ([TCA final] = 33.3%) optimally extracted EBD from the rat plasma protein-EBD complex in vitro and in vivo, and 1:2 and 1:3 weight-volume ratios of 50% TCA optimally extracted extravasated EBD from the rat brain and liver, respectively, in vivo. This procedure is particularly useful in the detection of EBD extravasation into the BBB-intact brain, but it can also be applied to detect dye extravasation into tissues where vascular permeability is less limiting.

Evaluation of the Efficacy, Potential for Vector Transmission, and Duration of Immunity of MP-12, an Attenuated Rift Valley Fever Virus Vaccine Candidate, in Sheep

DTIC Science & Technology

2015-08-01

testing by virus isolation (VI). Positive-control mosquitoes were sampled whole on the day of the spiked blood meal. b NA, not applicable. Miller et al ...percent inhibition of the negative control, calculated as 1 (test serum OD/negative serum OD) 100. Miller et al . 934 cvi.asm.org August 2015 Volume 22...Trop Med Hyg 44:278 –282. 34. Morrill JC, Mebus CA, Peters CJ. 1997. Safety of a mutagen-attenuated Miller et al . 936 cvi.asm.org August 2015 Volume 22

Blood Collection

NASA Technical Reports Server (NTRS)

1999-01-01

The method that is used for the collection, storage and real-time analysis of blood and other bodily fluids has been licensed to DBCD, Inc. by NASA. The result of this patent licensing agreement has been the development of a commercial product that can provide serum or plasma from whole blood volumes of 20 microliters to 4 milliliters. The device has a fibrous filter with a pore size of less than about 3 microns, and is coated with a mixture of mannitol and plasma fraction protein. The coating causes the cellular fraction to be trapped by the small pores, leaving the cellular fraction intact on the fibrous filter while the acellular fraction passes through the filter for collection in unaltered form from the serum sample collection chamber. The method used by this product is useful to NASA for blood analysis on manned space missions.

Decrease of pulmonary blood flow detected by phase contrast MRI is correlated with a decrease in lung volume and increase of lung fibrosis area determined by computed tomography in interstitial lung disease.

PubMed

Tsuchiya, Nanae; Yamashiro, Tsuneo; Murayama, Sadayuki

2016-09-01

Lung volume and pulmonary blood flow decrease in patients with interstitial lung disease (ILD). The purpose of this study was to assess the relationship between pulmonary blood flow and lung volume in ILD patients. This research was approved by the institutional review board. Twenty-seven patients (9 men, 18 women; mean age, 59 years; range, 24-79 years) with ILD were included. Blood flow was assessed in the pulmonary trunk and the left and right pulmonary arteries by phase contrast magnetic resonance imaging (MRI). Lung volume and the computed tomography (CT) visual score that indicates the severity of ILD were assessed on the left and right sides by thin-section CT scanning. Lung volume was automatically measured by lung analysis software (VINCENT Ver. 4). The CT visual score was measured by averaging the proportion of abnormal lung area at five anatomic levels. Pearson's correlation coefficient was used to determine the relationship between pulmonary blood flow and lung volume. Pulmonary blood flow showed a significant correlation with lung volume (both: r=0.52, p=0.006; left: r=0.61, p=0.001; right: r=0.54, p=0.004) and CT visual score (both: r=-0.39, p=0.04; left: r=-0.48, p=0.01; right: r=-0.38, p=0.04). Partial correlation analysis, controlled for age, height and weight, showed a significant correlation between pulmonary blood flow and lung volume (both: r=0.43, p=0.03; left: r=0.55, p=0.005; right: r=0.48, p=0.01) and CT visual score (both: r=-0.58, p=0.003; left: r=-0.51, p=0.01; right: r=-0.64, p=0.001). In ILD, reduced pulmonary blood flow is associated with reduced lung volume and increased abnormal lung area. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Self-aliquoting micro-grooves in combination with laser ablation-ICP-mass spectrometry for the analysis of challenging liquids: quantification of lead in whole blood.

PubMed

Nischkauer, Winfried; Vanhaecke, Frank; Limbeck, Andreas

2016-08-01

We present a technique for the fast screening of the lead concentration in whole blood samples using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). The whole blood sample is deposited on a polymeric surface and wiped across a set of micro-grooves previously engraved into the surface. The engraving of the micro-grooves was accomplished with the same laser system used for LA-ICP-MS analysis. In each groove, a part of the liquid blood is trapped, and thus, the sample is divided into sub-aliquots. These aliquots dry quasi instantly and are then investigated by means of LA-ICP-MS. For quantification, external calibration against aqueous standard solutions was relied on, with iron as an internal standard to account for varying volumes of the sample aliquots. The (208)Pb/(57)Fe nuclide ratio used for quantification was obtained via a data treatment protocol so far only used in the context of isotope ratio determination involving transient signals. The method presented here was shown to provide reliable results for Recipe ClinChek® Whole Blood Control levels I-III (nos. 8840-8842), with a repeatability of typically 3 % relative standard deviation (n = 6, for Pb at 442 μg L(-1)). Spiked and non-spiked real whole blood was analysed as well, and the results were compared with those obtained via dilution and sectorfield ICP-MS. A good agreement between both methods was observed. The detection limit (3 s) for lead in whole blood was established to be 10 μg L(-1) for the laser ablation method presented here. Graphical Abstract Micro-grooves are filled with whole blood, dried, and analyzed by laser ablation ICP-mass spectrometry. Notice that the laser moves in perpendicular direction with regard to the micro-grooves.

Red Cell Properties after Different Modes of Blood Transportation

PubMed Central

Makhro, Asya; Huisjes, Rick; Verhagen, Liesbeth P.; Mañú-Pereira, María del Mar; Llaudet-Planas, Esther; Petkova-Kirova, Polina; Wang, Jue; Eichler, Hermann; Bogdanova, Anna; van Wijk, Richard; Vives-Corrons, Joan-Lluís; Kaestner, Lars

2016-01-01

Transportation of blood samples is unavoidable for assessment of specific parameters in blood of patients with rare anemias, blood doping testing, or for research purposes. Despite the awareness that shipment may substantially alter multiple parameters, no study of that extent has been performed to assess these changes and optimize shipment conditions to reduce transportation-related artifacts. Here we investigate the changes in multiple parameters in blood of healthy donors over 72 h of simulated shipment conditions. Three different anticoagulants (K3EDTA, Sodium Heparin, and citrate-based CPDA) for two temperatures (4°C and room temperature) were tested to define the optimal transportation conditions. Parameters measured cover common cytology and biochemistry parameters (complete blood count, hematocrit, morphological examination), red blood cell (RBC) volume, ion content and density, membrane properties and stability (hemolysis, osmotic fragility, membrane heat stability, patch-clamp investigations, and formation of micro vesicles), Ca2+ handling, RBC metabolism, activity of numerous enzymes, and O2 transport capacity. Our findings indicate that individual sets of parameters may require different shipment settings (anticoagulants, temperature). Most of the parameters except for ion (Na+, K+, Ca2+) handling and, possibly, reticulocytes counts, tend to favor transportation at 4°C. Whereas plasma and intraerythrocytic Ca2+ cannot be accurately measured in the presence of chelators such as citrate and EDTA, the majority of Ca2+-dependent parameters are stabilized in CPDA samples. Even in blood samples from healthy donors transported using an optimized shipment protocol, the majority of parameters were stable within 24 h, a condition that may not hold for the samples of patients with rare anemias. This implies for as short as possible shipping using fast courier services to the closest expert laboratory at reach. Mobile laboratories or the travel of the patients to the specialized laboratories may be the only option for some groups of patients with highly unstable RBCs. PMID:27471472

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.

An ultra-high-throughput spiral microfluidic biochip for the enrichment of circulating tumor cells.

PubMed

Warkiani, Majid Ebrahimi; Khoo, Bee Luan; Tan, Daniel Shao-Weng; Bhagat, Ali Asgar S; Lim, Wan-Teck; Yap, Yoon Sim; Lee, Soo Chin; Soo, Ross A; Han, Jongyoon; Lim, Chwee Teck

2014-07-07

The detection and characterization of rare circulating tumor cells (CTCs) from the blood of cancer patients can potentially provide critical insights into tumor biology and hold great promise for cancer management. The ability to collect a large number of viable CTCs for various downstream assays such as quantitative measurements of specific biomarkers or targeted somatic mutation analysis is increasingly important in medical oncology. Here, we present a simple yet reliable microfluidic device for the ultra-high-throughput, label-free, size-based isolation of CTCs from clinically relevant blood volumes. The fast processing time of the technique (7.5 mL blood in less than 10 min) and the ability to collect more CTCs from larger blood volumes lends itself to a broad range of potential genomic and transcriptomic applications. A critical advantage of this protocol is the ability to return all fractions of blood (i.e., plasma (centrifugation), CTCs and white blood cells (WBCs) (size-based sorting)) that can be utilized for diverse biomarker studies or time-sensitive molecular assays such as RT-PCR. The clinical use of this biochip was demonstrated by detecting CTCs from 100% (10/10) of blood samples collected from patients with advanced-stage metastatic breast and lung cancers. The CTC recovery rate ranged from 20 to 135 CTCs mL(-1) and obtained under high purity (of 1 CTC out of every 30-100 WBCs which gives ∼4 log depletion of WBCs). They were identified with immunofluorescence assays (pan-cytokeratin+/CD45-) and molecular probes such as HER2/neu.

Solubility of Haloether Anesthetics in Human and Animal Blood

PubMed Central

Soares, Joao H. N.; Brosnan, Robert J.; Fukushima, Fabíola B.; Hodges, Joanne; Liu, Hong

2012-01-01

Background Anesthetic blood solubility predicts pharmacokinetics for inhaled agents and is essential for determination of blood anesthetic concentrations from end-tidal gas concentrations using Henry’s Law. Though used to model anesthetic effects in humans, there are limited interspecies solubility comparisons that include modern haloethers. This study aimed to measure hematocrit-adjusted blood:gas anesthetic partition coefficients (λB:G) for desflurane, sevoflurane, isoflurane, and methoxyflurane in humans and animals. Methods Whole blood was collected from 20 rats, 8 horses, and 4 each of cats, cattle, humans, dogs, goats, pigs, rabbits, and sheep. Plasma or cell volume was removed to adjust all samples to a packed cell volume of 40%. A single agent calibration gas headspace was added to blood in a glass syringe and was mixed and equilibrated at 37°C for 2 hours. Agent concentrations in the calibration gas and syringe headspace were measured using gas chromatography. Anesthetic solubility in saline, citrate-phosphate-dextrose-adenine, and olive oil were similarly measured. Results Except for goats, all animal species had at least one λB:G measurement that differed significantly from humans. For each agent, λB:G positively correlated with serum triglyceride concentrations, but this only explained 25% of interspecies variability. Desflurane was significantly less soluble in blood than sevoflurane in some species (e.g., humans) but not in others (e.g., rabbits). Conclusions Anesthetic partition coefficients differ significantly between humans and most animals for haloether anesthetics. Because of their similar λB:G values, goats may be a better animal model for inhaled anesthetic pharmacokinetics in people. PMID:22510863

Solubility of haloether anesthetics in human and animal blood.

PubMed

Soares, Joao H N; Brosnan, Robert J; Fukushima, Fabíola B; Hodges, Joanne; Liu, Hong

2012-07-01

Anesthetic blood solubility predicts pharmacokinetics for inhaled agents and is essential for determination of blood anesthetic concentrations from end-tidal gas concentrations using Henry's Law. Though used to model anesthetic effects in humans, there are limited interspecies solubility comparisons that include modern haloethers. This study aimed to measure hematocrit-adjusted blood:gas anesthetic partition coefficients (λ B:G) for desflurane, sevoflurane, isoflurane, and methoxyflurane in humans and animals. Whole blood was collected from 20 rats, 8 horses, and 4 each of cats, cattle, humans, dogs, goats, pigs, rabbits, and sheep. Plasma or cell volume was removed to adjust all samples to a packed cell volume of 40%. A single-agent calibration gas headspace was added to blood in a glass syringe and was mixed and equilibrated at 37°C for 2 h. Agent concentrations in the calibration gas and syringe headspace were measured using gas chromatography. Anesthetic solubility in saline, citrate-phosphate-dextrose-adenine, and olive oil were similarly measured. Except for goats, all animal species had at least one λ B:G measurement that differed significantly from humans. For each agent, λ B:G positively correlated with serum triglyceride concentrations, but this only explained 25% of interspecies variability. Desflurane was significantly less soluble in blood than sevoflurane in some species (e.g., humans) but not in others (e.g., rabbits). Anesthetic partition coefficients differ significantly between humans and most animals for haloether anesthetics. Because of their similar λ B:G values, goats may be a better animal model for inhaled anesthetic pharmacokinetics in people.

Development and performance test of an online blood sampling system for determination of the arterial input function in rats.

PubMed

Roehrbacher, Friedrich; Bankstahl, Jens P; Bankstahl, Marion; Wanek, Thomas; Stanek, Johann; Sauberer, Michael; Muellauer, Julia; Schroettner, Thales; Langer, Oliver; Kuntner, Claudia

2015-12-01

For positron emission tomography (PET) kinetic modelling, an accurate determination of the arterial input function is required. In this study, a blood sampling system was developed and tested using different radiotracers in rats. The detector consists of pairs of lutetium yttrium oxyorthosilicate (LYSO) detectors, photomultiplier tubes and lead shield assembled within a steel casing working in coincidence mode. Rats were cannulated with microtubes in the femoral artery and vein for arterial blood sampling as well as administration of the PET tracers. Connected PTFE microtubes were centred between the LYSO crystals using a special holder. To enhance sensitivity, three layers with two coils were used. A flexible tube pump was used to ensure a constant blood flow. Performance of the detector was assessed with [(18)F]fludeoxyglucose (FDG), [(18)F]ciprofloxacin, (R)-[(11)C]verapamil, [(11)C]tariquidar, [(11)C]mephobarbital and [(11)C]MC113. Obtained input function curves were compared with manual samples drawn every 5 s during the first 3 min and further on at 5, 10, 20, 30, 40, 50 and 60 min after radiotracer injection. After manual sampling, an arterio/venous shunt was established. Shape and area-under-the-curve (AUC; Bq/μl*h) of the input functions were evaluated. The developed detector system provided an absolute sensitivity of 6.5%. Maximum peak values agreed well between manual samples and the detector with a mean difference of -0.4% ± 7.0% (max 12.0%, min -9.9%). AUC values also exhibited an excellent correlation (R = 0.996) between manual sampling and detector measurements with a mean difference of 9.3% ± 9.7% (max 24.1%, min -3.2%). The system was able to measure peak blood activity concentration levels of 110 to 2,000 Bq/μl which corresponds to injected activities from 5.5 to 100 MBq depending on the used radiotracer, applied volume and weight of the animal. This study demonstrates that the developed blood sampling system can be used for in vivo small animal PET studies in rats in a reliable way. The usage of the systems enhances the accuracy of the input curve as handling of small blood samples especially with low activity (as for C-11) is prone to measurement errors. Additionally, the radiation dose of the experimenters can be reduced, as it is not required anymore to continuously draw samples where the personal is in close contact to the radioactive animals and blood.

Altered tryptophan catabolite concentrations in major depressive disorder and associated changes in hippocampal subfield volumes.

PubMed

Doolin, Kelly; Allers, Kelly A; Pleiner, Sina; Liesener, Andre; Farrell, Chloe; Tozzi, Leonardo; O'Hanlon, Erik; Roddy, Darren; Frodl, Thomas; Harkin, Andrew; O'Keane, Veronica

2018-05-19

Tryptophan depletion is a well-replicated biological finding in Major Depressive Disorder (MDD). The kynurenine pathway (KP) and its rate-limiting tryptophan degrading enzyme, indolamine 2,3 dioxygenase (IDO), have been implicated in the pathogenesis of depression. IDO expression is driven by inflammatory cytokines, providing a putative link between inflammation and neuropathology. This study examined circulating concentrations of C-reactive protein (CRP), plasma tryptophan, kynurenine (KYN), kynurenic acid (KYNA) and quinolinic acid (QUIN) and whole blood mRNA expression of IDO in patients with major depressive disorder (MDD) compared with healthy controls (HC). A diagnosis of major depression was made according to DSM-IV. Depression severity was assessed using the Hamilton depression (HAM-D) rating scale. 74 MDD patients, 39 with a first presentation of MDD (fpMDD) and 35 with chronic or recurrent episodes (rMDD), and 37 HC were recruited to the study. Whole blood and plasma samples were collected. Expression of markers in whole blood were measured by PCR, circulating CRP by ELISA and KP metabolites by LC-MS/MS. Hippocampal cornu ammonis (CA) and subiculum volumes were determined by MRI and calculated using FreeSurfer. Tryptophan concentrations were significantly reduced in MDD compared to HC. There was a positive correlation between QUIN and both CRP concentrations and whole blood IDO1 in MDD. KYNA concentrations were reduced in MDD patients presenting with a first episode (fpMDD) compared to those presenting with recurrent depression (rMDD) and HC. By contrast QUIN concentrations were elevated in rMDD compared to fpMDD and HC. KYNA/QUIN was reduced in MDD and rMDD but not fpMDD compared to HC. Hippocampal subfield volumes were smaller in MDD patients than HC for CA1 (left only), CA2/3 (left and right) and CA4 (right only). CRP and CA1 volumes were negatively correlated bilaterally in MDD patients. KYNA and subiculum volume were positively correlated bilaterally. This study found evidence of KP metabolism imbalance in MDD patients in addition to tryptophan reduction and mild immune activation. Relationships between CRP and KYNA with some hippocampal subfield volumes in MDD patients suggest that this inflammatory signature may be associated with reduced hippocampal subfield volumes in depression. Copyright © 2018 Elsevier Ltd. All rights reserved.

The Influence of Acute Handling Stress on Some Blood Parameters in Cultured Sea Bream (Sparus Aurata Linnaeus, 1758)

PubMed Central

Fazio, Francesco; Ferrantelli, Vincenzo; Fortino, Gianluca; Giangrosso, Giuseppe; Faggio, Caterina

2015-01-01

The effect of acute handling stress on haematological profile, blood glucose and lactate (secondary stress markers) in cultured sea bream Sparus aurata was evaluated. Sixty six Sparus aurata were used and equally divided into two groups (A and B). Group A was not subjected to stress, Group B was subjected to acute handling stress. From each fish, biometric data and blood samples were collected to evaluate haematological profile, blood glucose and lactate. Unpaired t-test Student was applied to evaluate possible differences in parameters between the two groups. Red blood cells, haematocrit, haemoglobin, white blood cells (WBC), glucose and lactate showed an increase (P<0.05) in Group B compared with Group A, while mean corpuscular volume decreased (P<0.05) in Group B. The results highlight the role of studied parameters in monitoring the stressful conditions of aquaculture production which affect animal welfare and fish products quality. PMID:27800375

The endocrine system in space flight

NASA Technical Reports Server (NTRS)

Leach, C. S.; Johnson, P. C.; Cintron, N. M.

1988-01-01

A trial natriuretic factor (ANF), a hormone recently shown to regulate sodium and water excretion, has been measured in blood specimens obtained during flight. After 30 or 42 h of weightlessness, mean ANF was elevated. After 175 or 180 h, ANF has increased by 59 percent, and it changed little between that time and soon after landing. There is probably an increase in ANF early inflight associated with the fluid shift, followed by a compensatory decrease in blood volume. Increased renal blood flow may cause the later ANF decrease. Erythropoietin (Ep), a hormone involved in the control of red blood cell proudction, was measured in blood samples taken during the first Spacelab mission and was significantly decreased on the second day of flight, suggesting also an increase in renal blood flow. Spacelab-2 investigators report that the active vitamin D metabolite 1 alpha, 25-dihydroxyvitamin D-3 increased early in the flight, indicating that a stimulus for increased bone resorption occurs by 30 h after launch.

Causes and consequences of reduced blood volume in space flight - A multi-discipline modeling study

NASA Technical Reports Server (NTRS)

Leonard, J. I.

1983-01-01

A group of mathematical models of various physiological systems have been developed and applied to studying problems associated with adaptation to weightlessness. One biomedical issue which could be addressed by at least three of these models from varying perspectives was the reduction in blood volume that universally occurs in astronauts. Accordingly, models of fluid-electrolyte, erythropoiesis, and cardiovascular regulation were employed to study the causes and consequences of blood volume loss during space flight. This analysis confirms the notion that alterations of blood volume are central to an understanding of adaptation to prolonged space flight. More importantly, the modeling studies resulted in specific hypotheses accounting for plasma volume and red cell mass losses and testable predictions concerning the behavior of the circulatory system.

Blood rheology effect of submaximal exercise on young subjects.

PubMed

Romagnoli, Marco; Alis, Rafael; Martinez-Bello, Vladimir; Sanchis-Gomar, Fabian; Aranda, Rafael; Gómez-Cabrera, Mari-Carmen

2014-01-01

Nowadays cardiac and metabolic diseases are a matter of concern. Exercise is a valid treatment and method of prevention for not only adults, but also young subjects. Physical activity causes transient blood rheology impairment in adults. However little is known about the effects of exercise on blood flow characteristics in young subjects. The aim of the study was to assess the effects of a light aerobic exercise session on blood rheology in young subjects. Ten young subjects (aged 12-16 years) performed 1 hour of submaximal aerobic exercise (70% HRmax). Blood samples were drawn just before and after exercise. We determined blood and plasma viscosity, fibrinogen, erythrocyte deformability and aggregability. No changes in blood viscosity (p > 0.05), erythrocyte aggregation (p > 0.05) and fibrinogen (p > 0.05) were observed. Hematocrit (p = 0.025) and plasma viscosity (p = 0.018) rose with exercise, while erythrocyte elongation index lowered (p < 0.001). Plasma volume slightly reduced which may explain the lack of changes in blood viscosity. The results of the present study indicate a similar hemorheological response to submaximal exercise in both young people and adults.

Combining MRI With PET for Partial Volume Correction Improves Image-Derived Input Functions in Mice

NASA Astrophysics Data System (ADS)

Evans, Eleanor; Buonincontri, Guido; Izquierdo, David; Methner, Carmen; Hawkes, Rob C.; Ansorge, Richard E.; Krieg, Thomas; Carpenter, T. Adrian; Sawiak, Stephen J.

2015-06-01

Accurate kinetic modelling using dynamic PET requires knowledge of the tracer concentration in plasma, known as the arterial input function (AIF). AIFs are usually determined by invasive blood sampling, but this is prohibitive in murine studies due to low total blood volumes. As a result of the low spatial resolution of PET, image-derived input functions (IDIFs) must be extracted from left ventricular blood pool (LVBP) ROIs of the mouse heart. This is challenging because of partial volume and spillover effects between the LVBP and myocardium, contaminating IDIFs with tissue signal. We have applied the geometric transfer matrix (GTM) method of partial volume correction (PVC) to 12 mice injected with 18F - FDG affected by a Myocardial Infarction (MI), of which 6 were treated with a drug which reduced infarction size [1]. We utilised high resolution MRI to assist in segmenting mouse hearts into 5 classes: LVBP, infarcted myocardium, healthy myocardium, lungs/body and background. The signal contribution from these 5 classes was convolved with the point spread function (PSF) of the Cambridge split magnet PET scanner and a non-linear fit was performed on the 5 measured signal components. The corrected IDIF was taken as the fitted LVBP component. It was found that the GTM PVC method could recover an IDIF with less contamination from spillover than an IDIF extracted from PET data alone. More realistic values of Ki were achieved using GTM IDIFs, which were shown to be significantly different (p <; 0.05) between the treated and untreated groups.

Assessment and Management of Volume Overload and Congestion in Chronic Heart Failure: Can Measuring Blood Volume Provide New Insights?

PubMed

Miller, Wayne L

2017-01-01

Volume overload and fluid congestion remain primary clinical challenges in the assessment and management of patients with chronic heart failure (HF). The pathophysiology of volume regulation is complex, and the simple concept of passive intravascular fluid accumulation is not adequate. The dynamics of interstitial and intravascular fluid compartment interactions and fluid redistribution from venous splanchnic beds to the central pulmonary circulation need to be taken into account in strategies of volume management. Clinical bedside evaluations and right heart hemodynamic assessments can alert of changes in volume status, but only the quantitative measurement of total blood volume can help identify the heterogeneity in plasma volume and red blood cell mass that are features of volume overload in chronic HF. The quantitative assessment of intravascular volume is an effective tool to help guide individualized, appropriate therapy. Not all volume overload is the same, and the measurement of intravascular volume identifies heterogeneity to guide tailored therapy.

The Role of Blood Osmolality and Volume in Regulating Vasopressin Secretion in the Rat

PubMed Central

Dunn, Fredrick L.; Brennan, Thomas J.; Nelson, Averial E.; Robertson, Gary L.

1973-01-01

A sensitive and specific radioimmunoassay for plasma arginine vasopressin (AVP) has been used to study the effects of blood osmolality and volume in regulating AVP secretion in unanesthetized rats. Under basal conditions, plasma AVP and osmolality were relatively constant, averaging 2.3±0.9 (SD) pg/ml and 294±1.4 mosmol/kg, respectively. Fluid restriction, which increased osmolality and decreased volume, resulted in a progressive rise in plasma AVP to about 10 times basal levels after 96 h. A 2-3-fold increase in plasma AVP occurred as early as 12 h, when osmolality and volume had each changed by less than 2%. Intraperitoneal injections of hypertonic saline, which had no effect on blood volume, also produced a rise in plasma AVP that was linearly correlated with the rise in osmolality (r > 0.9) and quantitatively similar to that found during fluid restriction (plasma AVP increased 2-4-fold with each 1% increase in osmolality). Intraperitoneal injection of polyethylene glycol, which decreased blood volume without altering osmolality, also increased plasma AVP but this response followed an exponential pattern and did not become significant until volume had decreased by 8% or more. At these levels of hypovolemia, the osmoregulatory system continued to function but showed a lower threshold and increase sensitivity to osmotic stimulation. We conclude that AVP secretion is regulated principally by blood osmolality but that the responsiveness of this mechanism may be significantly altered by modest changes in blood volume. PMID:4750450

21 CFR 864.6400 - Hematocrit measuring device.

Code of Federal Regulations, 2011 CFR

2011-04-01

..., racks, and a sealer and a holder. The device is used to measure the packed red cell volume in blood to determine whether the patient's total red cell volume is normal or abnormal. Abnormal states include anemia...). The packed red cell volume is produced by centrifuging a given volume of blood. (b) Classification...

21 CFR 864.6400 - Hematocrit measuring device.

Code of Federal Regulations, 2014 CFR

2014-04-01

..., racks, and a sealer and a holder. The device is used to measure the packed red cell volume in blood to determine whether the patient's total red cell volume is normal or abnormal. Abnormal states include anemia...). The packed red cell volume is produced by centrifuging a given volume of blood. (b) Classification...

21 CFR 864.6400 - Hematocrit measuring device.

Code of Federal Regulations, 2010 CFR

2010-04-01

..., racks, and a sealer and a holder. The device is used to measure the packed red cell volume in blood to determine whether the patient's total red cell volume is normal or abnormal. Abnormal states include anemia...). The packed red cell volume is produced by centrifuging a given volume of blood. (b) Classification...

21 CFR 864.6400 - Hematocrit measuring device.

Code of Federal Regulations, 2012 CFR

2012-04-01

..., racks, and a sealer and a holder. The device is used to measure the packed red cell volume in blood to determine whether the patient's total red cell volume is normal or abnormal. Abnormal states include anemia...). The packed red cell volume is produced by centrifuging a given volume of blood. (b) Classification...

21 CFR 864.6400 - Hematocrit measuring device.

Code of Federal Regulations, 2013 CFR

2013-04-01

..., racks, and a sealer and a holder. The device is used to measure the packed red cell volume in blood to determine whether the patient's total red cell volume is normal or abnormal. Abnormal states include anemia...). The packed red cell volume is produced by centrifuging a given volume of blood. (b) Classification...

Establishment of reference intervals for complete blood count parameters during normal pregnancy in Beijing.

PubMed

Li, Aiwei; Yang, Shuo; Zhang, Jie; Qiao, Rui

2017-11-01

To observe the changes of complete blood count (CBC) parameters during pregnancy and establish appropriate reference intervals for healthy pregnant women. Healthy pregnant women took the blood tests at all trimesters. All blood samples were processed on Sysmex XE-2100. The following CBC parameters were analyzed: red blood cell count (RBC), hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), white blood cell count (WBC), and leukocyte differential count. Reference intervals were established using the 2.5th and 97.5th percentile of the distribution. Complete blood count parameters showed dynamic changes during trimesters. RBC, Hb, Hct declined at trimester 1, reaching their lowest point at trimester 2, and began to rise again at trimester 3. WBC, neutrophil count (Neut), monocyte count (MONO), RDW, and PDW went up from trimester 1 to trimester 3. On the contrary, MCHC, lymphocyte count (LYMPH), PLT, and MPV gradually descended during pregnancy. There were statistical significances in all CBC parameters between pregnant women and normal women, regardless of the trimesters (P<.001). The median obtained were (normal vs pregnancy) as follows: RBC 4.50 vs 3.94×10 12 /L, Hb 137 vs 120 g/L, WBC 5.71 vs 9.06×10 9 /L, LYMPH% 32.2 vs 18.0, Neut% 58.7 vs 75.0, and PLT 251 vs 202×10 9 /L. The changes of CBC parameters during pregnancy are described, and reference intervals for Beijing pregnant women are demonstrated in this study. © 2017 Wiley Periodicals, Inc.

Analysis of benzodiazepines and their metabolites using DBS cards and LC-MS/MS.

PubMed

Lee, Heesang; Park, Yujin; Jo, Jiyeong; In, Sangwhan; Park, Yonghoon; Kim, Eunmi; Pyo, Jaesung; Choe, Sanggil

2015-10-01

Dried Blood Spot (DBS) has been used a blood extraction method for inherited metabolic disorder screening since 1960s. With introduction of LC-MS/MS, not only DBS could be used to analysis drugs in small blood volume, but in various fields, such as toxicology, drug therapeutic monitoring, drug diagnostic screening, and illicit drugs. In toxicology field, many drugs (e.g. benzodiazepines, acetaminophen, small molecule drugs) have been tested with DBS. Compared with earlier blood extraction methods (SPE and LLE), DBS has lots of advantages; lower blood volume (less than 50μL), shorter analysis time caused by a more concise analysis procedure and lower cost. We optimized the DBS procedure and LC-MS/MS conditions for 18 benzodiazepines, seven benzodiazepine metabolites, and one z-drug (zolpidem) analysis in blood. 30μL of whole blood was spotted on FTA DMPK card C and dried for 2h in a desiccator. A 6-mm disk was punched and vortexed for 1min in a centrifuge tube with 300μL methanol/acetonitrile mixture (1:1, v/v). After evaporation, redissolved in 100μL mobile phase of LC-MS/MS and 5μL was injected. In the analysis for 26 target compounds in blood, all of the method validation parameters - LLOD, LLOQ, accuracy (intra- and inter-assay), and precision (intra- and inter-assay) - were satisfied with method validation criteria, within 15%. The results of matrix effect, recovery, and process efficiency were good. We developed a fast and reliable sample preparation method using DBS for 26 benzodiazepines, benzodiazepine metabolites, and z-drug (zolpidem). Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

[State of the art in fluid and volume therapy : A user-friendly staged concept].

PubMed

Rehm, M; Hulde, N; Kammerer, T; Meidert, A S; Hofmann-Kiefer, K

2017-03-01

Adequate fluid therapy is highly important for the perioperative outcome of our patients. Both, hypovolemia and hypervolemia can lead to an increase in perioperative complications and can impair the outcome. Therefore, perioperative infusion therapy should be target-oriented. The main target is to maintain the patient's preoperative normovolemia by using a sophisticated, rational infusion strategy.Perioperative fluid losses should be discriminated from volume losses (surgical blood loss or interstitial volume losses containing protein). Fluid losses as urine or perspiratio insensibilis (0.5-1.0 ml/kg/h) should be replaced by balanced crystalloids in a ratio of 1:1. Volume therapy step 1: Blood loss up to a maximum value of 20% of the patient's blood volume should be replaced by balanced crystalloids in a ratio of 4(-5):1. Volume therapy step 2: Higher blood losses should be treated by using iso-oncotic, preferential balanced colloids in a ratio of 1:1. For this purpose hydroxyethyl starch can also be used perioperatively if there is no respective contraindication, such as sepsis, burn injuries, critically ill patients, renal impairment or renal replacement therapy, and severe coagulopathy. Volume therapy step 3: If there is an indication for red cell concentrates or coagulation factors, a differentiated application of blood and blood products should be performed.

Radiation-Induced Changes in Serum Lipidome of Head and Neck Cancer Patients

PubMed Central

Jelonek, Karol; Pietrowska, Monika; Ros, Malgorzata; Zagdanski, Adam; Suchwalko, Agnieszka; Polanska, Joanna; Marczyk, Michal; Rutkowski, Tomasz; Skladowski, Krzysztof; Clench, Malcolm R.; Widlak, Piotr

2014-01-01

Cancer radiotherapy (RT) induces response of the whole patient’s body that could be detected at the blood level. We aimed to identify changes induced in serum lipidome during RT and characterize their association with doses and volumes of irradiated tissue. Sixty-six patients treated with conformal RT because of head and neck cancer were enrolled in the study. Blood samples were collected before, during and about one month after the end of RT. Lipid extracts were analyzed using MALDI-oa-ToF mass spectrometry in positive ionization mode. The major changes were observed when pre-treatment and within-treatment samples were compared. Levels of several identified phosphatidylcholines, including (PC34), (PC36) and (PC38) variants, and lysophosphatidylcholines, including (LPC16) and (LPC18) variants, were first significantly decreased and then increased in post-treatment samples. Intensities of changes were correlated with doses of radiation received by patients. Of note, such correlations were more frequent when low-to-medium doses of radiation delivered during conformal RT to large volumes of normal tissues were analyzed. Additionally, some radiation-induced changes in serum lipidome were associated with toxicity of the treatment. Obtained results indicated the involvement of choline-related signaling and potential biological importance of exposure to clinically low/medium doses of radiation in patient’s body response to radiation. PMID:24747595

Erythrocytes as Volume Markers in Experimental PD Show that Albumin Transport in the Extracellular Space Depends on PD Fluid Osmolarity

PubMed Central

Braide, Magnus; Delbro, Dick; Waniewski, Jacek

2016-01-01

♦ Background: Macromolecules, when used as intraperitoneal volume markers, have the disadvantage of leaking into the surrounding tissue. Therefore, 51Cr-labeled erythrocytes were evaluated as markers of intraperitoneal volume and used in combination with 125I-labeled bovine serum albumin to study albumin transport into peritoneal tissues in a rat model of peritoneal dialysis (PD). ♦ Methods: Single dwells of 20 mL of lactate-buffered filter-sterilized PD fluid at glucose concentrations of 0.5%, 2.5%, and 3.9% were performed for 1 or 4 hours. Tissue biopsies from abdominal muscle, diaphragm, liver, and intestine, and blood and dialysate samples, were analyzed for radioactivity. ♦ Results: The dialysate distribution volume of labeled erythrocytes, measured after correction for lymphatic clearance to blood, was strongly correlated with, but constantly 3.3 mL larger than, drained volumes. Erythrocyte activity of rinsed peritoneal tissue biopsies corresponded to only 1 mL of dialysate, supporting our utilization of erythrocytes as markers of intraperitoneal volume. The difference between the distribution volumes of albumin and erythrocytes was analyzed to represent the albumin loss into the peritoneal tissues, which increased rapidly during the first few minutes of the dwell and then leveled out at 2.5 mL. It resumed when osmotic ultrafiltration turned into reabsorption and, at the end of the dwell, it was significantly lower for the highest osmolarity PD fluid (3.9% glucose). Biopsy data showed the lowest albumin accumulation and edema formation in abdominal muscle for the 3.9% fluid. ♦ Conclusion: Labeled erythrocytes are acceptable markers of intraperitoneal volume and, combined with labeled albumin, provided novel kinetic data on albumin transport in peritoneal tissues. PMID:26493751

Audit of clinical-laboratory practices in haematology and blood transfusion at Muhimbili National Hospital in Tanzania.

PubMed

Makubi, Abel N; Meda, Collins; Magesa, Alex; Minja, Peter; Mlalasi, Juliana; Salum, Zubeda; Kweka, Rumisha E; Rwehabura, James; Quaresh, Amrana; Magesa, Pius M; Robert, David; Makani, Julie; Kaaya, Ephata

2012-10-01

In Tanzania, there is paucity of data for monitoring laboratory medicine including haematology. This therefore calls for audits of practices in haematology and blood transfusion in order to provide appraise practice and devise strategies that would result in improved quality of health care services. This descriptive cross-sectional study which audited laboratory practice in haematology and blood transfusion at Muhimbili National Hospital (MNH) aimed at assessing the pre-analytical stage of laboratory investigations including laboratory request forms and handling specimen processing in the haematology laboratory and assessing the chain from donor selection, blood component processing to administration of blood during transfusion. A national standard checklist was used to audit the laboratory request forms (LRF), phlebotomists' practices on handling and assessing the from donor selection to administration 6f blood during transfusion. Both interview and observations were used. A total of 195 LRF were audited and 100% of had incomplete information such as patients' identification numbers, time sample ordered, reason for request, summary of clinical assessment and differential diagnoses. The labelling of specimens was poorly done by phlebotomists/clinicians in 82% of the specimens. Also 65% (132/202) of the blood samples delivered in the haematology laboratory did not contain the recommended volume of blood. There was no laboratory request form specific for ordering blood and there were no guidelines for indication of blood transfusion in the wards/ clinics. The blood transfusion laboratory section was not participating in external quality assessment and the hospital transfusion committee was not in operation. It is recommended that a referral hospital like MNH should have a transfusion committee to provide an active forum to facilitate communication between those involved with transfusion, monitor, coordinate and audit blood transfusion practices as per national guidelines.

Determination of cyanide in whole blood by capillary gas chromatography with cryogenic oven trapping.

PubMed

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

1998-11-15

Cyanide, one of the most important toxic substances, has been found measurable with high sensitivity by capillary gas chromatography (GC) with cryogenic oven trapping upon injection of headspace (HS) vapor samples. The entire amount of cyanide in the HS sample could be cryogenically trapped prior to on-line GC analysis. A 0.5-mL volume of blood in the presence or absence of cyanide and propionitrile (internal standard, IS) was added to a vial containing 0.25 mL of distilled water, 0.3 g of Na2-SO4, 0.2 mL of 50% H3PO4, and 0.1 g of ascorbic acid (when needed), and the mixture was heated at 70 degrees C for 15 min. A 5-mL volume of the HS vapor was introduced into a GC capillary column in the splitless mode at -30 degrees C oven temperature that was programmed up to 160 degrees C for GC analysis with nitrogen-phosphorus detection. A sharp peak was obtained for cyanide under the present conditions, and backgrounds were very clean. The extraction efficiencies of cyanide and IS were 2.89-3.22 (100 or 500 ng/mL) and 2.42%, respectively. The calibration curve showed good linearity in the range of 25-1000 ng/mL and the detection limit was approximately 2 ng/mL. The coefficients of intraday and interday variations were 2.9 and 11.8%, respectively. The mean blood cyanide level measured for actual fire victims was 687 +/- 597 ng/mL (mean +/- SD, n = 9). Endogenous blood cyanide concentration for healthy subjects was 8.41 +/- 3.09 ng/mL (mean +/- SD, n = 6).

A dye-assisted paper-based point-of-care assay for fast and reliable blood grouping.

PubMed

Zhang, Hong; Qiu, Xiaopei; Zou, Yurui; Ye, Yanyao; Qi, Chao; Zou, Lingyun; Yang, Xiang; Yang, Ke; Zhu, Yuanfeng; Yang, Yongjun; Zhou, Yang; Luo, Yang

2017-03-15

Fast and simultaneous forward and reverse blood grouping has long remained elusive. Forward blood grouping detects antigens on red blood cells, whereas reverse grouping identifies specific antibodies present in plasma. We developed a paper-based assay using immobilized antibodies and bromocresol green dye for rapid and reliable blood grouping, where dye-assisted color changes corresponding to distinct blood components provide a visual readout. ABO antigens and five major Rhesus antigens could be detected within 30 s, and simultaneous forward and reverse ABO blood grouping using small volumes (100 μl) of whole blood was achieved within 2 min through on-chip plasma separation without centrifugation. A machine-learning method was developed to classify the spectral plots corresponding to dye-based color changes, which enabled reproducible automatic grouping. Using optimized operating parameters, the dye-assisted paper assay exhibited comparable accuracy and reproducibility to the classical gel-card assays in grouping 3550 human blood samples. When translated to the assembly line and low-cost manufacturing, the proposed approach may be developed into a cost-effective and robust universal blood-grouping platform. Copyright © 2017, American Association for the Advancement of Science.

The effect of vagal afferent on total vascular compliance in rats.

PubMed

Kinoshita, T

1993-04-01

This study was designed to investigate the effect of vagal afferent stimulation on total vascular compliance (TVC). Rats were anesthetized with sodium pentobarbital and artificially ventilated, TVC was determined together with stressed and unstressed blood volumes by measuring mean circulatory filling pressure (Pmcf) at three different levels of circulating blood volume. Measurements was repeated with the intact vagus, after vagotomy and during stimulation of vagal afferents. Vagotomy caused no change in TVC, Pmcf, and stressed and unstressed blood volumes. On the other hand, electrical stimulation of the vagal afferents for 30 sec increased TVC from 3.03 +/- 0.51 to 3.39 +/- 0.44 ml.mmHg(-1).kg(-1) (P < 0.05) and decreased Pmcf from 7.83 +/- 1.40 to 7.22 +/- 1.21 mmHg (P < 0.05). Neither stressed nor unstressed blood volume was changed by vagal stimulation. These results indicate that excitation of vagal afferent causes venodilation and increases TVC without changing stressed and unstressed blood volumes.

Simplifying [18F]GE-179 PET: are both arterial blood sampling and 90-min acquisitions essential?

PubMed

McGinnity, Colm J; Riaño Barros, Daniela A; Trigg, William; Brooks, David J; Hinz, Rainer; Duncan, John S; Koepp, Matthias J; Hammers, Alexander

2018-06-11

The NMDA receptor radiotracer [ 18 F]GE-179 has been used with 90-min scans and arterial plasma input functions. We explored whether (1) arterial blood sampling is avoidable and (2) shorter scans are feasible. For 20 existing [ 18 F]GE-179 datasets, we generated (1) standardised uptake values (SUVs) over eight intervals; (2) volume of distribution (V T ) images using population-based input functions (PBIFs), scaled using one parent plasma sample; and (3) V T images using three shortened datasets, using the original parent plasma input functions (ppIFs). Correlations with the original ppIF-derived 90-min V T s increased for later interval SUVs (maximal ρ = 0.78; 80-90 min). They were strong for PBIF-derived V T s (ρ = 0.90), but between-subject coefficient of variation increased. Correlations were very strong for the 60/70/80-min original ppIF-derived V T s (ρ = 0.97-1.00), which suffered regionally variant negative bias. Where arterial blood sampling is available, reduction of scan duration to 60 min is feasible, but with negative bias. The performance of SUVs was more consistent across participants than PBIF-derived V T s.

Condition of cardiovascular systems of astronauts during flight of Soyuz orbital station

NASA Technical Reports Server (NTRS)

Degtyarev, V. A.; Popov, I. I.; Batenchuk-Tusko, T. V.; Kolmykova, N. D.; Lapshina, N. A.; Kirillova, Z. A.; Doroshev, V. G.; Kukushkin, Y. A.

1975-01-01

Extensive studies of blood circulation functions during manned space flight demonstrated a pronounced tendency toward an increase in minute volume of the blood and a decrease in pulse wave propagation rate. Individual blood circulation indices had large amplitude fluctuations. Physical work loads caused slow recovery of heart rate, arterial pressure and minute blood volume.

Physiologic mechanisms of circulatory and body fluid losses in weightlessness identified by mathematical modeling

NASA Technical Reports Server (NTRS)

Simanonok, K. E.; Srinivasan, R. S.; Charles, J. B.

1993-01-01

Central volume expansion due to fluid shifts in weightlessness is believed to activate adaptive reflexes which ultimately result in a reduction of the total circulating blood volume. However, the flight data suggests that a central volume overdistention does not persist, in which case some other factor or factors must be responsible for body fluid losses. We used a computer simulation to test the hypothesis that factors other than central volume overdistention are involved in the loss of blood volume and other body fluid volumes observed in weightlessness and in weightless simulations. Additionally, the simulation was used to identify these factors. The results predict that atrial volumes and pressures return to their prebedrest baseline values within the first day of exposure to head down tilt (HDT) as the blood volume is reduced by an elevated urine formation. They indicate that the mechanisms for large and prolonged body fluid losses in weightlessness is red cell hemoconcentration that elevates blood viscosity and peripheral resistance, thereby lowering capillary pressure. This causes a prolonged alteration of the balance of Starling forces, depressing the extracellular fluid volume until the hematocrit is returned to normal through a reduction of the red cell mass, which also allows some restoration of the plasma volume. We conclude that the red cell mass becomes the physiologic driver for a large 'undershoot' of the body fluid volumes after the normalization of atrial volumes and pressures.

Mobile phone exposure influences some erythrocytes parameters in vitro. A novel source of preanalytical variability?

PubMed

Danese, Elisa; Lippi, Giuseppe; Brocco, Giorgio; Montagnana, Martina; Salvagno, Gian Luca

2016-06-01

The effect of radiofrequency exposure on human health and health care equipment is a matter of ongoing debate. This study was planned to investigate the influence of radiofrequency (RF) waves emitted by a commercial mobile phone on red blood cells (RBC) in vitro. The study population consisted of 16 ostensibly healthy volunteers. Two whole blood specimens were collected from each volunteer. One sample was placed in a plastic rack, 1 cm distant from the chassis of a commercial mobile phone which was activated by a remote phone call lasting 30 min. The other blood sample was placed in another plastic rack, but was kept distant from any type of RF source. The main RBC parameters including RBC count, hematocrit (Ht), hemoglobin, mean corpuscular platelet volume (MPV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and RBC distribution width (RDW-CV) were assessed with an Advia 2120. The exposure of whole blood to the mobile phone call significantly increased Ht, hemoglobin, MCV and MCH, whereas the RBC count, MCHC and RDW-CV remained unchanged. A significant correlation was observed between variation of Ht and those of hemoglobin (p=0.008), MCV (p=0.009) or MCH (p=0.037), as well as between hemoglobin and MCV (p=0.048). Increased values were found in 13/16 (81%) samples for both Ht and hemoglobin, 14/16 (88%) samples for MCH and 16/16 (100%) samples for MCV. These results suggest that close mobile phone exposure may be an unappreciated and possibly underestimated cause of preanalytical bias in RBC testing.

Quantitative determination of n-propane, iso-butane, and n-butane by headspace GC-MS in intoxications by inhalation of lighter fluid.

PubMed

Bouche, Marie-Paule L A; Lambert, Willy E; Van Bocxlaer, Jan F P; Piette, Michel H; De Leenheer, André P

2002-01-01

This report describes a fully elaborated and validated method for quantitation of the hydrocarbons n-propane, iso-butane, and n-butane in blood samples. The newly developed analytical procedure is suitable for both emergency cases and forensic medicine investigations. Its practical applicability is illustrated with a forensic blood sample after acute inhalative intoxication with lighter fluid; case history and toxicological findings are included. Identification and quantitation of the analytes were performed using static headspace extraction combined with gas chromatography-mass spectrometry. In order to reconcile the large gas volumes injected (0.5 mL) with the narrowbore capillary column and thus achieve preconcentration, cold trapping on a Tenax sorbent followed by flash desorption was applied. Adequate retention and separation were achieved isothermally at 35 degrees C on a thick-film capillary column. Sample preparation was kept to a strict minimum and involved simply adding 2.5 microL of a liquid solution of 1,1,2-trichlorotrifluoroethane in t-butyl-methylether as an internal standard to aliquots of blood in a capped vial. Standards were created by volumetric dilution departing from a gravimetrically prepared calibration gas mixture composed of 0.3% of n-propane, 0.7% of iso-butane, and 0.8% of n-butane in nitrogen. In the forensic blood sample, the following concentrations were measured: 90.0 microg/L for n-propane, 246 microg/L for iso-butane, and 846 microg/L for n-butane.

Anesthesia and blood sampling of wild big brown bats (Eptesicus fuscus) with an assessment of impacts on survival

USGS Publications Warehouse

Wimsatt, J.; O'Shea, T.J.; Ellison, L.E.; Pearce, R.D.; Price, V.R.

2005-01-01

We anesthetized and blood sampled wild big brown bats (Eptesicus fuscus) in Fort Collins, Colorado (USA) in 2001 and 2002 and assessed effects on survival. Inhalant anesthesia was delivered into a specially designed restraint and inhalation capsule that minimized handling and bite exposures. Bats were immobilized an average of 9.1±5.1 (SD) min (range 1–71, n=876); blood sample volumes averaged 58±12 μl (range 13–126, n=718). We randomly selected control (subject to multiple procedures before release) and treatment (control procedures plus inhalant anesthesia and 1% of body weight blood sampling) groups in 2002 to assess treatment effects on daily survival over a 14-day period for adult female and volant juvenile bats captured at maternity roosts in buildings. We monitored survival after release using passive integrated transponder tag detection hoops placed at openings to selected roosts. Annual return rates of bats sampled in 2001 were used to assess long-term outcomes. Comparison of 14-day maximum-likelihood daily survival estimates from control (86 adult females, 92 volant juveniles) and treated bats (187 adult females, 87 volant juveniles) indicated no adverse effect from anesthesia and blood sampling (juveniles: χ2=22.22, df=27, P>0.05; adults: χ2=9.72, df=18, P>0.05). One-year return rates were similar among adult female controls (81%, n=72, 95% confidence interval [CI] =70–91%), females treated once (82%, n=276, 95% CI=81–84%), and females treated twice (84%, n=50, 95% CI=74–94%). Lack of an effect was also noted in 1-yr return rates of juvenile female controls (55%, n=29, 95% CI=37–73%), juveniles treated once (66%, n=113, 95% CI=58–75%), and juveniles treated twice (71%, n=17, 95% CI=49–92%). These data suggest that anesthesia and blood sampling for health monitoring did not measurably affect survival of adult female and volant juvenile big brown bats.

The accuracy of noninvasive hemoglobin monitoring using the radical-7 pulse CO-Oximeter in children undergoing neurosurgery.

PubMed

Park, Yong-Hee; Lee, Ji-Hyun; Song, Hyun-Gul; Byon, Hyo-Jin; Kim, Hee-Soo; Kim, Jin-Tae

2012-12-01

The most common method for determining the hemoglobin concentration is to draw blood from a patient. However, the Radical-7 Pulse CO-Oximeter (Masimo Corporation, Irvine, CA) can noninvasively provide continuous hemoglobin concentration (SpHb). In our study we compared noninvasive measurements of SpHb with simultaneous laboratory measurements of total hemoglobin in arterial blood samples taken from children (tHb). Arterial blood samples were analyzed using a laboratory CO-oximeter, and SpHb was simultaneously recorded in pediatric patients undergoing neurosurgery. When patients met the criteria for hypovolemia, 10 mL/kg of colloids or red blood cells were administered over 10 minutes. SpHb and tHb data were collected before and after intravascular volume resuscitation. The relationship between SpHb and tHb was assessed using a 4-quadrant plot, linear regression, mixed-effect model, and modified Bland-Altman analyses. One hundred nineteen paired samples were analyzed. The correlation coefficient between SpHb and tHb was 0.53 (P < 0.001), whereas that of change in SpHb versus change in tHb was 0.75 (P < 0.001). The average difference (bias) between tHb and SpHb was 0.90 g/dL (95% confidence interval [CI], 0.48-1.32 g/dL) and 1 standard deviation of the difference (sd) was 1.35 g/dL. The concordance rate (a measure of the number of data points that are in 1 of the 2 quadrants of agreement) determined using a 4-quadrant plot was 93%. The correlation coefficient between SpHb and tHb after intravascular volume resuscitation was 0.58 (P < 0.001), whereas that of changes in SpHb and tHb was 0.87 (P < 0.001). The bias immediately after volume resuscitation was 1.18 g/dL (95% CI, 0.81-1.55 g/dL), and sd was 1.28 g/dL with a concordance rate of 94.4%. The bias was -0.03 g/dL when tHb was ≥11 g/dL, which was significantly lower in comparison with biases when tHb <9 g/dL (1.24 g/dL) and tHb was 9-11 g/dL (1.17 g/dL) (P = 0.004). The Radical-7 Pulse CO-Oximeter can be useful as a trend monitor in children during surgery even immediately after intravascular volume expanders are administered. However, it is advisable to confirm the baseline hemoglobin level and to consider the influence of tHb level on the bias. In addition, one should be cautious with regard to using SpHb alone when making transfusion decisions.

Mineral elements and essential trace elements in blood of seals of the North Sea measured by total-reflection X-ray fluorescence analysis

NASA Astrophysics Data System (ADS)

Griesel, S.; Mundry, R.; Kakuschke, A.; Fonfara, S.; Siebert, U.; Prange, A.

2006-11-01

Mineral and essential trace elements are involved in numerous physiological processes in mammals. Often, diseases are associated with an imbalance of the electrolyte homeostasis. In this study, the concentrations of mineral elements (P, S, K, Ca) and essential trace elements (Fe, Cu, Zn, Se, Rb, Sr) in whole blood of harbor seals ( Phoca vitulina) were determined using total-reflection X-ray fluorescence spectrometry (TXRF). Samples from 81 free-ranging harbor seals from the North Sea and two captive seals were collected during 2003-2005. Reference ranges and element correlations for health status determination were derived for P, S, K, Ca, Fe, Cu, and Zn level in whole blood. Grouping the seals by age, gender and sample location the concentration levels of the elements were compared. The blood from two captive seals with signs of diseases and four free-ranging seals showed reduced element levels of P, S, and Ca and differences in element correlation of electrolytes were ascertained. Thus, simultaneous measurements of several elements in only 500 μL volumes of whole blood provide the possibility to obtain information on both, the electrolyte balance and the hydration status of the seals. The method could therefore serve as an additional biomonitoring tool for the health assessment.

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.

A machine for haemodialysing very small infants.

PubMed

Everdell, Nicholas L; Coulthard, Malcolm G; Crosier, Jean; Keir, Michael J

2005-05-01

Babies weighing under 6 kg are difficult to dialyse, especially those as small as 1 kg. Peritoneal dialysis is easier than haemodialysis, but is not always possible, and clears molecules less efficiently. Two factors complicate haemodialysis. First, extracorporeal circuits are large relative to a baby's blood volume, necessitating priming with fresh or modified blood. Second, blood flow from infants' access vessels is disproportionately low (Poiseuille's law), causing inadequate dialysis, or clotting within the circuit. These problems are minimised by using single lumen access, a very small circuit, and a reservoir syringe to separate the sampling and dialyser blood flow rates. Its manual operation is tedious, so we developed a computer-controlled, pressure-monitored machine to run it, including adjusting the blood withdrawal rate from poorly sampling lines. We have dialysed four babies weighing 0.8-3.4 kg, with renal failure or metabolic disorders. The circuits did not require priming. Clearances of creatinine, urea, potassium, phosphate and ammonia were mean (SD) 0.54 (0.22) ml/min using one dialyser, and 0.98 (0.22) ml/min using two in parallel. Ammonia clearance in a 2.4 kg baby had a 9 h half-life. Ultrafiltration up to 45 ml/h was achieved easily. This device provided infants with immediate, effective and convenient haemodialysis, typically delivered for prolonged periods.

Closed-Loop Control of Fluid Therapy for Treatment of Hypovolemia

DTIC Science & Technology

2008-04-01

index) (Table 2). Invasive bladder catheterization and blood sampling add the use of urinary output, lactate, central venous oxygen saturation, he...for guiding burn resuscitation Time delay, renal failure, and drugs complicate interpretation Central venous oxygenation ScvO2 Global perfusion index... Central venous pressure (CVP) Sensitive to volume change Pre-injury baseline values vary Arterial pressure by transducer Real-time continuous

Human Finger-Prick Induced Pluripotent Stem Cells Facilitate the Development of Stem Cell Banking

PubMed Central

Tan, Hong-Kee; Toh, Cheng-Xu Delon; Ma, Dongrui; Yang, Binxia; Liu, Tong Ming; Lu, Jun; Wong, Chee-Wai; Tan, Tze-Kai; Li, Hu; Syn, Christopher; Tan, Eng-Lee; Lim, Bing; Lim, Yoon-Pin; Cook, Stuart A.

2014-01-01

Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients can be a good model for studying human diseases and for future therapeutic regenerative medicine. Current initiatives to establish human iPSC (hiPSC) banking face challenges in recruiting large numbers of donors with diverse diseased, genetic, and phenotypic representations. In this study, we describe the efficient derivation of transgene-free hiPSCs from human finger-prick blood. Finger-prick sample collection can be performed on a “do-it-yourself” basis by donors and sent to the hiPSC facility for reprogramming. We show that single-drop volumes of finger-prick samples are sufficient for performing cellular reprogramming, DNA sequencing, and blood serotyping in parallel. Our novel strategy has the potential to facilitate the development of large-scale hiPSC banking worldwide. PMID:24646489

Comparison of Blood Loss in Laser Lipolysis vs Traditional Liposuction.

PubMed

Abdelaal, Mohammed Mahmoud; Aboelatta, Yasser Abdallah

2014-08-01

Laser-assisted liposuction has been associated with reduced blood loss. However, this clinical finding has not been evaluated objectively. In this study, the authors objectively estimated the blood loss volume associated with laser lipolysis vs traditional liposuction in various anatomic regions. In this prospective study, 56 patients underwent equal amounts of traditional and laser-assisted liposuction at 2 contralateral anatomic sites. Blood loss volumes were calculated from the lipoaspirates by measuring hemoglobin and red blood cell content. The data were analyzed statistically with repeated-measures analysis of variance and the Mann-Whitney U test. Laser lipolysis can reduce blood loss by more than 50% compared with traditional liposuction. Laser lipolysis resulted in significant reductions in mean blood loss volumes in the abdomen, flanks, back, and breast. The authors provide objective evidence that laser lipolysis significantly reduces blood loss compared with traditional liposuction. 3. © 2014 The American Society for Aesthetic Plastic Surgery, Inc.

Tranexamic Acid Reduced the Percent of Total Blood Volume Lost During Adolescent Idiopathic Scoliosis Surgery.

PubMed

Jones, Kristen E; Butler, Elissa K; Barrack, Tara; Ledonio, Charles T; Forte, Mary L; Cohn, Claudia S; Polly, David W

2017-01-01

Multilevel posterior spine fusion is associated with significant intraoperative blood loss. Tranexamic acid is an antifibrinolytic agent that reduces intraoperative blood loss. The goal of this study was to compare the percent of total blood volume lost during posterior spinal fusion (PSF) with or without tranexamic acid in patients with adolescent idiopathic scoliosis (AIS). Thirty-six AIS patients underwent PSF in 2011-2014; the last half (n=18) received intraoperative tranexamic acid. We retrieved relevant demographic, hematologic, intraoperative and outcomes information from medical records. The primary outcome was the percent of total blood volume lost, calculated from estimates of intraoperative blood loss (numerator) and estimated total blood volume per patient (denominator, via Nadler's equations). Unadjusted outcomes were compared using standard statistical tests. Tranexamic acid and no-tranexamic acid groups were similar (all p>0.05) in mean age (16.1 vs. 15.2 years), sex (89% vs. 83% female), body mass index (22.2 vs. 20.2 kg/m2), preoperative hemoglobin (13.9 vs. 13.9 g/dl), mean spinal levels fused (10.5 vs. 9.6), osteotomies (1.6 vs. 0.9) and operative duration (6.1 hours, both). The percent of total blood volume lost (TBVL) was significantly lower in the tranexamic acid-treated vs. no-tranexamic acid group (median 8.23% vs. 14.30%, p = 0.032); percent TBVL per level fused was significantly lower with tranexamic acid than without it (1.1% vs. 1.8%, p=0.048). Estimated blood loss (milliliters) was similar across groups. Tranexamic acid significantly reduced the percentage of total blood volume lost versus no tranexamic acid in AIS patients who underwent PSF using a standardized blood loss measure.Level of Evidence: 3. Institutional Review Board status: This medical record chart review (minimal risk) study was approved by the University of Minnesota Institutional Review Board.

Clinical utility of an optimised multiplex real-time PCR assay for the identification of pathogens causing sepsis in Vietnamese patients.

PubMed

Tat Trung, Ngo; Van Tong, Hoang; Lien, Tran Thi; Van Son, Trinh; Thanh Huyen, Tran Thi; Quyen, Dao Thanh; Hoan, Phan Quoc; Meyer, Christian G; Song, Le Huu

2018-02-01

For the identification of bacterial pathogens, blood culture is still the gold standard diagnostic method. However, several disadvantages apply to blood cultures, such as time and rather large volumes of blood sample required. We have previously established an optimised multiplex real-time PCR method in order to diagnose bloodstream infections. In the present study, we evaluated the diagnostic performance of this optimised multiplex RT-PCR in blood samples collected from 110 septicaemia patients enrolled at the 108 Military Central Hospital, Hanoi, Vietnam. Positive results were obtained by blood culture, the Light Cylcler-based SeptiFast ® assay and our multiplex RT-PCR in 35 (32%), 31 (28%), and 31 (28%) samples, respectively. Combined use of the three methods confirmed 50 (45.5%) positive cases of bloodstream infection, a rate significantly higher compared to the exclusive use of one of the three methods (P=0.052, 0.012 and 0.012, respectively). The sensitivity, specificity and area under the curve (AUC) of our assay were higher compared to that of the SeptiFast ® assay (77.4%, 86.1% and 0.8 vs. 67.7%, 82.3% and 0.73, respectively). Combined use of blood culture and multiplex RT-PCR assay showed a superior diagnostic performance, as the sensitivity, specificity, and AUC reached 83.3%, 100%, and 0.95, respectively. The concordance between blood culture and the multiplex RT-PCR assay was highest for Klebsiella pneumonia (100%), followed by Streptococcus spp. (77.8%), Escherichia coli (66.7%), Staphylococcus spp. (50%) and Salmonella spp. (50%). In addition, the use of the newly established multiplex RT-PCR assay increased the spectrum of identifiable agents (Acintobacter baumannii, 1/32; Proteus mirabilis, 1/32). The combination of culture and the multiplex RT-PCR assay provided an excellent diagnostic accomplishment and significantly supported the identification of causative pathogens in clinical samples obtained from septic patients. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Erythrocyte volume, folate levels, and the presence of methylenetetrahydrofolate reductase polymorphism.

PubMed

García-García, Inés; García-Fragoso, Lourdes; Renta, Jessicca; Arce, Sylvia; Cadilla, Carmen L

2002-03-01

Homozygosity for a common polymorphism in the 5,10 methylenetetrahydrofolate reductase (MTHFR) gene (C677T) has been associated to an increased risk of neural tube defects as well as derangements in folate, homocysteine, and hematological parameters. This study analyzed the relationship between folate levels, the erythrocyte volume, and the presence of homozygosity for the C677T polymorphism in a group of 126 Puerto Rican healthy women of childbearing age. Blood samples were analyzed for erythrocyte mean corpuscular volume (MCV), mean erythrocyte hemoglobin content (MCH), folate, and RBC folate. Homozygosity for the C677T mutation was determined by PCR. Thirty-two percent (32%) of women used a folic acid supplement during the three months prior to sampling. Mean folate and RBC folate levels were within the normal range. Individuals homozygous for the MTHFR C677T polymorphism had no elevation of MCV (p = 0.70) or MCH (p = 0.68). Women in the lower quartile of folate levels did not show differences in their MCV or MCH. In this sample of Puerto Rican women, homozygosity for the C677T MTHFR polymorphism was not associated to elevations of MCV or MCH even in the presence of lower folate levels.

Multicenter evaluation of the Sepsityper™ extraction kit and MALDI-TOF MS for direct identification of positive blood culture isolates using the BD BACTEC™ FX and VersaTREK(®) diagnostic blood culture systems.

PubMed

Schieffer, K M; Tan, K E; Stamper, P D; Somogyi, A; Andrea, S B; Wakefield, T; Romagnoli, M; Chapin, K C; Wolk, D M; Carroll, K C

2014-04-01

(i) Evaluation of delayed time to blood culture extraction by the Sepsityper kit and impact of shipping pellets off-site for MALDI-TOF MS analysis. (ii) Comparison of Sepsityper and laboratory-developed extraction methods from a literature review. Using two blood culture systems (BD BACTEC and VersaTREK), we extracted 411 positive blood cultures using the Sepsityper kit to mimic a potential protocol for institutions without a MALDI-TOF MS. Extracted pellets were shipped and analysed on the Bruker UltraflexIII. Successful extraction of 358 (87·1%) samples was determined by the presence of detectable proteins. MALDI-TOF MS correctly identified 332 (80·8%) samples. Delayed time to extraction did not affect Sepsityper extraction or MALDI-TOF MS accuracy. The extracted pellets remain stable and provide accurate results by MALDI-TOF MS when shipped at room temperature to off-site reference laboratories. This is the first study to show that institutions without a MALDI-TOF MS can take advantage of this innovative technology by shipping a volume of blood to an off-site laboratory for extraction and MALDI-TOF MS analysis. We also performed a literature review to compare various extraction methods. © 2014 The Society for Applied Microbiology.

Creatinine generation from kinetic modeling with or without postdialysis serum creatinine measurement: results from the HEMO study.

PubMed

Daugirdas, John T; Depner, Thomas A

2017-11-01

A convenient method to estimate the creatinine generation rate and measures of creatinine clearance in hemodialysis patients using formal kinetic modeling and standard pre- and postdialysis blood samples has not been described. We used data from 366 dialysis sessions characterized during follow-up month 4 of the HEMO study, during which cross-dialyzer clearances for both urea and creatinine were available. Blood samples taken at 1 h into dialysis and 30 min and 60 min after dialysis were used to determine how well a two-pool kinetic model could predict creatinine concentrations and other kinetic parameters, including the creatinine generation rate. An extrarenal creatinine clearance of 0.038 l/kg/24 h was included in the model. Diffusive cross-dialyzer clearances of urea [230 (SD 37 mL/min] correlated well (R2 = 0.78) with creatinine clearances [164 (SD 30) mL/min]. When the effective diffusion volume flow rate was set at 0.791 times the blood flow rate for the cross-dialyzer clearance measurements at 1 h into dialysis, the mean calculated volume of creatinine distribution averaged 29.6 (SD 7.2) L], compared with 31.6 (SD 7.0) L for urea (P < 0.01). The modeled creatinine generation rate [1183 (SD 463) mg/day] averaged 100.1 % (SD 29; median 99.3) of that predicted in nondialysis patients by an anthropometric equation. A simplified method for modeling the creatinine generation rate using the urea distribution volume and urea dialyzer clearance without use of the postdialysis serum creatinine measurement gave results for creatinine generation rate [1187 (SD 475) mg/day; that closely matched the value calculated using the formally modeled value, R2 = 0.971]. Our analysis confirms previous findings of similar distribution volumes for creatinine and urea. After taking extra-renal clearance into consideration, the creatinine generation rate in dialysis patients is similar to that in nondialysis patients. A simplified method based on urea clearance and urea distribution volume not requiring a postdialysis serum creatinine measurement can be used to yield creatinine generation rates that closely match those determined from standard modeling. © The Author 2017. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.

Blood Glucose Measurement in the Intensive Care Unit: What Is the Best Method?

PubMed Central

Le, Huong T.; Harris, Neil S.; Estilong, Abby J.; Olson, Arvid; Rice, Mark J.

2013-01-01

Abnormal glucose measurements are common among intensive care unit (ICU) patients for numerous reasons and hypoglycemia is especially dangerous because these patients are often sedated and unable to relate the associated symptoms. Additionally, wide swings in blood glucose have been closely tied to increased mortality. Therefore, accurate and timely glucose measurement in this population is critical. Clinicians have several choices available to assess blood glucose values in the ICU, including central laboratory devices, blood gas analyzers, and point-of-care meters. In this review, the method of glucose measurement will be reviewed for each device, and the important characteristics, including accuracy, cost, speed of result, and sample volume, will be reviewed, specifically as these are used in the ICU environment. Following evaluation of the individual measurement devices and after considering the many features of each, recommendations are made for optimal ICU glucose determination. PMID:23567008

Multicenter Clinical Evaluation of BacT/Alert Virtuo Blood Culture System.

PubMed

Jacobs, Michael R; Mazzulli, Tony; Hazen, Kevin C; Good, Caryn E; Abdelhamed, Ayman M; Lo, Pauline; Shum, Bianche; Roman, Katharine P; Robinson, Danielle C

2017-08-01

BacT/Alert Virtuo is an advanced, automated blood culture system incorporating improved automation and an enhanced detection algorithm to shorten time to detection. A multicenter study of the investigational Virtuo system (bioMérieux, Inc., Durham, NC) compared to BacT/Alert 3D (BTA3D) for detection of bacteremia/fungemia in four bottle types, SA and FA Plus (aerobic) and SN and FN Plus (anaerobic), was performed in a clinical setting with patient samples in a matched system design clinical trial. Blood was added to paired aerobic or anaerobic bottles, with the volume in each bottle in each pair required to be ≤10 ml and with the volumes required to be within 30% of each other. Of 5,709 bottle sets (52.5% aerobic pairs and 47.5% anaerobic pairs), 430 (7.5%) were positive for bacterial or fungal growth, with 342 (6.0%) clinically significant and 83 (1.5%) contaminated. A total of 3,539 sets (62.0%) were volume compliant, with 203 sets (5.7%) clinically significant. The positivity rates for volume-compliant bottle pairs determined by the two systems were comparable, with 68.7% of clinically significant isolates detected by both instruments, 15.7% by Virtuo only, and 15.7% by BTA3D only. Virtuo detected microbial growth nearly 2 h sooner overall than BTA3D (mean, 15.9 h versus 17.7 h). Shorter time to detection by Virtuo was related to organism group, with the time to detection being significantly shorter for enteric Gram-negative bacilli and enterococci (means, 3.6 h and 2.3 h shorter, respectively). This large clinical study demonstrated that the Virtuo blood culture system produced results comparable to those seen with the long-established BTA3D system, with significantly shorter time to detection. Copyright © 2017 Jacobs et al.

Evaluation of whole genome amplified DNA to decrease material expenditure and increase quality.

PubMed

Bækvad-Hansen, Marie; Bybjerg-Grauholm, Jonas; Poulsen, Jesper B; Hansen, Christine S; Hougaard, David M; Hollegaard, Mads V

2017-06-01

The overall aim of this study is to evaluate whole genome amplification of DNA extracted from dried blood spot samples. We wish to explore ways of optimizing the amplification process, while decreasing the amount of input material and inherently the cost. Our primary focus of optimization is on the amount of input material, the amplification reaction volume, the number of replicates and amplification time and temperature. Increasing the quality of the amplified DNA and the subsequent results of array genotyping is a secondary aim of this project. This study is based on DNA extracted from dried blood spot samples. The extracted DNA was subsequently whole genome amplified using the REPLIg kit and genotyped on the PsychArray BeadChip (assessing > 570,000 SNPs genome wide). We used Genome Studio to evaluate the quality of the genotype data by call rates and log R ratios. The whole genome amplification process is robust and does not vary between replicates. Altering amplification time, temperature or number of replicates did not affect our results. We found that spot size i.e. amount of input material could be reduced without compromising the quality of the array genotyping data. We also showed that whole genome amplification reaction volumes can be reduced by a factor of 4, without compromising the DNA quality. Whole genome amplified DNA samples from dried blood spots is well suited for array genotyping and produces robust and reliable genotype data. However, the amplification process introduces additional noise to the data, making detection of structural variants such as copy number variants difficult. With this study, we explore ways of optimizing the amplification protocol in order to reduce noise and increase data quality. We found, that the amplification process was very robust, and that changes in amplification time or temperature did not alter the genotyping calls or quality of the array data. Adding additional replicates of each sample also lead to insignificant changes in the array data. Thus, the amount of noise introduced by the amplification process was consistent regardless of changes made to the amplification protocol. We also explored ways of decreasing material expenditure by reducing the spot size or the amplification reaction volume. The reduction did not affect the quality of the genotyping data.

Potential Use of Salivary Markers for Longitudinal Monitoring of Inflammatory Immune Responses to Vaccination

PubMed Central

Garssen, Johan; Sandalova, Elena

2016-01-01

Vaccination, designed to trigger a protective immune response against infection, is a trigger for mild inflammatory responses. Vaccination studies can address the question of inflammation initiation, levels, and resolution as well as its regulation for respective studied pathogens. Such studies largely based on analyzing the blood components including specific antibodies and cytokines were usually constrained by number of participants and volume of collected blood sample. Hence, blood-based studies may not be able to cover the full dynamic range of inflammation responses induced by vaccination. In this review, the potential of using saliva in addition to blood for studying the kinetics of inflammatory response studies was assessed. Saliva sampling is noninvasive and has a great potential to be used for studies aimed at analysing the magnitude, time course, and variance in immune responses, including inflammation after vaccination. Based on a literature survey of inflammatory biomarkers that can be determined in saliva and an analysis of how these biomarkers could help to understand the mechanisms and dynamics of immune reactivity and inflammation, we propose that the saliva-based approach might have potential to add substantial value to clinical studies, particularly in vulnerable populations such as infants, toddlers, and ill individuals. PMID:27022211

Transcapillary protein flux following blood volume modification in dog.

PubMed

Miki, K; Nose, H; Tanaka, Y; Morimoto, T

1984-01-01

The net fluid and protein movements between intravascular and interstitial space following blood withdrawal and retransfusion of 15% of circulating blood volume were measured in dogs using a continuous monitoring method of circulating blood volume. A significant (p less than 0.01) increase in transvascular fluid shift was observed after the start of blood withdrawal and a new equilibrium state was reached within 15 to 20 min. Associated with the fluid shift, a significant increase in plasma protein of about 1 g was observed. On the other hand, blood retransfusion caused significant (p less than 0.01) increases in the shift of fluid and protein from intravascular space to interstitial space. The magnitudes of the shift of fluid and protein were almost identical in both blood withdrawal and retransfusion. The Kedem-Katchalsky transport equation was applied to the results obtained in the present study in order to analyze the relative role of diffusion and convection and to estimate the reflection coefficient for protein. A significant (p less than 0.01) linear relationship was observed between fluid and protein movement. These results suggest that the convective process is involved in the shift of protein between intravascular and interstitial space observed after blood volume modification.

[Clinical research of minimal extracorporeal circulation in perioperative blood conservation of coronary artery bypass graft].

PubMed

Liu, Yan; Cui, Hu-jun; Tao, Liang; Chen, Xu-fa

2011-04-01

To analyze the clinical effect of minimal extracorporeal circulation (MECC) in blood conservation perioperatively coronary artery bypass graft (CABG). The data of 120 cases received simple CABG since August 2006 to October 2009 was analyzed retrospectively. All the patients were divided to three groups according to the mode of circulation support in-operation: MECC, conventional extracorporeal circulation (cECC) or off-pump, 40 cases in each group. Jostra MECC system with normal temperature was used in MECC group, and common membrane oxygenator with moderate hypo-temperature was used in cECC group. Collect the data of coagulation and the blood cytological examination perioperatively, the draining volume during the first 24 h after operation, and consumption of blood products perioperatively. Standard and logistic EuroSCORE were higher in MECC group than the others (P < 0.01). The operative time and the number of distal anastomosis of off-pump group were less than MECC and cECC groups (P < 0.05), while no difference between MECC group and cECC group. Intrinsic coagulation (activated partial thromboplastin time) were much more prolonged early postoperatively in cECC group, and higher than in MECC group and off-pump group at 2 h, 6 h and 12 h postoperatively (P < 0.05), but no difference in extrinsic coagulation (prothrombin time) among three group. Adjusted by hematocrit of the same sample, free hemoglobin level rose up during the ECC procedure and reached the maximum at the end of ECC in cECC group and MECC group, but the levels were more higher in cECC group than in MECC group (P < 0.05). The draining volume during the first 24 h after operation of cECC group was larger than MECC group and off-pump group (P < 0.05). Although the decreased platelet count perioperatively and more consumed of the blood products in cECC group, but no difference among the three groups. MECC could reduce the ruin to blood cell and interfere to coagulation function during the conventional ECC procedure, decrease the postoperative draining volume and requirement of blood products.

The regulation of fluid and electrolyte metabolism in weightlessness

NASA Technical Reports Server (NTRS)

Leach, C. S.; Johnson, P. C.; Cintron, N. M.

1986-01-01

Endocrine and biochemical changes in astronauts caused by weightlessness are discussed. Translocation of fluid from the extremities to the head and chest at the onset of weightlessness is thought to lead to the establishment of a lower blood volume as an adaptation to microgravity. Results of Skylab experiments indicate that several other regulatory systems have lower homeostatic set points during space flight. Inflight blood samples from three Spacelab flights show increased antidiuretic hormone throughout these short flights and decreased aldosterone and cortisol after 3 days. Results help to explain blood hypoosmolality and hyponatremia but do not explain what happens between the onset of weightlessness and hormone changes. Other factors such as natriuretic peptides and changes in renal function are being studied to elucidate the physiologic adaptation mechanisms.

Plasma biochemical and PCV ranges for healthy, wild, immature hawksbill (Eretmochelys imbricata) sea turtles.

PubMed

Whiting, S D; Guinea, M L; Fomiatti, K; Flint, M; Limpus, C J

2014-06-14

In recent years, the use of blood chemistry as a diagnostic tool for sea turtles has been demonstrated, but much of its effectiveness relies on reference intervals. The first comprehensive blood chemistry values for healthy wild hawksbill (Eretmochelys imbricata) sea turtles are presented. Nineteen blood chemistry analytes and packed cell volume were analysed for 40 clinically healthy juvenile hawksbill sea turtles captured from a rocky reef habitat in northern Australia. We used four statistical approaches to calculate reference intervals and to investigate their use with non-normal distributions and small sample sizes, and to compare upper and lower limits between methods. Eleven analytes were correlated with curved carapace length indicating that body size should be considered when designing future studies and interpreting analyte values. British Veterinary Association.

Automated processing of human bone marrow grafts for transplantation.

PubMed

Zingsem, J; Zeiler, T; Zimmermanm, R; Weisbach, V; Mitschulat, H; Schmid, H; Beyer, J; Siegert, W; Eckstein, R

1993-01-01

Prior to purging or cryopreservation, we concentrated 21 bone marrow (BM) harvests using a modification of the 'grancollect-protocol' of the Fresenius AS 104 cell separator with the P1-Y set. Within 40-70 min, the initial marrow volume of 1,265 ml (+/- 537 ml) was processed two to three times. A mean of 47% (+/- 21%) of the initial mononuclear cells was recovered in a mean volume of 128 ml (+36 ml). The recovery of clonogenic cells, measured by CFU-GM assays, was 68% (+/- 47%). Red blood cells in the BM concentrates were reduced to 7% (+/- 4%) of the initial number. The procedure was efficient and yielded a BM cell fraction suitable for purging, cryopreservation and transplantation. At this time, 10 of the 21 patients whose BM was processed using this technique have been transplanted. Seven of these 10 patients have been grafted using the BM alone. Three of the 10 patients showed reduced cell viability and colony growth in the thawed BM samples, and therefore obtained BM and peripheral blood-derived stem cells. All transplanted patients showed an evaluable engraftment, achieving 1,000 granulocytes per microliter of peripheral blood in a mean of 18 days.

Blood salvage produces higher total blood product costs in single-level lumbar spine surgery.

PubMed

Canan, Chelsea E; Myers, John A; Owens, Roger Kirk; Crawford, Charles H; Djurasovic, Mladen; Burke, Lauren O; Bratcher, Kelly R; McCarthy, Kathryn J; Carreon, Leah Y

2013-04-15

Retrospective review. To determine the incremental cost-effectiveness of cell saver for single-level posterior lumbar decompression and fusion (PLDF). Intraoperative cell salvage is used during surgery to reduce the need for perioperative allogeneic blood transfusion. Although the use of cell saver may be beneficial in certain circumstances, its utility has not been clearly established for the common procedure of an adult single-level PLDF. Randomly selected adult patients treated with a single-level PLDF between July 2010 and June 2011 at a single institution were identified. Patients who had a combined anterior and posterior approach were excluded. The final study sample for analysis consisted of 180 patients. Hospital records were reviewed to determine whether: (1) cell saver was available during surgery, (2) recovered autologous blood was infused, and (3) the patient received intra- or postoperative allogeneic transfusions. Estimated blood loss, levels fused, volume(s) transfused, and all related complications were recorded. Costs included the cost of allogeneic blood transfusion, setting up the cell saver recovery system, and infusing autologous blood from cell saver, whereas effectiveness measures were allogeneic blood transfusions averted and quality adjusted life years. The incremental cost-effectiveness ratio was $55,538 per allogeneic transfusion averted, with a decrease in the transfusion rate from 40.0% to 38.7% associated with the cell saver approach. This translated into an incremental cost-effectiveness ratio of $5,555,380 per quality adjusted life years gained, which is well above the threshold for an intervention to be considered cost-effective ($100,000 per quality adjusted life years gained). The use of cell saver during a single-level PLDF does not significantly reduce the need for allogeneic blood transfusion and is not cost-effective. The high cost of cell saver in combination with the low complication rate of allogeneic blood transfusion, suggest that cell saver should not be used for single-level PLDF. Further studies are needed to evaluate the necessity for cell saver among other types of spinal surgery.

Body water compartments during bed rest: Evaluation of analytical methods

NASA Technical Reports Server (NTRS)

Young, H. L.; Juhos, L.; Castle, B. L.; Yusken, J.; Greenleaf, J. E.

1973-01-01

Nine healthy young men were studied to determine the reproducibility and interchangeability of the use of radio-iodinated human serum albumin and Evans Blue dye for estimating plasma volume, sodium bromide for extracellular fluid volume, and deuterium oxide for total body water volume. All subjects were tested in a semibasal condition and allowed to rest for at least 30 min. after arriving at the laboratory. The results indicate that there was uniform distribution of I131 and Evans Blue dye 10 min. after injection and of NaBr and D2O 3 hours after oral ingestion; the buildup of residual tracer did not interfere appreciably with the measurement of either or Evans Blue spaces when they are administered at equal intervals, and the buildup of background tracer after ingestion of NaBr and D2O once per week for three consecutive weeks did not affect the accuracy of the measurement. It was found that I131 and Evans Blue may be used interchangeably for estimating plasma volume; for estimating bromide and D2O spaces, one 3-hour equilibrium blood sample gives results similar to the extrapolation of multiple samples.

[Full blood count reference values in children of 8 to 12 years old residing at 2,760 m above sea level].

PubMed

Armando García-Miranda, L; Contreras, I; Estrada, J A

2014-04-01

To determine reference values for full blood count parameters in a population of children 8 to 12 years old, living at an altitude of 2760 m above sea level. Our sample consisted of 102 individuals on whom a full blood count was performed. The parameters included: total number of red blood cells, platelets, white cells, and a differential count (millions/μl and %) of neutrophils, lymphocytes, monocytes, eosinophils and basophils. Additionally, we obtained values for hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, concentration of corpuscular hemoglobin and red blood cell distribution width. The results were statistically analyzed with a non-parametric test, to divide the sample in quartiles and obtain the lower and upper limits for our intervals. Moreover, the values for the intervals obtained from this analysis were compared to intervals obtained estimating+- 2 standard deviations above and below from our mean values. Our results showed significant differences compared to normal interval values reported for the adult Mexican population in most of the parameters studied. The full blood count is an important laboratory test used routinely for the initial assessment of a patient. Values of full blood counts in healthy individuals vary according to gender, age and geographic location; therefore, each population should have its own reference values. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

Patient-Specific Instrumentation Affects Perioperative Blood Loss in Total Knee Arthroplasty.

PubMed

Cucchi, Davide; Menon, Alessandra; Zanini, Beatrice; Compagnoni, Riccardo; Ferrua, Paolo; Randelli, Pietro

2018-05-23

Patient-specific instrumentation (PSI) may contribute to reduced blood loss related to total knee arthroplasty (TKA). The purpose of this study was to compare the estimated hemoglobin (Hb) and red blood cell volume (RBC) losses in two groups of patients undergoing TKA with PSI and conventional instrumentation. Pre- and postoperative blood samples were collected from 22 patients randomly assigned to receive a PSI-assisted or conventional TKA. Post- to preoperative Hb difference was calculated and RBC loss was estimated according to Sehat et al. A significant difference in Hb reduction in favor of the PSI group was registered on the last day of stay ( p  = 0.0084) and significant treatment effect ( p  = 0.027) on Hb reduction after intervention was found with a regression model for longitudinal measurements. This study demonstrated that PSI leads to a significant trend in earlier Hb regain. These promising results suggest a beneficial effect of PSI in blood loss reduction. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Measuring Intracranial Pressure And Volume Noninvasively

NASA Technical Reports Server (NTRS)

Cantrell, John H.; Yost, William T.

1994-01-01

Ultrasonic technique eliminates need to drill into brain cavity. Intracranial dynamics instrument probes cranium ultrasonically to obtain data for determination of intracranial pressure (ICP) and pressure-volume index (PVI). Instrument determines sensitivity of skull to changes in pressure and by use of mechanical device to exert external calibrated pressure on skull. By monitoring volume of blood flowing into jugular vein, one determines change of volume of blood in cranial system. By measuring response of skull to increasing pressure (where pressure increased by tilting patient known amount) and by using cranial blood pressure, one determines intial pressure in cerebrospinal fluid. Once PVI determined, ICP determined.

Hypervolemia and plasma vasopressin response during water immersion in men

NASA Technical Reports Server (NTRS)

Greenleaf, J. E.; Morse, J. T.; Barnes, P. R.; Silver, J.; Keil, L. C.

1983-01-01

Immersion studies were performed on seven mildly dehydrated male subjects to examine the effect of suppression of plasma vasopressin (PVP) on diuresis in water immersion. The water was kept at close to 34.5 C and the subjects remained in the water for 4 hr after sitting for 2 hr. Na and K levels in the serum and urine were analyzed, as were osmolality, red blood cell count, renin activity, total protein, albumin amounts, hematocrit, and hemoglobin. Plasma volume was monitored from samples drawn at specified intervals during immersion. The plasma volume increased significantly 30 min after immersion, but no PVP was observed. The dehydration induced elevated serum osmotic concentrations. It is concluded that the hydration condition before immersion and the volume of fluid intake during immersion affects the hemodilution during immersion.

Optimising methods of red cell sedimentation from cord blood to maximise nucleated cell recovery prior to cryopreservation.

PubMed

Madkaikar, M; Gupta, M; Ghosh, K; Swaminathan, S; Sonawane, L; Mohanty, D

2007-01-01

Human cord blood is now an established source of stem cells for haematopoietic reconstitution. Red blood cell (RBC) depletion is required to reduce the cord blood unit volume for commercial banking. Red cell sedimentation using hydroxy ethyl starch (HES) is a standard procedure in most cord blood banks. However, while standardising the procedure for cord blood banking, a significant loss of nucleated cells (NC) may be encountered during standard HES sedimentation protocols. This study compares four procedures for cord blood processing to obtain optimal yield of nucleated cells. Gelatin, dextran, 6% HES and 6% HES with an equal volume of phosphate-buffered saline (PBS) were compared for RBC depletion and NC recovery. Dilution of the cord blood unit with an equal volume of PBS prior to sedimentation with HES resulted in maximum NC recovery (99% [99.5 +/- 1.3%]). Although standard procedures using 6% HES are well established in Western countries, they may not be applicable in India, as a variety of factors that can affect RBC sedimentation (e.g., iron deficiency, hypoalbuminaemia, thalassaemia trait, etc.) may reduce RBC sedimentation and thus reduce NC recovery. While diluting cord blood with an equal volume of PBS is a simple method to improve the NC recovery, it does involve an additional processing step.

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

Effect of exercise training on cardiopulmonary baroreflex control of forearm vascular resistance in humans

NASA Technical Reports Server (NTRS)

Mack, G. W.; Convertino, V. A.; Nadel, E. R.

1993-01-01

We studied the stimulus-response characteristics of cardiopulmonary baroreflex control of forearm vascular resistance (FVR) in four groups of male volunteer subjects: i) unfit, ii) physically fit, iii) before and after 10 wk of endurance training (chronic blood volume expansion), and iv) before and after acute blood volume expansion. We assessed the relationship between reflex stimulus, i.e., changes in central venous pressure and response, i.e., FVR, during unloading of cardiopulmonary mechanoreceptors with lower body negative pressure (LBNP, 0 to -20 mm Hg). The slope of the linear relationship between FVR and CVP, the index of the responsiveness of this baroreflex, was significantly diminished (> 50%) in the fit subjects compared with the unfit. The slope of the FVR-CVP relationship was inversely correlated with the subject's total blood volume, suggesting that blood volume expansion was related to the attenuated CP baroreflex. In the exercise training study, maximal oxygen consumption and blood volume increased following 10 wk of endurance training (N = 14) but were unchanged in the time control group (N = 7). The slope of the FVR-CVP relationship was significantly reduced (32%) following 10 wk of training but was unchanged in the time control group. The reduction in slope of the FVR-CVP relationship was inversely related to the increase in blood volume associated with exercise training. Acute blood volume expansion 8 ml.kg-1 body weight with 5% human serum albumin solution) significantly reduced the slope of the FVR-CVP relationship. These data support the hypothesis that the attenuated forearm vascular reflex in physically fit individuals is related to a training-induced hypervolemia.(ABSTRACT TRUNCATED AT 250 WORDS).

Effects of pretransport handling stress on physiological and behavioral response of ostriches.

PubMed

Bejaei, M; Cheng, K M

2014-05-01

Ostrich (Struthio camelus) production is a relatively young industry and there has been little research on ostrich welfare during pretransport handling and the transportation process. A heavy body with a high center of gravity makes ostriches' handling and transportation problems different from other livestock. The main objective of this study was to investigate the effects of the pretransport holding time duration on ostrich behavior and physiological responses. A second objective was to identify and validate behavioral indicator(s) that could be used to identify stressed birds during pretransport handling. Prior to shipping, twenty-four 2.5-yr-old ostriches were moved into a holding pen. Birds were then individually restrained, hooded, and walked from the holding pen (approximately 12 min/bird) to a sampling pen (visually isolated from the holding pen) where they were weighed and a 10-mL blood sample obtained. A second blood sample was taken from each bird after a 1,100-km transportation. Blood samples were analyzed for concentrations of blood metabolites, enzymes, corticosterone, and white blood cell and differential counts. Behavioral responses and physical damages of ostriches were also recorded before and after transport. Results indicated that birds that spent longer time in the pretransport holding pen had higher pretransport plasma concentrations of aspartate aminotransferase, alanine aminotransferase, sodium, and packed cell volume. Immobile sitting behavior, observed in 5 out of the last 11 birds handled, was positively correlated with higher pretransport handling stress, higher posttransport aspartate aminotransferase, alanine aminotransferase, creatine phosphokinase, and glucose concentrations, and transport losses. Knowledge of pretransport handling impacts on ostrich stress and availability of behavioral indicators (e.g., immobile sitting response) could be used to improve handing processes, thereby decreasing potential weight loss, injury, and mortality.

Picric acid capped silver nanoparticles as a probe for colorimetric sensing of creatinine in human blood and cerebrospinal fluid samples.

PubMed

Parmar, Ankita K; Valand, Nikunj N; Solanki, Kalpesh B; Menon, Shobhana K

2016-02-21

Creatinine is the most important parameter to be determined in the diagnosis of renal, muscular and thyroid function. The most common method for the determination of creatinine is Jaffe's reaction, a routine practice for blood and urine analysis. However, in cases of icteric and haemolyzed blood samples, interference occurs during the estimation of creatinine by other constituents present in the blood like bilirubin, creatine, and urea, which lead to wrong diagnosis. To overcome such difficulty, we have developed a silver nanoparticle (Ag NPs) based sensor for the selective determination of creatinine. In this study, a new approach has been given to the traditional Jaffe's reaction, by coating Ag NPs with picric acid (PA) to form an assembly that can selectively detect creatinine. The Ag NPs based sensor proficiently and selectively recognizes creatinine due to the ability of picric acid to bind with it and form a complex. The nanoassembly and the interactions were investigated by transmission electron microscopy (TEM), dynamic light scattering (DLS) analysis, UV-Vis spectroscopy, FT-IR spectroscopy and ESI-MS, which demonstrated the binding affinity of creatinine with PA-capped Ag NPs. A linear correlation was obtained in the range of 0.01 μM-1 μM with an R(2) value of 0.9998 and a lower detection limit of 8.4 nM. The sensor was successfully applied to different types of blood and CSF samples for the determination of creatinine, and the results were compared to that of the Jaffe's method. With the advantages of high sensitivity, selectivity and low sample volume, this method is potentially suitable for the on-site monitoring of creatinine.

Thyroid Volume and Its Relation to Anthropometric Measures in a Healthy Cuban Population

PubMed Central

Turcios, Silvia; Lence-Anta, Juan J.; Santana, Jose-Luis; Pereda, Celia M.; Velasco, Milagros; Chappe, Mae; Infante, Idalmis; Bustillo, Marlene; García, Anabel; Clero, Enora; Maillard, Stephane; Rodriguez, Regla; Xhaard, Constance; Ren, Yan; Rubino, Carole; Ortiz, Rosa M.; de Vathaire, Florent

2015-01-01

Objectives The aim of this study was to describe the thyroid volume in healthy adults by ultrasound and to correlate this volume with some anthropometric measures and other differentiated thyroid cancer risk factors. Study Design Thyroid volume and anthropometric measures were recorded in a sample of 100 healthy adults, including 21 men and 79 women aged 18-50 years, living in a non-iodine-deficient area of Havana city. Results The average thyroid volume was 6.6 ± 0.26 ml; it was higher in men (7.3 ml) than in women (6.4 ml; p = 0.15). In the univariate analysis, thyroid volume was correlated with all anthropometric measures, but in the multivariate analysis, body surface area was found to be the only significant anthropometric parameter. Thyroid volume was also higher in current or former smokers and in persons with blood group AB or B. Conclusion Specific reference values of thyroid volume as a function of body surface area could be used for evaluating thyroid volume in clinical practice. The relation between body surface area and thyroid volume is coherent with what is known about the relation of thyroid volume to thyroid cancer risk, but the same is not true about the relation between thyroid volume and smoking habit. PMID:25960963

Optoelectronic system for the determination of blood volume in pneumatic heart assist devices.

PubMed

Konieczny, Grzegorz; Pustelny, Tadeusz; Setkiewicz, Maciej; Gawlikowski, Maciej

2015-12-10

The following article describes the concept of optical measurement of blood volume in ventricular assist devices (VAD's) of the pulsatile type. The paper presents the current state of art in blood volume measurements of such devices and introduces a newly developed solution in the optic domain. The objective of the research is to overcome the disadvantages of the previously developed acoustic method-the requirement of additional sensor chamber. The idea of a compact measurement system has been introduced, followed by laboratory measurements. Static tests of the system have been presented, followed by dynamic measurements on a physical model of the human ventricular system. The results involving the measurements of blood chamber volume acquired by means of an optical system have been compared with the results acquired by means of the Transonic T410 ultrasound flow rate sensor (11PLX transducer, uncertainty ±5 %). Preliminary dynamic measurements conducted on the physical model of the human cardiovascular system show that the proposed optical measurement system may be used to measure the transient blood chamber volumes of pulsatile VAD's with the uncertainties (standard mean deviation) lower than 10 %. The results show that the noninvasive measurements of the temporary blood chamber volume in the POLVAD prosthesis with the use of the developed optical system allows us to carry out accurate static and dynamic measurements.

Fluid shifts, vasodilatation and ambulatory blood pressure reduction during long duration spaceflight

PubMed Central

Norsk, Peter; Asmar, Ali; Damgaard, Morten; Christensen, Niels Juel

2015-01-01

Acute weightlessness in space induces a fluid shift leading to central volume expansion. Simultaneously, blood pressure is either unchanged or decreased slightly. Whether these effects persist for months in space is unclear. Twenty-four hour ambulatory brachial arterial pressures were automatically recorded at 1–2 h intervals with portable equipment in eight male astronauts: once before launch, once between 85 and 192 days in space on the International Space Station and, finally, once at least 2 months after flight. During the same 24 h, cardiac output (rebreathing method) was measured two to five times (on the ground seated), and venous blood was sampled once (also seated on the ground) for determination of plasma catecholamine concentrations. The 24 h average systolic, diastolic and mean arterial pressures (mean ± se) in space were reduced by 8 ± 2 mmHg (P = 0.01; ANOVA), 9 ± 2 mmHg (P < 0.001) and 10 ± 3 mmHg (P = 0.006), respectively. The nightly blood pressure dip of 8 ± 3 mmHg (P = 0.015) was maintained. Cardiac stroke volume and output increased by 35 ± 10% and 41 ± 9% (P < 0.001); heart rate and catecholamine concentrations were unchanged; and systemic vascular resistance was reduced by 39 ± 4% (P < 0.001). The increase in cardiac stroke volume and output is more than previously observed during short duration flights and might be a precipitator for some of the vision problems encountered by the astronauts. The spaceflight vasodilatation mechanism needs to be explored further. PMID:25774397

Combination of MRI hippocampal volumetry and arterial spin labeling MR perfusion at 3-Tesla improves the efficacy in discriminating Alzheimer's disease from cognitively normal elderly adults.

PubMed

Mak, Henry Ka-Fung; Qian, Wenshu; Ng, Kwok Sing; Chan, Queenie; Song, You-Qiang; Chu, Leung Wing; Yau, Kelvin Kai-Wing

2014-01-01

Structural magnetic resonance imaging has been employed for evaluation of medial temporal atrophy in patients with Alzheimer's disease (AD). Arterial spin labeling (ASL) technique could detect cerebral perfusion abnormalities in AD. We hypothesized that combination of hippocampal volumetry and cerebral blood flow yield higher accuracy than either method alone in discriminating AD patients from cognitively normal elderly adults. 13 AD patients and 15 healthy controls were studied using a 3-tesla scanner. Standardized T1W 3D volumetric Fast Field Echo and QUASAR ASL sequences were employed for cerebral volumetry and perfusion respectively. Manual Right and left hippocampal volumetry was performed manually by ANALYZE software, with total intracranial volume normalization. ASL data were analyzed by institutional specially-design software to calculate cerebral blood flow of region-of-interests placed at the middle and posterior cingulate gyri. Right and left hippocampal volumes and middle and posterior cingulate gyri cerebral blood flows were significantly lower in the patients than in the controls (independent-samples t-tests, p < 0.05), and prediction accuracies of 89.3%, 82.1%, 75.0% and 71.4% were achieved for each of the above parameters, respectively. In distinguishing patients from controls using corresponding optimized cut-off values, various combinations of these parameters were used to create the Receiver Operating Characteristic curves. The highest area under curve value was 0.944, by combining cerebral blood flow at the middle cingulate gyrus, normalized right and left hippocampal volumes. A 'one-stop-shop' magnetic resonance study of combined hippocampal volumetry and cerebral perfusion has improved efficacy in discriminating AD patients from cognitively normal elderly adults.

Parametrically defined cerebral blood vessels as non-invasive blood input functions for brain PET studies

NASA Astrophysics Data System (ADS)

Asselin, Marie-Claude; Cunningham, Vincent J.; Amano, Shigeko; Gunn, Roger N.; Nahmias, Claude

2004-03-01

A non-invasive alternative to arterial blood sampling for the generation of a blood input function for brain positron emission tomography (PET) studies is presented. The method aims to extract the dimensions of the blood vessel directly from PET images and to simultaneously correct the radioactivity concentration for partial volume and spillover. This involves simulation of the tomographic imaging process to generate images of different blood vessel and background geometries and selecting the one that best fits, in a least-squares sense, the acquired PET image. A phantom experiment was conducted to validate the method which was then applied to eight subjects injected with 6-[18F]fluoro-L-DOPA and one subject injected with [11C]CO-labelled red blood cells. In the phantom study, the diameter of syringes filled with an 11C solution and inserted into a water-filled cylinder were estimated with an accuracy of half a pixel (1 mm). The radioactivity concentration was recovered to 100 ± 4% in the 8.7 mm diameter syringe, the one that most closely approximated the superior sagittal sinus. In the human studies, the method systematically overestimated the calibre of the superior sagittal sinus by 2-3 mm compared to measurements made in magnetic resonance venograms on the same subjects. Sources of discrepancies related to the anatomy of the blood vessel were found not to be fundamental limitations to the applicability of the method to human subjects. This method has the potential to provide accurate quantification of blood radioactivity concentration from PET images without the need for blood samples, corrections for delay and dispersion, co-registered anatomical images, or manually defined regions of interest.

Direct detection of fibrinogen in human plasma using electric-double-layer gated AlGaN/GaN high electron mobility transistors

NASA Astrophysics Data System (ADS)

Regmi, Abiral; Sarangadharan, Indu; Chen, Yen-Wen; Hsu, Chen-Pin; Lee, Geng-Yen; Chyi, Jen-Inn; Shiesh, Shu-Chu; Lee, Gwo-Bin; Wang, Yu-Lin

2017-08-01

Fibrinogen found in blood plasma is an important protein biomarker for potentially fatal diseases such as cardiovascular diseases. This study focuses on the development of an assay to detect plasmatic fibrinogen using electrical double layer gated AlGaN/GaN high electron mobility transistor biosensors without complex sample pre-treatment methods used in the traditional assays. The test results in buffer solution and clinical plasma samples show high sensitivity, specificity, and dynamic range. The sensor exhibits an ultra-low detection limit of 0.5 g/l and a detection range of 0.5-4.5 g/l in 1× PBS with 1% BSA. The concentration dependent sensor signal in human serum samples demonstrates the specificity to fibrinogen in a highly dense matrix of background proteins. The sensor does not require complicated automation, and quantitative results are obtained in 5 min with <5 μl sample volume. This sensing technique is ideal for speedy blood based diagnostics such as POC (point of care) tests, homecare tests, or personalized healthcare.

Sweat: a sample with limited present applications and promising future in metabolomics.

PubMed

Mena-Bravo, A; Luque de Castro, M D

2014-03-01

Sweat is a biofluid with present scant use as clinical sample. This review tries to demonstrate the advantages of sweat over other biofluids such as blood or urine for routine clinical analyses and the potential when related to metabolomics. With this aim, critical discussion of sweat samplers and equipment for analysis of target compounds in this sample is made. Well established routine analyses in sweat as is that to diagnose cystic fibrosis, and the advantages and disadvantages of sweat versus urine or blood for doping control have also been discussed. Methods for analytes such as essential metals and xenometals, ethanol and electrolytes in sweat in fact constitute target metabolomics approaches or belong to any metabolomics subdiscipline such as metallomics, ionomics or xenometabolomics. The higher development of biomarkers based on genomics or proteomics as omics older than metabolomics is discussed and also the potential role of metabolomics in systems biology taking into account its emergent implementation. Normalization of the volume of sampled sweat constitutes a present unsolved shortcoming that deserves investigation. Foreseeable trends in this area are outlined. Copyright © 2013 Elsevier B.V. All rights reserved.

Treatment of carprofen overdose with therapeutic plasma exchange in a dog.

PubMed

Kjaergaard, Astrid B; Davis, Jennifer L; Acierno, Mark J

2018-06-13

To report the use of therapeutic plasma exchange (TPE) in a dog with carprofen toxicosis. A 6-year-old female neutered Bichon Frise weighing 6.9 kg was examined after it had ingested 72 mg/kg carprofen. Mild dehydration without azotemia and with a urine specific gravity of 1.050 was noted at presentation. Treatment consisted of induction of emesis, symptomatic medical therapy, and TPE. The TPE achieved 1.5 plasma volume exchanges over 3 hours. Blood samples and effluent samples were collected every 30 minutes during TPE and additional blood samples were collected 11 and 35 hours after treatment. Carprofen concentrations in these samples were determined by high-pressure liquid chromatography. A 51% reduction in serum carprofen concentration was achieved following TPE. This report describes the successful reduction of plasma carprofen concentration in a dog using TPE. Although recent studies suggest that this particular dog may not have received a toxic dose, a 51% reduction of plasma carprofen concentration was achieved over 180 minutes, and TPE may be beneficial for treatment of dogs that have ingested higher doses. © Veterinary Emergency and Critical Care Society 2018.

Comparative Study on Compositions and Functional Properties of Porcine, Chicken and Duck Blood

PubMed Central

2017-01-01

Hematological, chemical and functional characteristics of porcine, chicken and duck blood were evaluated. A porcine blood sample showed the most abundant red blood cell, hemoglobin concentration, packed cell volume and plasma protein content as well as its freeze-dried blood possessed the highest contents of protein, fat, Cu and Cr with the highest percentage of heme iron (p<0.05). Unlike porcine blood, chicken blood showed a well balance in some essential amino acids, specifically for a higher isoleucine content (p<0.05). Furthermore, it possessed the highest contents of carbohydrate, Zn and non-heme iron (p<0.05). The most rapid response to form a strong gel, especially at 70°C and 80°C, was found in chicken blood, followed by duck and porcine blood, respectively. The result of emulsion activity index (EAI) and emulsion stability index (ESI) at the low protein concentration indicated that chicken blood had the most superior emulsion properties (p<0.05). Regarding duck blood, it exhibited the highest content of Mg and Mn (p<0.05). Moreover, duck blood had similar foaming properties to porcine blood in which they showed higher values than chicken blood (p<0.05). Specific characteristics of blood were therefore diminished by animal species in which this information could be used as food supplementation or product development based on their potential applications. PMID:28515647

Acute effects of firefighting on cardiac performance.

PubMed

Fernhall, Bo; Fahs, Christopher A; Horn, Gavin; Rowland, Thomas; Smith, Denise

2012-02-01

This study examined standard echocardiographic measures of cardiac size and performance in response to a 3-h firefighting training exercise. Forty experienced male personnel completed a standardized 3 h live firefighting exercise. Before and after the firefighting activities, participants were weighed, height, heart rate, blood pressure and blood samples were obtained, and echocardiographic measurements were made. Firefighting produced significant decreases in left ventricular diastolic dimension, stroke volume, fractional shortening, and mitral E velocity, tachycardia, a rise in core temperature, and a reduction in calculated plasma volume. On tissue Doppler imaging, there were no changes in systolic contractile function, but a decreased lateral wall diastolic velocity was observed. These findings show that 3 h of live firefighting produced cardiac changes consistent with cardiac fatigue, coupled with a decrease in systemic arterial compliance. These data show that live firefighting produces significant cardiovascular changes and future work is needed to evaluate if these changes are related to the increase in cardiovascular risk during live firefighting.

Chronic fatigue syndrome: illness severity, sedentary lifestyle, blood volume and evidence of diminished cardiac function.

PubMed

Hurwitz, Barry E; Coryell, Virginia T; Parker, Meela; Martin, Pedro; Laperriere, Arthur; Klimas, Nancy G; Sfakianakis, George N; Bilsker, Martin S

2009-10-19

The study examined whether deficits in cardiac output and blood volume in a CFS (chronic fatigue syndrome) cohort were present and linked to illness severity and sedentary lifestyle. Follow-up analyses assessed whether differences in cardiac output levels between CFS and control groups were corrected by controlling for cardiac contractility and TBV (total blood volume). The 146 participants were subdivided into two CFS groups based on symptom severity data, severe (n=30) and non-severe (n=26), and two healthy non-CFS control groups based on physical activity, sedentary (n=58) and non-sedentary (n=32). Controls were matched to CFS participants using age, gender, ethnicity and body mass. Echocardiographic measures indicated that the severe CFS participants had 10.2% lower cardiac volume (i.e. stroke index and end-diastolic volume) and 25.1% lower contractility (velocity of circumferential shortening corrected by heart rate) than the control groups. Dual tag blood volume assessments indicated that the CFS groups had lower TBV, PV (plasma volume) and RBCV (red blood cell volume) than control groups. Of the CFS subjects with a TBV deficit (i.e. > or = 8% below ideal levels), the mean+/-S.D. percentage deficit in TBV, PV and RBCV were -15.4+/-4.0, -13.2+/-5.0 and -19.1+/-6.3% respectively. Lower cardiac volume levels in CFS were substantially corrected by controlling for prevailing TBV deficits, but were not affected by controlling for cardiac contractility levels. Analyses indicated that the TBV deficit explained 91-94% of the group differences in cardiac volume indices. Group differences in cardiac structure were offsetting and, hence, no differences emerged for left ventricular mass index. Therefore the findings indicate that lower cardiac volume levels, displayed primarily by subjects with severe CFS, were not linked to diminished cardiac contractility levels, but were probably a consequence of a co-morbid hypovolaemic condition. Further study is needed to address the extent to which the cardiac and blood volume alterations in CFS have physiological and clinical significance.

Changes in scalp and cortical blood flow during hyperventilation measured with diffusing-wave spectroscopy

NASA Astrophysics Data System (ADS)

Li, Jun; Ninck, Markus; Gisler, Thomas

2009-07-01

Changes in scalp and cortical blood flow induced by voluntary hyperventilation are investigated by near-infrared diffusing-wave spectroscopy. The temporal intensity autocorrelation function g(2) (τ) of multiply scattered light is recorded from the forehead of subjects during hyperventilation. Blood flow within the sampled tissue volume is estimated by the mean decay rate of g(2) (τ) . Data measured from six subjects show that the pattern of the hemodynamic response during 50 s hyperventilation is rather complicated: within the first 10 s, in three subjects an initial increase in blood flow is observed; from 10 s to 20 s, the mean blood flow is smaller than its baseline value for all six subjects; for the duration from 20 s to 30 s, the blood flow increases again. However, after 30 s the change is not consistent across subjects. Further study on one of these subjects by using two receivers probing the blood flow in the cortex and in the superficial layers simultaneously, reveals that during hyperventilation, the direction of change in blood flow within the scalp is opposite to the one in the brain. This helps to understand the complicated hemodynamic response observed in our measurements.

Elevated central venous pressure: A consequence of exercise training-induced hypervolemia

NASA Technical Reports Server (NTRS)

Convertino, Victor A.; Mack, Gary W.; Nadel, Ethan R.

1990-01-01

Resting plasma volumes, and arterial and central venous pressures (CVP) were measured in 16 men before and after exercise training to determine if training-induced hypervolemia could be explained by a change in total vascular capacitance. In addition, resting levels of plasma vasopressin (AVP), atrial natriuretic peptide (ANP), aldosterone (ALD), and norepinephrine (NE) were measured before and after training. The same measurements of vacular volume, pressures, and plasma hormones were measured in 8 subjects who did not undergo exercise and acted as controls. The exercise training program consisted of 10 weeks of controlled cycle exercise for 30 min/d, 4 d/wk at 75 to 80 percent of maximal oxygen uptake (VO2max). A training effect was verified by a 20 percent increase in VO2max, a resting bradycardia, and a 370 ml (9 percent) increase in blood volume. Mean arterial blood pressure was unaltered by exercise training, but resting CVP increased. The percent change in blood volume from before to after training was linearly related to the percent change in CVP. As a consequence of elevations in both blood volume and CVP, the volume-to-pressure ratio was essentially unchanged following exercise training. Plasma AVP, ANP, ALD, and NE were unaltered. Results indicate that elevated CVP is a consequence of training-induced hypervolemia without alteration in total effective venous capacitance. This may represent a resetting of the pressure-volume stimulus-response relation for regulation of blood volume.

Measurement of Human Blood and Plasma Volumes

NASA Technical Reports Server (NTRS)

Greenleaf, J. E.; Szalkay, H. G. H.

1987-01-01

Report reviews techniques for measuring blood-plasma volume in humans. Common technique of using radioactive iodine isotope to label plasma albumin involves unwarranted risks from low-level radiation. Report emphasizes techniques using Evans-blue-dye (T-1824) labeling of albumin, hematocrit or hemoglobin/hematocrit measurements, or blood densitometry. In Evans-blue-dye technique, plasma volume determined from decrease in dye concentration occurring after small amount of dye solution injected into circulatory system. Subjection of Evans blue dye to test for carcinogenicity gave negative results.

WE-FG-206-05: New Arterial Spin Labeling Method for Simultaneous Estimation of Arterial Cerebral Blood Volume, Cerebral Blood Flow and Arterial Transit Time

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

Johnston, M; Whitlow, C; Jung, Y

Purpose: To demonstrate the feasibility of a novel Arterial Spin Labeling (ASL) method for simultaneously measuring cerebral blood flow (CBF), arterial transit time (ATT), and arterial cerebral blood volume (aCBV) without the use of a contrast agent. Methods: A series of multi-TI ASL images were acquired from one healthy subject on a 3T Siemens Skyra, with the following parameters: PCASL labeling with variable TI [300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000] ms, labeling bolus 1400 ms when TI allows, otherwise 100 ms less than TI, TR was minimized for each TI, two sincmore » shaped pre-saturation pulses were applied in the imaging plane immediately before 2D EPI acquisition. 64×64×24 voxels, 5 mm slice thickness, 1 mm gap, full brain coverage, 6 averages per TI, no crusher gradients, 11 ms TE, scan time of 4:56. The perfusion weighted time-series was created for each voxel and fit to a novel model. The model has two components: 1) the traditional model developed by Buxton et al., accounting for CBF and ATT, and 2) a box car function characterizing the width of the labeling bolus, with variable timing and height in proportion to the aCBV. All three parameters were fit using a nonlinear fitting routine that constrained all parameters to be positive. The main purpose of the high-temporal resolution TI sampling for the first second of data acquisition was to precisely estimate the blood volume component for better detection of arrival time and magnitude of signal. Results: Whole brain maps of CBF, ATT, and aCBV were produced, and all three parameters maps are consistent with similar maps described in the literature. Conclusion: Simultaneous mapping of CBF, ATT, and aCBV is feasible with a clinically tractable scan time (under 5 minutes).« less

Uncontrolled Hemorrhage Differs From Volume- or Pressure-Matched Controlled Hemorrhage in Swine

DTIC Science & Technology

2007-10-01

differences between these models, we evaluated the relationship between blood volume loss and blood pressure in controlled versus uncontrolled hemorrhage...aortotomy; (2) group P, controlled hemorrhage matched to the blood pressure profile of group U; or (3) group V, controlled hemorrhage matched to the...hemorrhage and received no fluid resuscitation. Group U resulted in a blood loss of 17.6 T 0.7 mL kgj1 and a reduction in blood pressure to 28 T 3 mmHg at

Medical Laboratory Technician--Hematology, Serology, Blood Banking, and Immunohematology (AFSC 90470).

ERIC Educational Resources Information Center

Thompson, Joselyn H.

This three-volume student text is designed for use by Air Force personnel enrolled in a self-study extension course for medical laboratory technicians. Covered in the individual volumes are hematology (the physiology of blood, complete blood counts and related studies, erythrocyte studies, leukocyte and thrombocyte maturation, and blood…

The Effects of Acute High-Intensity Interval Training on Hematological Parameters in Sedentary Subjects.

PubMed

Belviranli, Muaz; Okudan, Nilsel; Kabak, Banu

2017-07-19

The objective of the study was to determine the effects of acute high-intensity interval training (HIIT) on hematological parameters in sedentary men. Ten healthy, non-smoker, and sedentary men aged between 18 and 24 years participated in the study. All subjects performed four Wingate tests with 4 min intervals between the tests. Blood samples were collected at pre-exercise, immediately after, 3 and 6 h after the fourth Wingate test. Hematological parameters were analyzed in these samples. The results showed that hematocrit percentage, hemoglobin values, red cell count, mean cell volume, platelet count, total white cell count, and counts of the white cell subgroups increased immediately after the acute HIIT and their values began to return to resting levels 3 h after exercise, and completely returned to resting levels 6 h after exercise. In conclusion, acute HIIT causes an inflammatory response in blood.

[The physiologic significance of 2,3-diphosphoglycerate and changes in its erythrocyte levels in an experiment].

PubMed

Béder, I; Orgonásová, M; Brozman, B; Horecký, J; Mataseje, A

1990-05-01

The regulatory effect of 2,3-DPG on oxygen transport and binding to hemoglobin was analyzed. Under conditions of substitution hemodilution by isooncotic 3.4% Rheodextran (Spofa), changes in the content of 2,3-DPG in arterial and venous blood were enzymatically determined over several days. Reference values of 2,3-DPG were obtained in the studied series of dogs (2.05 +/- 0.74 x 10(-6) mol.ml-1 in whole blood 4.69 +/- 1.52 x 10(-6) mol.g-1 of erythrocyte volume, and 13.39 +/- 2.82 x 10(-6) mol.ml-1 of hemoglobin). In anesthetized animals the content of 2,3-DPG in arterial blood was significantly higher (6.28 +/- 0.84 x 10(-6) mol.ml-1 of erythrocyte volume) than in venous blood (6.01 +/- 0.80 x 10(-6) mol.ml-1 of erythrocyte volume). At substitution hemodilution the 2,3-DPG content in erythrocytes of venous blood decreased from 5.46 +/- 0.67 to 4.97 +/- 1.31 x 10(-6) mol.ml-1 of erythrocyte volume. The subsequent increase to 6.04 +/- 0.71 x 10(-6) mol.ml-1 of erythrocyte volume was achieved by nonlinear increase over the following days, at persisting low hemoglobin content in blood.

Volume-dependent hemodynamic effects of blood collection in canine donors - evaluation of 13% and 15% of total blood volume depletion.

PubMed

Ferreira, Rui R F; Gopegui, Rafael R; De Matos, Augusto J F

2015-03-01

There is no consensus regarding the blood volume that could be safely donated by dogs, ranging from 11 to 25% of its total blood volume (TBV). No previous studies evaluated sedated donors. To evaluate the hemodynamic effects of blood collection from sedated and non-sedated dogs and to understand if such effects were volume-dependent. Fifty three donations of 13% of TBV and 20 donations of 15% TBV were performed in dogs sedated with diazepam and ketamine. Additionally, a total of 30 collections of 13% TBV and 20 collections of 15% TBV were performed in non-sedated dogs. Non-invasive arterial blood pressures and pulse rates were registered before and 15 min after donation. Post-donation pulse rates increased significantly in both sedated groups, with higher differences in the 15% TBV collections. Systolic arterial pressures decreased significantly in these groups, while diastolic pressures increased significantly in 13% TBV donations. Non-sedated groups revealed a slight, but significant, SBP decrease. No clinical signs related to donations were registered. These results suggest that the collection of 15% TBV in sedated donors induces hemodynamic variations that may compromise the harmlessness of the procedure, while it seems to be a safe procedure in non-sedated dogs.

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

[Improving venous tone and capillary sealing. Effect of a combination of Ruscus extract and hesperidine methyl chalcone in healthy probands in heat stress].

PubMed

Rudofsky, G

1989-06-30

The drug combination of Ruscus-extract and hesperidine methyl chalcone (HMC) involves two basic mechanisms in the treatment of venous diseases: increase in venous tonicity and edema protection. This was shown in a double-blind study on 20 healthy volunteers by comparing the effectiveness of the individual substances, the combination and a placebo on the venous hemodynamics and the volume of the foot. Ruscus-extract augments the tonicity of the venous wall. This is expressed by a decrease in venous capacity (p less than 0.01), a reduction in the blood pool in the lower leg under orthostatic conditions, and a decrease in tissue volume of the foot and ankle (p less than 0.01). HMC lowers the capillary filtration rate (p less than 0.01) but augmented the blood pool. The increase in blood volume can be explained by dehydration of the tissue of the lower leg lowering the pressure of tissue on the venous system and increasing the blood pool in the limb. After administration of the combination, the blood volume was between the Ruscus and HMC volumes, while the effects on filtration rate, venous capacity and tissue volume corresponded to the changes seen after administration of HMC and Ruscus extract alone.

Observational study of changes in epidural pressure and elastance during epidural blood patch in obstetric patients.

PubMed

Pratt, S D; Kaczka, D W; Hess, P E

2014-05-01

During an epidural blood patch, we inject blood until the patient describes mild back pressure, often leading to injection of more than 20 mL of blood. We undertook this study to measure the epidural pressures generated during an epidural blood patch and to identify the impact of volume on epidural elastance in obstetric patients. This study was performed in postpartum patients who presented for an epidural blood patch with symptoms consistent with a postdural puncture headache. After identification of the epidural space using loss of resistance to air or saline, we measured static epidural pressure after each 5-mL injection of blood. Models were then fitted to the data and the epidural elastance and compliance calculated. Eighteen blood patches were performed on 17 patients. The mean final volume injected was 18.9±7.8 mL [range 6-38 mL]. The mean final pressure generated was 13.1±13.4 mmHg [range 2-56 mmHg]. A curvilinear relationship existed between volume injected and pressure, which was described by two models: (1) pressure=0.0254×(mL injected)(2)+0.0297 mL, or (2) pressure=0.0679×mL(1.742). The value for r2 was approximately 0.57 for both models. We found no correlation between the final pressure generated and the success of the epidural blood patch. We found a curvilinear relationship between the volume of blood injected during an epidural blood patch and the pressure generated in the epidural space. However, there was a large variation in both the volume of blood and the epidural pressure generated. The clinical importance of this finding is not known. A larger study would be required to demonstrate whether pressure is a predictor of success. Copyright © 2014 Elsevier Ltd. All rights reserved.

Are You Bleeding? Validation of a Machine-learning Algorithm for Determination of Blood Volume Status: Application to Remote Triage

DTIC Science & Technology

2014-01-09

workloads were determined by matching heart rate responses from each LBNP level. Heart rate and stroke volume (SV) were measured via Finom- eter. ECG, heat...learning algorithm for the assessment of central blood volume via pulse pressure [a noninvasive surrogate of stroke volume (SV)]. These data...30 130 125 120 115 110 105 100 Actual - Finometer Predicted - Algorithm Fig. 3. Comparison of average stroke volume (SV) derived from the

Development and first evaluation of a novel multiplex real-time PCR on whole blood samples for rapid pathogen identification in critically ill patients with sepsis.

PubMed

van de Groep, Kirsten; Bos, Martine P; Savelkoul, Paul H M; Rubenjan, Anna; Gazenbeek, Christel; Melchers, Willem J G; van der Poll, Tom; Juffermans, Nicole P; Ong, David S Y; Bonten, Marc J M; Cremer, Olaf L

2018-04-26

Molecular tests may enable early adjustment of antimicrobial therapy and be complementary to blood culture (BC) which has imperfect sensitivity in critically ill patients. We evaluated a novel multiplex real-time PCR assay to diagnose bloodstream pathogens directly in whole blood samples (BSI-PCR). BSI-PCR included 11 species- and four genus-specific PCRs, a molecular Gram-stain PCR, and two antibiotic resistance markers. We collected 5 mL blood from critically ill patients simultaneously with clinically indicated BC. Microbial DNA was isolated using the Polaris method followed by automated DNA extraction. Sensitivity and specificity were calculated using BC as reference. BSI-PCR was evaluated in 347 BC-positive samples (representing up to 50 instances of each pathogen covered by the test) and 200 BC-negative samples. Bacterial species-specific PCR sensitivities ranged from 65 to 100%. Sensitivity was 26% for the Gram-positive PCR, 32% for the Gram-negative PCR, and ranged 0 to 7% for yeast PCRs. Yeast detection was improved to 40% in a smaller set-up. There was no overall association between BSI-PCR sensitivity and time-to-positivity of BC (which was highly variable), yet Ct-values were lower for true-positive versus false-positive PCR results. False-positive results were observed in 84 (4%) of the 2200 species-specific PCRs in 200 culture-negative samples, and ranged from 0 to 6% for generic PCRs. Sensitivity of BSI-PCR was promising for individual bacterial pathogens, but still insufficient for yeasts and generic PCRs. Further development of BSI-PCR will focus on improving sensitivity by increasing input volumes and on subsequent implementation as a bedside test.

Systematic Review of the Use of Dried Blood Spots for Monitoring HIV Viral Load and for Early Infant Diagnosis

PubMed Central

Smit, Pieter W.; Sollis, Kimberly A.; Fiscus, Susan; Ford, Nathan; Vitoria, Marco; Essajee, Shaffiq; Barnett, David; Cheng, Ben; Crowe, Suzanne M.; Denny, Thomas; Landay, Alan; Stevens, Wendy; Habiyambere, Vincent; Perriens, Joseph H.; Peeling, Rosanna W.

2014-01-01

Background Dried blood spots (DBS) have been used as alternative specimens to plasma to increase access to HIV viral load (VL) monitoring and early infant diagnosis (EID) in remote settings. We systematically reviewed evidence on the performance of DBS compared to plasma for VL monitoring and EID. Methods and Findings Thirteen peer reviewed HIV VL publications and five HIV EID papers were included. Depending on the technology and the viral load distribution in the study population, the percentage of DBS samples that are within 0.5 log of VL in plasma ranged from 52–100%. Because the input sample volume is much smaller in a blood spot, there is a risk of false negatives with DBS. Sensitivity of DBS VL was found to be 78–100% compared to plasma at VL below 1000 copies/ml, but this increased to 100% at a threshold of 5000 copies/ml. Unlike a plasma VL test which measures only cell free HIV RNA, a DBS VL also measures proviral DNA as well as cell-associated RNA, potentially leading to false positive results when using DBS. The systematic review showed that specificity was close to 100% at DBS VL above 5000 copies/ml, and this threshold would be the most reliable for predicting true virologic failure using DBS. For early infant diagnosis, DBS has a sensitivity of 100% compared to fresh whole blood or plasma in all studies. Conclusions Although limited data are available for EID, DBS offer a highly sensitive and specific sampling strategy to make viral load monitoring and early infant diagnosis more accessible in remote settings. A standardized approach for sampling, storing, and processing DBS samples would be essential to allow successful implementation. Trial Registration PROSPERO Registration #: CRD42013003621. PMID:24603442

Quantifying Mesoscale Neuroanatomy Using X-Ray Microtomography

PubMed Central

Gray Roncal, William; Prasad, Judy A.; Fernandes, Hugo L.; Gürsoy, Doga; De Andrade, Vincent; Fezzaa, Kamel; Xiao, Xianghui; Vogelstein, Joshua T.; Jacobsen, Chris; Körding, Konrad P.

2017-01-01

Methods for resolving the three-dimensional (3D) microstructure of the brain typically start by thinly slicing and staining the brain, followed by imaging numerous individual sections with visible light photons or electrons. In contrast, X-rays can be used to image thick samples, providing a rapid approach for producing large 3D brain maps without sectioning. Here we demonstrate the use of synchrotron X-ray microtomography (µCT) for producing mesoscale (∼1 µm 3 resolution) brain maps from millimeter-scale volumes of mouse brain. We introduce a pipeline for µCT-based brain mapping that develops and integrates methods for sample preparation, imaging, and automated segmentation of cells, blood vessels, and myelinated axons, in addition to statistical analyses of these brain structures. Our results demonstrate that X-ray tomography achieves rapid quantification of large brain volumes, complementing other brain mapping and connectomics efforts. PMID:29085899

Advanced Techniques in Pulmonary Function Test Analysis Interpretation and Diagnosis

PubMed Central

Gildea, T.J.; Bell, C. William

1980-01-01

The Pulmonary Functions Analysis and Diagnostic System is an advanced clinical processing system developed for use at the Pulmonary Division, Department of Medicine at the University of Nebraska Medical Center. The system generates comparative results and diagnostic impressions for a variety of routine and specialized pulmonary functions test data. Routine evaluation deals with static lung volumes, breathing mechanics, diffusing capacity, and blood gases while specialized tests include lung compliance studies, small airways dysfunction studies and dead space to tidal volume ratios. Output includes tabular results of normal vs. observed values, clinical impressions and commentary and, where indicated, a diagnostic impression. A number of pulmonary physiological and state variables are entered or sampled (A to D) with periodic status reports generated for the test supervisor. Among the various physiological variables sampled are respiratory frequency, minute ventilation, oxygen consumption, carbon dioxide production, and arterial oxygen saturation.

The effects of platelet apheresis on blood saving and coagulation in bilateral total hip replacement: A prospective study on 60 patients.

PubMed

Qu, Zhijun; Wang, Geng; Xu, Chengshi; Zhang, Dazhi; Qu, Xiangdong; Zhou, Haibin; Ma, Jun

2016-10-01

Preoperative platelet rich plasma (PRP) harvest has been used in cardiopulmonary surgery for more than 10 years. There is no previous study dealing with PRP in bilateral total hip replacement. This study was to investigate the effects of PRP on blood saving and blood coagulation function in patients with bilateral total hip replacement. A prospective, randomized, clinical trial was conducted. Sixty patients were enrolled, including 30 patients undergoing PRP in the PRP group and 30 controls. The surgery time, total transfusion volume, blood loss, allogenic blood transfusion, autologous blood transfusion, urine volume, drainage volume, some blood parameters (including Fibrinogen, D-dimer, Prothrombin time, international normalizedratio, activated partial thromboplastin time, Platelet, Haemoglobin B), thrombelastogram (TEG) and blood-gas parameters were studied in the perioperative stage. The measurement data were analyzed statistically. There was no statistical difference between the two groups in baseline characteristics, surgery time, total transfusion volume, blood loss, autologous blood transfusion, etc. Allogenic blood transfusion in the PRP group was less than the control group with statistical difference (p = 0.024). Fibrinogen in the PRP group was higher than the control group (p = 0.008). Among the TEG indicators, activated clotting time and coagulation time K in the PRP group were less than the control group. Clotting rate and maximum amplitude in the PRP group were higher. The blood-gas parameters presented no statistical difference. The results suggested that PRP probably played a positive role in blood coagulation function as well as blood saving in patients with bilateral total hip replacement. Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.

Liquid chromatography-mass spectrometry platform for both small neurotransmitters and neuropeptides in blood, with automatic and robust solid phase extraction

NASA Astrophysics Data System (ADS)

Johnsen, Elin; Leknes, Siri; Wilson, Steven Ray; Lundanes, Elsa

2015-03-01

Neurons communicate via chemical signals called neurotransmitters (NTs). The numerous identified NTs can have very different physiochemical properties (solubility, charge, size etc.), so quantification of the various NT classes traditionally requires several analytical platforms/methodologies. We here report that a diverse range of NTs, e.g. peptides oxytocin and vasopressin, monoamines adrenaline and serotonin, and amino acid GABA, can be simultaneously identified/measured in small samples, using an analytical platform based on liquid chromatography and high-resolution mass spectrometry (LC-MS). The automated platform is cost-efficient as manual sample preparation steps and one-time-use equipment are kept to a minimum. Zwitter-ionic HILIC stationary phases were used for both on-line solid phase extraction (SPE) and liquid chromatography (capillary format, cLC). This approach enabled compounds from all NT classes to elute in small volumes producing sharp and symmetric signals, and allowing precise quantifications of small samples, demonstrated with whole blood (100 microliters per sample). An additional robustness-enhancing feature is automatic filtration/filter back-flushing (AFFL), allowing hundreds of samples to be analyzed without any parts needing replacement. The platform can be installed by simple modification of a conventional LC-MS system.

Prevalence and effect of Theileria orientalis infection in homebred calves in the Gloucester region of New South Wales, Australia.

PubMed

Swilks, E; Jenkins, C; Poynting, A; Collins, D; Krebs, G L

2017-06-01

To examine the prevalence of Theileria orientalis in the Gloucester area of New South Wales and its effect on individual animals. Blood samples (EDTA and clotted blood) were collected from a total of 55 calves and their dams from 6 properties over a 16-week period. A total of 202 and 190 blood samples were collected from the calves and dams, respectively, and were examined via blood film for the presence of intraerythrocytic T. orientalis piroplasms. Packed cell volume (PCV) was measured to determine infection resulting in anaemia. The presence of antibodies against the T. orientalis major piroplasm surface protein (MPSP) was tested using ELISA. The overall prevalence of T. orientalis infection in both dams and calves from all herds examined was 95%. Mean peak parasitaemia was observed in calves between 6 and 9 weeks of age (P = 0.051), coinciding with a decline in mean PCV. Only 3 (6%) of the blood samples collected from the dams were positive for Theileria-associated antibodies and no significant relationship (P > 0.05) was found between the presence of antibodies in the dams and PCV levels in the calves. There was no evidence that passive transfer of antibodies from dams to calves protected the calves against a decline in PCV. This study confirmed a high prevalence of low-level Theileria infection, but low MPSP seroconversion rates, in dams and calves in an area where the disease has been endemic for a number of years. © 2017 Australian Veterinary Association.

The intravascular volume effect of Ringer's lactate is below 20%: a prospective study in humans.

PubMed

Jacob, Matthias; Chappell, Daniel; Hofmann-Kiefer, Klaus; Helfen, Tobias; Schuelke, Anna; Jacob, Barbara; Burges, Alexander; Conzen, Peter; Rehm, Markus

2012-05-16

Isotonic crystalloids play a central role in perioperative fluid management. Isooncotic preparations of colloids (for example, human albumin or hydroxyethyl starch) remain nearly completely intravascular when infused to compensate for acute blood losses. Recent data were interpreted to indicate a comparable intravascular volume effect for crystalloids, challenging the occasionally suggested advantage of using colloids to treat hypovolemia. General physiological knowledge and clinical experience, however, suggest otherwise. In a prospective study, double-tracer blood volume measurements were performed before and after intended normovolemic hemodilution in ten female adults, simultaneously substituting the three-fold amount of withdrawn blood with Ringer's lactate. Any originated deficits were substituted with half the volume of 20% human albumin, followed by a further assessment of blood volume. To assess significance between the measurements, repeated measures analysis of variance (ANOVA) according to Fisher were performed. If significant results were shown, paired t tests (according to Student) for the singular measurements were taken. P < 0.05 was considered to be significant. A total of 1,097 ± 285 ml of whole blood were withdrawn (641 ± 155 ml/m(2) body surface area) and simultaneously replaced by 3,430 ± 806 ml of Ringer's lactate. All patients showed a significant decrease in blood volume after hemodilution (-459 ± 185 ml; P < 0.05) that did not involve relevant hemodynamical changes, and a significant increase in interstitial water content (+2,157 ± 606 ml; P < 0.05). The volume effect of Ringer's lactate was 17 ± 10%. The infusion of 245 ± 64 ml of 20% human albumin in this situation restored blood volume back to baseline values, the volume effect being 184 ± 63%. Substitution of isolated intravascular deficits in cardiopulmonary healthy adults with the three-fold amount of Ringer's lactate impedes maintenance of intravascular normovolemia. The main side effect was an impressive interstitial fluid accumulation, which was partly restored by the intravenous infusion of 20% human albumin. We recommend to substitute the five-fold amount of crystalloids or to use an isooncotic preparation in the face of acute bleeding in patients where edema prevention might be advantageous.

Comparison of the quantification of acetaminophen in plasma, cerebrospinal fluid and dried blood spots using high-performance liquid chromatography-tandem mass spectrometry

PubMed Central

Taylor, Rachel R.; Hoffman, Keith L.; Schniedewind, Björn; Clavijo, Claudia; Galinkin, Jeffrey L.; Christians, Uwe

2013-01-01

Acetaminophen (paracetamol, N-(4-hydroxyphenyl) acetamide) is one of the most commonly prescribed drugs for the management of pain in children. Quantification of acetaminophen in pre-term and term neonates and small children requires the availability of highly sensitive assays in small volume blood samples. We developed and validated an LC-MS/MS assay for the quantification of acetaminophen in human plasma, cerebro-spinal fluid (CSF) and dried blood spots (DBS). Reconstitution in water (DBS only) and addition of a protein precipitation solution containing the deuterated internal standard were the only manual steps. Extracted samples were analyzed on a Kinetex 2.6 μm PFP column using an acetonitrile/formic acid gradient. The analytes were detected in the positive multiple reaction mode. Alternatively, DBS were automatically processed using direct desorption in a sample card and preparation (SCAP) robotic autosampler in combination with online extraction. The range of reliable response in plasma and CSF was 3.05-20,000 ng/ml (r2 > 0.99) and 27.4-20,000 ng/ml (r2 > 0.99) for DBS (manual extraction and automated direct desorption). Inter-day accuracy was always within 85-115% and inter-day precision for plasma, CSF and manually extracted DBS were less than 15%. Deming regression analysis comparing 167 matching pairs of plasma and DBS samples showed a correlation coefficient of 0.98. Bland Altman analysis indicated a 26.6% positive bias in DBS, most likely reflecting the blood: plasma distribution ratio of acetaminophen. DBS are a valid matrix for acetaminophen pharmacokinetic studies. PMID:23670126

Use of Noncontrast Computed Tomography and Computed Tomographic Perfusion in Predicting Intracerebral Hemorrhage After Intravenous Alteplase Therapy.

PubMed

Batchelor, Connor; Pordeli, Pooneh; d'Esterre, Christopher D; Najm, Mohamed; Al-Ajlan, Fahad S; Boesen, Mari E; McDougall, Connor; Hur, Lisa; Fainardi, Enrico; Shankar, Jai Jai Shiva; Rubiera, Marta; Khaw, Alexander V; Hill, Michael D; Demchuk, Andrew M; Sajobi, Tolulope T; Goyal, Mayank; Lee, Ting-Yim; Aviv, Richard I; Menon, Bijoy K

2017-06-01

Intracerebral hemorrhage is a feared complication of intravenous alteplase therapy in patients with acute ischemic stroke. We explore the use of multimodal computed tomography in predicting this complication. All patients were administered intravenous alteplase with/without intra-arterial therapy. An age- and sex-matched case-control design with classic and conditional logistic regression techniques was chosen for analyses. Outcome was parenchymal hemorrhage on 24- to 48-hour imaging. Exposure variables were imaging (noncontrast computed tomography hypoattenuation degree, relative volume of very low cerebral blood volume, relative volume of cerebral blood flow ≤7 mL/min·per 100 g, relative volume of T max ≥16 s with all volumes standardized to z axis coverage, mean permeability surface area product values within T max ≥8 s volume, and mean permeability surface area product values within ipsilesional hemisphere) and clinical variables (NIHSS [National Institutes of Health Stroke Scale], onset to imaging time, baseline systolic blood pressure, blood glucose, serum creatinine, treatment type, and reperfusion status). One-hundred eighteen subjects (22 patients with parenchymal hemorrhage versus 96 without, median baseline NIHSS score of 15) were included in the final analysis. In multivariable regression, noncontrast computed tomography hypoattenuation grade ( P <0.006) and computerized tomography perfusion white matter relative volume of very low cerebral blood volume ( P =0.04) were the only significant variables associated with parenchymal hemorrhage on follow-up imaging (area under the curve, 0.73; 95% confidence interval, 0.63-0.83). Interrater reliability for noncontrast computed tomography hypoattenuation grade was moderate (κ=0.6). Baseline hypoattenuation on noncontrast computed tomography and very low cerebral blood volume on computerized tomography perfusion are associated with development of parenchymal hemorrhage in patients with acute ischemic stroke receiving intravenous alteplase. © 2017 American Heart Association, Inc.

[Comparison of MPure-12 Automatic Nucleic Acid Purification and Chelex-100 Method].

PubMed

Shen, X; Li, M; Wang, Y L; Chen, Y L; Lin, Y; Zhao, Z M; Que, T Z

2017-04-01

To explore the forensic application value of MPure-12 automatic nucleic acid purification （MPure-12 Method） for DNA extraction by extracting and typing DNA from bloodstains and various kinds of biological samples with different DNA contents. Nine types of biological samples, such as bloodstains, semen stains, and saliva were collected. DNA were extracted using MPure-12 method and Chelex-100 method, followed by PCR amplification and electrophoresis for obtaining STR-profiles. The samples such as hair root, chutty, butt, muscular tissue, saliva stain, bloodstain and semen stain were typed successfully by MPure-12 method. Partial alleles were lacked in the samples of saliva, and the genotyping of contact swabs was unsatisfactory. Additional, all of the bloodstains （20 μL, 15 μL, 10 μL, 5 μL, 1 μL） showed good typing results using Chelex-100 method. But the loss of alleles occurred in 1 μL blood volume by MPure-12 method. MPure-12 method is suitable for DNA extraction of a certain concentration blood samples．Chelex-100 method may be better for the extraction of trace blood samples．This instrument used in nucleic acid extraction has the advantages of simplicity of operator, rapidity, high extraction efficiency, high rate of reportable STR-profiles and lower man-made pollution. Copyright© by the Editorial Department of Journal of Forensic Medicine

Rehabilitation with dental prosthesis can increase cerebral regional blood volume.

PubMed

Miyamoto, Ikuya; Yoshida, Kazuya; Tsuboi, Yoichi; Iizuka, Tadahiko

2005-12-01

Treatment with denture for edentulous people is highly important for maintaining quality of life. However, its effect on the brain is unknown. In this experimental study, we hypothesized that dental prosthesis can recover not only the physical condition of mastication system but also the regional brain activity. We evaluated functional brain imaging of edentulous subjects fixed by dental implant prosthesis with clenching tasks by multi-channel near-infrared optical topography. Results revealed a significantly (P<0.001; paired t-test) increased cerebral regional blood volume during maximum voluntary clenching task by implant-retained prosthesis. There were no statistically significant differences between patients with and without prosthesis in the latency to the maximum regional blood volume after the task. Conclusively, clenching can be effective for increasing cerebral blood volume; accordingly maintenance of normal chewing might prevent the brain from degenerating.

Effect of pre-donation fluid intake on fluid shift from interstitial to intravascular compartment in blood donors.

PubMed

Deepika, Chenna; Murugesan, Mohandoss; Shastry, Shamee

2018-02-01

Fluid shifts from interstitial to intravascular space during blood donation helps in compensating the lost blood volume. We aimed to determine the volume of fluid shift following donation in donors with and without pre-donation fluid intake. We studied the fluid shift in 325 blood donors prospectively. Donors were divided in groups- with no fluid intake (GI) and either water (GII) or oral rehydrating fluids (GIII) before donation. Fluid shift following donation was calculated based on the difference between the pre and post donation blood volume. The influence of oral fluid intake, age, gender and body mass index (BMI) on volume of fluid shift was analyzed. The fluid shift was significant between donors without fluids (GI: 127 ± 81 ml) and donors with fluid intake (GII & III: 96 ± 45 ml) (p < 0.05). The difference was not significant between donors with water intake (GII: 106 ± 52 ml) and oral rehydrating fluid intake (GIII: 87 ± 41 ml). The shifted fluid volume increased with increasing BMI and decreased with increasing age in females. The fluid shift increased in females than in males. The age, gender, BMI and VVR did not significantly contribute to the volume of fluid shift following donation. As per our observation, the oral fluids before donation might not contribute to increase in fluid shift in blood donors after donation. Copyright © 2017 Elsevier Ltd. All rights reserved.

State of the art in fluid and volume therapy : A user-friendly staged concept. English version.

PubMed

Rehm, M; Hulde, N; Kammerer, T; Meidert, A S; Hofmann-Kiefer, K

2017-04-10

Adequate intraoperative infusion therapy is essential for the perioperative outcome of a patient. Both hypo- and hypervolemia can lead to an increased rate of perioperative complications and to a worse outcome. Perioperative infusion therapy should therefore be needs-based. The primary objective is the maintenance of preoperative normovolemia using a rational infusion strategy. Perioperative fluid losses should be differentiated from volume losses due to surgical bleeding or protein losses into the interstitial space. Fluid loss via urine excretion or insensible perspiration (0.5-1.0 ml/kg/h) should be replaced with balanced, isooncotic, crystalloid infusion solutions in a ratio of 1:1. Volume therapy stage 1: intraoperative volume losses up to a blood loss corresponding to 20% of the patient's total blood volume are compensated for by balanced crystalloids in a ratio of 4-5:1. Stage 2: blood losses exceeding this level are to be treated with isooncotic colloids (preferably balanced) in a 1:1 ratio. In this regard taking into consideration the contraindications, e. g., sepsis, burns, critical illness (usually patients in the intensive care unit), impaired renal function or renal replacement therapy, intracranial hemorrhage, or severe coagulopathy, artificial colloids such as hydroxyethyl starch (HES) can be used perioperatively for volume replacement. Stage 3: if an allogeneic blood transfusion is indicated, blood and blood products are applied in a differentiated manner.

Optimal method for collection of umbilical cord blood: an Egyptian trial for a public cord blood bank.

PubMed

Bassiouny, M R; El-Chennawi, F; Mansour, A K; Yahia, S; Darwish, A

2015-06-01

Umbilical cord blood (UCB) contains stem cells and can be used as an alternative to bone marrow transplantation. Engraftment is dependent on the total nucleated cell (TNC) and CD34+ cell counts of the cord blood units. This study was designed to evaluate the effect of the method of collection of the UCB on the yield of the cord blood units. Informed consent was obtained from 100 eligible mothers for donation of cord blood. Both in utero and ex utero methods were used for collection. The cord blood volume was measured. The TNC and the CD34+ cell counts were enumerated. We have found that in utero collection gave significantly larger volumes of cord blood and higher TNC counts than ex utero collection. There was no significant difference between both methods regarding the CD34+ cell counts. This study revealed a significant correlation between the volume of the collected cord blood and both TNC and CD34+ cell counts. It is better to collect cord blood in utero before placental delivery to optimize the quality of the cord blood unit. © 2015 AABB.

Regulation of Blood Volume During Spaceflight

NASA Technical Reports Server (NTRS)

Alfrey, Clarence P.

1997-01-01

The effects of spaceflight on erythropoiesis and blood volume in the rat were studied during the 14-day NASA Spacelab Life Sciences 2 (SLS-2) Shuttle mission. Measurements included red blood cell mass (RBCM), plasma volume (PV), iron utilization and iron utilization in response to an injection of erythropoietin. Red blood cell (RBC) survival, splenic sequestration and erythrocyte morphology were also evaluated. At landing, the RBCM adjusted for body weight was significantly lower in the flight animals than in the ground controls. While the PV was also decreased, the change was not statistically significant. Incorporation of iron into circulating RBCs was normal when measured after five days of spaceflight and the rat responded normally to the single in-flight injection of erythropoietin. No change in RBC morphology could be attributed to spaceflight. A normal survival was found for the RBC population that was represented by Cr-51 labeled RBCS. These results demonstrate that rats, like humans, return from spaceflight with a decreased RBCM and total blood volume.

Shortened dental arch and cerebral regional blood volume: an experimental pilot study with optical topography.

PubMed

Miyamoto, Ikuya; Yoshida, Kazuya; Bessho, Kazuhisa

2009-04-01

A shortened dental arch without posterior occlusal support has been thought to maintain sufficient oral function. The mechanism of occlusal adaptation with a shortened dental arch is unclear. For a better understanding of the effects of molar teeth on brain function, the authors combined experimentally-shortened dental arches and a neuro-imaging technique. Regional cerebral blood volume was measured using near-infrared optical topography during maximum voluntary clenching tasks from 10 subjects on individually fabricated oral appliances, which can create experimentally complete and shortened dental arches. Results suggested that clenching on the complete dental arch showed a significantly higher brain blood volume than that on the shortened dental arch. Moreover, there were no differences between the two splints in the latency to the maximum oxyhemoglobin concentration. These findings suggest that occlusal status is closely related to brain blood flow and lack of occlusal molar support rapidly reduces cerebral blood volume in the maximum voluntary clenching condition.

Effect of blood activity on dosimetric calculations for radiopharmaceuticals

NASA Astrophysics Data System (ADS)

Zvereva, Alexandra; Petoussi-Henss, Nina; Li, Wei Bo; Schlattl, Helmut; Oeh, Uwe; Zankl, Maria; Graner, Frank Philipp; Hoeschen, Christoph; Nekolla, Stephan G.; Parodi, Katia; Schwaiger, Markus

2016-11-01

The objective of this work was to investigate the influence of the definition of blood as a distinct source on organ doses, associated with the administration of a novel radiopharmaceutical for positron emission tomography-computed tomography (PET/CT) imaging—(S)-4-(3-18F-fluoropropyl)-L-glutamic acid (18F-FSPG). Personalised pharmacokinetic models were constructed based on clinical PET/CT images from five healthy volunteers and blood samples from four of them. Following an identifiability analysis of the developed compartmental models, person-specific model parameters were estimated using the commercial program SAAM II. Organ doses were calculated in accordance to the formalism promulgated by the Committee on Medical Internal Radiation Dose (MIRD) and the International Commission on Radiological Protection (ICRP) using specific absorbed fractions for photons and electrons previously derived for the ICRP reference adult computational voxel phantoms. Organ doses for two concepts were compared: source organ activities in organs parenchyma with blood as a separate source (concept-1); aggregate activities in perfused source organs without blood as a distinct source (concept-2). Aggregate activities comprise the activities of organs parenchyma and the activity in the regional blood volumes (RBV). Concept-1 resulted in notably higher absorbed doses for most organs, especially non-source organs with substantial blood contents, e.g. lungs (92% maximum difference). Consequently, effective doses increased in concept-1 compared to concept-2 by 3-10%. Not considering the blood as a distinct source region leads to an underestimation of the organ absorbed doses and effective doses. The pronounced influence of the blood even for a radiopharmaceutical with a rapid clearance from the blood, such as 18F-FSPG, suggests that blood should be introduced as a separate compartment in most compartmental pharmacokinetic models and blood should be considered as a distinct source in dosimetric calculations. Hence, blood samples should be included in all pharmacokinetic and dosimetric studies for new tracers if possible.

Mature red blood cells: from optical model to inverse light-scattering problem.

PubMed

Gilev, Konstantin V; Yurkin, Maxim A; Chernyshova, Ekaterina S; Strokotov, Dmitry I; Chernyshev, Andrei V; Maltsev, Valeri P

2016-04-01

We propose a method for characterization of mature red blood cells (RBCs) morphology, based on measurement of light-scattering patterns (LSPs) of individual RBCs with the scanning flow cytometer and on solution of the inverse light-scattering (ILS) problem for each LSP. We considered a RBC shape model, corresponding to the minimal bending energy of the membrane with isotropic elasticity, and constructed an analytical approximation, which allows rapid simulation of the shape, given the diameter and minimal and maximal thicknesses. The ILS problem was solved by the nearest-neighbor interpolation using a preliminary calculated database of 250,000 theoretical LSPs. For each RBC in blood sample we determined three abovementioned shape characteristics and refractive index, which also allows us to calculate volume, surface area, sphericity index, spontaneous curvature, hemoglobin concentration and content.

Mature red blood cells: from optical model to inverse light-scattering problem

PubMed Central

Gilev, Konstantin V.; Yurkin, Maxim A.; Chernyshova, Ekaterina S.; Strokotov, Dmitry I.; Chernyshev, Andrei V.; Maltsev, Valeri P.

2016-01-01

We propose a method for characterization of mature red blood cells (RBCs) morphology, based on measurement of light-scattering patterns (LSPs) of individual RBCs with the scanning flow cytometer and on solution of the inverse light-scattering (ILS) problem for each LSP. We considered a RBC shape model, corresponding to the minimal bending energy of the membrane with isotropic elasticity, and constructed an analytical approximation, which allows rapid simulation of the shape, given the diameter and minimal and maximal thicknesses. The ILS problem was solved by the nearest-neighbor interpolation using a preliminary calculated database of 250,000 theoretical LSPs. For each RBC in blood sample we determined three abovementioned shape characteristics and refractive index, which also allows us to calculate volume, surface area, sphericity index, spontaneous curvature, hemoglobin concentration and content. PMID:27446656

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco.

PubMed

Ait Lbacha, H; Alali, S; Zouagui, Z; El Mamoun, L; Rhalem, A; Petit, E; Haddad, N; Gandoin, C; Boulouis, H-J; Maillard, R

2017-02-01

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases. © 2015 Blackwell Verlag GmbH.

Optimizing direct amplification of forensic commercial kits for STR determination.

PubMed

Caputo, M; Bobillo, M C; Sala, A; Corach, D

2017-04-01

Direct DNA amplification in forensic genotyping reduces analytical time when large sample sets are being analyzed. The amplification success depends mainly upon two factors: on one hand, the PCR chemistry and, on the other, the type of solid substrate where the samples are deposited. We developed a workflow strategy aiming to optimize times and cost when starting from blood samples spotted onto diverse absorbent substrates. A set of 770 blood samples spotted onto Blood cards, Whatman ® 3 MM paper, FTA™ Classic cards, and Whatman ® Grade 1 was analyzed by a unified working strategy including a low-cost pre-treatment, a PCR amplification volume scale-down, and the use of the 3500 Genetic Analyzer as the analytical platform. Samples were analyzed using three different commercial multiplex STR direct amplification kits. The efficiency of the strategy was evidenced by a higher percentage of high-quality profiles obtained (over 94%), a reduced number of re-injections (average 3.2%), and a reduced amplification failure rate (lower than 5%). Average peak height ratio among different commercial kits was 0.91, and the intra-locus balance showed values ranging from 0.92 to 0.94. A comparison with previously reported results was performed demonstrating the efficiency of the proposed modifications. The protocol described herein showed high performance, producing optimal quality profiles, and being both time and cost effective. Copyright © 2017 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

Leukodepletion as a Point-of-Care Method for Monitoring HIV-1 Viral Load in Whole Blood

PubMed Central

Titchmarsh, Logan; Zeh, Clement; Verpoort, Thierry; Allain, Jean-Pierre

2014-01-01

In order to limit the interference of HIV-1 cellular nucleic acids in estimating viral load (VL), the feasibility of leukodepletion of a small whole-blood (WB) volume to eliminate only leukocyte cell content was investigated, using a selection of filters. The efficacy of leukocyte filtration was evaluated by counting, CD45 quantitative PCR, and HIV-1 DNA quantification. Plasma HIV-1 was tested by real-time reverse transcription (RT)-PCR. A specific, miniaturized filter was developed and tested for leukocyte and plasma virus retention, WB sample dilution, and filtration parameters in HIV-1-spiked WB samples. This device proved effective to retain >99.9% of white blood cells in 100 μl of WB without affecting plasma VL. The Samba sample preparation chemistry was adapted to use a leukodepleted WB sample for VL monitoring using the point-of-care Samba-1 semiautomated system. The clinical performance of the assay was evaluated by testing 207 consecutive venous EDTA WB samples from HIV-1-infected patients attending a CD4 testing clinic. Most patients were on antiretroviral treatment (ART), but their VL status was unknown. Compared to the Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test, the new Samba assay had a concordance of 96.5%. The use of the Samba system with a VL test for WB might contribute to HIV-1 ART management and reduce loss-to-follow-up rates in resource-limited settings. PMID:25428162

A randomized, controlled, double-blind crossover study on the effects of 2-L infusions of 0.9% saline and plasma-lyte® 148 on renal blood flow velocity and renal cortical tissue perfusion in healthy volunteers.

PubMed

Chowdhury, Abeed H; Cox, Eleanor F; Francis, Susan T; Lobo, Dileep N

2012-07-01

We compared the effects of intravenous infusions of 0.9% saline ([Cl] 154 mmol/L) and Plasma-Lyte 148 ([Cl] 98 mmol/L, Baxter Healthcare) on renal blood flow velocity and perfusion in humans using magnetic resonance imaging (MRI). Animal experiments suggest that hyperchloremia resulting from 0.9% saline infusion may affect renal hemodynamics adversely, a phenomenon not studied in humans. Twelve healthy adult male subjects received 2-L intravenous infusions over 1 hour of 0.9% saline or Plasma-Lyte 148 in a randomized, double-blind manner. Crossover studies were performed 7 to 10 days apart. MRI scanning proceeded for 90 minutes after commencement of infusion to measure renal artery blood flow velocity and renal cortical perfusion. Blood was sampled and weight recorded hourly for 4 hours. Sustained hyperchloremia was seen with saline but not with Plasma-Lyte 148 (P < 0.0001), and fall in strong ion difference was greater with the former (P = 0.025). Blood volume changes were identical (P = 0.867), but there was greater expansion of the extravascular fluid volume after saline (P = 0.029). There was a significant reduction in mean renal artery flow velocity (P = 0.045) and renal cortical tissue perfusion (P = 0.008) from baseline after saline, but not after Plasma-Lyte 148. There was no difference in concentrations of urinary neutrophil gelatinase-associated lipocalin after the 2 infusions (P = 0.917). This is the first human study to demonstrate that intravenous infusion of 0.9% saline results in reductions in renal blood flow velocity and renal cortical tissue perfusion. This has implications for intravenous fluid therapy in perioperative and critically ill patients. NCT01087853.

The Aachen miniaturized heart-lung machine--first results in a small animal model.

PubMed

Schnoering, Heike; Arens, Jutta; Sachweh, Joerg S; Veerman, Melanie; Tolba, Rene; Schmitz-Rode, Thomas; Steinseifer, Ulrich; Vazquez-Jimenez, Jaime F

2009-11-01

Congenital heart surgery most often incorporates extracorporeal circulation. Due to foreign surface contact and the administration of foreign blood in many children, inflammatory response and hemolysis are important matters of debate. This is particularly an issue in premature and low birth-weight newborns. Taking these considerations into account, the Aachen miniaturized heart-lung machine (MiniHLM) with a total static priming volume of 102 mL (including tubing) was developed and tested in a small animal model. Fourteen female Chinchilla Bastard rabbits were operated on using two different kinds of circuits. In eight animals, a conventional HLM with Dideco Kids oxygenator and Stöckert roller pump (Sorin group, Milan, Italy) was used, and the Aachen MiniHLM was employed in six animals. Outcome parameters were hemolysis and blood gas analysis including lactate. The rabbits were anesthetized, and a standard median sternotomy was performed. The ascending aorta and the right atrium were cannulated. After initiating cardiopulmonary bypass, the aorta was cross-clamped, and cardiac arrest was induced by blood cardioplegia. Blood samples for hemolysis and blood gas analysis were drawn before, during, and after cardiopulmonary bypass. After 1 h aortic clamp time, all animals were weaned from cardiopulmonary bypass. Blood gas analysis revealed adequate oxygenation and perfusion during cardiopulmonary bypass, irrespective of the employed perfusion system. The use of the Aachen MiniHLM resulted in a statistically significant reduced decrease in fibrinogen during cardiopulmonary bypass. A trend revealing a reduced increase in free hemoglobin during bypass in the MiniHLM group could also be observed. This newly developed Aachen MiniHLM with low priming volume, reduced hemolysis, and excellent gas transfer (O(2) and CO(2)) may reduce circuit-induced complications during heart surgery in neonates.

Comparison of capillary electrophoresis and high performance liquid chromatography for detection and quantification of hemoglobin New York.

PubMed

You-Qiong, Li; Hui-Ping, Huang; Zhi-Zhong, Chen; Lin, Zhao; Liang, Liang; Gui-Fang, Qin; Yun, Mo

2016-01-01

Hemoglobin (Hb) New York [β113 (G15) Val→Glu, GTG→GAG], also known as Hb Kaohsiung, is one of the most common Hb variants in South China. Currently, most used screening methods for hemoglobinopathies in South China are high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). However, there is no study comparing the performance of CE and HPLC in the detection of Hb New York. In total 15 samples (including 13 adult blood samples and 2 cord blood samples) with heterozygous Hb New York were analyzed by CE and HPLC. Levels of Hb New York, HbA2, Hb, mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were compared within the two groups. All 15 cases (100%) were detected by CE, whereas none was detected by HPLC. Mean values of Hb New York and HbA2 in adult heterozygous group were (43.82±2.47)% and (2.85±0.31)%, respectively; mean values for cord blood group were (7.95±0.78)% and (0.30±0.14)%. CE allows the detection of Hb New York, while this variant is not separated from Hb A on HPLC. CE may be the preferred method for hemoglobinopathy screening in areas with high prevalence of Hb New York.

Isovolemic hemodilution alters the ratio of whole-body to large-vessel hematocrit (F-cell ratio). A prospective, randomized study comparing the volume effects of hydroxyethyl starch 200,000/0.62 and albumin.

PubMed

Haller, M; Brechtelsbauer, H; Akbulut, C; Fett, W; Briegel, J; Finsterer, U

1995-04-01

To evaluate potential changes in the ratio of whole-body/large-vessel hematocrit (f-cell ratio) during isovolemic hemodilution and to compare the volume effects of 2 different plasma exchange solutions (hydroxyethyl starch 200,000/0.62 6% and human albumin 5%). Prospective, randomized, controlled trial. Operating theater in a university hospital. 24 gynecological patients scheduled for elective surgery. Isovolemic hemodilution was performed using 2 different plasma exchange solutions. Plasma volume was determined using dye dilution technique before and after hemodilution. The volume of withdrawn blood was measured from the change in weight of the blood bags taking into account the specific gravity of blood. The volume of administered plasma exchange solutions exceeded the amount of withdrawn blood by 80 +/- 47 ml (p < 0.001). Plasma volume was 3,067 +/- 327 ml before and 3,517 +/- 458 ml after hemodilution. Using red cell volumes calculated from measured plasma volumes and peripheral hematocrit, a deficit of 249 +/- 133 ml (p < 0.0001) in red cells after hemodilution appeared with the measured withdrawn red cell volumes taken into account. This finding can be explained by a change in the f-cell ratio during isovolemic hemodilution. The volume effect of the exchange solutions was 1.05 for hydroxyethyl starch and 0.95 for albumin. The results demonstrate that a change in the f-cell ratio occurs during isovolemic hemodilution. The estimation of red cell volume or plasma volume changes by using either the hematocrit or plasma or red cell volume determinations together with the hematocrit may lead to erroneous results.

A comprehensive Guyton model analysis of physiologic responses to preadapting the blood volume as a countermeasure to fluid shifts

NASA Technical Reports Server (NTRS)

Simanonok, K. E.; Srinivasan, R. S.; Myrick, E. E.; Blomkalns, A. L.; Charles, J. B.

1994-01-01

The Guyton model of fluid, electrolyte, and circulatory regulation is an extensive mathematical model capable of simulating a variety of experimental conditions. It has been modified for use at NASA to simulate head-down tilt, a frequently used analog of weightlessness. Weightlessness causes a headward shift of body fluids that is believed to expand central blood volume, triggering a series of physiologic responses resulting in large losses of body fluids. We used the modified Guyton model to test the hypothesis that preadaptation of the blood volume before weightless exposure could counteract the central volume expansion caused by fluid shifts, and thereby attenuate the circulatory and renal responses that result in body fluid losses. Simulation results show that circulatory preadaptation, by a procedure resembling blood donation immediately before head-down bedrest, is effective in damping the physiologic responses to fluid shifts and reducing body fluid losses. After 10 hours of head-down tilt, preadaptation also produces higher blood volume, extracellular volume, and total body water for 20 to 30 days of bedrest, compared with non-preadapted control. These results indicate that circulatory preadaptation before current Space Shuttle missions may be beneficial for the maintenance of reentry and postflight orthostatic tolerance in astronauts. This paper presents a comprehensive examination of the simulation results pertaining to changes in relevant physiologic variables produced by blood volume reduction before a prolonged head-down tilt. The objectives were to study and develop the countermeasure theoretically, to aid in planning experimental studies of the countermeasure, and to identify potentially disadvantageous physiologic responses that may be caused by the countermeasure.

Small-volume resuscitation from hemorrhagic shock with polymerized human serum albumin.

PubMed

Messmer, Catalina; Yalcin, Ozlem; Palmer, Andre F; Cabrales, Pedro

2012-10-01

Human serum albumin (HSA) is used as a plasma expander; however, albumin is readily eliminated from the intravascular space. The objective of this study was to establish the effects of various-sized polymerized HSAs (PolyHSAs) during small-volume resuscitation from hemorrhagic shock on systemic parameters, microvascular hemodynamics, and functional capillary density in the hamster window chamber model. Polymerized HSA size was controlled by varying the cross-link density (ie, molar ratio of glutaraldehyde to HSA). Hemorrhage was induced by controlled arterial bleeding of 50% of the animal's blood volume (BV), and hypovolemic shock was maintained for 1 hour. Resuscitation was implemented in 2 phases, first, by infusion of 3.5% of the BV of hypertonic saline (7.5% NaCl) then followed by infusion of 10% of the BV of each PolyHSA. Resuscitation provided rapid recovery of blood pressure, blood gas parameters, and microvascular perfusion. Polymerized HSA at a glutaraldehyde-to-HSA molar ratio of 60:1 (PolyHSA(60:1)) provided superior recovery of blood pressure, microvascular blood flow, and functional capillary density, and acid-base balance, with sustained volume expansion in relation to the volume infused. The high molecular weight of PolyHSA(60:1) increased the hydrodynamic radius and solution viscosity. Pharmacokinetic analysis of PolyHSA(60:1) indicates reduced clearance and increased circulatory half-life compared with monomeric HSA and other PolyHSA formulations. In conclusion, HSA molecular size and solution viscosity affect central hemodynamics, microvascular blood flow, volume expansion, and circulation persistence during small-volume resuscitation from hemorrhagic shock. In addition, PolyHSA can be an alternative to HSA in pathophysiological situations with compromised vascular permeability. Copyright © 2012 Elsevier Inc. All rights reserved.

Hemolysis area: A new parameter of erythrocyte osmotic fragility for screening of thalassemia trait

PubMed Central

Tatu, Thanusak; Sweatman, Denis

2018-01-01

BACKGROUND: One-tube osmotic fragility test (OFT) is widely used for screening thalassemia traits. Interobserver variation may occur with 0.36% NaCl-based OFT due to the naked eye result reading style. PURPOSE: The purpose of this study was to establish and evaluate the novel numerical OFT-based parameter, so-called hemolysis area (HA), in screening thalassemia traits. MATERIALS AND METHODS: The portable spectrophotometer was invented capable of calculating the HA values. The HA values were then compared among 69, 156, and 19 blood samples having positive, negative, and suspicious 0.36% NaCl-based OFT results, respectively; 109 and 135 blood samples having mean corpuscular volume (MCV) ≤80 fL and >80 fL, respectively; and 138 and 106 blood samples having mean corpuscular hemoglobin (MCH) ≤27 pg and >27 pg, respectively. In addition, the HA values were compared in 166 blood samples having different globin gene genotypes. Finally, the HA cutoff value was determined by receiver operation curve (ROC) analysis. RESULTS: The HA values in samples having positive, suspicious, and negative 0.36% NaCl-based OFT were 33.3 ± 14.4, 42.9 ± 10.5, and 65.3 ± 13.4, respectively; in sample having MCV ≤80 fL and >80 fL were 43.1 ± 19.6 and 63.8 ± 14.5, respectively; and in samples having MCH ≤27 pg and >27 pg were 46.7 ± 20.1 and 64.8 ± 14.2, respectively. The HA values in normal, hemoglobin E, SEA-α thalassemia 1, and β-thalassemia traits were 67.1 ± 12.6, 36.4 ± 13.9, 20.2 ± 4.8, and 18.6 ± 1.1, respectively. All were significantly different. ROC analysis established 52.4 as the HA cutoff that had comparable effectiveness to the conventional screening tests. CONCLUSION: The new HA value was effective and could be an alternative choice for screening thalassemia traits. PMID:29692590

Assessing intravascular volume by difference in pulse pressure in pigs submitted to graded hemorrhage.

PubMed

Pestel, Gunther J; Hiltebrand, Luzius B; Fukui, Kimiko; Cohen, Delphine; Hager, Helmut; Kurz, Andrea M

2006-10-01

We assessed changes in intravascular volume monitored by difference in pulse pressure (dPP%) after stepwise hemorrhage in an experimental pig model. Six pigs (23-25 kg) were anesthetized (isoflurane 1.5 vol%) and mechanically ventilated to keep end-tidal CO2 (etCO2) at 35 mmHg. A PA-catheter and an arterial catheter were placed via femoral access. During and after surgery, animals received lactated Ringer's solution as long as they were considered volume responders (dPP>13%). Then animals were allowed to stabilize from the induction of anesthesia and insertion of catheters for 30 min. After stabilization, baseline measurements were taken. Five percent of blood volume was withdrawn, followed by another 5%, and then in 10%-increments until death from exsanguination occurred. After withdrawal of 5% of blood volume, all pigs were considered volume responders (dPP>13%); dPP rose significantly from 6.1+/-3.3% to 19.4+/-4.2%. The regression analysis of stepwise hemorrhage revealed a linear relation between blood loss (hemorrhage in %) and dPP (y=0.99*x+14; R2=0.7764; P<.0001). In addition, dPP was the only parameter that changed significantly between baseline and a blood loss of 5% (P<0.01), whereas cardiac output, stroke volume, heart rate, MAP, central venous pressure, pulmonary artery occlusion pressure, and systemic vascular resistance, respectively, remained unchanged. We conclude that in an experimental hypovolemic pig model, dPP correlates well with blood loss.

Higher Donor Apheresis Blood Volumes Are Associated with Reduced Relapse Risk and Improved Survival in Reduced-Intensity Allogeneic Transplantations with Unrelated Donors.

PubMed

Crisalli, Lisa M; Hinkle, Joanne T; Walling, Christopher C; Sell, Mary; Frey, Noelle V; Hexner, Elizabeth O; Loren, Alison W; Luger, Selina M; Stadtmauer, Edward A; Porter, David L; Reshef, Ran

2018-06-01

Allogeneic hematopoietic stem cell transplantation (HSCT) with reduced-intensity conditioning (RIC) offers a curative option for patients with hematologic malignancies who are unable to undergo myeloablative conditioning, but its success is limited by high rates of relapse. Several studies have suggested a role for T cell doses in peripheral blood stem cell grafts in RIC HSCT. Because T cell dose is typically not known until after the collection, and apheresis blood volume is easily modifiable, we hypothesized that higher donor apheresis blood volumes would improve transplantation outcomes through an effect on graft composition. Thus, we analyzed the relationships between apheresis volume, graft composition, and transplantation outcomes in 142 consecutive patients undergoing unrelated donor allogeneic RIC HSCT. We found that apheresis volume ≥15 L was associated with a significantly decreased risk of relapse (adjusted hazard ratio [aHR], .48; 95% confidence interval [CI], .28 to .84]; P = .01) and improved relapse-free survival (aHR, .56; 95% CI, .35 to .89; P = .02) and overall survival (aHR, .55; 95% CI, .34 to .91; P = .02). A high apheresis volume was not associated with increased rates of acute or chronic graft-versus-host disease. These results demonstrate that an apheresis volume of at least 15 L is independently predictive of improved transplantation outcomes after RIC allogeneic HSCT. Copyright © 2018 The American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved.

The effect of vitamin C on plasma volume in the early stage of sepsis in the rat.

PubMed

Bark, Björn P; Grände, Per-Olof

2014-12-01

Previous experimental studies have shown that vitamin C has several beneficial effects in sepsis and burns, such as decreased tissue oedema, improved endothelial barrier function and decreased transcapillary leakage of plasma markers. It has still not been investigated, though, if vitamin C has any impact specifically on plasma volume. The present study aims at testing the hypothesis that vitamin C decreases plasma volume loss in sepsis. Anaesthetized male adult Sprague-Dawley rats were used in this prospective randomized study. All experiments were carried out at a university hospital laboratory. Sepsis was induced by caecal ligation and incision. After 3 h, vitamin C was given either as a bolus dose (66 mg/kg) followed by a continuous infusion (33 mg/kg/h) (n = 9), or as a single bolus dose (200 mg/kg) (n = 9). A sham group (n = 9) underwent the same surgical procedure, but no vitamin C was given. Plasma volume was measured ((125)I-dilution technique) at baseline, at 3 h after end of initiation of sepsis and at the end of the experiment 3 h later. Arterial blood samples for analyses of electrolytes, blood gases, haematocrit and lactate were taken at the same time points. There were no significant differences in plasma volumes or the physiological parameters analysed between any of the three groups at any time point. There was a significantly larger urine production in the single bolus dose group (200 mg/kg) compared to the sham group. Vitamin C treatment did not decrease the loss of plasma volume in the septic rat. The diuretic effect of vitamin C was in accordance with previous studies.

CHF: circulatory homeostasis gone awry.

PubMed

Weber, Karl T; Burlew, Brad S; Davis, Richard C; Newman, Kevin P; D'Cruz, Ivan A; Hawkins, Ralph G; Wall, Barry M; Parker, Robert B

2002-01-01

The role of the renin-angiotensin-aldosterone system (RAAS) is integral to salt and water retention, particularly by the kidneys. Over time, positive sodium balance leads first to intra- and then to extravascular volume expansion, with subsequent symptomatic heart failure. This report examines the role of the RAAS in regulating a less well recognized component essential to circulatory homeostasis--central blood volume. The regulation of central blood volume draws on integrative cardiorenal physiology and a key role played by the RAAS in its regulation. In presenting insights into the role of the RAAS in regulating central blood volume, this review also addresses other sodium-retaining states with a predisposition to edema formation, such as cirrhosis and nephrosis. (c)2002 CHF, Inc

Clarification of the circulatory patho-physiology of anaesthesia - implications for high-risk surgical patients.

PubMed

Wolff, Christopher B; Green, David W

2014-12-01

The paper examines the effects of anaesthesia on circulatory physiology and their implications regarding improvement in perioperative anaesthetic management. Changes to current anaesthetic practice, recommended recently, such as the use of flow monitoring in high risk patients, are already beginning to have an impact in reducing complications but not mortality [1]. Better understanding of the patho-physiology should help improve management even further. Analysis of selected individual clinical trials has been used to illustrate particular areas of patho-physiology and how changes in practice have improved outcome. There is physiological support for the importance of achieving an appropriate rate of oxygen delivery (DO2), particularly following induction of anaesthesia. It is suggested that ensuring adequate DO2 during anaesthesia will avoid development of oxygen debt and hence obviate the need to induce a high, compensatory, DO2 in the post-operative period. In contrast to the usual assumptions underlying strategies requiring a global increase in blood flow [1] by a stroke volume near maximization strategy, blood flow control actually resides entirely at the tissues not at the heart. This is important as the starting point for understanding failed circulatory control as indicated by 'volume dependency'. Local adjustments in blood flow at each individual organ - auto-regulation - normally ensure the appropriate local rate of oxygen supply, i.e. local DO2. Inadequate blood volume leads to impairment of the regulation of blood flow, particularly in the individual tissues with least capable auto-regulatory capability. As demonstrated by many studies, inadequate blood flow first occurs in the gut, brain and kidney. The inadequate blood volume which occurs with induction of anaesthesia is not due to blood volume loss, but probably results from redistribution due to veno-dilation. The increase in venous capacity renders the existing blood volume inadequate to maintain venous return and pre-load. Blood volume shifted to the veins will, necessarily, also reduce the arterial volume. As a result stroke volume and cardiac output fall below normal with little or no change in peripheral resistance. The resulting pre-load dependency is often successfully treated with colloid infusion and, in some studies, 'inotropic' agents, particularly in the immediate post-operative phase. Treatment during the earliest stage of anaesthesia can avoid the build up of oxygen debt and may be supplemented by drugs which maintain or restore venous tone, such as phenylephrine; an alternative to volume expansion. Interpretation of circulatory patho-physiology during anaesthesia confirms the need to sustain appropriate oxygen delivery. It also supports reduction or even elimination of supplementary crystalloid maintenance infusion, supposedly to replace the "mythical" third space loss. As a rational evidence base for future research it should allow for further improvements in anaesthetic management. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

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

NASA Astrophysics Data System (ADS)

Tsuchiya, Kazuyoshi; Nakanishi, Naoyuki; Nakamachi, Eiji

2005-02-01

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

Blood filling and flow in lungs during change in body position in space

NASA Technical Reports Server (NTRS)

Pogodin, A. S.; Mazhbich, B. I.

1980-01-01

In the horizontal position (supine and lateral), in the upright position (head up and head down) and during change of the cat body position in space, quantitative responses of regional blood volume and blood flow in the lungs (ml/100 cu cm) revealed presence of the gradient in the gravitation direction. Blood volume and blood flow of different lung portions changed qualitatively and quantitatively in different ways. These changes occurred only in the direction producing the equality of regional hydrostatical and hemodynamic loads in the lungs at either horizontal level.

A Method for Electrochemical Detection of Brain Derived Neurotrophic Factor (BDNF) in plasma.

PubMed

Bockaj, Marina; Fung, Barnabas; Tsoulis, Michael; Foster, Lauren Warren; Soleymani, Leyla

2018-06-22

Currently, a blood test for the diagnosis of endometriosis, a common estrogen-dependent gynecological disease, does not exist. Recent studies suggest that circulating concentrations of brain derived neurotrophic factor (BDNF) have potential for the diagnosis of endometriosis. However, at present BDNF can only be measured by ELISA which requires a clinic visit, a routine blood sample, and laboratory testing. Therefore, we developed a point-of-care device (EndoChip) for use with small blood volumes that can be collected through a finger prick. Specifically, the presented device is a polymer-based chip with a wrinkled nanoporous gold film acting as the electrode/sensing layer, allowing for the electrochemical detection of BDNF in plasma. Increasing concentrations of BDNF (0.25 - 2.0 ng/ml) induced significant differences in redox current. The biosensor produces a signal readout in a matter of seconds, and is ideal for realizing multiplexing. Blood samples were collected from women (n=20) with chronic pelvic pain undergoing a diagnostic laparoscopy. Plasma BDNF concentrations measured by commercial ELISA were positively correlated (r2=0.8216; p<0.001) with results from the EndoChip. Our results demonstrate a quick and reliable method for point-of-care quantification of circulating concentrations of BDNF and a promising diagnostic tool for endometriosis.

Robot-assisted versus open radical prostatectomy: the differential effect of regionalization, procedure volume and operative approach.

PubMed

Sammon, Jesse D; Karakiewicz, Pierre I; Sun, Maxine; Sukumar, Shyam; Ravi, Praful; Ghani, Khurshid R; Bianchi, Marco; Peabody, James O; Shariat, Shahrokh F; Perrotte, Paul; Hu, Jim C; Menon, Mani; Trinh, Quoc-Dien

2013-04-01

The use of robot-assisted radical prostatectomy has increased rapidly despite the absence of randomized, controlled trials showing the superiority of this approach. While recent studies suggest an advantage for perioperative complication rates, they fail to account for the volume-outcome relationship. We compared perioperative outcomes after robot-assisted and open radical prostatectomy, while considering the impact of this established relationship. Using the NIS (Nationwide Inpatient Sample), we abstracted data on patients treated with radical prostatectomy in 2009. Univariable and multivariable logistic regression analyses were done to compare the rates of blood transfusion, intraoperative and postoperative complications, prolonged length of stay, increased hospital charges and mortality between robot-assisted and open radical prostatectomy overall and across volume quartiles. An estimated 77,616 men underwent radical prostatectomy, including a robot-assisted and an open procedure in 63.9% and 36.1%, respectively. Low volume centers averaged 26.2 robot-assisted and 5.2 open cases, while very high volume centers averaged 578.8 robot-assisted and 150.2 open cases. Overall, patients treated with the robot-assisted procedure experienced a lower rate of adverse outcomes than those treated with the open procedure for all measured categories. Across equivalent volume quartiles robot-assisted radical prostatectomy outcomes were generally favorable. However, the open procedure at high volume centers resulted in a lower postoperative complication rate (OR 0.59, 95% CI 0.46-0.75), elevated hospital charges (OR 0.75, 95% CI 0.64-0.87) and a comparable blood transfusion rate (OR 1.38, 95% CI 0.93-2.02) relative to the robot-assisted procedure at low volume centers. Regionalization has occurred to a greater extent for robot-assisted than for open radical prostatectomy with an associated benefit in overall outcomes. Nonetheless, low volume institutions experienced inferior outcomes relative to the highest volume centers irrespective of approach. These findings demonstrate the importance of accounting for hospital volume when examining the benefit of a surgical technique. Copyright © 2013 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

Changes in haematology measurements with the Sysmex XT-2000iV during storage of feline blood sampled in EDTA or EDTA plus CTAD.

PubMed

Granat, Fanny; Geffré, Anne; Bourgès-Abella, Nathalie; Braun, Jean-Pierre; Trumel, Catherine

2013-06-01

In veterinary medicine a complete blood cell count (CBC) cannot always be performed within 24 h as usually recommended, particularly for specimens shipped to a reference laboratory. This raises the question of the stability of the variables, especially in ethylenediamine tetra-acetic acid (EDTA) feline blood specimens, known to be prone to in vitro platelet aggregation. Citrate, theophylline, adenosine and dipyridamole (CTAD) has been reported to limit platelet aggregation in feline blood specimens. The aim of this study was to measure the stability of the haematological variables and the platelet aggregation score in EDTA and EDTA plus CTAD (EDCT) feline blood specimens during 48 h of storage at room temperature. Forty-six feline EDTA and EDCT blood specimens were analysed with a Sysmex XT-2000iV analyser, and the platelet count and score of platelet aggregation were estimated immediately and after 24 and 48 h of storage. A significant increase in mean corpuscular volume, haematocrit, reticulocyte and eosinophil counts, and a significant decrease in mean corpuscular haemoglobin concentration and monocyte count were observed. Haemoglobin, mean corpuscular haemoglobin, and red blood cell, white blood cell, neutrophil and lymphocyte counts remained stable. Changes in reticulocyte indexes with time (low fluorescence ratio, medium fluorescence ratio, high fluorescence ratio and immature reticulocyte fraction) were not significant. Changes were generally more pronounced in EDTA than in EDCT. Platelet aggregation decreased markedly in initially highly aggregated EDTA specimens, and increased slightly in initially non- or mildly-aggregated EDTA or EDCT specimens. Platelet counts increased and decreased, or remained stable, respectively. CTAD can reduce storage-induced changes of the haematological variables in feline samples, thus improving the reliability of a CBC and limiting clinical misinterpretations.

Free-breathing black-blood CINE fast-spin echo imaging for measuring abdominal aortic wall distensibility: a feasibility study

NASA Astrophysics Data System (ADS)

Lin, Jyh-Miin; Patterson, Andrew J.; Chao, Tzu-Cheng; Zhu, Chengcheng; Chang, Hing-Chiu; Mendes, Jason; Chung, Hsiao-Wen; Gillard, Jonathan H.; Graves, Martin J.

2017-05-01

The paper reports a free-breathing black-blood CINE fast-spin echo (FSE) technique for measuring abdominal aortic wall motion. The free-breathing CINE FSE includes the following MR techniques: (1) variable-density sampling with fast iterative reconstruction; (2) inner-volume imaging; and (3) a blood-suppression preparation pulse. The proposed technique was evaluated in eight healthy subjects. The inner-volume imaging significantly reduced the intraluminal artifacts of respiratory motion (p  =  0.015). The quantitative measurements were a diameter of 16.3  ±  2.8 mm and wall distensibility of 2.0  ±  0.4 mm (12.5  ±  3.4%) and 0.7  ±  0.3 mm (4.1  ±  1.0%) for the anterior and posterior walls, respectively. The cyclic cross-sectional distensibility was 35  ±  15% greater in the systolic phase than in the diastolic phase. In conclusion, we developed a feasible CINE FSE method to measure the motion of the abdominal aortic wall, which will enable clinical scientists to study the elasticity of the abdominal aorta.

Specialized physiological studies in support of manned space flight

NASA Technical Reports Server (NTRS)

1977-01-01

Breath-by-breath measurements of pulmonary capillary O2 transfer and ventilation with a box-balloon spirometer and mass spectrometer were made on 3 subjects before, during and after 10 min of lower body negative pressure (LBNP) at -20, -40 and -60 Torr with arterial samples at -60 Torr. Deficits in blood O2 stores (O2B) were noted during LBNP with repayment of O2B during recovery being related to the intensity of LBNP stress. Concurrent calf volume, measured with a Hg strain gauge, decreased towards baseline well before the peak rise in pulmonary capillary O2 transfer after the release of LBNP which signified that O2B changes were related to blood volume shifts. The return of O2-depleted pooled blood to the central circulation during the first min. of recovery caused significant stress-related hyperpnea which peaked near 30 sec. and returned to near baseline after the first min. Three-compartment lung model analyses indicated an increase in the ventilated unperfused lung fraction from 0.09 to 0.17 with the effective compartment decreasing from 0.83 to 0.77. It appears that the 30% increase in ventilation equivalent was primarily the result of less effective lung perfusion during the LBNP.

Contemporary bloodletting in cardiac surgical care.

PubMed

Koch, Colleen G; Reineks, Edmunds Z; Tang, Anne S; Hixson, Eric D; Phillips, Shannon; Sabik, Joseph F; Henderson, J Michael; Blackstone, Eugene H

2015-03-01

Health care providers are seldom aware of the frequency and volume of phlebotomy for laboratory testing, bloodletting that often leads to hospital-acquired anemia. Our objectives were to examine the frequency of laboratory testing in patients undergoing cardiac surgery, calculate cumulative phlebotomy volume from time of initial surgical consultation to hospital discharge, and propose strategies to reduce phlebotomy volume. From January 1, 2012 to June 30, 2012, 1,894 patients underwent cardiac surgery at Cleveland Clinic; 1,867 had 1 hospitalization and 27 had 2. Each laboratory test was associated with a test name and blood volume. Phlebotomy volume was estimated separately for the intensive care unit (ICU), hospital floors, and cumulatively. A total of 221,498 laboratory tests were performed, averaging 115 tests per patient. The most frequently performed tests were 88,068 blood gas analyses, 39,535 coagulation tests, 30,421 complete blood counts, and 29,374 metabolic panels. Phlebotomy volume differed between ICU and hospital floors, with median volumes of 332 mL and 118 mL, respectively. Cumulative median volume for the entire hospital stay was 454 mL. More complex procedures were associated with higher overall phlebotomy volume than isolated procedures; eg, combined coronary artery bypass grafting (CABG) and valve procedure median volume was 653 mL (25th/75th percentiles, 428 of 1,065 mL) versus 448 mL (284 of 658 mL) for isolated CABG and 338 mL (237 of 619) for isolated valve procedures. We were astonished by the extent of bloodletting, with total phlebotomy volumes approaching amounts equivalent to 1 to 2 red blood cell units. Implementation of process improvement initiatives can potentially reduce phlebotomy volumes and resource utilization. Copyright © 2015 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.

Renal function, renal volume, and blood pressure in infants with antecedent of antenatal steroids.

PubMed

Carballo-Magdaleno, Deyanira; Guízar-Mendoza, Juan M; Amador-Licona, Norma; Domínguez-Domínguez, Víctor

2011-10-01

Steroids have been used for more than 20 years in preterm infants to induce pulmonary maturity; however, some long-term effects have been reported, such as insulin resistance and elevation of blood pressure. The aim of our study was to compare renal volume, renal function, and blood pressure in infants between 12-36 months of age with and without antecedent of antenatal steroid treatment. This was a cross-sectional study comprised of three groups of infants (n = 30, respectively): preterm infants with and without antecedent of receiving antenatal steroids, respectively, and full-term infants. Blood pressure, renal volume, glomerular filtration rate, and tubular function were measured. Blood pressure and cystatin C levels and glomerular filtration rate were higher in both groups of preterm infants than in the control group (p < 0.01). However, no difference in any of the tested variables between the steroid and non-steroid group of preterm infants. Renal volume was similar in preterm and control infants. Based on these results, we conclude that prematurity independent of antenatal steroid use is associated with higher cystatin C and blood pressure levels and a higher glomerular filtration rate in infants between 12-36 months of age.

An estimation of intracranial blood flow in the new-born infant.

PubMed Central

Cross, K W; Dear, P R; Hathorn, M K; Hyams, A; Kerslake, D M; Milligan, D W; Rahilly, P M; Stothers, J K

1979-01-01

1. A non-invasive method for the estimation of the intracranial blood flow of the new-born infant is described, and results obtained with it are presented. 2. The technique is a novel application of the principle of blood flow measurement by venous occlusion plethysmography. It is possible to apply a plethysmographic technique to the neonatal cranium because the presence of open sutures between the component bones permits small, but readily measurable, changes in intracranial volume to occur. 3. Skull volume changes are calculated from changes in the occipito-frontal circumference of the cranium as recorded and measured with a mercury-in-Silastic strain gauge. 4. The jugular veins in the baby's neck are occluded by finger pressure and there follows an increase in skull volume, which is rapid at first, but which decreases exponentially as venous drainage diverts to non-occluded channels such as the vertebral venous plexus. At the instant of jugular occlusion the rate of skull volume increase is representative of the rate of flow in the jugular vessels prior to occlusion, and so provides an index of the relative magnitude of the intracranial blood flow. The method thus allows changes in intracranial blood flow to be followed. When occlusion is released cranial volume decreases, initially rapidly, but slowing exponentially as resting volume is regained. 5. A theoretical model of the events occurring during the inflow and outflow phases has been developed, and a formula derived which allows an estimation to be made of the flow of blood through uncompressed channels. The measured value of jugular blood flow can then be augmented to an estimate of total intracranial flow. 6. The mean cerebral blood flow of sixteen normal babies was estimated to be 40 ml. 100 g-1.min-1 (S.D. = +/- 11.63). Images Fig. 2 Fig. 6 PMID:458665

Blood pressure, brain structure, and cognition: opposite associations in men and women.

PubMed

Cherbuin, Nicolas; Mortby, Moyra E; Janke, Andrew L; Sachdev, Perminder S; Abhayaratna, Walter P; Anstey, Kaarin J

2015-02-01

Research on associations between blood pressure, brain structure, and cognitive function has produced somewhat inconsistent results. In part, this may be due to differences in age ranges studied and because of sex differences in physiology and/or exposure to risk factors, which may lead to different time course or patterns in cardiovascular disease progression. The aim of this study was to investigate the impact of sex on associations between blood pressure, regional cerebral volumes, and cognitive function in older individuals. In this cohort study, brachial blood pressure was measured twice at rest in 266 community-based individuals free of dementia aged 68-73 years who had also undergone a brain scan and a neuropsychological assessment. Associations between mean blood pressure (MAP), regional brain volumes, and cognition were investigated with voxel-wise regression analyses. Positive associations between MAP and regional volumes were detected in men, whereas negative associations were found in women. Similarly, there were sex differences in the brain-volume cognition relationship, with a positive relationship between regional brain volumes associated with MAP in men and a negative relationship in women. In this cohort of older individuals, higher MAP was associated with larger regional volume and better cognition in men, whereas opposite findings were demonstrated in women. These effects may be due to different lifetime risk exposure or because of physiological differences between men and women. Future studies investigating the relationship between blood pressure and brain structure or cognitive function should evaluate the potential for differential sex effects. © American Journal of Hypertension, Ltd 2014. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Evaluation of the BD BACTEC FX blood volume monitoring system as a continuous quality improvement measure.

PubMed

Coorevits, L; Van den Abeele, A-M

2015-07-01

The yield of blood cultures is proportional to the volume of blood cultured. We evaluated an automatic blood volume monitoring system, recently developed by Becton Dickinson within its BACTEC EpiCenter module, that calculates mean volumes of negative aerobic bottles and generates boxplots and histograms. First, we evaluated the filling degree of 339 aerobic glass blood cultures by calculating the weight-based volume for each bottle. A substantial amount of the bottles (48.3%) were inadequately filled. Evaluation of the accuracy of the monitoring system showed a mean bias of -1.4 mL (-15.4%). Additional evaluation, using the amended software on 287 aerobic blood culture bottles, resulted in an acceptable mean deviation of -0.3 mL (-3.3%). The new software version was also tested on 200 of the recently introduced plastic bottles, which will replace the glass bottles in the near future, showing a mean deviation of +2.8 mL (+26.7%). In conclusion, the mean calculated volumes can be used for the training of a single phlebotomist. However, filling problems appear to be masked when using them for phlebotomist groups or on wards. Here, visual interpretation of boxplots and histograms can serve as a useful tool to observe the spread of the filling degrees and to develop a continuous improvement program. Re-adjustment of the software has proven to be necessary for use with plastic bottles. Due to our findings, BD has developed further adjustments to the software for validated use with plastic bottles, which will be released soon.

Photosensitizer quantitation in vivo by flourescence microsampling

NASA Astrophysics Data System (ADS)

Pogue, Brian W.; Burke, Gregory C.; Lee, Claudia C.; Hoopes, P. Jack

2000-06-01

Photodynamic therapy can provide a reliable method of tumor destruction when the appropriate dosimetry is applied. Current dosimetry practice involves quantification of the drug and light doses applied to the tumor, but it would be desirable to monitor in vivo light and drug levels to provide the most accurate determination of dosimetry. In vivo measurements can be used to minimize variations in treatment response due to inter-animal variability, by providing animal-specific or patient-specific treatment planning. This study reports on the development of a micro-sampling method to measure fluorescence from tissue, which is not significantly affected by the tissue optical properties. The system measures fluorescence from the surface of a tissue, using a fiber bundle composed of individual 100 micron fibers which ar all spaced apart by 700 microns from one another at the tissue contact end. This design provides sampling of the fluorescence at multiple sites to increase the signal intensity, while maintaining a micro- sampling of the tissue volume just below the surface. The calibration studies here indicate that the 1/e sampling depth is near 60 microns when measured in optical phantoms, which are similar to typical tissue properties. The probe fluorescence signal is independent of blood concentration up to a maximum of 10% blood by volume, which is similar to most tumor tissue. Animal tests indicate that the sensitivity to drug concentration is essentially the same in when measured in murine liver and muscle tissues, both in vivo and ex vivo. These preliminary calibration results suggest that the probe can be used to measure photosensitizer uptake in vivo non- invasively and rapidly via conversion of fluorescence intensity to photosensitizer concentration.

Image-derived input function in PET brain studies: blood-based methods are resistant to motion artifacts.

PubMed

Zanotti-Fregonara, Paolo; Liow, Jeih-San; Comtat, Claude; Zoghbi, Sami S; Zhang, Yi; Pike, Victor W; Fujita, Masahiro; Innis, Robert B

2012-09-01

Image-derived input function (IDIF) from carotid arteries is an elegant alternative to full arterial blood sampling for brain PET studies. However, a recent study using blood-free IDIFs found that this method is particularly vulnerable to patient motion. The present study used both simulated and clinical [11C](R)-rolipram data to assess the robustness of a blood-based IDIF method (a method that is ultimately normalized with blood samples) with regard to motion artifacts. The impact of motion on the accuracy of IDIF was first assessed with an analytical simulation of a high-resolution research tomograph using a numerical phantom of the human brain, equipped with internal carotids. Different degrees of translational (from 1 to 20 mm) and rotational (from 1 to 15°) motions were tested. The impact of motion was then tested on the high-resolution research tomograph dynamic scans of three healthy volunteers, reconstructed with and without an online motion correction system. IDIFs and Logan-distribution volume (VT) values derived from simulated and clinical scans with motion were compared with those obtained from the scans with motion correction. In the phantom scans, the difference in the area under the curve (AUC) for the carotid time-activity curves was up to 19% for rotations and up to 66% for translations compared with the motionless simulation. However, for the final IDIFs, which were fitted to blood samples, the AUC difference was 11% for rotations and 8% for translations. Logan-VT errors were always less than 10%, except for the maximum translation of 20 mm, in which the error was 18%. Errors in the clinical scans without motion correction appeared to be minor, with differences in AUC and Logan-VT always less than 10% compared with scans with motion correction. When a blood-based IDIF method is used for neurological PET studies, the motion of the patient affects IDIF estimation and kinetic modeling only minimally.

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

PubMed

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

2017-08-01

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

Characterization of Tumor Cells Using a Medical Wire for Capturing Circulating Tumor Cells: A 3D Approach Based on Immunofluorescence and DNA FISH

PubMed Central

Gallerani, Giulia; Cocchi, Claudia; Bocchini, Martine; Piccinini, Filippo; Fabbri, Francesco

2017-01-01

Circulating tumor cells (CTCs) are associated with poor survival in metastatic cancer. Their identification, phenotyping, and genotyping could lead to a better understanding of tumor heterogeneity and thus facilitate the selection of patients for personalized treatment. However, this is hampered because of the rarity of CTCs. We present an innovative approach for sampling a high volume of the patient blood and obtaining information about presence, phenotype, and gene translocation of CTCs. The method combines immunofluorescence staining and DNA fluorescent-in-situ-hybridization (DNA FISH) and is based on a functionalized medical wire. This wire is an innovative device that permits the in vivo isolation of CTCs from a large volume of peripheral blood. The blood volume screened by a 30-min administration of the wire is approximately 1.5-3 L. To demonstrate the feasibility of this approach, epithelial cell adhesion molecule (EpCAM) expression and the chromosomal translocation of the ALK gene were determined in non-small-cell lung cancer (NSCLC) cell lines captured by the functionalized wire and stained with an immuno-DNA FISH approach. Our main challenge was to perform the assay on a 3D structure, the functionalized wire, and to determine immuno-phenotype and FISH signals on this support using a conventional fluorescence microscope. The results obtained indicate that catching CTCs and analyzing their phenotype and chromosomal rearrangement could potentially represent a new companion diagnostic approach and provide an innovative strategy for improving personalized cancer treatments. PMID:29286485

Clinical investigation of the protective effects of palm vitamin E tocotrienols on brain white matter.

PubMed

Gopalan, Yogheswaran; Shuaib, Ibrahim Lutfi; Magosso, Enrico; Ansari, Mukhtar Alam; Abu Bakar, Mohd Rizal; Wong, Jia Woei; Khan, Nurzalina Abdul Karim; Liong, Wei Chuen; Sundram, Kalyana; Ng, Bee Hong; Karuthan, Chinna; Yuen, Kah Hay

2014-05-01

Previous cell-based and animal studies showed mixed tocotrienols are neuroprotective, but the effect is yet to be proven in humans. Thus, the present study aimed to evaluate the protective activity of mixed tocotrienols in humans with white matter lesions (WMLs). WMLs are regarded as manifestations of cerebral small vessel disease, reflecting varying degrees of neurodegeneration and tissue damage with potential as a surrogate end point in clinical trials. A total of 121 volunteers aged ≥35 years with cardiovascular risk factors and MRI-confirmed WMLs were randomized to receive 200 mg mixed tocotrienols or placebo twice a day for 2 years. The WML volumes were measured from MRI images taken at baseline, 1 year, and 2 years using a validated software and were compared. Fasting blood samples were collected for full blood chemistry investigation. According to per-protocol (88 volunteers) and intention-to-treat (121 volunteers) analyses, the mean WML volume of the placebo group increased after 2 years, whereas that of the tocotrienol-supplemented group remained essentially unchanged. The mean WML volume change between the 2 groups was not significantly different (P=0.150) at the end of 1 year but was significant at the end of 2 years for both per-protocol and intention-to-treat analyses (P=0.019 and P=0.018). No significant difference was observed in the blood chemistry parameters between the 2 groups. Mixed tocotrienols were found to attenuate the progression of WMLs. http://www.clinicaltrials.gov. Unique identifier: NCT00753532.

Control of rectal gland secretion by blood acid-base status in the intact dogfish shark (Squalus acanthias).

PubMed

Wood, Chris M; Munger, R Stephen; Thompson, Jill; Shuttleworth, Trevor J

2007-05-14

In order to address the possible role of blood acid-base status in controlling the rectal gland, dogfish were fitted with indwelling arterial catheters for blood sampling and rectal gland catheters for secretion collection. In intact, unanaesthetized animals, isosmotic volume loading with 500 mmol L-1 NaCl at a rate of 15 mL kg-1 h-1 produced a brisk, stable rectal gland secretion flow of about 4 mL kg-1 h-1. Secretion composition (500 mmol L-1 Na+ and Cl-; 5 mmol L-1 K+; <1 mmol L-1 Ca2+, Mg2+, SO(4)2-, or phosphate) was almost identical to that of the infusate with a pH of about 7.2, HCO3- mmol L-1<1 mmol L-1 and a PCO2 (1 Torr) close to PaCO2. Experimental treatments superimposed on the infusion caused the expected disturbances in systemic acid-base status: respiratory acidosis by exposure to high environmental PCO2, metabolic acidosis by infusion of HCl, and metabolic alkalosis by infusion of NaHCO3. Secretion flow decreased markedly with acidosis and increased with alkalosis, in a linear relationship with extracellular pH. Secretion composition did not change, apart from alterations in its acid-base status, and made negligible contribution to overall acid-base balance. An adaptive control of rectal gland secretion by systemic acid-base status is postulated-stimulation by the "alkaline tide" accompanying the volume load of feeding and inhibition by the metabolic acidosis accompanying the volume contraction of exercise.

Influence of Gravity on Blood Volume and Flow Distribution

NASA Technical Reports Server (NTRS)

Pendergast, D.; Olszowka, A.; Bednarczyk, E.; Shykoff, B.; Farhi, L.

1999-01-01

In our previous experiments during NASA Shuttle flights SLS 1 and 2 (9-15 days) and EUROMIR flights (30-90 days) we observed that pulmonary blood flow (cardiac output) was elevated initially, and surprisingly remained elevated for the duration of the flights. Stroke volume increased initially and then decreased, but was still above 1 Gz values. As venous return was constant, the changes in SV were secondary to modulation of heart rate. Mean blood pressure was at or slightly below 1 Gz levels in space, indicating a decrease in total peripheral resistance. It has been suggested that plasma volume is reduced in space, however cardiac output/venous return do not return to 1 Gz levels over the duration of flight. In spite of the increased cardiac output, central venous pressure was not elevated in space. These data suggest that there is a change in the basic relationship between cardiac output and central venous pressure, a persistent "hyperperfusion" and a re-distribution of blood flow and volume during space flight. Increased pulmonary blood flow has been reported to increase diffusing capacity in space, presumably due to the improved homogeneity of ventilation and perfusion. Other studies have suggested that ventilation may be independent of gravity, and perfusion may not be gravity- dependent. No data for the distribution of pulmonary blood volume were available for flight or simulated microgravity. Recent studies have suggested that the pulmonary vascular tree is influenced by sympathetic tone in a manner similar to that of the systemic system. This implies that the pulmonary circulation is dilated during microgravity and that the distribution of blood flow and volume may be influenced more by vascular control than by gravity. The cerebral circulation is influenced by sympathetic tone similarly to that of the systemic and pulmonary circulations; however its effects are modulated by cerebral autoregulation. Thus it is difficult to predict if cerebral perfusion is increased and if there is edema in space. Anecdotal evidence suggests there may be cerebral edema early in flight. Cerebral artery velocity has been shown to be elevated in simulated microgravity. The elevated cerebral artery velocity during simulated microgravity may reflect vasoconstriction of the arteries and not increased cerebral blood flow. The purpose of our investigations was to evaluate the effects of alterations in simulated gravity (+/-), resulting in changes in cardiac output (+/-), and on the blood flow and volume distribution in the lung and brain of human subjects. The first hypothesis of these studies was that blood flow and volume would be affected by gravity, but their distribution in the lung would be independent of gravity and due to vasoactivity changing vascular resistance in lung vessels. The vasodilitation of the lung vasculature (lower resistance) along with increased "compliance" of the heart could account for the absence of increased central venous pressure in microgravity. Secondly, we postulate that cerebral blood velocity is increased in microgravity due to large artery vasoconstriction, but that cerebral blood flow would be reduced due to autoregulation.

Development of a new score to estimate clinical East Coast Fever in experimentally infected cattle.

PubMed

Schetters, Th P M; Arts, G; Niessen, R; Schaap, D

2010-02-10

East Coast Fever is a tick-transmitted disease in cattle caused by Theileria parva protozoan parasites. Quantification of the clinical disease can be done by determining a number of variables, derived from parasitological, haematological and rectal temperature measurements as described by Rowlands et al. (2000). From a total of 13 parameters a single ECF-score is calculated that allows categorization of infected cattle in five different classes that correlate with the severity of clinical signs. This score is complicated not only by the fact that it requires estimation of 13 parameters but also because of the subsequent mathematics. The fact that the values are normalised over a range of 0-10 for each experiment makes it impossible to compare results from different experiments. Here we present an alternative score based on the packed cell volume and the number of piroplasms in the circulation and that is calculated using a simple equation; ECF-score=PCV(relday0)/log(PE+10). In this equation the packed cell volume is expressed as a value relative to that of the day on infection (PCV(relday0)) and the number of piroplasms is expressed as the logarithmic value of the number of infected red blood cells (=PE) in a total of 1000 red blood cells. To allow PE to be 0, +10 is added in the denominator. We analysed a data set of 54 cattle from a previous experiment and found a statistically significant linear correlation between the ECF-score value reached during the post-infection period and the Rowlands' score value. The new score is much more practical than the Rowlands score as it only requires daily blood sampling. From these blood samples both PCV and number of piroplasms can be determined, and the score can be calculated daily. This allows monitoring the development of ECF after infection, which was hitherto not possible. In addition, the new score allows for easy comparison of results from different experiments.

Cardiovascular and metabolic adaptations in horses competing in cross-country events.

PubMed

Muñoz, A; Riber, C; Santisteban, R; Rubio, M D; Agüera, E I; Castejón, F M

1999-01-01

The cardiovascular and metabolic response to two cross-country events (CC*: preliminary level and CC*** advanced level) were analysed in 8 male eventing horses (4 Anglo-Hunter and 4 Anglo-Arabian). This study focused on the establishment of the main metabolic pathways involved in the muscle energy resynthesis during the competitions. Heart rate (HR) was recorded throughout the CC events. Jugular venous blood samples were withdrawn before the warm-up period, immediately after the competitions and at 5 and 10 min in the recuperation period. The following haematological parameters were studied: red blood cells (RBC), packed cell volume (PCV), haemoglobin concentration (Hb), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), white blood cells (WBC), and number and percentages of lymphocytes (LYM) and granulocytes and monocytes (GRAN). One fraction of blood was centrifuged and, in plasma, lactate (LA), total plasma protein (TPP) and the rate of LA disappearance were determined. The competitions induced significant increases in RBC, Hb, PCV, MCV and TPP. Plasma LA response exceeded the anaerobic threshold of 4 mmol/l, reaching a maximum level of 13.3 mmol/l. HR ranged from 140 to more than 200 bpm, peaking at 230 bpm, revealing a limitation in the oxygen supply to the working muscles. It was concluded that muscle energy resynthesis during a CC event is provided both through oxidative processes and glycolysis with LA formation. Therefore, both stamina and power exercises are required for eventing horses.

The Importance of Mean Neutrophil Volume (MNV) Values in Differential Diagnosis of Bacterial Sepsis.

PubMed

Şafak, Birol; Baykan, Özgür; Kılınç, Osman; Özer Yıldırım, Diğdem

2017-01-01

Bloodstream infections are a significant cause of morbidity and mortality in hospitalized patients. Blood cultures and other laboratory tests are used for diagnosis. Among these tests, the mean neutrophil volume (MNV) value is reported as a potential indicator that supports the diagnosis of sepsis. Our study identified the MNV values of patients via microorganisms cultivated from blood cultures and examined the role of these MNV values in the early diagnosis of bloodstream infections. Our study surveyed retrospectively 148 adult patient blood culture samples that had been sent to our laboratory. BACTEC 9050 (Becton Dickinson, USA) and BACTEC FX 40 (Becton Dickinson, USA) devices were used in the blood culture isolation procedures. The average MNV value was found to be 159.0 (+11.3) in patients whose sepsis originated from Gram-negative bacteria, and the average MNV value was measured as 152.4 (+14.5) among patients whose sepsis originated from Gram-positive bacteria. When comparing groups of patients having Gram-negative bacteria and patients having Gram-positive bacteria, a statistically significant difference (p = 0.041) in the MNV values was observed. The MNV value was found to be statistically significant in discrimination of Gram-negative and Gram-positive sepsis. Considering these findings, measuring the MNV values can help initiate proper antibiotic treatment more quickly, and we think that this will help lower the mortality rate. However, these findings should be supported with further studies. Copyright © 2017 National Medical Association. Published by Elsevier Inc. All rights reserved.

The effect of long-term, high-volume aerobic exercise training on postprandial lipemia and oxidative stress.

PubMed

Bloomer, Richard J; Fisher-Wellman, Kelsey H; Bell, Heather K

2010-04-01

We have previously found no effect of moderate-volume aerobic exercise training (approximately 3 hrs*wk(-1)) on postprandial oxidative stress. It is possible that a higher volume of exercise is needed to impact postprandial oxidative stress in young, otherwise healthy individuals. Our purpose was to compare blood triglycerides (TAGs) and oxidative stress biomarkers in 10 healthy untrained and 10 healthy highly aerobically trained (eg, >or= 40 miles running*wk(-1) or >or= 150 miles cycling*wk(-1)) men and women following ingestion of a lipid meal. Blood samples were collected before (in a 10-hour fasted state), and 1, 2, 4, and 6 hours after ingestion of a lipid load (heavy whipping cream at 1 g*kg(-1)). Blood samples were analyzed for TAGs, malondialdehyde (MDA), hydrogen peroxide (H(2)O(2)), and nitrate/nitrite (NOx). No training status or interaction effects were noted for TAGs, MDA, H2O2, or NOx (P > 0.05). However, a time effect was noted for TAGs (P = 0.01), with values higher at 2 hours (67 +/- 6 mg*dL(-1)) compared with premeal (41 +/- 6 mg*dL(-1)). A time effect was also noted for H2O2 (P = 0.0001), with values higher at 2 hours (24 +/- 3 micromol*L(-1)), 4 hours (23 +/- 3 micromol*L(-1)), and 6 hours (21 +/- 3 mumol.L(-1)) compared with premeal (7 +/- 2 micromol*L(-1)). The time effect for MDA approached significance (P = 0.07), with values peaking at 4 hours post-meal (1.59 +/- 0.16 micromol*L(-1)) compared with premeal (0.99 +/- 0.15 micromol*L(-1)). These data indicate that aerobic exercise training (even when performed at a relatively high volume) does not attenuate postprandial lipemia or oxidative stress as compared with no exercise when healthy men and women consume a lipid load in the form of heavy whipping cream. Fasting TAG values may be most important in this regard. It is possible that long-term exercise may be capable of attenuating postprandial lipemia or oxidative stress in older individuals, those with chronic disease, or those with elevated fasting TAG values. Future work is needed to confirm these hypotheses.

Plasma volume, osmolality, vasopressin, and renin activity during graded exercise in man

NASA Technical Reports Server (NTRS)

Convertino, V. A.; Keil, L. C.; Bernauer, E. M.; Greenleaf, J. E.

1981-01-01

The influence of work intensity on plasma volume, osmolality, vasopressin and renin activity and the interrelationships between these responses are investigated. Plasma volume, renin activity and osmotic, sodium and arginine vasopressin concentrations were measured in venous blood samples taken from 15 healthy male subjects before and after six minutes of bicycle ergometer exercise at 100, 175 and 225 W. Plasma volume is found to decrease significantly with increasing work intensity, while increases in Na(+) concentration, osmolality and vasopressin are only observed to be significant when the work intensity exceeds 40% maximal aerobic capacity and plasma resin activity increased linearly at all work levels. In addition, significant correlations are observed between plasma volume and osmolality and sodium changes, and between vasopressin and osmolality and sodium content changes. Data thus support the hypotheses that (1) vasopressin may be the primary controlling endocrine for fluid and electrolyte levels following exercise; (2) an exercise intensity greater than 40% maximal aerobic capacity is required to stimulate vasopressin release through changes in plasma osmolality; and (3) the stimulation of the renin-angiotensin system is a more general stress response.

Reduced left ventricular filling following blood volume extraction does not result in compensatory augmentation of cardiac mechanics.

PubMed

Lord, Rachel; MacLeod, David; George, Keith; Oxborough, David; Shave, Rob; Stembridge, Mike

2018-04-01

What is the central question of this study? A reduction in left ventricular (LV) filling, and concomitant increase in heart rate, augments LV mechanics to maintain stroke volume (SV); however, the impact of reduced LV filling in isolation on SV and LV mechanics is currently unknown. What is the main finding and its importance? An isolated decrease in LV filling did not provoke a compensatory increase in mechanics to maintain SV; in contrast, LV mechanics and SV were reduced. These data indicate that when LV filling is reduced without changes in heart rate, LV mechanics do not compensate to maintain SV. An acute non-invasive reduction in preload has been shown to augment cardiac mechanics to maintain stroke volume and cardiac output. Such interventions induce concomitant changes in heart rate, whereas blood volume extraction reduces preload without changes in heart rate. Therefore, the purpose of this study was to determine whether a preload reduction in isolation resulted in augmented stroke volume achieved via enhanced cardiac mechanics. Nine healthy volunteers (four female, age 29 ± 11 years) underwent echocardiography for the assessment of left ventricular (LV) volumes and mechanics in a supine position at baseline and end extraction after the controlled removal of 25% of total blood volume (1062 ± 342 ml). Arterial blood pressure was monitored continuously by a pressure transducer attached to an indwelling radial artery catheter. Heart rate and total peripheral resistance were unchanged from baseline to end extraction, but systolic blood pressure was reduced (from 148 to 127 mmHg). From baseline to end extraction there were significant reductions in left ventricular end-diastolic volume (from 89 to 71 ml) and stroke volume (from 56 to 37 ml); however, there was no change in LV twist, basal or apical rotation. In contrast, LV longitudinal strain (from -20 to -17%) and basal circumferential strain (from -22 to -19%) were significantly reduced from baseline to end extraction. In conclusion, a reduction in preload during blood volume extraction does not result in compensatory changes in stroke volume or cardiac mechanics. Our data suggest that LV strain is dependent on LV filling and consequent geometry, whereas LV twist could be mediated by heart rate. © 2018 The Authors. Experimental Physiology © 2018 The Physiological Society.

EFFECT ON PERFUSION VALUES OF SAMPLING INTERVAL OF CT PERFUSION ACQUISITIONS IN NEUROENDOCRINE LIVER METASTASES AND NORMAL LIVER

PubMed Central

Ng, Chaan S.; Hobbs, Brian P.; Wei, Wei; Anderson, Ella F.; Herron, Delise H.; Yao, James C.; Chandler, Adam G.

2014-01-01

Objective To assess the effects of sampling interval (SI) of CT perfusion acquisitions on CT perfusion values in normal liver and liver metastases from neuroendocrine tumors. Methods CT perfusion in 16 patients with neuroendocrine liver metastases were analyzed by distributed parameter modeling to yield tissue blood flow, blood volume, mean transit time, permeability, and hepatic arterial fraction, for tumor and normal liver. CT perfusion values for the reference sampling interval of 0.5s (SI0.5) were compared with those of SI datasets of 1s, 2s, 3s and 4s, using mixed-effects model analyses. Results Increases in SI beyond 1s were associated with significant and increasing departures of CT perfusion parameters from reference values at SI0.5 (p≤0.0009). CT perfusion values deviated from reference with increasing uncertainty with increasing SIs. Findings for normal liver were concordant. Conclusion Increasing SIs beyond 1s yield significantly different CT perfusion parameter values compared to reference values at SI0.5. PMID:25626401

Medullary cystic kidney disease

MedlinePlus

... Tests that may be done include: 24-hour urine volume and electrolytes Blood urea nitrogen (BUN) Complete blood count (CBC) Creatinine blood test Creatinine clearance -- blood and urine Uric acid blood test Urine specific gravity (will ...

Integrated printed circuit board device for cell lysis and nucleic acid extraction.

PubMed

Marshall, Lewis A; Wu, Liang Li; Babikian, Sarkis; Bachman, Mark; Santiago, Juan G

2012-11-06

Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 μm × 300 μm × 3.7 cm microfluidic channel connecting two 15 μL reservoirs. We demonstrated this device by extracting pathogenic nucleic acids from 1 μL dispensed volume of whole blood spiked with Plasmodium falciparum. We dispensed whole blood directly onto an on-chip reservoir, and the system's integrated heaters simultaneously lysed and mixed the sample. We used isotachophoresis to extract the nucleic acids into a secondary buffer via isotachophoresis. We analyzed the convective mixing action with micro particle image velocimetry (micro-PIV) and verified the purity and amount of extracted nucleic acids using off-chip quantitative polymerase chain reaction (PCR). We achieved a clinically relevant limit of detection of 500 parasites per microliter. The system has no moving parts, and the process is potentially compatible with a wide range of on-chip hybridization or amplification assays.

Pulse oximeter plethysmographic waveform changes in awake, spontaneously breathing, hypovolemic volunteers.

PubMed

McGrath, Susan P; Ryan, Kathy L; Wendelken, Suzanne M; Rickards, Caroline A; Convertino, Victor A

2011-02-01

The primary objective of this study was to determine whether alterations in the pulse oximeter waveform characteristics would track progressive reductions in central blood volume. We also assessed whether changes in the pulse oximeter waveform provide an indication of blood loss in the hemorrhaging patient before changes in standard vital signs. Pulse oximeter data from finger, forehead, and ear pulse oximeter sensors were collected from 18 healthy subjects undergoing progressive reduction in central blood volume induced by lower body negative pressure (LBNP). Stroke volume measurements were simultaneously recorded using impedance cardiography. The study was conducted in a research laboratory setting where no interventions were performed. Pulse amplitude, width, and area under the curve (AUC) features were calculated from each pulse wave recording. Amalgamated correlation coefficients were calculated to determine the relationship between the changes in pulse oximeter waveform features and changes in stroke volume with LBNP. For pulse oximeter sensors on the ear and forehead, reductions in pulse amplitude, width, and area were strongly correlated with progressive reductions in stroke volume during LBNP (R(2) ≥ 0.59 for all features). Changes in pulse oximeter waveform features were observed before profound decreases in arterial blood pressure. The best correlations between pulse features and stroke volume were obtained from the forehead sensor area (R(2) = 0.97). Pulse oximeter waveform features returned to baseline levels when central blood volume was restored. These results support the use of pulse oximeter waveform analysis as a potential diagnostic tool to detect clinically significant hypovolemia before the onset of cardiovascular decompensation in spontaneously breathing patients.