Fetal scalp blood sampling during labor

DEFF Research Database (Denmark)

Chandraharan, Edwin; Wiberg, Nana

2014-01-01

Fetal cardiotocography is characterized by low specificity; therefore, in an attempt to ensure fetal well-being, fetal scalp blood sampling has been recommended by most obstetric societies in the case of a non-reassuring cardiotocography. The scientific agreement on the evidence for using fetal...... scalp blood sampling to decrease the rate of operative delivery for fetal distress is ambiguous. Based on the same studies, a Cochrane review states that fetal scalp blood sampling increases the rate of instrumental delivery while decreasing neonatal acidosis, whereas the National Institute of Health...... and Clinical Excellence guideline considers that fetal scalp blood sampling decreases instrumental delivery without differences in other outcome variables. The fetal scalp is supplied by vessels outside the skull below the level of the cranial vault, which is likely to be compressed during contractions...

Impact of blood sampling in very preterm infants

DEFF Research Database (Denmark)

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

2000-01-01

; the groups were then subdivided into critically ill or not. Diagnostic blood sampling and blood transfusion events were recorded. In total, 1905 blood samples (5,253 analysis) were performed, corresponding to 0.7 samples (1.9 analysis) per day per infant. The highest frequencies were found during the first....../kg. For the extremely preterm infants a significant correlation between sampled and transfused blood volume was found (mean 37.1 and 33.3 ml/kg, respectively, r = + 0.71, p = 0.0003). The most frequently requested analyses were glucose, sodium and potassium. Few blood gas analyses were requested (1.9/ infant). No blood...... losses attributable to excessive generous sampling were detected. The results show an acceptable low frequency of sampling and transfusion events for infants of GA 28-32 weeks. The study emphasizes the necessity of thorough reflection and monitoring of blood losses when ordering blood sampling...

21 CFR 868.1100 - Arterial blood sampling kit.

Science.gov (United States)

2010-04-01

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

Blood sampling from adrenal gland vein

International Nuclear Information System (INIS)

Sun Yong; Ni Caifang

2009-01-01

Adrenal gland vein sampling is an interventional method to get the blood samples from the adrenal gland vein. The blood is obtained via a catheter which is selectively inserted in the adrenal gland vein. This technique is mainly used to be diagnostic for primary hyperaldosteronism. A full knowledge of the anatomy and variations of the adrenal gland vein, serious preoperative preparation and skilled catheterization manipulation are necessary for obtaining sufficient blood sample and for reducing the occurrence of complications. Providing the physicians with definite diagnostic evidence and being technically feasible, adrenal gland vein sampling should become one of the routine examinations for clarifying the cause of primary hyperaldosteronism. (authors)

The quantitative regional cerebral blood flow measurement with autoradiography method using 123I-IMP SPECT. Evaluation of arterialized venous blood sampling as a substitute for arterial blood sampling

International Nuclear Information System (INIS)

Ohnishi, Takashi; Yano, Takao; Nakano, Shinichi; Jinnouchi, Seishi; Nagamachi, Shigeki; Flores, L. II; Nakahara, Hiroshi; Watanabe, Katsushi.

1996-01-01

The purpose of this study is validation of calibrating a standard input function in autoradiography (ARG) method by one point venous blood sampling as a substitute for that by one point arterial blood sampling. Ten and 20 minutes after intravenous constant infusion of 123 I-IMP, arterialized venous blood sampling from a dorsal vein were performed on 15 patients having ischemic cerebrovascular disease. And arterial blood sampling from radial artery was performed 10 min after 123 I-IMP infusion. The mean difference rates of integrated input function between calibrated standard input function by arterial blood sampling at 10 min and that by venous blood sampling were 4.1±3% and 9.3±5.4% at 10 and 20 min after 123 I-IMP infusion, respectively. The ratio of venous blood radioactivity to arterial blood radioactivity at 10 min after 123 I-IMP infusion was 0.96±0.02. There was an excellent correlation between ARG method CBF values obtained by arterial blood sampling at 10 min and those obtained by arterialized venous blood sampling at 10 min. In conclusion, a substitution by arterialized venous blood sampling from dorsal hand vein for artery can be possible. The optimized time for arterialized venous blood sampling was 10 min after 123 I-IMP infusion. (author)

Preanalytical Blood Sampling Errors in Clinical Settings

International Nuclear Information System (INIS)

Zehra, N.; Malik, A. H.; Arshad, Q.; Sarwar, S.; Aslam, S.

2016-01-01

Background: Blood sampling is one of the common procedures done in every ward for disease diagnosis and prognosis. Daily hundreds of samples are collected from different wards but lack of appropriate knowledge of blood sampling by paramedical staff and accidental errors make the samples inappropriate for testing. Thus the need to avoid these errors for better results still remains. We carried out this research with an aim to determine the common errors during blood sampling; find factors responsible and propose ways to reduce these errors. Methods: A cross sectional descriptive study was carried out at the Military and Combined Military Hospital Rawalpindi during February and March 2014. A Venous Blood Sampling questionnaire (VBSQ) was filled by the staff on voluntary basis in front of the researchers. The staff was briefed on the purpose of the survey before filling the questionnaire. Sample size was 228. Results were analysed using SPSS-21. Results: When asked in the questionnaire, around 61.6 percent of the paramedical staff stated that they cleaned the vein by moving the alcohol swab from inward to outwards while 20.8 percent of the staff reported that they felt the vein after disinfection. On contrary to WHO guidelines, 89.6 percent identified that they had a habit of placing blood in the test tube by holding it in the other hand, which should actually be done after inserting it into the stand. Although 86 percent thought that they had ample knowledge regarding the blood sampling process but they did not practice it properly. Conclusion: Pre analytical blood sampling errors are common in our setup. Eighty six percent participants though thought that they had adequate knowledge regarding blood sampling, but most of them were not adhering to standard protocols. There is a need of continued education and refresher courses. (author)

Uranium concentration in blood samples of Southern Iraqi leukemia patients using CR-39 track detector

International Nuclear Information System (INIS)

Al-Hamzawi, A.A.; Al-Qadisiyah University, Qadisiyah; Jaafar, M.S.; Tawfiq, N.F.

2014-01-01

The simple and effective technique of fission track etch has been applied to determine trace concentration of uranium in human blood samples taken from two groups of male and female participants: leukemia patients and healthy subjects group. The blood samples of leukemia patients and healthy subjects were collected from three key southern governorates namely, Basrah, Muthanna and Dhi-Qar. These governorates were the centers of intensive military activities during the 1991 and 2003 Gulf wars, and the discarded weapons are still lying around in these regions. CR-39 track detector was used for registration of induced fission tracks. The results show that the highest recorded uranium concentration in the blood samples of leukemia patients was 4.71 ppb (female, 45 years old, from Basrah) and the minimum concentration was 1.91 ppb (male, 3 years old, from Muthanna). For healthy group, the maximum uranium concentration was 2.15 ppb (female, 55 years old, from Basrah) and the minimum concentration was 0.86 ppb (male, 5 years old, from Dhi-Qar). It has been found that the uranium concentrations in human blood samples of leukemia patients are higher than those of the healthy group. These uranium concentrations in the leukemia patients group were significantly different (P < 0.001) from those in the healthy group. (author)

A simple method for measurement of cerebral blood flow using 123I-IMP SPECT with calibrated standard input function by one point blood sampling. Validation of calibration by one point venous blood sampling as a substitute for arterial blood sampling

International Nuclear Information System (INIS)

Ito, Hiroshi; Akaizawa, Takashi; Goto, Ryoui

1994-01-01

In a simplified method for measurement of cerebral blood flow using one 123 I-IMP SPECT scan and one point arterial blood sampling (Autoradiography method), input function is obtained by calibrating a standard input function by one point arterial blood sampling. A purpose of this study is validation of calibration by one point venous blood sampling as a substitute for one point arterial blood sampling. After intravenous infusion of 123 I-IMP, frequent arterial and venous blood sampling were simultaneously performed on 12 patients of CNS disease without any heart and lung disease and 5 normal volunteers. The radioactivity ratio of venous whole blood which obtained from cutaneous cubital vein to arterial whole blood were 0.76±0.08, 0.80±0.05, 0.81±0.06, 0.83±0.11 at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivities were always 20% lower than those of arterial blood radioactivity during 50 min. However, the ratio which obtained from cutaneous dorsal hand vein to artery were 0.93±0.02, 0.94±0.05, 0.98±0.04, 0.98±0.03, at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivity was consistent with artery. These indicate that arterio-venous difference of radioactivity in a peripheral cutaneous vein like a dorsal hand vein is minimal due to arteriovenous shunt in palm. Therefore, a substitution by blood sampling from cutaneous dorsal hand vein for artery will be possible. Optimized time for venous blood sampling evaluated by error analysis was 20 min after 123 I-IMP infusion, which is 10 min later than that of arterial blood sampling. (author)

On-chip sample preparation for complete blood count from raw blood.

Science.gov (United States)

Nguyen, John; Wei, Yuan; Zheng, Yi; Wang, Chen; Sun, Yu

2015-03-21

This paper describes a monolithic microfluidic device capable of on-chip sample preparation for both RBC and WBC measurements from whole blood. For the first time, on-chip sample processing (e.g. dilution, lysis, and filtration) and downstream single cell measurement were fully integrated to enable sample preparation and single cell analysis from whole blood on a single device. The device consists of two parallel sub-systems that perform sample processing and electrical measurements for measuring RBC and WBC parameters. The system provides a modular environment capable of handling solutions of various viscosities by adjusting the length of channels and precisely controlling mixing ratios, and features a new 'offset' filter configuration for increased duration of device operation. RBC concentration, mean corpuscular volume (MCV), cell distribution width, WBC concentration and differential are determined by electrical impedance measurement. Experimental characterization of over 100,000 cells from 10 patient blood samples validated the system's capability for performing on-chip raw blood processing and measurement.

Effect of Sample Storage Temperature and Time Delay on Blood Gases, Bicarbonate and pH in Human Arterial Blood Samples.

Science.gov (United States)

Mohammadhoseini, Elham; Safavi, Enayat; Seifi, Sepideh; Seifirad, Soroush; Firoozbakhsh, Shahram; Peiman, Soheil

2015-03-01

Results of arterial blood gas analysis can be biased by pre-analytical factors, such as time interval before analysis, temperature during storage and syringe type. To investigate the effects of samples storage temperature and time delay on blood gases, bicarbonate and PH results in human arterial blood samples. 2.5 mL arterial blood samples were drawn from 45 patients via an indwelling Intraarterial catheter. Each sample was divided into five equal samples and stored in multipurpose tuberculin plastic syringes. Blood gas analysis was performed on one of five samples as soon as possible. Four other samples were divided into two groups stored at 22°C and 0°C. Blood gas analyses were repeated at 30 and 60 minutes after sampling. PaO2 of the samples stored at 0°C was increased significantly after 60 minutes (P = 0.007). The PaCO2 of the samples kept for 30 and 60 minutes at 22°C was significantly higher than primary result (P = 0.04, P samples stored at 22°C, pH decreased significantly after 30 and 60 minutes (P = 0.017, P = 0.001). There were no significant differences in other results of samples stored at 0°C or 22°C after 30 or 60 minutes. In samples stored in plastic syringes, overestimation of PaO2 levels should be noted if samples cooled before analysis. In samples stored in plastic syringes, it is not necessary to store samples in iced water when analysis delayed up to one hour.

A simple method for regional cerebral blood flow measurement by one-point arterial blood sampling and 123I-IMP microsphere model (part 2). A study of time correction of one-point blood sample count

International Nuclear Information System (INIS)

Masuda, Yasuhiko; Makino, Kenichi; Gotoh, Satoshi

1999-01-01

In our previous paper regarding determination of the regional cerebral blood flow (rCBF) using the 123 I-IMP microsphere model, we reported that the accuracy of determination of the integrated value of the input function from one-point arterial blood sampling can be increased by performing correction using the 5 min: 29 min ratio for the whole-brain count. However, failure to carry out the arterial blood collection at exactly 5 minutes after 123 I-IMP injection causes errors with this method, and there is thus a time limitation. We have now revised out method so that the one-point arterial blood sampling can be performed at any time during the interval between 5 minutes and 20 minutes after 123 I-IMP injection, with addition of a correction step for the sampling time. This revised method permits more accurate estimation of the integral of the input functions. This method was then applied to 174 experimental subjects: one-point blood samples collected at random times between 5 and 20 minutes, and the estimated values for the continuous arterial octanol extraction count (COC) were determined. The mean error rate between the COC and the actual measured continuous arterial octanol extraction count (OC) was 3.6%, and the standard deviation was 12.7%. Accordingly, in 70% of the cases, the rCBF was able to be estimated within an error rate of 13%, while estimation was possible in 95% of the cases within an error rate of 25%. This improved method is a simple technique for determination of the rCBF by 123 I-IMP microsphere model and one-point arterial blood sampling which no longer shows a time limitation and does not require any octanol extraction step. (author)

Ocular Blood Flow Measurements in Healthy White Subjects Using Laser Speckle Flowgraphy.

Directory of Open Access Journals (Sweden)

Nikolaus Luft

Full Text Available To assess the feasibility and reliability of Laser Speckle Flowgraphy (LSFG to measure ocular perfusion in a sample of healthy white subjects and to elucidate the age-dependence of the parameters obtained.This cross-sectional study included 80 eyes of 80 healthy, non-smoking white subjects of Western European descent between 19 and 79 years of age. A commercial LSFG instrument was applied to measure ocular blood flow at the optic nerve head (ONH three successive times before and after pharmacological pupil dilation. The mean blur rate (MBR, a measure of relative blood flow velocity, was obtained for different regions of the ONH. Eight parameters of ocular perfusion derived from the pulse-waveform analysis of MBR including blowout time (BOT and falling rate (FR were also recorded.Artifact-free LSFG images meeting the quality criteria for automated image analysis were obtainable in 93.8% without pupil dilation and in 98.8% with pharmacological pupil dilation. Measurements of MBR showed excellent repeatability with intraclass correlation coefficients ≥ 0.937 and were barely affected by pupil dilation. The majority of pulse-waveform derived variables exhibited equally high repeatability. MBR-related blood flow indices exhibited significant age dependence (p<0.001. FR (r = 0.747, p<0.001 and BOT (r = -0.714, p<0.001 most strongly correlated with age.LSFG represents a reliable method for the quantitative assessment of ocular blood flow in white subjects. Our data affirms that the LSFG-derived variables FR and BOT may be useful biomarkers for age-related changes in ocular perfusion.

The effects of storage temperature and duration of blood samples on DNA and RNA qualities.

Science.gov (United States)

Huang, Lien-Hung; Lin, Pei-Hsien; Tsai, Kuo-Wang; Wang, Liang-Jen; Huang, Ying-Hsien; Kuo, Ho-Chang; Li, Sung-Chou

2017-01-01

DNA and RNA samples from blood are the common examination target for non-invasive physical tests and/or biomedical studies. Since high-quality DNA and RNA samples guarantee the correctness of these tests and/or studies, we investigated the effects of storage temperature and storage duration of whole blood on DNA and RNA qualities. Subjects were enrolled to donate blood samples which were stored for different durations and at different temperatures, followed by the examinations on RNA quality, qPCR, DNA quality and DNA methylation. For RNA, we observed obvious quality decline with storage duration longer than 24 hours. Storage at low temperature does not keep RNA samples from degradation. And, storing whole blood samples in freezer dramatically damage RNA. For DNA, quality decline was not observed even with storage duration for 15 days. However, DNA methylation significantly altered with storage duration longer than three days. Storage duration within 24 hours is critical for collecting high-quality RNA samples for next-generation sequencing (NGS) assays (RIN≧8). If microarray assays are expected (RIN≧7), storage duration within 32 hours is acceptable. Although DNA is resistant within 15 days when kept in whole blood, DNA quantity dramatically decreases owing to WBC lysis. In addition, duration for more than three days significantly alter DNA methylation status, globally and locally. Our result provides a reference for dealing with blood samples.

Reproducibility of blood pressure variation in older ambulatory and bedridden subjects.

Science.gov (United States)

Tsuchihashi, Takuya; Kawakami, Yasunobu; Imamura, Tsuyoshi; Abe, Isao

2002-06-01

We investigated the influence of ambulation on the reproducibility of circadian blood pressure variation in older nursing home residents. Ambulatory blood pressure monitoring was performed twice in 37 older nursing home residents. Nursing home in Japan. Subjects included 18 ambulatory nursing home residents who had no limitation on physical activity and 19 bedridden residents who did not participate in physical activity. Twenty-four-hour, daytime, and nighttime blood pressure levels and their variability. The 24-hour and daytime variability of systolic blood pressure (SBP) was significantly greater in ambulatory than in bedridden subjects, whereas nighttime variability was similar. Significant correlations in SBP averaged for the whole day, daytime, and nighttime were observed between the two examinations in ambulatory (r =.80-.83) and bedridden (r =.83-.91) subjects, but the variabilities of SBP for the whole day and during the daytime of the first measurement were correlated with those of the second measurement in bedridden (r =.67 and r =.47, respectively) but not in ambulatory (r =.39 and r =.28, respectively) subjects. Significant correlations were found between the nocturnal SBP changes at two occasions in both ambulatory (r =.50) and bedridden (r =.51) subjects, but the dipper versus nondipper profiles, defined as reduction in SBP of greater than 10% versus not, showed low reproducibility in ambulatory subjects; five ambulatory (28%) and one bedridden (5%) subjects showed divergent profiles between the two examinations. The reproducibility of blood pressure variation in nursing home residents is influenced by ambulation.

Characterization of the disposition of fostamatinib in Japanese subjects including pharmacokinetic assessment in dry blood spots: results from two phase I clinical studies.

Science.gov (United States)

Martin, Paul; Cheung, S Y Amy; Yen, Mark; Han, David; Gillen, Michael

2016-01-01

The aims of the present study were to characterize the pharmacokinetics of fostamatinib in two phase I studies in healthy Japanese subjects after single- and multiple-dose administration, and to evaluate the utility of dried blood spot (DBS) sampling. In study A, 40 Japanese and 16 white subjects were randomized in a double-blind parallel group study consisting of seven cohorts, which received either placebo or a fostamatinib dose between 50 and 200 mg after single and multiple dosing. Pharmacokinetics of R406 (active metabolite of fostamatinib) in plasma and urine was assessed, and safety was intensively monitored. Study B was an open-label study that assessed fostamatinib 100 and 200 mg in 24 Japanese subjects. In addition to plasma and urine sampling (as for study A), pharmacokinetics was also assessed in blood. Mean maximum plasma concentration (C max) and area under total plasma concentration–time curve (AUC) increased with increasing dose in Japanese subjects. Steady state was achieved in 5–7 days for all doses. C max and AUC were both higher in Japanese subjects administered a 150-mg single dose than in white subjects. This difference was maintained for steady state exposure by day 10. Overall, R406 blood concentrations were consistent and ∼2.5-fold higher than in plasma. Minimal (blood cells, and DBS sampling was a useful method for assessing R406 pharmacokinetics.

Measurement of regional cerebral blood flow using one-point arterial blood sampling and microsphere model with 123I-IMP. Correction of one-point arterial sampling count by whole brain count ratio

International Nuclear Information System (INIS)

Makino, Kenichi; Masuda, Yasuhiko; Gotoh, Satoshi

1998-01-01

The experimental subjects were 189 patients with cerebrovascular disorders. 123 I-IMP, 222 MBq, was administered by intravenous infusion. Continuous arterial blood sampling was carried out for 5 minutes, and arterial blood was also sampled once at 5 minutes after 123 I-IMP administration. Then the whole blood count of the one-point arterial sampling was compared with the octanol-extracted count of the continuous arterial sampling. A positive correlation was found between the two values. The ratio of the continuous sampling octanol-extracted count (OC) to the one-point sampling whole blood count (TC5) was compared with the whole brain count ratio (5:29 ratio, Cn) using 1-minute planar SPECT images, centering on 5 and 29 minutes after 123 I-IMP administration. Correlation was found between the two values. The following relationship was shown from the correlation equation. OC/TC5=0.390969 x Cn-0.08924. Based on this correlation equation, we calculated the theoretical continuous arterial sampling octanol-extracted count (COC). COC=TC5 x (0.390969 x Cn-0.08924). There was good correlation between the value calculated with this equation and the actually measured value. The coefficient improved to r=0.94 from the r=0.87 obtained before using the 5:29 ratio for correction. For 23 of these 189 cases, another one-point arterial sampling was carried out at 6, 7, 8, 9 and 10 minutes after the administration of 123 I-IMP. The correlation coefficient was also improved for these other point samplings when this correction method using the 5:29 ratio was applied. It was concluded that it is possible to obtain highly accurate input functions, i.e., calculated continuous arterial sampling octanol-extracted counts, using one-point arterial sampling whole blood counts by performing correction using the 5:29 ratio. (K.H.)

Multidrug resistant Salmonellae isolated from blood culture samples ...

African Journals Online (AJOL)

This study investigates the prevalence of R-plasmids in Salmonella sp. isolated from blood samples of suspected typhoid patients in Warri, Nigeria. A total of 136 blood samples were collected between May and December,2009 and screened for the presence of Salmonellae using standard blood culture techniques of which ...

Blood oxygen saturation determined by transmission spectrophotometry of hemolyzed blood samples

Science.gov (United States)

Malik, W. M.

1967-01-01

Use of the Lambert-Beer Transmission Law determines blood oxygen saturation of hemolyzed blood samples. This simplified method is based on the difference in optical absorption properties of hemoglobin and oxyhemoglobin.

Non-terminal blood sampling techniques in guinea pigs.

Science.gov (United States)

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.

The relationship between blood viscosity and blood pressure in a random sample of the population aged 55 to 74 years.

Science.gov (United States)

Fowkes, F G; Lowe, G D; Rumley, A; Lennie, S E; Smith, F B; Donnan, P T

1993-05-01

Blood viscosity is elevated in hypertensive subjects, but the association of viscosity with arterial blood pressure in the general population, and the influence of social, lifestyle and disease characteristics on this association, are not established. In the Edinburgh Artery Study, 1592 men and women aged 55-74 years selected randomly from the general population attended a university clinic. A fasting blood sample was taken for the measurement of blood viscosity and its major determinants (haematocrit, plasma viscosity and fibrinogen). Systolic pressure was related univariately to blood viscosity (P viscosity (P index. Diastolic pressure was related univariately to blood viscosity (P viscosity (P viscosity and systolic pressure was confined to males. Blood viscosity was associated equally with systolic and diastolic pressures in males, and remained independently related on multivariate analysis adjusting for age, sex, body mass index, social class, smoking, alcohol intake, exercise, angina, HDL and non-HDL cholesterol, diabetes mellitus, plasma viscosity, fibrinogen, and haematocrit.

A method to quantitate cerebral blood flow using a rotating gamma camera and iodine-123 iodoamphetamine with one blood sampling

International Nuclear Information System (INIS)

Iida, Hidehiro; Itoh, Hiroshi; Bloomfield, P.M.; Munaka, Masahiro; Higano, Shuichi; Murakami, Matsutaro; Inugami, Atsushi; Eberl, S.; Aizawa, Yasuo; Kanno, Iwao; Uemura, Kazuo

1994-01-01

A method has been developed to quantitate regional cerebral blood blow (rCBF) using iodine-123-labelled N-isopropyl-p-iodoamphetamine (IMP). This technique requires only two single-photon emission tomography (SPET) scans and one blood sample. Based on a two-compartment model, radioactivity concentrations in the brain for each scan time are calculated. A standard input function has been generated by combining the input functions from 12 independent studies prior to this work to avoid frequent arterial blood sampling, and one blood sample is taken at 10 min following IMP administration for calibration of the standard arterial input function. This calibration time was determined such that the integration of the first 40 min of the calibrated, combined input function agreed best with those from 12 individual input functions (the difference was 5.3% on average). This method was applied to eight subjects (two normals and six patients with cerebral infarction), and yielded rCBF values which agreed well with those obtained by a positron emission tomography H 2 15 O autoradiography method. This method was also found to provide rCBF values that were consistent with those obtained by the non-linear least squares fitting technique and those obtained by conventional microsphere model analysis. The optimum SPET scan times were found to be 40 and 180 min for the early and delayed scans, respectively. These scan times allow the use of a conventional rotating gamma camera for clinical purposes. V d values ranged between 10 and 40 ml/g depending on the pathological condition, thereby suggesting the importance of measuring V d for each ROI. In conclusion, optimization of the blood sampling time and the scanning time enabled quantitative measurement of rCBF with two SPET scans and one blood sample. (orig.)

Measurement of cerebral blood flow the blood sampling method using 99mTc-ECD. Simultaneous scintigram scanning of arterial blood samples and the brain with a gamma camera

International Nuclear Information System (INIS)

Hachiya, Takenori; Inugami, Atsushi; Iida, Hidehiro; Mizuta, Yoshihiko; Kawakami, Takeshi; Inoue, Minoru

1999-01-01

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

Research results: preserving newborn blood samples.

Science.gov (United States)

Lewis, Michelle Huckaby; Scheurer, Michael E; Green, Robert C; McGuire, Amy L

2012-11-07

Retention and use, without explicit parental permission, of residual dried blood samples from newborn screening has generated public controversy over concerns about violations of family privacy rights and loss of parental autonomy. The public debate about this issue has included little discussion about the destruction of a potentially valuable public resource that can be used for research that may yield improvements in public health. The research community must advocate for policies and infrastructure that promote retention of residual dried blood samples and their use in biomedical research.

Platelet-rich fibrin prepared from stored whole-blood samples.

Science.gov (United States)

Isobe, Kazushige; Suzuki, Masashi; Watanabe, Taisuke; Kitamura, Yutaka; Suzuki, Taiji; Kawabata, Hideo; Nakamura, Masayuki; Okudera, Toshimitsu; Okudera, Hajime; Uematsu, Kohya; Nakata, Koh; Tanaka, Takaaki; Kawase, Tomoyuki

2017-12-01

In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics. Blood samples were collected from non-smoking, healthy male donors (aged 27-67 years, N = 6), and PRF clots were prepared immediately or after storage for 1-2 days. Fibrin fiber was examined by scanning electron microscopy. Bioactivity was evaluated by means of a bioassay system involving human periosteal cells, whereas PDGF-BB concentrations were determined by an enzyme-linked immunosorbent assay. Addition of optimal amounts of a 10% CaCl 2 solution restored the coagulative ability of whole-blood samples that contained an anticoagulant (acid citrate dextrose) and were stored for up to 2 days at ambient temperature. In PRF clots prepared from the stored whole-blood samples, the thickness and cross-links of fibrin fibers were almost identical to those of freshly prepared PRF clots. PDGF-BB concentrations in the PRF extract were significantly lower in stored whole-blood samples than in fresh samples; however, both extracts had similar stimulatory effects on periosteal-cell proliferation. Quality of PRF clots prepared from stored whole-blood samples is not reduced significantly and can be ensured for use in regenerative therapy. Therefore, the proposed method enables a more flexible treatment schedule and choice of a more suitable platelet concentrate immediately before treatment, not after blood collection.

Impact of partial pressure of oxygen in blood samples on the performance of systems for self-monitoring of blood glucose.

Science.gov (United States)

Schmid, Christina; Baumstark, Annette; Pleus, Stefan; Haug, Cornelia; Tesar, Martina; Freckmann, Guido

2014-03-01

The partial pressure of oxygen (pO2) in blood samples can affect glucose measurements with oxygen-sensitive systems. In this study, we assessed the influence of different pO2 levels on blood glucose (BG) measurements with five glucose oxidase (GOD) systems and one glucose dehydrogenase (GDH) system. All selected GOD systems were indicated by the manufacturers to be sensitive to increased oxygen content of the blood sample. Venous blood samples of 16 subjects (eight women, eight men; mean age, 52 years; three with type 1 diabetes, four with type 2 diabetes, and nine without diabetes) were collected. Aliquots of each sample were adjusted to the following pO2 values: ≤45 mm Hg, approximately 70 mm Hg, and ≥150 mm Hg. For each system, five consecutive measurements on each sample were performed using the same test strip lot. Relative differences between the mean BG value at a pO2 level of approximately 70 mm Hg, which was considered to be similar to pO2 values in capillary blood samples, and the mean BG value at pO2 levels ≤45 mm Hg and ≥150 mm Hg were calculated. The GOD systems showed mean relative differences between 11.8% and 44.5% at pO2 values ≤45 mm Hg and between -14.6% and -21.2% at pO2 values ≥150 mm Hg. For the GDH system, the mean relative differences were -0.3% and -0.2% at pO2 values ≤45 mm Hg and ≥150 mm Hg, respectively. The magnitude of the pO2 impact on BG measurements seems to vary among the tested oxygen-sensitive GOD systems. The pO2 range in which oxygen-sensitive systems operate well should be provided in the product information.

Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads.

Science.gov (United States)

Langenskiöld, Cecilia; Mellgren, Karin; Abrahamsson, Jonas; Bemark, Mats

2016-06-24

In many studies it would be advantageous if blood samples could be collected and analyzed using flow cytometry at a later stage. Ideally, sample collection should involve little hands-on time, allow for long-term storage, and minimally influence the samples. Here we establish a flow cytometry antibody panel that can be used to determine granulocytes, monocytes, and lymphocyte subset concentrations in fresh and frozen whole blood using TruCount technology. The panel can be used on fresh whole-blood samples as well as whole-blood samples that have been frozen after mixing with 10% DMSO. Concentrations in frozen and fresh sample is highly correlated both when frozen within 4 h and the day after collection (r ≥ 0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. Using this method whole-blood samples can be frozen using a simple preparation method, and stored long-term before accurate determination of cell concentration. This allows for standardized analysis of the samples at a reference laboratory in multi-center studies. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

Stability and reliability of glycated haemoglobin measurements in blood samples stored at -20°C.

Science.gov (United States)

Venkataraman, Vijayachandrika; Anjana, Ranjit Mohan; Pradeepa, Rajendra; Deepa, Mohan; Jayashri, Ramamoorthy; Anbalagan, Viknesh Prabu; Akila, Bridgitte; Madhu, Sri Venkata; Lakshmy, Ramakrishnan; Mohan, Viswanathan

2016-01-01

To validate the stability of glycated haemoglobin (HbA1c) measurements in blood samples stored at -20°C for up to one month. The study group comprised 142 type 2 diabetic subjects visiting a tertiary centre for diabetes at Chennai city in south India. The HbA1c assay was done on a fasting blood sample using the Bio-Rad Variant machine on Day 0 (day of blood sample collection). Several aliquots were stored at -20°C and the assay was repeated on the 3rd, 7th, 15th, and 30th day after the sample collection. Bland-Altman plots were constructed and variation in the HbA1c levels on the different days was compared with the day 0 level. The median differences between HbA1c levels measured on Day 0 and the 3rd, 7th, 15th, and 30th day after blood collection were 0.0%, 0.2%, 0.3% and 0.5% respectively. Bland-Altman plot analysis showed that the differences between the day '0' and the different time points tend to get larger with time, but these were not clinically significant. HbA1c levels are relatively stable up to 2weeks, if blood samples are stored at -20°C. Copyright © 2016 Elsevier Inc. All rights reserved.

Effect of postmortem sampling technique on the clinical significance of autopsy blood cultures.

Science.gov (United States)

Hove, M; Pencil, S D

1998-02-01

Our objective was to investigate the value of postmortem autopsy blood cultures performed with an iodine-subclavian technique relative to the classical method of atrial heat searing and antemortem blood cultures. The study consisted of a prospective autopsy series with each case serving as its own control relative to subsequent testing, and a retrospective survey of patients coming to autopsy who had both autopsy blood cultures and premortem blood cultures. A busy academic autopsy service (600 cases per year) at University of Texas Medical Branch Hospitals, Galveston, Texas, served as the setting for this work. The incidence of non-clinically relevant (false-positive) culture results were compared using different methods for collecting blood samples in a prospective series of 38 adult autopsy specimens. One hundred eleven adult autopsy specimens in which both postmortem and antemortem blood cultures were obtained were studied retrospectively. For both studies, positive culture results were scored as either clinically relevant or false positives based on analysis of the autopsy findings and the clinical summary. The rate of false-positive culture results obtained by an iodine-subclavian technique from blood drawn soon after death were statistically significantly lower (13%) than using the classical method of obtaining blood through the atrium after heat searing at the time of the autopsy (34%) in the same set of autopsy subjects. When autopsy results were compared with subjects' antemortem blood culture results, there was no significant difference in the rate of non-clinically relevant culture results in a paired retrospective series of antemortem blood cultures and postmortem blood cultures using the iodine-subclavian postmortem method (11.7% v 13.5%). The results indicate that autopsy blood cultures obtained using the iodine-subclavian technique have reliability equivalent to that of antemortem blood cultures.

Total reflection x-ray analysis of metals in blood samples

International Nuclear Information System (INIS)

Nakamura, Takuya; Matsui, Hiroshi; Kawamata, Masaya

2009-01-01

The sample preparation for TXRF (total reflection X-ray fluorescence) quantitative analysis of trace elements in human blood samples was investigated. In the TXRF analysis, a solution sample is dropped and dried on a flat substrate, and then the dried residue is measured. In this case, the dried residue should be flat not to disturb X-ray total reflection on the substrate. In addition, it is required to simply measure the whole blood sample by TXRF method, although a serum is analyzed in many cases. Thus, we studied the optimum conditions of the sample preparation of the whole blood by adding the pure water to apply Hemolysis phenomenon, where blood cells are destroyed due to different of the osmotic pressure, leading to flat residue. It was found that the best S/B ratio was obtained when the whole blood was diluted 8 times with pure water. Moreover, it was investigated the influence of the surface chemical condition of the glass substrate on the shape of the dried reside of the blood sample. When the surface of the glass substrate was hydrophilic, the shape of the dried residues was not uniform, as a result, the quantitative data of TXRF analysis gave a large deviation. On the other hand, when the surface of the glass was hydrophobic, the shape of the residue was almost uniform, as a result, a good reproducibility was obtained. Another problem was an outer ring of the dried residue of the blood. This uneven ring absorbs the primary X-rays, caused to low determined quantitative data. Thus, we tried the heating way of the dropped blood sample at a high temperature of 200 degrees. In this case, the blood sample was dried immediately, and a flat homogeneous dried residue was obtained without the outer ring. Using the optimized conditions for sample preparation, human blood sample was quantitatively measured by TXRF and ICP-AES. A good agreement was obtained in TXRF and ICP-AES determinations; however, the measurement of Cl and Br will be an advantage of TXRF, because

Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry.

Science.gov (United States)

Silva-Malta, M C F; Araujo, N C Fidélis; Vieira, O V Neves; Schmidt, L Cayres; Gonçalves, P de Cassia; Martins, M Lobato

2015-10-01

In this study, we present a strategy for RHD gene screening based on real-time polymerase chain reaction (PCR) using dried blood spots of pooled samples. Molecular analysis of blood donors may be used to detect RHD variants among the presumed D-negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost. RHD gene detection based on real-time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD-negative. Positive results were re-evaluated by retesting single samples using real-time PCR and conventional multiplex PCR to amplify five RHD-specific exons. PCR-sequence-specific primers was used to amplify RHDψ allele. We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D-negative blood donors, 58 (3.74%) had the RHD gene. The non-functional RHDψ allele was detected in 47 samples (3.02%). The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry. © 2015 British Blood Transfusion Society.

Student measurement of blood pressure using a simulator arm compared with a live subject's arm.

Science.gov (United States)

Lee, Jennifer J; Sobieraj, Diana M; Kuti, Effie L

2010-06-15

To compare accuracy of blood pressure measurements using a live subject and a simulator arm, and to determine students' preferences regarding measurement. This was a crossover study comparing blood pressure measurements from a live subject and a simulator arm. Students completed an anonymous survey instrument defining opinions on ease of measurement. Fifty-seven students completed blood pressure measurements on live subjects while 72 students completed blood pressure measurements using the simulator arm. There were no significant systematic differences between the 2 measurement techniques. Systolic blood pressure measurements from a live subject arm were less likely to be within 4 mm Hg compared with measurements of a simulator arm. Diastolic blood pressure measurements were not significantly different between the 2 techniques. Accuracy of student measurement of blood pressure using a simulator arm was similar to the accuracy with a live subject. There was no difference in students' preferences regarding measurement techniques.

Neonatal blood gas sampling methods | Goenka | South African ...

African Journals Online (AJOL)

There is little published guidance that systematically evaluates the different methods of neonatal blood gas sampling, where each method has its individual benefits and risks. This review critically surveys the available evidence to generate a comparison between arterial and capillary blood gas sampling, focusing on their ...

Combination syringe provides air-free blood samples

Science.gov (United States)

Pool, S. L.

1970-01-01

Standard syringe and spinal needle are combined in unique manner to secure air-free blood samples. Combination syringe obtains air free samples because air bubbles become insignificant when samples greater than 1 cc are drawn.

Stability of carboxyhemoglobin in stored and mailed blood samples.

Science.gov (United States)

Hampson, Neil B

2008-02-01

Elevated blood carboxyhemoglobin (COHb) levels are used to confirm a clinical diagnosis of exposure to carbon monoxide (CO) and, in some instances, assess severity of poisoning. However, many hospital laboratories cannot measure COHb because they do not have CO-oximeters. In such instances, blood samples are often sent to outside laboratories or with a transported patient for measurement at the receiving hospital. This study was conducted to assess the stability of COHb in stored and mailed blood samples anticoagulated with heparin. Adult human blood was drawn into standard sample tubes anticoagulated with sodium heparin. Carbon monoxide gas was infused to raise the COHb level to 25% to 35%. Samples were then refrigerated or stored at room temperature, and serial COHb determinations were performed for 28 days. Additional samples were measured after being mailed locally or across the United States and back. No significant changes in COHb levels were seen in samples stored either in refrigeration or at room temperature over a period of 28 days or in samples shipped without refrigeration locally or across the United States. Carboxyhemoglobin levels in whole blood samples anticoagulated with heparin are stable with or without refrigeration for up to 4 weeks. If COHb measurement capability is not available, such samples may be shipped or transported with patients with confidence that the COHb level will be stable when measured at a later time.

Blood donors and factors impacting the blood donation decision: motives for donating blood in Turkish sample.

Science.gov (United States)

Karacan, Eda; Cengiz Seval, Guldane; Aktan, Zeynep; Ayli, Meltem; Palabiyikoglu, Refia

2013-12-01

Donations in Turkey are insufficient to cover the high transfusion needs arising from large numbers of thalassemia and sickle cell anemia patients and increasing demands for blood due to advanced surgery and cancer treatment. The most acceptable means to get blood is voluntary blood donation and the blood donor system in Turkey mostly depends on a combination of voluntary and involuntary donors. The main aim of this study is to explore the motivations of Turkish voluntary blood donors toward blood donation and to determine predictors of blood donation motivation. A cross-sectional sample survey of active blood donors in Ankara, Turkey was conducted. The sample consisted of 189 male volunteer blood donor adults. Donors filled in a self-administered questionnaire including the measures of demographic information, empathetic concern, altruism, social responsibility and blood donation motivation questionnaire during donation. Factor analysis of Blood Donation Motivation Measure with varimax rotation revealed a three-factor solution named as "values and moral duty", "positive feelings and esteem" and "self-benefit and external reasons". The results with regression analyses showed that only social responsibility had an significant effect independent of age, income, and education on blood donation motivation. These result reflects that blood donation motivation not only linked to a high degree of altruistic reasons, but also to a combination of some self-regarding motives. Additionally, feelings of empathy or altruism may be less strong at the time the decision to help, other factors may have a larger influence on helping decisions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Dynamic 2-[18F]fluoro-2-deoxy-d-glucose positron emission tomography of liver tumours without blood sampling

International Nuclear Information System (INIS)

Keiding, S.; Munk, O.L.; Schioett, K.M.; Hansen, S.B.

2000-01-01

Positron emission tomography (PET) using 2-[ 18 F]fluoro-2-deoxy-d-glucose (FDG) is a useful diagnostic tool for the detection of tumours. Using dynamic FDG PET, net metabolic clearance of FDG, K, can be calculated by Gjedde-Patlak analysis of the time course of the radioactivity concentrations in tissue and arterial blood. We examined whether time-activity curves (TACs) based on arterial blood sampling could be replaced by TACs obtained from the descending aorta in dynamic PET scans of patients with liver tumours. The study was performed in two parts, using data from dynamic liver scans with arterial blood sampling in human subjects: First, data from four patients with no liver tumours and five patients with liver tumours were used as a training group. Volumes of interest were defined in the descending aorta (aorta VOIs) by four different methods. K values were calculated based on the corresponding TACs and compared with those based on TACs of the arterial blood sample radioactivity concentrations. The aorta VOI which gave K values that were in best agreement with the K values based on the arterial blood sample measurements was called the AORTA-VOI. Use of the AORTA-VOI was subsequently tested in a test group of 19 tumour patients by comparing the K values from the AORTA-VOI with the K values based on the arterial blood sample measurements. The AORTA-VOI consisted of the sum of small regions of interest (ROIs) drawn in the centre of the aorta (approximately six pixels of 2.4 x 2.4 mm per transaxial slice of 3.1 mm thickness) in as many transaxial slices as possible (30-40 slices). There were no statistically significant differences between the two sets of K values. The ratio of K values in tumour tissue to K values in reference tissue was 2.1-9.7:1 (mean, 5.4:1) based on the AORTA TACs, and 2.1-8.4:1 (mean, 4.6:1) based on blood sample TACs (P>0.3). We conclude that arterial blood sampling can be replaced by the present AORTA-VOI in the calculation of the net

Congener Production in Blood Samples During Preparation and Storage

DEFF Research Database (Denmark)

Felby, Søren; Nielsen, Erik

1995-01-01

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

High plasma corticosterone levels persist during frequent automatic blood sampling in rats

DEFF Research Database (Denmark)

Abelson, Klas S P; Adem, Bashir; Royo, Felix

2005-01-01

Corticosterone levels in blood may be used as a marker of stress in rodents, provided that the blood sampling procedure itself is non-stressful. Automated blood sampling equipment (Accusampler) allows blood sampling without any interference with the animal and might be useful as a tool for an on......-line measurement of stress markers in blood. However, the impact of the blood sampling itself on the corticosterone levels in blood is unknown. The present study was designed to evaluate whether the frequency of blood sampling influences the plasma corticosterone levels in male and female rats. During anaesthesia...... the importance of considering the frequency of blood withdrawal during automated blood sampling. This parameter may have an impact on the experimental results when using blood corticosterone levels as a stress marker, but also during any in vivo study where blood is collected, since high corticosterone levels...

Automated nucleic acid amplification testing in blood banks: An additional layer of blood safety

Directory of Open Access Journals (Sweden)

Pragati Chigurupati

2015-01-01

Full Text Available Context: A total of 30 million blood components are transfused each year in India. Blood safety thus becomes a top priority, especially with a population of around 1.23 billion and a high prevalence rate of human immunodeficiency virus (HIV, hepatitis B virus (HBV and hepatitis C virus (HCV in general population. Nucleic acid amplification testing (NAT in blood donor screening has been implemented in many developed countries to reduce the risk of transfusion-transmitted viral infections (TTIs. NAT takes care of the dynamics of window period of viruses and offers the safest blood pack for donation. Aims: The aim of this study is to show the value of NAT in blood screening. Settings and Design: Dhanavantari Blood Bank, Rajahmundry, Andhra Pradesh, India. Subjects and Methods: Over a period of 1 year from January 2012 to December 2012, a total number of 15,000 blood donor samples were subjected to tests for HIV, HBV, and HCV by enzyme-linked immunosorbent assay (ELISA method and 8000 ELISA nonreactive samples were subjected for NAT using multiplex polymerase chain reaction technology. Results: Of the 15,000 donors tested, 525 were seroreactive. In 8000 ELISA negative blood samples subjected to NAT, 4 donor samples were reactive for HBV. The NAT yield was 1 in 2000. Conclusions: NAT could detect HIV, HBV, and HCV cases in blood donor samples those were undetected by serological tests. NAT could interdict 2500 infectious donations among our approximate 5 million annual blood donations.

Blood Sample Transportation by Pneumatic Transportation Systems

DEFF Research Database (Denmark)

Nybo, Mads; Lund, Merete E; Titlestad, Kjell

2018-01-01

BACKGROUND: Pneumatic transportation systems (PTSs) are increasingly used for transportation of blood samples to the core laboratory. Many studies have investigated the impact of these systems on different types of analyses, but to elucidate whether PTSs in general are safe for transportation...... analysis, and the hemolysis index). CONCLUSIONS: Owing to their high degree of heterogeneity, the retrieved studies were unable to supply evidence for the safety of using PTSs for blood sample transportation. In consequence, laboratories need to measure and document the actual acceleration forces...

A femoral arteriovenous shunt facilitates arterial whole blood sampling in animals

International Nuclear Information System (INIS)

Weber, Bruno; Burger, Cyrill; Buck, Alfred; Biro, Peter

2002-01-01

In this study we evaluated on-line continuous blood sampling in a femoral arteriovenous (a-v) shunt for use in quantitative tracer studies using gamma-emitting radionuclides in animals. The shunt consisted of 40 cm polyethylene tubing (PE-50) guided through a coincidence probe. Two three-way valves allowed blood pressure measurements and tracer injection. Blood flow in the shunt and the impulse response function (IRF) were assessed using heparinized human blood mixed with fluorine-18 fluorodeoxyglucose (FDG). In vivo experiments were performed in eight male rats (300-350 g) anaesthetized with halothane. In three rats, manual blood sampling was performed in parallel with on-line sampling. In another five animals, the arterial whole blood activity was recorded on-line for 40 min. For the experiments 150-180 MBq FDG was injected over 35 s. Blood flow in the shunt was 23.6, 29.2 and 42.8 ml/h at 100, 120 and 160 mmHg, respectively. The IRF was characterized by minimal dispersion (1-2 s FWHM). Deconvolution of the measured arterial input curves with the IRF changed the measured curve only minimally. Whole blood radioactivity concentration derived from manual and on-line sampling were in excellent agreement. The curves derived from on-line sampling were of high statistical quality. In conclusion, a femoral a-v shunt allows multiple manipulations such as measurement of the arterial whole blood activity, continuous blood pressure monitoring, injection of the tracer and collection of blood samples if necessary. It is not associated with blood loss if the collection of blood samples is not required. It is more convenient to use than manual sampling, the peak of the input curve is never missed and the input curves are of high statistical quality. (orig.)

Studies of U in the blood of two population samples

International Nuclear Information System (INIS)

Segovia, N.; Olguin, M.E.; Romero, M.

1986-01-01

The present work, attempts to establish the statistical distribution of blood uranium in a population of the same community, similar in age and in living patterns. U traces were evaluated by a fission track technique both in whole blood and plasma samples. Dried samples were compressed into pellets and irradiated in a nuclear reactor using the external detector method. For U quantification, standard U samples were used. A comparative sampling of U content in blood samples from a group of radiation exposed workers and another of leukemia patients was also carried out. Results from the sampling groups are reported and discussed. (author)

High-throughput miRNA profiling of human melanoma blood samples

Directory of Open Access Journals (Sweden)

Rass Knuth

2010-06-01

Full Text Available Abstract Background MicroRNA (miRNA signatures are not only found in cancer tissue but also in blood of cancer patients. Specifically, miRNA detection in blood offers the prospect of a non-invasive analysis tool. Methods Using a microarray based approach we screened almost 900 human miRNAs to detect miRNAs that are deregulated in their expression in blood cells of melanoma patients. We analyzed 55 blood samples, including 20 samples of healthy individuals, 24 samples of melanoma patients as test set, and 11 samples of melanoma patients as independent validation set. Results A hypothesis test based approch detected 51 differentially regulated miRNAs, including 21 miRNAs that were downregulated in blood cells of melanoma patients and 30 miRNAs that were upregulated in blood cells of melanoma patients as compared to blood cells of healthy controls. The tets set and the independent validation set of the melanoma samples showed a high correlation of fold changes (0.81. Applying hierarchical clustering and principal component analysis we found that blood samples of melanoma patients and healthy individuals can be well differentiated from each other based on miRNA expression analysis. Using a subset of 16 significant deregulated miRNAs, we were able to reach a classification accuracy of 97.4%, a specificity of 95% and a sensitivity of 98.9% by supervised analysis. MiRNA microarray data were validated by qRT-PCR. Conclusions Our study provides strong evidence for miRNA expression signatures of blood cells as useful biomarkers for melanoma.

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

Science.gov (United States)

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.

Response of growth hormone (GH), FFA, blood sugar and insulin to exercise in obese patients and normal subjects

NARCIS (Netherlands)

Schwarz, F.; Haar, D.J. ter; Riet, H.G. van; Thijssen, J.H.H.

1969-01-01

Ergometer tests with a constant workload of 600 Kg./min. during 30 minutes were done on eight normal subjects, eight severely obese patients, and two women who had formerly been obese. Arterial blood was sampled three times before, four times during and three times after exercise. The incidence and

Sources of variability of resting cerebral blood flow in healthy subjects

DEFF Research Database (Denmark)

Henriksen, Otto Mølby; Kruuse, Christina Rostrup; Olesen, Jes

2013-01-01

Measurements of cerebral blood flow (CBF) show large variability among healthy subjects. The aim of the present study was to investigate the relative effect of established factors influencing CBF on the variability of resting CBF. We retrospectively analyzed spontaneous variability in 430 CBF...... measurements acquired in 152 healthy, young subjects using (133)Xe single-photon emission computed tomography. Cerebral blood flow was correlated positively with both end-tidal expiratory PCO2 (PETCO2) and female gender and inversely with hematocrit (Hct). Between- and within-subject CO2 reactivity...... when Hct was also accounted for. The present study confirms large between-subject variability in CBF measurements and that gender, Hct, and PETCO2 explain only a small part of this variability. This implies that a large fraction of CBF variability may be due to unknown factors such as differences...

Automated blood-sample handling in the clinical laboratory.

Science.gov (United States)

Godolphin, W; Bodtker, K; Uyeno, D; Goh, L O

1990-09-01

The only significant advances in blood-taking in 25 years have been the disposable needle and evacuated blood-drawing tube. With the exception of a few isolated barcode experiments, most sample-tracking is performed through handwritten or computer-printed labels. Attempts to reduce the hazards of centrifugation have resulted in air-tight lids or chambers, the use of which is time-consuming and cumbersome. Most commonly used clinical analyzers require serum or plasma, distributed into specialized containers, unique to that analyzer. Aliquots for different tests are prepared by handpouring or pipetting. Moderate to large clinical laboratories perform so many different tests that even multi-analyzers performing multiple analyses on a single sample may account for only a portion of all tests ordered for a patient. Thus several aliquots of each specimen are usually required. We have developed a proprietary serial centrifuge and blood-collection tube suitable for incorporation into an automated or robotic sample-handling system. The system we propose is (a) safe--avoids or prevents biological danger to the many "handlers" of blood; (b) small--minimizes the amount of sample taken and space required to adapt to the needs of satellite and mobile testing, and direct interfacing with analyzers; (c) serial--permits each sample to be treated according to its own "merits," optimizes throughput, and facilitates flexible automation; and (d) smart--ensures quality results through monitoring and intelligent control of patient identification, sample characteristics, and separation process.

Measurement of regional cerebral blood flow using one-point venous blood sampling and causality model. Evaluation by comparing with conventional continuous arterial blood sampling method

International Nuclear Information System (INIS)

Mimura, Hiroaki; Sone, Teruki; Takahashi, Yoshitake

2008-01-01

Optimal setting of the input function is essential for the measurement of regional cerebral blood flow (rCBF) based on the microsphere model using N-isopropyl-4-[ 123 I]iodoamphetamine ( 123 I-IMP), and usually the arterial 123 I-IMP concentration (integral value) in the initial 5 min is used for this purpose. We have developed a new convenient method in which 123 I-IMP concentration in arterial blood sample is estimated from that in venous blood sample. Brain perfusion single photon emission computed tomography (SPECT) with 123 I-IMP was performed in 110 cases of central nervous system disorders. The causality was analyzed between the various parameters of SPECT data and the ratio of octanol-extracted arterial radioactivity concentration during the first 5 min (Caoct) to octanol-extracted venous radioactivity concentration at 27 min after intravenous injection of 123 I-IMP (Cvoct). A high correlation was observed between the measured and estimated values of Caoct/Cvoct (r=0.856) when the following five parameters were included in the regression formula: radioactivity concentration in venous blood sampled at 27 min (Cv), Cvoct, Cvoct/Cv, and total brain radioactivity counts that were measured by a four-head gamma camera 5 min and 28 min after 123 I-IMP injection. Furthermore, the rCBF values obtained using the input parameters estimated by this method were also highly correlated with the rCBF values measured using the continuous arterial blood sampling method (r=0.912). These results suggest that this method would serve as the new, convenient and less invasive method of rCBF measurement in clinical setting. (author)

Dynamic 2-[18F]fluoro-2-deoxy-D-glucose positron emission tomography of liver tumours without blood sampling

DEFF Research Database (Denmark)

Keiding, S; Munk, O L; Schiøtt, K M

2000-01-01

Positron emission tomography (PET) using 2-[18F]fluoro-2-deoxy-D-glucose (FDG) is a useful diagnostic tool for the detection of tumours. Using dynamic FDG PET, net metabolic clearance of FDG, K, can be calculated by Gjedde-Patlak analysis of the time course of the radioactivity concentrations...... in tissue and arterial blood. We examined whether time-activity curves (TACs) based on arterial blood sampling could be replaced by TACs obtained from the descending aorta in dynamic PET scans of patients with liver tumours. The study was performed in two parts, using data from dynamic liver scans......, and 2.1-8.4:1 (mean, 4.6:1) based on blood sample TACs (P>0.3). We conclude that arterial blood sampling can be replaced by the present AORTA-VOI in the calculation of the net metabolic clearance of FDG in dynamic PET studies of liver tumours in human subjects. Udgivelsesdato: 2000-Apr...

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

Science.gov (United States)

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

2013-10-01

In recent years, the role of donor blood has taken an important place in epidemiology of Leishmaniasis. According to the WHO, the numbers of patients considered as symptomatic are only 5-20% of individuals with asymptomatic leishmaniasis. In this study for detection of Leishmania infection in donor blood samples, 343 samples from the Capa Red Crescent Blood Center were obtained and primarily analyzed by microscopic and serological methods. Subsequently, the traditional culture (NNN), Immuno-chromatographic test (ICT) and Polymerase Chain Reaction (PCR) methods were applied to 21 samples which of them were found positive with at least one method. Buffy coat (BC) samples from 343 blood donors were analyzed: 15 (4.3%) were positive by a microculture method (MCM); and 4 (1.1%) by smear. The sera of these 343 samples included 9 (2.6%) determined positive by ELISA and 7 (2%) positive by IFAT. Thus, 21 of (6.1%) the 343 subjects studied by smear, MCM, IFAT and ELISA techniques were identified as positive for leishmaniasis at least one of the techniques and the sensitivity assessed. According to our data, the sensitivity of the methods are identified as MCM (71%), smear (19%), IFAT (33%), ELISA (42%), NNN (4%), PCR (14%) and ICT (4%). Thus, with this study for the first time, the sensitivity of a MCM was examined in blood donors by comparing MCM with the methods used in the diagnosis of leishmaniasis. As a result, MCM was found the most sensitive method for detection of Leishmania parasites in samples obtained from a blood bank. In addition, the presence of Leishmania parasites was detected in donor bloods in Istanbul, a non-endemic region of Turkey, and these results is a vital importance for the health of blood recipients. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of Glucose-6-Phosphate Dehydrogenase stability in stored blood samples.

Science.gov (United States)

Jalil, Norunaluwar; Azma, Raja Zahratul; Mohamed, Emida; Ithnin, Azlin; Alauddin, Hafiza; Baya, Siti Noor; Othman, Ainoon

2016-01-01

Glucose-6-Phosphate Dehydrogenase (G6PD) deficiency is the commonest cause of neonatal jaundice in Malaysia. Recently, OSMMR2000-D G6PD Assay Kit has been introduced to quantitate the level of G6PD activity in newborns delivered in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). As duration of sample storage prior to analysis is one of the matters of concern, this study was conducted to identify the stability of G6PD enzyme during storage. A total of 188 cord blood samples from normal term newborns delivered at UKMMC were selected for this study. The cord bloods samples were collected in ethylene-diamine-tetra-acetic acid (EDTA) tubes and refrigerated at 2-8 °C. In addition, 32 out of 188 cord blood samples were spotted on chromatography paper, air-dried and stored at room temperature. G6PD enzyme activities were measured daily for 7 days using the OSMMR2000-D G6PD Assay Kit on both the EDTA blood and dried blood samples. The mean value for G6PD activity was compared between days of analysis using Student Paired T-Test. In this study, 172 out of 188 cord blood samples showed normal enzyme levels while 16 had levels corresponding to severe enzyme deficiency. The daily mean G6PD activity for EDTA blood samples of newborns with normal G6PD activity showed a significant drop on the fourth day of storage (p samples with severely deficient G6PD activity, significant drop was seen on third day of storage (p = 0.002). Analysis of dried cord blood showed a significant reduction in enzyme activity as early as the second day of storage (p = 0.001). It was also noted that mean G6PD activity for spotted blood samples were lower compared to those in EDTA tubes for all days (p = 0.001). Thus, EDTA blood samples stored at 2-8 °C appeared to have better stability in terms of their G6PD enzyme level as compared to dried blood samples on filter paper, giving a storage time of up to 3 days.

Prevalence of dyslipidemia in normoglycemic subjects with newly diagnosed high blood pressure in Abuja, Nigeria.

Science.gov (United States)

Ojji, Dike B; Ajayi, Samuel O; Mamven, Manmak H; Atherton, John

2009-02-01

High blood pressure and dyslipidemia additively increases the risk of cardiovascular disease. There is a high prevalence of high blood pressure in Nigeria, but there are little data regarding the prevalence of dyslipidemia in subjects with high blood pressure. In this observational prospective study, we examined the prevalence of dyslipidemia in newly diagnosed normoglycemic subjects with high blood pressure. A total of 171 subjects presenting with high blood pressure for the first time in the cardiology and nephrology clinics at the University of Abuja Teaching Hospital were studied. Height, weight, and blood pressure were measured. Total cholesterol, low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were determined in fasting plasma. The total cholesterol/HDL-C and non-HDL-C values were calculated. These measures were then classified according to the 2001 report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults. Of the 171 subjects studied, 84 (49%) were male and 87 (51%) were female. Low HDL-C was present in 71 (45.8%), elevated LDL-C in 29 (17%), elevated total cholesterol in 19 (11.1%), and elevated triglyceride in 13 (7.6%), whereas eight (4.7%) of the study population had combined elevated total cholesterol and triglyceride. Female subjects had higher total cholesterol and lower HDL-C than male subjects, but these differences were not statistically significant. Obese subjects, compared to the nonobese, had significantly higher LDL-C and total cholesterol/HDL-C ratios in males and significantly higher triglyceride levels in females. Given the prevalence of dyslipidemia seen in this study, we suggest that fasting lipid measurements should be performed in all Nigerians with high blood pressure. These data suggest the need for health education and lifestyle modifications in hypertensive Nigerians to reduce both types of risk

Vessel Sampling and Blood Flow Velocity Distribution With Vessel Diameter for Characterizing the Human Bulbar Conjunctival Microvasculature.

Science.gov (United States)

Wang, Liang; Yuan, Jin; Jiang, Hong; Yan, Wentao; Cintrón-Colón, Hector R; Perez, Victor L; DeBuc, Delia C; Feuer, William J; Wang, Jianhua

2016-03-01

This study determined (1) how many vessels (i.e., the vessel sampling) are needed to reliably characterize the bulbar conjunctival microvasculature and (2) if characteristic information can be obtained from the distribution histogram of the blood flow velocity and vessel diameter. Functional slitlamp biomicroscope was used to image hundreds of venules per subject. The bulbar conjunctiva in five healthy human subjects was imaged on six different locations in the temporal bulbar conjunctiva. The histograms of the diameter and velocity were plotted to examine whether the distribution was normal. Standard errors were calculated from the standard deviation and vessel sample size. The ratio of the standard error of the mean over the population mean was used to determine the sample size cutoff. The velocity was plotted as a function of the vessel diameter to display the distribution of the diameter and velocity. The results showed that the sampling size was approximately 15 vessels, which generated a standard error equivalent to 15% of the population mean from the total vessel population. The distributions of the diameter and velocity were not only unimodal, but also somewhat positively skewed and not normal. The blood flow velocity was related to the vessel diameter (r=0.23, Psampling size of the vessels and the distribution histogram of the blood flow velocity and vessel diameter, which may lead to a better understanding of the human microvascular system of the bulbar conjunctiva.

Determination of toxic and essential elements in blood and milk from female subjects in a Lead contaminated community in Zamfara State, Nigeria

International Nuclear Information System (INIS)

Ojo, J.; Komolafe, O.; Obiajunwa, P.; Ogunba, B.; Adesanmi, C.; Kanoma, M.; Adedeji, S.; Horsvat, M.

2011-01-01

Occurrence of lead poisoning associated with gold mining and processing was reported around March 2010 in some communities of Zamfara State, leading to hundreds of fatalities, especially among children. We have evaluated the levels of lead and other elements (both toxic and essential) in human milk and whole blood samples obtained from 27 subjects at the village of Yargalma, in Anka Local Government Area of Zamfara State. Milk and blood samples were lyophilized at the Obafemi Awolowo University Teaching Hospital Complex, lIe-Ife, and were further sterilized by gamma irradiation (20 -30 kGy) at the National Radiation Centre, Abuja. Elemental analyses were carried out at the Josef Stefan Institute, Ljubljana, Slovenia for determination of Pb, Cd, As, Se, Zn and Cu by ICP-MS; and Hg by Direct Hg analyser. The significant contamination of the environment leading to the reported cases of Pb-poisoning in Zamfara gold mines have been confirmed by elevated levels of Pb measured in samples of blood and human milk. The levels of Pb in blood ranged from 7.9 - 86.8μg/dL (mean±SD: 55.6±19.6μg/dL) while levels of this toxic element in human milk ranged from 2 - 140 ng/ml (mean±SD: 45±37 ng/ml). These values, in asymptomatic subjects, are extremely high indeed. A number of interesting correlations between elemental levels in milk and blood of mothers, and the age of mother and period after birth have also been demonstrated. For instance Mothers age significantly correlates with blood Pb (confirming accumulation in bones which affect blood lead level in lactating mothers). On the other hand, Mother's age significantly correlate negatively with Blood Selenium. Levels of the essential element Zn in both blood and milk of mother significantly correlate negatively with the time elapsed after child delivery (Baby's Age) as is reported in literature. We hesitate to suggest that breast-feeding of babies be limited in these subjects, knowing the several other important nutrients

Home readings of blood pressure in assessment of hypertensive subjects

DEFF Research Database (Denmark)

Nielsen, P.E.; Myschetzky, P; Andersen, A R

1986-01-01

Out-patient clinic blood pressure (OPC-BP) was compared to home blood pressure (Home-BP) measured three times daily during a two week period in 122 consecutively referred hypertensive subjects. A semi-automatic device (TM-101) including a microphone for detection of Korotkoff-sounds, self......-deflation of cuff pressure and digital display of blood pressure was used. Mean difference between OPC-BP and Home-BP was systolic +13 mm Hg (range -21 - +100 mg Hg) and diastolic +5 mm Hg (range -27 - +36 mm Hg). Although a significant correlation could be demonstrated between Home-BP and OPC-BP, the inter...

Identifying the potential of changes to blood sample logistics using simulation

DEFF Research Database (Denmark)

Jørgensen, Pelle Morten Thomas; Jacobsen, Peter; Poulsen, Jørgen Hjelm

2013-01-01

of the simulation was to evaluate changes made to the transportation of blood samples between wards and the laboratory. The average- (AWT) and maximum waiting time (MWT) from a blood sample was drawn at the ward until it was received at the laboratory, and the distribution of arrivals of blood samples......, each of the scenarios was tested in terms of what amount of resources would give the optimal result. The simulations showed a big improvement potential in implementing a new technology/mean for transporting the blood samples. The pneumatic tube system showed the biggest potential lowering the AWT...

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

Science.gov (United States)

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

Blood sampling and hemolysis affect concentration of plasma metabolites

DEFF Research Database (Denmark)

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

2012-01-01

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

Extensive monitoring through multiple blood samples in professional soccer players

DEFF Research Database (Denmark)

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

2013-01-01

of the season. Leucocytes decreased with increased physical training. Lymphocytes decreased at the end of the season. VO2max decreased towards the end of the season whereas no significant changes were observed in the IE2 test.The regular blood samples from elite soccer players reveal significant changes......ABSTRACT: The aim of this study was to make a comprehensive gathering of consecutive detailed blood samples from professional soccer players, and to analyze different blood parameters in relation to seasonal changes in training and match exposure.Blood samples were collected five times during a six...... months period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, players were tested for body composition, VO2max and physical performance by the Yo-Yo intermittent endurance sub-max test (IE2).Multiple variations in blood parameters occurred during...

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

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Tsuchiya Ami

2012-06-01

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

Automated Blood Sample Preparation Unit (ABSPU) for Portable Microfluidic Flow Cytometry.

Science.gov (United States)

Chaturvedi, Akhil; Gorthi, Sai Siva

2017-02-01

Portable microfluidic diagnostic devices, including flow cytometers, are being developed for point-of-care settings, especially in conjunction with inexpensive imaging devices such as mobile phone cameras. However, two pervasive drawbacks of these have been the lack of automated sample preparation processes and cells settling out of sample suspensions, leading to inaccurate results. We report an automated blood sample preparation unit (ABSPU) to prevent blood samples from settling in a reservoir during loading of samples in flow cytometers. This apparatus automates the preanalytical steps of dilution and staining of blood cells prior to microfluidic loading. It employs an assembly with a miniature vibration motor to drive turbulence in a sample reservoir. To validate performance of this system, we present experimental evidence demonstrating prevention of blood cell settling, cell integrity, and staining of cells prior to flow cytometric analysis. This setup is further integrated with a microfluidic imaging flow cytometer to investigate cell count variability. With no need for prior sample preparation, a drop of whole blood can be directly introduced to the setup without premixing with buffers manually. Our results show that integration of this assembly with microfluidic analysis provides a competent automation tool for low-cost point-of-care blood-based diagnostics.

Non-terminal blood sampling techniques in Guinea pigs

DEFF Research Database (Denmark)

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

2014-01-01

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...

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

Directory of Open Access Journals (Sweden)

Li Ling Lee

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

SPIDIA-DNA: An External Quality Assessment for the pre-analytical phase of blood samples used for DNA-based analyses

Czech Academy of Sciences Publication Activity Database

Malentacchi, F.; Pazzagli, M.; Simi, L.; Orlando, C.; Wyrich, R.; Hartmann, C.C.; Verderio, P.; Pizzamiglio, S.; Ciniselli, C.M.; Tichopád, Aleš; Kubista, Mikael; Gelmini, S.

-, č. 424 (2013), s. 274-286 ISSN 0009-8981 Institutional research plan: CEZ:AV0Z50520701 Keywords : Pre-analytical phase * DNA quality * Blood samples Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.764, year: 2013

Effect of order of draw of blood samples during phlebotomy on routine biochemistry results.

Science.gov (United States)

Sulaiman, Raashda A; Cornes, Michael P; Whitehead, Simon J; Othonos, Nadia; Ford, Clare; Gama, Rousseau

2011-11-01

To investigate whether incorrect order of draw of blood samples during phlebotomy causes in vitro potassium ethylenediaminetetraacetic acid EDTA (kEDTA) contamination of blood samples. Serum kEDTA, potassium, calcium, magnesium, alkaline phosphatase, zinc and iron concentrations were measured in blood samples drawn before and after collecting blood into kEDTA containing sample tubes by an experienced phlebotomist using the Sarstedt Safety Monovette system. EDTA was undetectable in all samples. The concentrations of other analytes were similar in blood samples drawn before and after collection of the EDTA blood sample. Order of draw of blood samples using the Sarstedt Safety Monovette system has no effect on serum biochemistry results, when samples are taken by an experienced phlebotomist.

Studies of mercury, arsenic and selenium levels in human hair, nail and blood samples from the Wassa West District of Ghana, using neutron activation analysis and atomic absorption spectrophotometric technique

International Nuclear Information System (INIS)

Samlafu, V.B.

2011-06-01

This study examines Arsenic, mercury and selenium levels in human hair, nail and blood samples collected from Wassa West District, a mining impacted site in contrast with Akatsi and its environs, a non-mining impacted site using INAA facility at Ghana Research Reactor-1 (GHARR-1), Kwabenya, Accra. Other elements determined alongside the elements of interest were Na, K, Ca, Al, Mg, Mn, V, Cl, I, Br, Au, La, Cu, Zn, Sr, Ni and Ag. An alternative method was developed for the Preconcentration Neutron Activation Analysis using dithizone/CHCl 3 mixture as the chelating agent for very low levels of mercury and arsenic in human blood. Factors that affect dithizone complexation such as PH, and concentration were investigated. Selenium levels in all the matrices were determined by Hydride Generation Atomic Absorption Spectrophotometric (HGAAS) technique. Speciation of Se(IV) and Se(VI) was carried out on selected blood samples using HNO 3 /H 2 O 2 as digestion reagent. The precision of the analytical methods used were evaluated by Schewart control chart for internal quality assessment purposes. The results were within the acceptable limits of Xσ. Four certified reference materials namely GBW 09101, DOME-2, Lichen IAEA-336 and Peach leave SRM 1547 were also used to validate the analytical methods for external quality assessment. The results obtained in this work were in good agreement with the certified values. The levels of total mercury in hair ranged from 1.65 μg/g to 20.46 μg/g. Mercury in nail samples ranged from 0.97μg/g to 31.94 μg/g and blood samples ranged from 0.01μg/mL to 0.70 μg/mL in experimental subjects. Mercury levels in control subjects ranged from 0.24 μg/g to 7.15 μg/g in hair, 0.2 μg/g to 1.83 μg/g of mercury in nail and 0.01 μg/mL to 2.38 μg/mL of mercury in blood. Generally the decreasing order of accumulation of mercury in the three matrices in the experimental subjects were as follows nail > hair > blood, while the control subjects had the

Effect of whey on blood glucose and insulin responses to composite breakfast and lunch meals in type 2 diabetic subjects.

Science.gov (United States)

Frid, Anders H; Nilsson, Mikael; Holst, Jens Juul; Björck, Inger M E

2005-07-01

Whey proteins have insulinotropic effects and reduce the postprandial glycemia in healthy subjects. The mechanism is not known, but insulinogenic amino acids and the incretin hormones seem to be involved. The aim was to evaluate whether supplementation of meals with a high glycemic index (GI) with whey proteins may increase insulin secretion and improve blood glucose control in type 2 diabetic subjects. Fourteen diet-treated subjects with type 2 diabetes were served a high-GI breakfast (white bread) and subsequent high-GI lunch (mashed potatoes with meatballs). The breakfast and lunch meals were supplemented with whey on one day; whey was exchanged for lean ham and lactose on another day. Venous blood samples were drawn before and during 4 h after breakfast and 3 h after lunch for the measurement of blood glucose, serum insulin, glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide 1 (GLP-1). The insulin responses were higher after both breakfast (31%) and lunch (57%) when whey was included in the meal than when whey was not included. After lunch, the blood glucose response was significantly reduced [-21%; 120 min area under the curve (AUC)] after whey ingestion. Postprandial GIP responses were higher after whey ingestion, whereas no differences were found in GLP-1 between the reference and test meals. It can be concluded that the addition of whey to meals with rapidly digested and absorbed carbohydrates stimulates insulin release and reduces postprandial blood glucose excursion after a lunch meal consisting of mashed potatoes and meatballs in type 2 diabetic subjects.

Improving blood sample logistics using simulation

DEFF Research Database (Denmark)

Jørgensen, Pelle Morten Thomas; Jacobsen, Peter

2012-01-01

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

SPIDIA-RNA: First external quality assessment for the pre-analytical phase of blood samples used for RNA based analyses

Czech Academy of Sciences Publication Activity Database

Pazzagli, M.; Malentacchi, F.; Simi, L.; Wyrich, R.; Guenther, K.; Hartmann, C.; Verderio, P.; Pizzamiglio, S.; Ciniselli, C.M.; Tichopád, Aleš; Kubista, Mikael; Gelmini, S.

2013-01-01

Roč. 59, č. 1 (2013), s. 20-31 ISSN 1046-2023 Institutional research plan: CEZ:AV0Z50520701 Keywords : Pre-analytical phase * RNA quality * Blood samples Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.221, year: 2013

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

Science.gov (United States)

Chambers, Andrew G.; Percy, Andrew J.; Hardie, Darryl B.; Borchers, Christoph H.

2013-09-01

Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach.

Validation of curve-fitting method for blood retention of 99mTc-GSA. Comparison with blood sampling method

International Nuclear Information System (INIS)

Ha-Kawa, Sang Kil; Suga, Yutaka; Kouda, Katsuyasu; Ikeda, Koshi; Tanaka, Yoshimasa

1997-01-01

We investigated a curve-fitting method for the rate of blood retention of 99m Tc-galactosyl serum albumin (GSA) as a substitute for the blood sampling method. Seven healthy volunteers and 27 patients with liver disease underwent 99m Tc-GSA scanning. After normalization of the y-intercept as 100 percent, a biexponential regression curve for the precordial time-activity curve provided the percent injected dose (%ID) of 99m Tc-GSA in the blood without blood sampling. The discrepancy between %ID obtained by the curve-fitting method and that by the multiple blood samples was minimal in normal volunteers 3.1±2.1% (mean±standard deviation, n=77 sampling). Slightly greater discrepancy was observed in patients with liver disease (7.5±6.1%, n=135 sampling). The %ID at 15 min after injection obtained from the fitted curve was significantly greater in patients with liver cirrhosis than in the controls (53.2±11.6%, n=13; vs. 31.9±2.8%, n=7, p 99m Tc-GSA and the plasma retention rate for indocyanine green (r=-0.869, p 99m Tc-GSA and could be a substitute for the blood sampling method. (author)

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

Science.gov (United States)

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

2016-08-01

Since driving under the influence of drugs (DUID) is as dangerous as drink-driving, many countries regulate DUID by law. However, laws against the use of drugs while driving are not yet established in Korea. In order to investigate the type and frequency of drugs used by drivers in Korea, we analyzed controlled and non-controlled drugs in alcohol-positive blood samples. Total 275 blood samples were taken from Korean drivers, which were positive in roadside alcohol testing. The following analyses were performed: blood alcohol concentrations by GC; screening for controlled drugs by immunoassay and confirmation for positive samples by GC-MS. For the detection of DUID related drugs in blood samples, a total of 49 drugs were selected and were examined by GC-MS. For a rapid detection of these drugs, an automated identification software called "DrugMan" was used. Concentrations of alcohol in 275 blood samples ranged from 0.011 to 0.249% (average 0.119%). Six specimens showed positive results by immunoassay: one methamphetamine and five benzodiazepines I. By GC-MS confirmation, only benzodiazepines in four cases were identified, while methamphetamine and benzodiazepine in two cases were not detected from the presumptive positive blood samples. Using DrugMan, four drugs were detected; chlorpheniramine (5)*, diazepam (4), dextromethorphan (1) and doxylamine (1). In addition, ibuprofen (1), lidocaine (1) and topiramate (1) were also detected as general drugs in blood samples ('*' indicates frequency). The frequency of drug abuse by Korean drivers was relatively low and a total 14 cases were positive in 275 blood samples with a ratio of 5%. However it is necessary to analyze more samples including alcohol negative blood, and to expand the range of drug lists to get the detailed information. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Classical and additional antiphospholipid antibodies in blood samples of ischemic stroke patients and healthy controls.

Science.gov (United States)

Carmel-Neiderman, Narin-Nard; Tanne, David; Goren, Idan; Rotman-Pikielny, Pnina; Levy, Yair

2017-04-01

Classical antiphospholipid antibodies (aPLa) are found in 6-25% of blood samples from stroke patients. The frequency of novel aPLa antibodies in blood samples of CVA patients is not known. Enzyme-linked immunosorbent assays (ELISA) were performed on blood samples from 209 CVA patients (170 samples were obtained during the acute phase and 39 samples were from patients with complete carotid stenosis) and compared to 54 healthy controls. Subjects were tested for the presence of the classical aPL antibodies anticardiolipin (aCL) and anti-beta2-glycoprotein (aβ2gI), in addition to antiphosphatidylethanolamine (aPE), anti-phosphatidylserine (aPS), and Annexin V. All antibodies were tested for both IgM and IgG subclasses. Numeric analysis of the antibody titer levels (μ/ml) revealed a significantly higher subclinical titer by two standard deviations of many aPL autoantibodies among CVA patients (Pv < 0.05). However, according to the kit manufacturer's cutoff value, no positive antibodies were found except a trend toward higher percentage of positive aPS IgG titer in the CVA group compared to controls (6.2 vs. %0; P = 0.077). According to the manufacturer's cutoff, significantly higher levels of positive antibodies were not found among stroke patients. However, the absolute ELISA values of stroke patients were significantly higher. These results suggest that lower cutoff values than those used for APS diagnosis should be used for risk stratification of CVA among healthy individuals.

An automated blood sampling system used in positron emission tomography

International Nuclear Information System (INIS)

Eriksson, L.; Bohm, C.; Kesselberg, M.

1988-01-01

Fast dynamic function studies with positron emission tomography (PET), has the potential to give accurate information of physiological functions of the brain. This capability can be realised if the positron camera system accurately quantitates the tracer uptake in the brain with sufficiently high efficiency and in sufficiently short time intervals. However, in addition, the tracer concentration in blood, as a function of time, must be accurately determined. This paper describes and evaluates an automated blood sampling system. Two different detector units are compared. The use of the automated blood sampling system is demonstrated in studies of cerebral blood flow, in studies of the blood-brain barrier transfer of amino acids and of the cerebral oxygen consumption. 5 refs.; 7 figs

Nocturnal blood pressure in normotensive subjects and those with white coat, primary, and secondary hypertension.

OpenAIRE

Middeke, M.; Schrader, J.

1994-01-01

OBJECTIVE--To compare the mean nocturnal blood pressure of patients with various forms of renal and endocrine hypertension with that in patients with primary and white coat hypertension, and normal blood pressure. DESIGN--Ambulatory monitoring of blood pressure over 24 hours in a prospective study. SETTING--Two German centres for outpatients with hypertension and kidney diseases. SUBJECTS--176 normotensive subjects, 490 patients with primary hypertension including mild and severe forms, 42 wi...

Effect of supplementation of tender coconut water on blood pressure of primary hypertensive subjects

Directory of Open Access Journals (Sweden)

Gullapalli HS, Avinash P Tekade, Namrata H Gullapalli

2013-04-01

Full Text Available Background: Hypertension is a major health problem worldwide. Increased vascular resistance, sodium retention & sympathetic over activity contributes to the blood pressure elevation. Plant foods may be beneficial in decreasing blood pressure (BP. Recently much attention has been focused on plant foods that may be beneficial in preventing Hypertension, metabolic syndrome and possibly reduce the risk of various diseases. This clinical study was conducted to test the effectiveness of a structured intervention on BP of primary hypertensive subjects. Aim: To study the effect of Tender Coconut Water (TCW on BP of Primary hypertensive subjects. Methods and Material: 70 subjects were selected randomly sample for 6 weeks of the intervention program. Among them 40 subjects were selected as the experimental group and 30 300ml/day for 6 weeks whereas the control group was instructed to follow the same routine without modifications. One initial, two mid intervention (after every 15 days and one final (post intervention BP recorded for both the groups. The obtained data was statistically analyzed. Results: The mean systolic BP of experimental group and control group were decreased from 145.8 mm Hg and 141mm of Hg to 135.3 mm of Hg and 140 mm of Hg respectively. The mean diastolic BP of experimental group and control group were decreased from 93.7 mm H g and 90.9 mmHg to 86.9 mm of Hg and 89.7 mm of Hg respectively. Conclusion: Irrespective of cause of hypertension TCW has beneficial effect on BP. TCW contains high amount of potassium which causes vasodilatation and also improve the endothelial function.

Preanalytic Factors Associated With Hemolysis in Emergency Department Blood Samples.

Science.gov (United States)

Phelan, Michael P; Reineks, Edmunds Z; Schold, Jesse D; Hustey, Frederic M; Chamberlin, Janelle; Procop, Gary W

2018-02-01

- Hemolysis of emergency department blood samples is a common occurrence and has a negative impact on health care delivery. - To determine the effect of preanalytic factors (straight stick, intravenous [IV] line, needle gauge, location of blood draw, syringe versus vacuum tube use, tourniquet time) on hemolysis in emergency department blood samples. - A single 65 000-visit emergency department's electronic health record was queried for emergency department potassium results and blood draw technique for all samples obtained in calendar year 2014, resulting in 54 531 potassium results. Hemolyzed potassium was measured by hemolysis index. Comparisons of hemolysis by sampling technique were conducted by χ 2 tests. - Overall hemolysis was 10.0% (5439 of 54 531). Hemolysis among samples obtained from straight stick was significantly less than among those obtained with IV line (5.4% [33 of 615] versus 10.2% [4821 of 47 266], P < .001). For IV-placed blood draws, antecubital location had a statistically significant lower overall hemolysis compared with other locations: 7.4% (2117 of 28 786) versus 14.6% (2622 of 17 960) ( P < .001). For blood drawn with a syringe compared with vacuum, hemolysis was 13.0% (92 of 705) and 11.0% (1820 of 16 590), respectively ( P = .09, not significant). For large-gauge IV blood draws versus smaller-gauge IV lines, a lower hemolysis was also observed (9.3% [3882 of 41 571] versus 16.7% [939 of 5633]) ( P < .001). For IV-drawn blood with tourniquet time less than 60 seconds, hemolysis was 10.3% (1362 of 13 162) versus 13.9% for more than 60 seconds (532 of 3832), P < .001. - This study confirmed previous findings that straight stick and antecubital location are significantly associated with reduced hemolysis and indicated that shorter tourniquet time and larger gauge for IV draws were significantly associated with lower hemolysis.

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

DEFF Research Database (Denmark)

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

2014-01-01

weight following blood sampling, but the body weight loss was higher in mice subjected to facial vein phlebotomy. The food consumption was not significantly different between the two groups. At gross necropsy, subcutaneous hematomas were found in both groups and the histopathological analyses revealed...

Effects of Transport and Storage Conditions on Gene Expression in Blood Samples.

Science.gov (United States)

Malentacchi, Francesca; Pizzamiglio, Sara; Wyrich, Ralf; Verderio, Paolo; Ciniselli, Chiara; Pazzagli, Mario; Gelmini, Stefania

2016-04-01

Inappropriate handling of blood samples might induce or repress gene expression and/or lead to RNA degradation affecting downstream analysis. In particular, sample transport is a critical step for biobanking or multicenter studies because of uncontrolled variables (i.e., unstable temperature). We report the results of a pilot study implemented within the EC funded SPIDIA project, aimed to investigate the role of transport and storage of blood samples containing and not containing an RNA stabilizer. Blood was collected from a single donor both in EDTA and in PAXgene Blood RNA tubes. Half of the samples were sent to a second laboratory both at room temperature and at 4°C, whereas the remaining samples were stored at room temperature and at 4°C. Gene expression of selected genes (c-FOS, IL-1β, IL-8, and GAPDH) known to be induced or repressed by ex vivo blood handling and of blood-mRNA quality biomarkers identified and validated within the SPIDIA project, which allow for monitoring changes in unstabilized blood samples after collection and during transport and storage, were analyzed by RT-qPCR. If the shipment of blood in tubes not containing RNA stabilizer is not performed under a stable condition, gene profile studies can be affected by the effects of transport. Moreover, also controlled temperature shipment (4°C) can influence the expression of specific genes if blood is collected in tubes not containing a stabilizer. The use of dedicated biomarkers or time course experiments should be performed in order to verify potential bias on gene expression analysis due to sample shipment and storage conditions. Alternatively, the use of RNA stabilizer containing tubes can represent a reliable option to avoid ex vivo RNA changes.

Performance of an app measuring spot quality in dried blood spot sampling

NARCIS (Netherlands)

Veenhof, Herman

2016-01-01

Introduction: The Dried Blood Spot sampling (DBS) method gives patients and health care workers the opportunity for remote sampling using a drop of blood from a fingerprick on a sampling card which can be send to the laboratory by mail. Laboratory analysts frequently reject DBS samples because of

A technique for extracting blood samples from mice in fire toxicity tests

Science.gov (United States)

Bucci, T. J.; Hilado, C. J.; Lopez, M. T.

1976-01-01

The extraction of adequate blood samples from moribund and dead mice has been a problem because of the small quantity of blood in each animal and the short time available between the animals' death and coagulation of the blood. These difficulties are particularly critical in fire toxicity tests because removal of the test animals while observing proper safety precautions for personnel is time-consuming. Techniques for extracting blood samples from mice were evaluated, and a technique was developed to obtain up to 0.8 ml of blood from a single mouse after death. The technique involves rapid exposure and cutting of the posterior vena cava and accumulation of blood in the peritoneal space. Blood samples of 0.5 ml or more from individual mice have been consistently obtained as much as 16 minutes after apparent death. Results of carboxyhemoglobin analyses of blood appeared reproducible and consistent with carbon monoxide concentrations in the exposure chamber.

5D.07: THE IMPACT OF FLAVONOL-RICH DARK CHOCOLATE ON BLOOD PRESSURE AND VASCULAR FUNCTION IN HEALTHY SUBJECTS.

Science.gov (United States)

Heuten, H; Van Ackeren, K; Hoymans, V; Wouters, K; Goovaerts, I; Conraads, V; Vrints, C

2015-06-01

Flavanoids may have a beneficial effect on blood pressure (BP) and endothelial function. There is however, limited data on this effect during a longer period (8 weeks) and no data on the effect on EPC (endothelial progenitor cells) in healthy subjects. Healthy, non-smoking, male and female volunteers aged 35-65 year with no history of diabetes or cardiovascular disease and with normal or mild hypertensive blood pressure (chocolate (High-DC). Group 2 (n = 26): daily consuming 20 gram of low-flavanol dark chocolate (Low-DC). At week 0,4,6,7 and 8 blood pressure was assessed in all subjects, and endothelial function (FMD, flow mediated dilation) in a subgroup. A blood sample was taken in each subject at week 0 and 8 for measuring glucose, lipids and EPC. Baseline characteristics were comparable between both groups. There was a decrease in systolic and diastolic blood pressure over time in both groups, however at 8 weeks there was no statistically significant difference between groups (delta SBP -2.17  + /-8.53 mmHg in gr 1 versus -4.06 + /-8.05 mmHg in gr 2, p = 0.4; delta DBP -3.97  + /-7.1 mmHg in gr 1 versus -4.67  + /-5.99 mmHg in gr 2, p = 0.7).FMD was performed in 9 subjects from each group, no significant difference was noted between both groups over time (delta FMD gr 1: -3.50  + /- 6.00 % versus gr 2: + 0.12  + /- 2.51 %, p = 0.06). EPC values did not differ between groups at baseline (T0) and at the end of the study (T8) (ISHAGE count (T8-T0): gr1: 3.23 (-68.01 - 41.71) versus gr 2: -9.23 (-57.59 - 17.28), p = 0.4). Glucose and lipids were comparable between both groups at baseline and at the end of the study (p = ns). In this study, no beneficial effect was noticed in favour of the consumption of flavanol-rich dark chocolate during 8 weeks on blood pressure or vascular function, in healthy subjects.

Thermal activation of catalytic microjets in blood samples using microfluidic chips.

Science.gov (United States)

Soler, Lluís; Martínez-Cisneros, Cynthia; Swiersy, Anka; Sánchez, Samuel; Schmidt, Oliver G

2013-11-21

We demonstrate that catalytic microjet engines can out-swim high complex media composed of red blood cells and serum. Despite the challenge presented by the high viscosity of the solution at room temperature, the catalytic microjets can be activated at physiological temperature and, consequently, self-propel in diluted solutions of blood samples. We prove that these microjets self-propel in 10× diluted blood samples using microfluidic chips.

Thermal activation of catalytic microjets in blood samples using microfluidic chips†

Science.gov (United States)

Soler, Lluís; Martínez-Cisneros, Cynthia; Swiersy, Anka; Sánchez, Samuel; Schmidt, Oliver G.

2014-01-01

We demonstrate that catalytic microjet engines can out-swim high complex media composed of red blood cells and serum. Despite the challenge presented by the high viscosity of the solution at room temperature, the catalytic microjets can be activated at physiological temperature and, consequently, self-propel in diluted solutions of blood samples. We prove that these microjets self-propel in 10× diluted blood samples using microfluidic chips. PMID:24089195

Na2EDTA anticoagulant impaired blood samples from the teleost Piaractus mesopotamicus

Directory of Open Access Journals (Sweden)

Thaís Heloisa Vaz Farias

2016-05-01

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

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

Science.gov (United States)

Braniff, Heather; DeCarlo, Ann; Haskamp, Amy Corey; Broome, Marion E

2014-01-01

Aiming to minimize pain in a hospitalized child, the purpose of this observational study was to describe characteristics of blood samples collected from pre-existing peripheral intravenous (PIV) catheters in pediatric patients. One hundred and fifty blood samples were reviewed for number of unusable samples requiring a specimen to be re-drawn. Success of the blood draw and prevalence of the loss of the PIV following blood collection was also measured. Findings included one clotted specimen, success rate of 91.3%, and 1.3% of PIVs becoming non-functional after collection. Obtaining blood specimens from a pre-existing PIV should be considered in a pediatric patient. Copyright © 2014 Elsevier Inc. All rights reserved.

Calculation of Blood Dose in Patients Treated With 131I Using MIRD, Imaging, and Blood Sampling Methods.

Science.gov (United States)

Piruzan, Elham; Haghighatafshar, Mahdi; Faghihi, Reza; Entezarmahdi, Seyed Mohammad

2016-03-01

Radioiodine therapy is known as the most effective treatment of differentiated thyroid carcinoma (DTC) to ablate remnant thyroid tissue after surgery. In patients with DTC treated with radioiodine, internal radiation dosimetry of radioiodine is useful for radiation risk assessment. The aim of this study is to describe a method to estimate the absorbed dose to the blood using medical internal radiation dosimetry methods. In this study, 23 patients with DTC with different administrated activities, 3.7, 4.62, and 5.55 GBq after thyroidectomy, were randomly selected. Blood dosimetry of treated patients was performed with external whole body counting using a dual-head gamma camera imaging device and also with blood sample activity measurements using a dose calibrator. Absorbed dose to the blood was measured at 2, 6, 12, 24, 48, and 96 hours after the administration of radioiodine with the 2 methods. Based on the results of whole body counting and blood sample activity dose rate measurements, 96 hours after administration of 3.7, 4.62, and 5.55 GBq of radioiodine, absorbed doses to patients' blood were 0.65 ± 0.20, 0.67 ± 0.18, 0.79 ± 0.51 Gy, respectively. Increasing radioiodine activity from 3.7 to 5.55 GBq increased blood dose significantly, while there was no significant difference in blood dose between radioiodine dosages of 3.7 and 4.62 GBq. Our results revealed a significant correlation between the blood absorbed dose and blood sample activity and between the blood absorbed dose and whole body counts 24 to 48 hours after the administration of radioiodine.

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

DEFF Research Database (Denmark)

Seemann, T. L.; Nybo, M.

2015-01-01

Background: An important preanalytical factor is the blood sampling procedure and its adherence to the guidelines, i.e. CLSI and ISO 15189, in order to ensure a consistent quality of the blood collection. Therefore, it is critically important to introduce quality control on this part of the process....... As suggested by the EFLM working group on the preanalytical phase we introduced continuous quality control of the blood sampling procedure using a structured observation scheme to monitor the quality of blood sampling performed on an everyday basis. Materials and methods: Based on our own routines the EFLM....... Conclusion: It is possible to establish a continuous quality control on blood sampling. It has been well accepted by the staff and we have already been able to identify critical areas in the sampling process. We find that continuous auditing increase focus on the quality of blood collection which ensures...

Bacterial Isolates from Blood Samples of Patients in University of ...

African Journals Online (AJOL)

Bacterial Isolates from Blood Samples of Patients in University of Benin Teaching Hospital Benin City, Edo State, Nigeria. ... The thioglychollate broth was sub cultured onto blood agar plate for anaerobic incubation, while the brain heart infusion broth was sub cultured onto chocolate, blood agar and McConkey agar for ...

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

Energy Technology Data Exchange (ETDEWEB)

Radford, E.P.

1976-01-01

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

Whole blood solubilization and discoloration before LSC of yttrium samples

International Nuclear Information System (INIS)

Lima, Marina F.; Leonardo, Lucio

2009-01-01

Liquid scintillation counting of whole blood and animal tissues samples could be severely impaired owing to quenching by their compounds. The objective of this previous study is preparing one protocol of 90 Y measurement to apply in biodistribution and dosimetry studies of radiopharmaceuticals labeled with this isotope and other beta emitters in in vivo and ex vivo samples. The first parameters considered to choose a method were: the largest blood sample per collection (80μl-90μl), attending the collection limit of less than 7.5% of total circulating blood volume for in vivo samples. Other parameters were the use of EDTA and cyclohexydine as solubilization and lytic agents, HNO 3 and H 2 O 2 as mineralizing agents and NH 4 OH as neutralization agent. One samples batch was tested in a water bath under the lower temperature to prevent the volume lose of the ionic phase. Other samples batch was mineralized over a hot-plate at 120 deg C following the currently largest sample amounts processing procedure in our laboratory by using HNO 3 and H 2 O 2 . Results show the contribution of the blood fragments as quenching in the region A ( 3 and/or NH 4 OH in the hot-plate digestion. As expected, the measurements in the three spectral regions show the proteins and colored fragments were completely removed by the hot-plate digestion. The rate between efficiency and 90 Sr- 90 Y concentration had not significant differences in the range between 120 Bq and 1200 Bq. (author)

Astronaut Joseph Kerwin takes blood sample from Astronaut Charles Conrad

Science.gov (United States)

1973-01-01

Scientist-Astronaut Joseph P. Kerwin (right), Skylab 2 science pilot and a doctor of medicine, takes a blood sample from Astronaut Charles Conrad Jr., Sylab 2 commander, as seen in this reproduction taken from a color television transmission made by a TV camera aboard the Skylab 1 and 2 space station cluster in Earth orbit. The blood sampling was part of the Skylab Hematology and Immunology Experiment M110 series.

Human blood RNA stabilization in samples collected and transported for a large biobank

Science.gov (United States)

2012-01-01

Background The Norwegian Mother and Child Cohort Study (MoBa) is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems – the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes. Findings High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times) compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression. PMID:22988904

Sampling methods to the statistical control of the production of blood components.

Science.gov (United States)

Pereira, Paulo; Seghatchian, Jerard; Caldeira, Beatriz; Santos, Paula; Castro, Rosa; Fernandes, Teresa; Xavier, Sandra; de Sousa, Gracinda; de Almeida E Sousa, João Paulo

2017-12-01

The control of blood components specifications is a requirement generalized in Europe by the European Commission Directives and in the US by the AABB standards. The use of a statistical process control methodology is recommended in the related literature, including the EDQM guideline. The control reliability is dependent of the sampling. However, a correct sampling methodology seems not to be systematically applied. Commonly, the sampling is intended to comply uniquely with the 1% specification to the produced blood components. Nevertheless, on a purely statistical viewpoint, this model could be argued not to be related to a consistent sampling technique. This could be a severe limitation to detect abnormal patterns and to assure that the production has a non-significant probability of producing nonconforming components. This article discusses what is happening in blood establishments. Three statistical methodologies are proposed: simple random sampling, sampling based on the proportion of a finite population, and sampling based on the inspection level. The empirical results demonstrate that these models are practicable in blood establishments contributing to the robustness of sampling and related statistical process control decisions for the purpose they are suggested for. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of clinical biomarkers for pre-analytical quality control of blood samples.

Science.gov (United States)

Kang, Hyun Ju; Jeon, Soon Young; Park, Jae-Sun; Yun, Ji Young; Kil, Han Na; Hong, Won Kyung; Lee, Mee-Hee; Kim, Jun-Woo; Jeon, Jae-Pil; Han, Bok Ghee

2013-04-01

Pre-analytical conditions are key factors in maintaining the high quality of biospecimens. They are necessary for accurate reproducibility of experiments in the field of biomarker discovery as well as achieving optimal specificity of laboratory tests for clinical diagnosis. In research at the National Biobank of Korea, we evaluated the impact of pre-analytical conditions on the stability of biobanked blood samples by measuring biochemical analytes commonly used in clinical laboratory tests. We measured 10 routine laboratory analytes in serum and plasma samples from healthy donors (n = 50) with a chemistry autoanalyzer (Hitachi 7600-110). The analyte measurements were made at different time courses based on delay of blood fractionation, freezing delay of fractionated serum and plasma samples, and at different cycles (0, 1, 3, 6, 9) of freeze-thawing. Statistically significant changes from the reference sample mean were determined using the repeated-measures ANOVA and the significant change limit (SCL). The serum levels of GGT and LDH were changed significantly depending on both the time interval between blood collection and fractionation and the time interval between fractionation and freezing of serum and plasma samples. The glucose level was most sensitive only to the elapsed time between blood collection and centrifugation for blood fractionation. Based on these findings, a simple formula (glucose decrease by 1.387 mg/dL per hour) was derived to estimate the length of time delay after blood collection. In addition, AST, BUN, GGT, and LDH showed sensitive responses to repeated freeze-thaw cycles of serum and plasma samples. These results suggest that GGT and LDH measurements can be used as quality control markers for certain pre-analytical conditions (eg, delayed processing or repeated freeze-thawing) of blood samples which are either directly used in the laboratory tests or stored for future research in the biobank.

Are They Bloody Guilty? Blood Doping with Simulated Samples

Science.gov (United States)

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

2014-01-01

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

Estimations of cholesterol, triglycerides and fractionation of lipoproteins in serum samples of some Nigerian female subjects

Directory of Open Access Journals (Sweden)

E.I. Adeyeye

2011-04-01

Full Text Available Blood samples (serum were collected to determine some biochemical parameters: total glycerides (TG, total cholesterol (TC, high density lipoprotein-cholesterol (HDL-C, low density lipoprotein-cholesterol (LDL-C and very low density lipoprotein-cholesterol (VLDL-C in 53 female subjects in Warri, Delta State, Nigeria using the Reflotron® (an auto analyser, supported with the use of questionnaire to get information on age and sex. Age range of the subjects was 18–80 years. The TG levels in all the subjects were < 200 mg/dL; only one subject (1.89% had TC < 200 mg/dL; nine subjects (17.0% had HDL-C ≤ 35 mg/dL; for LDL-C only one subject (1.89% had a desirable level of < 130 mg/dL; for VLDL-C 29 subjects (54.7% had values 17.2 mg/dL and above. For therapeutic decision-making, TC/HDL-C and LDL-C/HDL-C, were calculated. In TC/HDL-C, three subjects (5.66% had values < 4.4 and in LDL-C/HDL-C, 41 subjects (77.4% had values < 4.5. Hence, TC, HDL-C, LDL-C, TC/HDL-C and slightly LDL-C/HDL-C and VLDL-C in the subjects could lead to increase coronary heart diseases. Results were matched for the age and sex of subjects.

Blood flow in the forearm in patients with Rheumatoid arthritis and healthy subjects under local thermotherapy

Directory of Open Access Journals (Sweden)

C. Mucha

2002-02-01

Full Text Available Objectives: Muscle blood flow in the forearm of patients with rheuma-toid arthritis and healthy volunteers following treatment with temperature increasingarm baths, mudpacks and short- or decimeter-wave diathermy was studied in thisinvestigation. The aim of the study was to find out the difference of reactive hyperemia between the different temperature methods as well as the influence on theconsensual reaction. Subjects: Eighty patients with rheumatoid arthritis, stage 3 according toSteinbrocker, as well as 80 healthy human subjects had been assigned numerically in the four therapy- and controlgroups. Patients with diseases influencing the peripheral blood flow were excluded. Design: Blood flow was measured by venous occlusion plethysmography in both forearms with the subjects lyingsupine. The application of the local heat therapies had been excluded on the left forearm. The forearm blood flow wasmonitored before heat therapy, directly after as well as in two further 10 minutes intervals. An analysis of variancewas used to determine the influence on blood flow of the response to the heat therapies in patients with rheumatoidarthritis and healthy subjects.Results: Under homogeneous starting conditions and a statistically uniformed high blood flow in rest the reactive values of blood flow on the left-hand side of application and the right consensual side showed high significant differencesbetween all methods of therapy. Differences between the patients and the healthy subjects only showed tendencies withpartially lower reactions, concerning the patients with rheumatoid arthritis. All methods of heat therapy caused a statistically provable consensual reaction that turned out smaller after diathermic methods. Here the post therapeuticreaction of the blood flow on the side of application was also lower or rather shorter. Conclusion: Greater differences of the blood flow in rest between the patients with rheumatoid arthritis and healthysubjects

EFFECT OF ADDING THE INTERNAL STANDARD TO BLOOD SAMPLES, PRIOR TO THE PREPARATION OF BLOOD SPOTS FOR ACYLCARNITINE ANALYSIS

OpenAIRE

Osorio, José Henry; Pourfarzam, Morteza

2010-01-01

Background: some general factors can influence when determining acylcarnitines through tandem mass spectrometry. Objective: to study the effect of adding the internal standard to blood samples before the preparation of filter paper cards compared with the addition of internal standard after having the filter paper cards prepared for determining acylcarnitines in blood for tandem mass spectrometry. Methodology: two groups of blood samples were prepared: group one without adding internal standa...

A method for estimating radioactive cesium concentrations in cattle blood using urine samples.

Science.gov (United States)

Sato, Itaru; Yamagishi, Ryoma; Sasaki, Jun; Satoh, Hiroshi; Miura, Kiyoshi; Kikuchi, Kaoru; Otani, Kumiko; Okada, Keiji

2017-12-01

In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the 'difficult-to-return zone'. Urine 137 Cs, specific gravity, electrical conductivity, pH, sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood 137 Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood 137 Cs] = [urinary 137 Cs]/([specific gravity] - 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher 137 Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood 137 Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle. © 2017 Japanese Society of Animal Science.

Exercise induced pulmonary, hepatic and splenic blood volume changes in diabetic subjects

International Nuclear Information System (INIS)

Mubashar, M.

1993-01-01

Exercise induced blood volume changes in visceral organs were determined by scintillation gamma camera imaging in 11 normal healthy male volunteers and 15 NIDDM male diabetics without clinical signs of neuropathy. After in-vivo labelling of red cells with Technetium-99m, the data was acquired in the supine position at rest and immediately after graded upright ergometer bicycle exercise. From rest to peak exercise, pulmonary blood volume increased 19% and 75% in normal volunteers of less than and more than 40 years of age respectively. A decrease of 18% and 42% was noted in the hepatic and splenic blood volume respectively, regardless of the age, in the normal subjects. In contrast to normals, the diabetic patients showed in response to peak exercise as compared to age-matched controls. A significant difference in the drop in pulmonary blood volume 82.37% and 90% was observed between diabetics of more than and less than 7 years duration respectively. The liver and spleen of the diabetic subjects revealed a lesser decrease of 87.6% and 71.33% respectively in response to peak stress in comparison to the age matched controls. The reduction in the hepatic and splenic blood volume was equally evident in diabetics of more than or less than 50 years of age and it was statistically nonsignificant. This study demonstrates that the normal pattern of redistribution of blood volume in response to maximum exercise in diabetics is altered such that there is restricted pulmonary perfusion and diminished vasoconstriction of the hepato splenic vascular bed and the changes in the pulmonary circulation are related to the duration of the diabetics rather than the age of the patient. (author)

Use of Capillary Blood Samples Leads to Higher Parasitemia Estimates and Higher Diagnostic Sensitivity of Microscopic and Molecular Diagnostics of Malaria than Venous Blood Samples.

Science.gov (United States)

Mischlinger, Johannes; Pitzinger, Paul; Veletzky, Luzia; Groger, Mirjam; Zoleko-Manego, Rella; Adegnika, Ayola A; Agnandji, Selidji T; Lell, Bertrand; Kremsner, Peter G; Tannich, Egbert; Mombo-Ngoma, Ghyslain; Mordmüller, Benjamin; Ramharter, Michael

2018-05-25

Diagnosis of malaria is usually based on samples of peripheral blood. However, it is unclear whether capillary (CAP) or venous (VEN) blood samples provide better diagnostic performance. Quantitative differences of parasitemia between CAP and VEN blood and diagnostic performance characteristics were investigated. Patients were recruited between September 2015 and February 2016 in Gabon. Light microscopy and qPCR quantified parasitemia of paired CAP and VEN samples, whose preparation followed the exact same methodology. CAP and VEN performance characteristics using microscopy were evaluated against a qPCR gold-standard. Microscopy revealed a median (IQR) parasites/L of 495 (853,243) in CAP and 429 (524,074) in VEN samples manifesting in a +16.6% (p=0.04) higher CAPparasitemia compared with VENparasitemia. Concordantly, qPCR demonstrated that -0.278 (p=0.006) cycles were required for signal detection in CAP samples. CAPsensitivity of microscopy relative to the gold-standard was 81.5% (77.485.6%) versus VENsensitivity of 73.4% (68.878.1%), while CAPspecificity and VENspecificity were 91%. CAPsensitivity and VENsensitivity dropped to 63.3% and 45.9%, respectively for a sub-population of low-level parasitemias while specificities were 92%. CAP sampling leads to higher parasitemias compared to VEN sampling and improves diagnostic sensitivity. These findings may have important implications for routine diagnostics, research and elimination campaigns of malaria.

Quantification of multiple elements in dried blood spot samples

DEFF Research Database (Denmark)

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

2017-01-01

BACKGROUND: Dried blood spots (DBS) is a unique matrix that offers advantages compared to conventional blood collection making it increasingly popular in large population studies. We here describe development and validation of a method to determine multiple elements in DBS. METHODS: Elements were...... in venous blood. Samples with different hematocrit were spotted onto filter paper to assess hematocrit effect. RESULTS: The established method was precise and accurate for measurement of most elements in DBS. There was a significant but relatively weak correlation between measurement of the elements Mg, K...

Adaptive control of theophylline therapy: importance of blood sampling times.

Science.gov (United States)

D'Argenio, D Z; Khakmahd, K

1983-10-01

A two-observation protocol for estimating theophylline clearance during a constant-rate intravenous infusion is used to examine the importance of blood sampling schedules with regard to the information content of resulting concentration data. Guided by a theory for calculating maximally informative sample times, population simulations are used to assess the effect of specific sampling times on the precision of resulting clearance estimates and subsequent predictions of theophylline plasma concentrations. The simulations incorporated noise terms for intersubject variability, dosing errors, sample collection errors, and assay error. Clearance was estimated using Chiou's method, least squares, and a Bayesian estimation procedure. The results of these simulations suggest that clinically significant estimation and prediction errors may result when using the above two-point protocol for estimating theophylline clearance if the time separating the two blood samples is less than one population mean elimination half-life.

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

Science.gov (United States)

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

2008-02-01

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

Extent of, and variables associated with, blood pressure variability among older subjects.

Science.gov (United States)

Morano, Arianna; Ravera, Agnese; Agosta, Luca; Sappa, Matteo; Falcone, Yolanda; Fonte, Gianfranco; Isaia, Gianluca; Isaia, Giovanni Carlo; Bo, Mario

2018-02-23

Blood pressure variability (BPV) may have prognostic implications for cardiovascular risk and cognitive decline; however, BPV has yet to be studied in old and very old people. Aim of the present study was to evaluate the extent of BPV and to identify variables associated with BPV among older subjects. A retrospective study of patients aged ≥ 65 years who underwent 24-h ambulatory blood pressure monitoring (ABPM) was carried out. Three different BPV indexes were calculated for systolic and diastolic blood pressure (SBP and DBP): standard deviation (SD), coefficient of variation (CV), and average real variability (ARV). Demographic variables and use of antihypertensive medications were considered. The study included 738 patients. Mean age was 74.8 ± 6.8 years. Mean SBP and DBP SD were 20.5 ± 4.4 and 14.6 ± 3.4 mmHg. Mean SBP and DBP CV were 16 ± 3 and 20 ± 5%. Mean SBP and DBP ARV were 15.7 ± 3.9 and 11.8 ± 3.6 mmHg. At multivariate analysis older age, female sex and uncontrolled mean blood pressure were associated with both systolic and diastolic BPV indexes. The use of calcium channel blockers and alpha-adrenergic antagonists was associated with lower systolic and diastolic BPV indexes, respectively. Among elderly subjects undergoing 24-h ABPM, we observed remarkably high indexes of BPV, which were associated with older age, female sex, and uncontrolled blood pressure values.

Determination of proguanil and metabolites in small sample volumes of whole blood stored on filter paper by high-performance liquid chromatography.

Science.gov (United States)

Kolawole, J A; Taylor, R B; Moody, R R

1995-12-01

A method is reported for the determination of proguanil and its two metabolites cycloguanil and 4-chlorophenylbiguanide in whole blood and plasma samples obtained by thumbprick and stored dry on filter paper. The sample preparation involves liquid extraction from the filter paper and subsequent solid-phase extraction using C8 Bond-Elut cartridges. Separation and quantification is by a previously reported ion-pairing high-performance liquid chromatographic system with ODS Hypersil as stationary phase and an 50:50 acetonitrile-pH 2 phosphate buffer mobile phase containing 200 mM sodium dodecylsulphate as ion-pairing agent. The analytical characteristics of the method are reported. Representative concentrations are shown as a function of time from a human subject after ingestion of a single 200-mg dose of proguanil hydrochloride. Typical ranges of concentration detected by the proposed method in human subjects were proguanil 12-900 ng/ml, cycloguanil 16-44 ng/ml and 4-chlorophenylbiguanide 1.5-10 ng/ml in whole blood.

Single injection 51Cr EDTA plasma clearance determination in children using capillary blood samples

International Nuclear Information System (INIS)

Broechner-Mortensen, J.; Christoffersen, J.

1977-01-01

The reliability of a determination of the total 51 Cr EDTA plasma clearance (e) (and with it the glomerular filtration rate), by a simplified single injection method (injected dose: 4.5 μCi per kg b.w.) using capillary blood samples (0.2 ml), was investigated in twenty children. Clearance values determined from capillary blood samples did not differ significantly from those measured simultaneously from venous blood samples, the mean ratio+-SD being 1.02+-0.06(n = 10). The reproducibility (total day-to-day variation) of E determined from capillary blood samples was 6.7% in children with decreased renal function (n = 3) and 6.9% in children with normal renal function (n = 7). The present data indicate that the use of capillary blood samples is an accurate and very precise approach for determination of E in children. (Auth.)

[Effects of high intensity interval training on blood pressure in hypertensive subjects].

Science.gov (United States)

Olea, María Angélica; Mancilla, Rodrigo; Martínez, Sergio; Díaz, Erik

2017-09-01

Exercise training may reduce blood pressure. To determine the effects of a high intensity interval training (HIIT) exercise protocol on systolic and diastolic blood pressure in hypertensive subjects. Eleven men and 27 women aged 46.4 ± 9.8 years were divided in two groups according to their blood pressure. Sixteen were classified as normotensive and 22 as hypertensive. All attended an exercise program with 3 sessions per week for a total of 24 sessions. Each session consisted of one minute of intense exercise performed on a stationary bike, followed by an inactive pause lasting two minutes. This cycle was repeated 10 times and it was thus called 1 * 2 * 10. Blood pressure, weight (kg) and body fat were assessed. In the hypertensive group, there was a significant reduction in systolic blood pressure from 145.4 ± 9.0 to 118.3 ± 15.6 mm Hg (p hypertension in a period of two months and 24 sessions.

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

Science.gov (United States)

Bölenius, Karin; Brulin, Christine; Grankvist, Kjell; Lindkvist, Marie; Söderberg, Johan

2012-01-19

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

Do the venous blood samples replicate malaria parasite densities found in capillary blood? A field study performed in naturally-infected asymptomatic children in Cameroon.

Science.gov (United States)

Sandeu, Maurice M; Bayibéki, Albert N; Tchioffo, Majoline T; Abate, Luc; Gimonneau, Geoffrey; Awono-Ambéné, Parfait H; Nsango, Sandrine E; Diallo, Diadier; Berry, Antoine; Texier, Gaétan; Morlais, Isabelle

2017-08-17

The measure of new drug- or vaccine-based approaches for malaria control is based on direct membrane feeding assays (DMFAs) where gametocyte-infected blood samples are offered to mosquitoes through an artificial feeder system. Gametocyte donors are identified by the microscopic detection and quantification of malaria blood stages on blood films prepared using either capillary or venous blood. However, parasites are known to sequester in the microvasculature and this phenomenon may alter accurate detection of parasites in blood films. The blood source may then impact the success of mosquito feeding experiments and investigations are needed for the implementation of DMFAs under natural conditions. Thick blood smears were prepared from blood obtained from asymptomatic children attending primary schools in the vicinity of Mfou (Cameroon) over four transmission seasons. Parasite densities were determined microscopically from capillary and venous blood for 137 naturally-infected gametocyte carriers. The effect of the blood source on gametocyte and asexual stage densities was then assessed by fitting cumulative link mixed models (CLMM). DMFAs were performed to compare the infectiousness of gametocytes from the different blood sources to mosquitoes. Prevalence of Plasmodium falciparum asexual stages among asymptomatic children aged from 4 to 15 years was 51.8% (2116/4087). The overall prevalence of P. falciparum gametocyte carriage was 8.9% and varied from one school to another. No difference in the density of gametocyte and asexual stages was found between capillary and venous blood. Attempts to perform DMFAs with capillary blood failed. Plasmodium falciparum malaria parasite densities do not differ between capillary and venous blood in asymptomatic subjects for both gametocyte and trophozoite stages. This finding suggests that the blood source should not interfere with transmission efficiency in DMFAs.

Long-term facial artery catheter implantation for serial arterial blood sampling and invasive arterial blood pressure measurement in horses.

Science.gov (United States)

Dias, Deborah Penteado Martins; Teixeira, Luisa Gouvêa; Canola, Paulo Aléscio; Albernaz, Raquel Mincarelli; Marques, José Antônio; Neto, José Corrêa de Lacerda

2012-06-01

The purpose of this investigation was to evaluate surgical catheter implantation in the facial artery of horses and the long-term maintenance of such arteries using heparin and ascorbic acid as filling solution. Nine horses were implanted with a polyurethane catheter. The catheters were flushed with a heparin/ascorbic acid solution every 8h and remained patent for 25 days. Arterial blood samples were collected twice a day, and one exercise test that included serial blood samples and arterial pressure recordings was performed on a treadmill. Polyurethane catheters surgically implanted in the facial artery can be kept patent by filling with a heparin/ascorbic acid solution and provide convenient invasive arterial access in horses which is suitable for use for serial blood sampling and blood pressure recordings, even during exercise on treadmill. Copyright © 2011 Elsevier Ltd. All rights reserved.

Relative Abundance of Proteins in Blood Plasma Samples from Patients with Chronic Cerebral Ischemia.

Science.gov (United States)

Kaysheva, Anna L; Kopylov, Artur T; Ponomarenko, Elena A; Kiseleva, Olga I; Teryaeva, Nadezhda B; Potapov, Alexander A; Izotov, Alexander А; Morozov, Sergei G; Kudryavtseva, Valeria Yu; Archakov, Alexander I

2018-03-01

A comparative protein profile analysis of 17 blood plasma samples from patients with ischemia and 20 samples from healthy volunteers was carried out using ultra-high resolution mass spectrometry. The analysis of measurements was performed using the proteomics search engine OMSSA. Normalized spectrum abundance factor (NSAF) in the biological samples was assessed using SearchGUI. The findings of mass spectrometry analysis of the protein composition of blood plasma samples demonstrate that the depleted samples are quite similar in protein composition and relative abundance of proteins. By comparing them with the control samples, we have found a small group of 44 proteins characteristic of the blood plasma samples from patients with chronic cerebral ischemia. These proteins contribute to the processes of homeostasis maintenance, including innate immune response unfolding, the response of a body to stress, and contribution to the blood clotting cascade.

Blood biomonitoring of metals in subjects living near abandoned mining and active industrial areas.

Science.gov (United States)

Madeddu, Roberto; Tolu, Paola; Asara, Yolande; Farace, Cristiano; Forte, Giovanni; Bocca, Beatrice

2013-07-01

A human blood biomonitoring campaign to detect the environmental exposure to metals (Cd, Cu, Cr, Mn, Pb and Zn) in 265 subjects was performed in the South-Western part of Sardinia (an Italian island) that is a particular area with a great history of coal and metal mining (Pb/Zn mainly) activities and large industrial structures (as metallurgy). Subjects living near the industrial plant area had geometric means (GM) of blood Cd (0.79 μg/l), Cu (971 μg/l), Mn (12.2 μg/l), and Pb (55.7 μg/l) significantly higher than controls (Cd, 0.47 μg/l; Cu, 900 μg/l; Mn 9.98 μg/l; Pb, 26.5 μg/l) and than people living nearby the past mining sites. Subjects living next to one dismissed mine were statistically higher in blood Cu (GM, 1,022 μg/l) and Pb (GM, 41.4 μg/l) concentrations than controls. No differences were observed in people living in the different mining sites, and this might be related to the decennial disclosure of mines and the adoption of environmental remediation programmes. Some interindividual variables influenced blood biomonitoring data, as smoke and age for Cd, gender for Cu, age, sex and alcohol for Pb, and age for Zn. Moreover, blood metal levels of the whole population were similar to reference values representative of the Sardinian population and acceptably safe according to currently available health guidelines.

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

Science.gov (United States)

Jørgensen, Pelle; Jacobsen, Peter; Poulsen, Jørgen Hjelm

2013-01-01

Using simulation as an approach to display and improve internal logistics at hospitals has great potential. This study shows how a simulation model displaying the morning blood-taking round at a Danish public hospital can be developed and utilized with the aim of improving the logistics. The focus of the simulation was to evaluate changes made to the transportation of blood samples between wards and the laboratory. The average- (AWT) and maximum waiting time (MWT) from a blood sample was drawn at the ward until it was received at the laboratory, and the distribution of arrivals of blood samples in the laboratory were used as the evaluation criteria. Four different scenarios were tested and compared with the current approach: (1) Using AGVs (mobile robots), (2) using a pneumatic tube system, (3) using porters that are called upon, or (4) using porters that come to the wards every 45 minutes. Furthermore, each of the scenarios was tested in terms of what amount of resources would give the optimal result. The simulations showed a big improvement potential in implementing a new technology/mean for transporting the blood samples. The pneumatic tube system showed the biggest potential lowering the AWT and MWT with approx. 36% and 18%, respectively. Additionally, all of the scenarios had a more even distribution of arrivals except for porters coming to the wards every 45 min. As a consequence of the results obtained in the study, the hospital decided to implement a pneumatic tube system.

Strength training reduces arterial blood pressure but not sympathetic neural activity in young normotensive subjects

Science.gov (United States)

Carter, Jason R.; Ray, Chester A.; Downs, Emily M.; Cooke, William H.

2003-01-01

The effects of resistance training on arterial blood pressure and muscle sympathetic nerve activity (MSNA) at rest have not been established. Although endurance training is commonly recommended to lower arterial blood pressure, it is not known whether similar adaptations occur with resistance training. Therefore, we tested the hypothesis that whole body resistance training reduces arterial blood pressure at rest, with concomitant reductions in MSNA. Twelve young [21 +/- 0.3 (SE) yr] subjects underwent a program of whole body resistance training 3 days/wk for 8 wk. Resting arterial blood pressure (n = 12; automated sphygmomanometer) and MSNA (n = 8; peroneal nerve microneurography) were measured during a 5-min period of supine rest before and after exercise training. Thirteen additional young (21 +/- 0.8 yr) subjects served as controls. Resistance training significantly increased one-repetition maximum values in all trained muscle groups (P < 0.001), and it significantly decreased systolic (130 +/- 3 to 121 +/- 2 mmHg; P = 0.01), diastolic (69 +/- 3 to 61 +/- 2 mmHg; P = 0.04), and mean (89 +/- 2 to 81 +/- 2 mmHg; P = 0.01) arterial blood pressures at rest. Resistance training did not affect MSNA or heart rate. Arterial blood pressures and MSNA were unchanged, but heart rate increased after 8 wk of relative inactivity for subjects in the control group (61 +/- 2 to 67 +/- 3 beats/min; P = 0.01). These results indicate that whole body resistance exercise training might decrease the risk for development of cardiovascular disease by lowering arterial blood pressure but that reductions of pressure are not coupled to resistance exercise-induced decreases of sympathetic tone.

Payload specialist Reinhard Furrer show evidence of previous blood sampling

Science.gov (United States)

1985-01-01

Payload specialist Reinhard Furrer shows evidence of previous blood sampling while Wubbo J. Ockels, Dutch payload specialist (only partially visible), extends his right arm after a sample has been taken. Both men show bruises on their arms.

Microbiological studies of blood specimen from presumptively ...

African Journals Online (AJOL)

Three hundred and fifteen blood samples were obtained from presumptively diagnosed typhoid patients who were referred for Widal Serological test at four diagnostic centres. The blood samples were subjected to bacteriological investigations. Salmonella and non-Salmonella organisms isolated were identified according ...

Plasma volume changes during hypoglycaemia: the effect of arterial blood sampling

DEFF Research Database (Denmark)

Hilsted, J; Bendtsen, Flemming; Christensen, N J

1990-01-01

To investigate whether previously reported changes in venous blood volume and composition induced by acute hypoglycaemia in humans are representative for the entire body we measured erythrocyte 51Cr content, haematocrit, plasma volume, intravascular albumin content and transcapillary escape rate...... hypoglycaemia. The magnitude of the changes in arterial and venous blood were not significantly different. These results indicate that the above changes in blood volume and composition are whole-body phenomena: furthermore, the major part of the changes are likely to occur in tissues other than upper extremity...... of albumin in arterial and venous blood in seven healthy subjects before and during insulin-induced hypoglycaemia. In both vascular sites blood 51Cr content and the haematocrit increased, plasma volume and intravascular albumin content decreased and the transcapillary escape rate of albumin increased during...

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

Directory of Open Access Journals (Sweden)

Bölenius Karin

2012-01-01

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

Intraosseous blood samples for point-of-care analysis: agreement between intraosseous and arterial analyses.

Science.gov (United States)

Jousi, Milla; Saikko, Simo; Nurmi, Jouni

2017-09-11

Point-of-care (POC) testing is highly useful when treating critically ill patients. In case of difficult vascular access, the intraosseous (IO) route is commonly used, and blood is aspirated to confirm the correct position of the IO-needle. Thus, IO blood samples could be easily accessed for POC analyses in emergency situations. The aim of this study was to determine whether IO values agree sufficiently with arterial values to be used for clinical decision making. Two samples of IO blood were drawn from 31 healthy volunteers and compared with arterial samples. The samples were analysed for sodium, potassium, ionized calcium, glucose, haemoglobin, haematocrit, pH, blood gases, base excess, bicarbonate, and lactate using the i-STAT® POC device. Agreement and reliability were estimated by using the Bland-Altman method and intraclass correlation coefficient calculations. Good agreement was evident between the IO and arterial samples for pH, glucose, and lactate. Potassium levels were clearly higher in the IO samples than those from arterial blood. Base excess and bicarbonate were slightly higher, and sodium and ionised calcium values were slightly lower, in the IO samples compared with the arterial values. The blood gases in the IO samples were between arterial and venous values. Haemoglobin and haematocrit showed remarkable variation in agreement. POC diagnostics of IO blood can be a useful tool to guide treatment in critical emergency care. Seeking out the reversible causes of cardiac arrest or assessing the severity of shock are examples of situations in which obtaining vascular access and blood samples can be difficult, though information about the electrolytes, acid-base balance, and lactate could guide clinical decision making. The analysis of IO samples should though be limited to situations in which no other option is available, and the results should be interpreted with caution, because there is not yet enough scientific evidence regarding the agreement of IO

[DNA quantification of blood samples pre-treated with pyramidon].

Science.gov (United States)

Zhu, Chuan-Hong; Zheng, Dao-Li; Ni, Rao-Zhi; Wang, Hai-Sheng; Ning, Ping; Fang, Hui; Liu, Yan

2014-06-01

To study DNA quantification and STR typing of samples pre-treated with pyramidon. The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

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

Science.gov (United States)

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.

Characteristics of a new fully programmable blood sampling device for monitoring blood radioactivity during PET

International Nuclear Information System (INIS)

Boellaard, R.; Lingen, A. van; Balen, S.C.M. van; Hoving, B.G.; Lammertsma, A.A.

2001-01-01

The first performance tests of a new fully programmable blood sampling device for monitoring blood radioactivity during positron emission tomography (PET) are described. Blood is withdrawn through 1-mm internal diameter tubing using an infusion pump which can be operated at rates varying from 0 to 600 ml/h. Activity in blood is measured by a 6-cm-thick bismuth germanate crystal connected to a photomultiplier tube and multichannel analyser (MCA) which are positioned within 6 cm lead shielding. Positioning of the tubing is an exact and simple procedure. The minimal readout time of the MCA is 1 s. Two independent energy windows can be set. Operation of the pump and MCA is fully controlled by a PC, i.e. sampling time, interval time and pump rate can be varied at any time during the PET scan by user-defined scripts. A number of characteristics of the new system were studied, such as sensitivity, dead time, linearity, effect of background radiation and pump rate as a function of input pressure. In addition, dispersion was measured as a function of pump rate. Finally, first clinical results were compared with manual samples. The sensitivity equalled 0.7 and 0.2 cps/Bq for 511- and 1022-keV 30% energy windows, respectively, and the system dead time was 500 ns. The system remained linear within 2% with activity concentrations up to 2.5 MBq/cc. Short-term reproducibility was better than 3% for a 1-h period. Long-term reproducibility was about 5% (1SD), which was mainly caused by variation in the diameter of the tubing. If the device was positioned in such a way that maximum shielding was directed towards the patient, the effects of background radiation from the patient on the measured activity concentration for clinically relevant conditions was minimal ( -1 were observed for pump rates higher than 300 ml/h, indicating that the system dispersion is small. Clinical data showed an excellent agreement to within 3% (1SD) between the results obtained with the new system and

The prevalence of toxoplasmosis in Imam Reza Hospital blood bank samples, Tehran, Iran.

Science.gov (United States)

Shaddel, M; Mirzaii Dizgah, I; Sharif, F

2014-10-01

The prevalence of toxoplasma gondii (T.g) infection in blood donors has been poorly studied. The aim of this study was to assess the prevalence of acute and chronic toxoplasmosis in blood products. A total of 223 blood products (101 fresh frozen plasma (FFP) and 122 packed cells (PC)) in Imam Reza hospital blood bank, Tehran, Iran were tested for specific T.g antibodies (IgG and IgM) by ELISA method. Positive IgG anti-T.g samples were further tested for IgM anti-T.g. A positive IgG test with the negative and positive IgM test was interpreted as a chronic and acute toxoplasmosis respectively. Of 223 samples 38.6% and 0.45% were positive for IgG anti-T.g and IgM anti-T.g levels respectively. Therefore, one and 85 samples were involved acute and chronic toxoplasmosis respectively. Twenty-six of fresh frozen plasma samples were positive for IgG anti-T.g and one of them was positive for IgM anti-T.g. Sixty packed cell samples were positive for IgG anti-T.g. Our study showed that there were chronic and acute toxoplasmosis in blood products and the prevalence of toxoplasmosis especially chronic form was high. Therefore screening of blood for T.g antibodies may be considered. Copyright © 2014 Elsevier Ltd. All rights reserved.

Elimination of heparin interference during microarray processing of fresh and biobank-archived blood samples.

Science.gov (United States)

Hebels, Dennie G A J; van Herwijnen, Marcel H M; Brauers, Karen J J; de Kok, Theo M C M; Chalkiadaki, Georgia; Kyrtopoulos, Soterios A; Kleinjans, Jos C S

2014-07-01

In the context of environmental health research, biobank blood samples have recently been identified as suitable for high-throughput omics analyses enabling the identification of new biomarkers of exposure and disease. However, blood samples containing the anti-coagulant heparin could complicate transcriptomic analysis because heparin may inhibit RNA polymerase causing inefficient cRNA synthesis and fluorophore labelling. We investigated the inhibitory effect of heparin and the influence of storage conditions (0 or 3 hr bench times, storage at room temperature or -80°C) on fluorophore labelling in heparinized fresh human buffy coat and whole blood biobank samples during the mRNA work-up protocol for microarray analysis. Subsequently, we removed heparin by lithium chloride (LiCl) treatment and performed a quality control analysis of LiCl-treated biobank sample microarrays to prove their suitability for downstream data analysis. Both fresh and biobank samples experienced varying degrees of heparin-induced inhibition of fluorophore labelling, making most samples unusable for microarray analysis. RNA derived from EDTA and citrate blood was not inhibited. No effect of bench time was observed but room temperature storage gave slightly better results. Strong correlations were observed between original blood sample RNA yield and the amount of synthesized cRNA. LiCl treatment restored sample quality to normal standards in both fresh and biobank samples and the previously identified correlations disappeared. Microarrays hybridized with LiCl-treated biobank samples were of excellent quality with no identifiable influence of heparin. We conclude that, to obtain high quality results, in most cases heparin removal is essential in blood-derived RNA samples intended for microarray analysis. Copyright © 2014 Wiley Periodicals, Inc.

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

DEFF Research Database (Denmark)

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

2006-01-01

for rats sampled from the tail vein, which showed fluctuations in body temperature in excess of 30 h after sampling. Increases in heart rate and blood pressure within the first hours after sampling indicated that periorbital puncture was the method that had the largest acute impact on the rats......Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO?), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were...... registered for three days after sampling. Initially blood pressure increased, but shortly after sampling it decreased, which led to increased heart rate. Sampling induced rapid fluctuations in body temperature, and an increase in body temperature. Generally, rats recovered from sampling within 2-3 h, except...

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

Science.gov (United States)

Wasniewski, Marine; Barrat, Jacques; Combes, Benoit; Guiot, Anne Laure; Cliquet, Florence

2014-08-01

The effectiveness of oral rabies vaccination in wildlife is usually evaluated by the detection of rabies antibodies. However, the assessment of rabies antibodies has several technical difficulties in the field, such as the collection, storage, transport and titration of blood samples, often of poor quality. The objective of this study was to assess the feasibility of collecting blood on a filter paper (FP) coupled with enzyme-linked immunosorbent assay (ELISA) titration of rabies antibodies in raccoon dogs and red foxes. The FP blood sampling method was found highly specific and repeatable in both species. Overall, results obtained with the FP sampling method were highly concordant with the conventional (venipuncture) sampling methods. Blood eluates from FP samples from foxes and raccoon dogs tested using ELISA showed concordance values of 92% and 95%, respectively, with serum samples tested using the seroneutralisation test and values of 95% and 91%, respectively, when the ELISA was used on both types of sample. The use of FP blood sampling coupled with the titration of rabies antibodies by ELISA provides a reliable alternative to conventional blood sampling and serum testing by seroneutralisation. This simple procedure is particularly attractive and cost-effective for assessing the effectiveness of oral rabies vaccination in field conditions. Copyright © 2014. Published by Elsevier B.V.

Radioimmunological assay of alpha-fetoprotein concentrations in blood serum samples from women in the first half of gestation

International Nuclear Information System (INIS)

Chomanski, M.; Grzes, A.

1977-01-01

In 66 unpregnant women and in 199 pregnant subjects (in the first half of pregnancy) the alpha-fetoprotein (α-FP) concentrations were determined in peripheric blood-serum samples. The α-FP was determined using radioimmunoo-assay technique where the addition of I 125 labelled protein was preceded by 3 hours lasting preincubation period. The free protein was separated from the bound by means of precipitation with a mixture of 96% ethanol and 34.5% ammonium acetate in a ratio 77.5 : 22.5. The patients were grouped into 5 subgroups according to the gestational age. The mean values of α-FP concentration demonstrated a steady increase along with the advancement of gestation. It is suggested that the concentration of this protein in the blood-serum samples may serve as a valuable parameter for the study of gestation development and the state of the fetus. (author)

Differential effects of sulindac and indomethacin on blood pressure in treated essential hypertensive subjects.

Science.gov (United States)

Puddey, I B; Beilin, L J; Vandongen, R; Banks, R; Rouse, I

1985-09-01

Attenuation of the effectiveness of antihypertensive therapy by non-steroidal anti-inflammatory (NSAI) drugs has been attributed to inhibition of systemic or renal vasodilator prostaglandin synthesis, or a combination of both. Indomethacin is a NSAI drug with both renal and extrarenal cyclo-oxygenase inhibition properties. Sulindac is a relatively selective cyclo-oxygenase inhibitor said not to affect urinary prostaglandin excretion. This study examines the relative effect on blood pressure of 4 weeks' treatment, with indomethacin 25 mg three times daily and sulindac 200 mg twice daily, in a randomized placebo controlled trial in 26 hypertensive subjects. In nine patients treated with indomethacin, supine blood pressure rose 11 mmHg systolic and 4 mmHg diastolic by the end of the first week, whereas nine subjects treated with sulindac showed a fall in blood pressure similar to the trend seen in placebo-treated subjects. Indomethacin treatment inhibited renal cyclo-oxygenase with a 78% reduction in urinary prostaglandin E2 excretion and 89% suppression of plasma renin activity. Neither measurement was affected by sulindac. Extrarenal cyclo-oxygenase activity was inhibited by both indomethacin and sulindac with serum thromboxane B2 decreasing by 96% and 69% respectively. The results suggest that the pressor effect of NSAI drugs is predominantly related to renal cyclo-oxygenase inhibition. the lack of effect of sulindac on blood pressure may make it a safer therapeutic option if NSAI drug therapy is necessary in the hypertensive patient.

Anxiety and cerebral blood flow during behavioral challenge. Dissociation of central from peripheral and subjective measures

International Nuclear Information System (INIS)

Zohar, J.; Insel, T.R.; Berman, K.F.; Foa, E.B.; Hill, J.L.; Weinberger, D.R.

1989-01-01

To investigate the relationship between anxiety and regional cerebral blood flow, we administered behavioral challenges to 10 patients with obsessive-compulsive disorder while measuring regional cerebral blood flow with the xenon 133 inhalation technique. Each patient was studied under three conditions: relaxation, imaginal flooding, and in vivo (actual) exposure to the phobic stimulus. Subjective anxiety, obsessive-compulsive ratings, and autonomic measures (heart rate, blood pressure) increased significantly, but respiratory rate and PCO 2 did not change across the three conditions. Regional cerebral blood flow increased slightly (in the temporal region) during imaginal flooding, but decreased markedly in several cortical regions during in vivo exposure, when anxiety was highest by subjective and peripheral autonomic measures. These results demonstrate that intense anxiety can be associated with decreased rather than increased cortical perfusion and that ostensibly related states of anxiety (eg, anticipatory and obsessional anxiety) may be associated with opposite effects on regional cerebral blood flow

Blood Sampling in Newborns: A Systematic Review of YouTube Videos.

Science.gov (United States)

Bueno, Mariana; Nishi, Érika Tihemi; Costa, Taine; Freire, Laís Machado; Harrison, Denise

Objective of this study was to conduct a systematic review of YouTube videos showing neonatal blood sampling, and to evaluate pain management and comforting interventions used. Selected videos were consumer- or professional-produced videos showing human newborns undergoing heel lancing or venipuncture for blood sampling, videos showing the entire blood sampling procedure (from the first attempt or puncture to the time of application of a cotton ball or bandage), publication date prior to October 2014, Portuguese titles, available audio. Search terms included "neonate," "newborn," "neonatal screening," and "blood collection." Two reviewers independently screened the videos and extracted the following data. A total of 13 140 videos were retrieved, of which 1354 were further evaluated, and 68 were included. Videos were mostly consumer produced (97%). Heel lancing was performed in 62 (91%). Forty-nine infants (72%) were held by an adult during the procedure. Median pain score immediately after puncture was 4 (interquartile range [IQR] = 0-5), and median length of cry throughout the procedure was 61 seconds (IQR = 88). Breastfeeding (3%) and swaddling (1.5%) were rarely implemented. Posted YouTube videos in Portuguese of newborns undergoing blood collection demonstrate minimal use of pain treatment, and maximal distress during procedures. Knowledge translation strategies are needed to implement effective measures for neonatal pain relief and comfort.

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

Science.gov (United States)

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.

Multispectral Imaging Analysis of Circulating Tumor Cells in Negatively Enriched Peripheral Blood Samples.

Science.gov (United States)

Miller, Brandon; Lustberg, Maryam; Summers, Thomas A; Chalmers, Jeffrey J

2017-01-01

A variety of biomarkers are present on cells in peripheral blood of patients with a variety of disorders, including solid tumor malignancies. While rare, characterization of these cells for specific protein levels with the advanced technology proposed, will lead to future validation studies of blood samples as "liquid biopsies" for the evaluation of disease status and therapeutic response. While circulating tumor cells (CTCs) have been isolated in the blood samples of patients with solid tumors, the exact role of CTCs as clinically useful predictive markers is still debated. Current commercial technology has significant bias in that a positive selection technology is used that preassumes specific cell surface markers (such as EpCAM) are present on CTCs. However, CTCs with low EpCAM expression have been experimentally demonstrated to be more likely to be missed by this method. In contrast, this application uses a previously developed, technology that performs a purely negative enrichment methodology on peripheral blood, yielding highly enriched blood samples that contain CTCs as well as other, undefined cell types. The focus of this contribution is the use of multispectral imaging of epifluorescent, microscopic images of these enriched cells in order to help develop clinically relevant liquid biopsies from peripheral blood samples.

Survivability of Existing Peripheral Intravenous Access Following Blood Sampling in a Pediatric Population.

Science.gov (United States)

O'Neil, Sheree W; Friesen, Mary Ann; Stanger, Debra; Trickey, Amber Williams

2018-03-07

Although pediatric patients report venipuncture as their most feared experience during hospitalization, blood sampling from peripheral intravenous accesses (PIVs) is not standard of care. Blood sampling from PIVs has long been considered by healthcare personnel to harm the access. In an effort to minimize painful procedures, pediatric nursing staff conducted a prospective, observational study to determine if blood sampling using existing PIVs resulted in the loss of the access. The ability to obtain the sample from the PIV was measured along with patient and PIV characteristics. Specimen collection using 100 existing PIVs was attempted on pediatric inpatients. Each PIV was observed for functionality, infiltration, occlusion, and dislodgement following collection and again in 4h. Frequencies of PIV loss and successful blood sampling were calculated. Patient age, PIV gauge, access site, and PIV age were evaluated for associations with successful sampling using chi-square tests, Fisher's exact tests, and logistic regression. PIV survivability was reported at 99%. The ability to obtain a complete specimen was reported at 76% and found to be significantly related to PIV age and site. Size of PIV and patient's age were not significantly related to successful sampling. Encouraging rates of PIV survivability and collectability suggest blood sampling from PIVs to be a valuable technique to minimize painful and distressful procedures. Nursing practice was changed in this pediatric department. Patients and families are saved the pain and distress of venipuncture. Nurses reported saving time and personal distress by avoiding the venipuncture procedure. Copyright © 2018 Elsevier Inc. All rights reserved.

Sample preparation prior to the LC-MS-based metabolomics/metabonomics of blood-derived samples.

Science.gov (United States)

Gika, Helen; Theodoridis, Georgios

2011-07-01

Blood represents a very important biological fluid and has been the target of continuous and extensive research for diagnostic, or health and drug monitoring reasons. Recently, metabonomics/metabolomics have emerged as a new and promising 'omics' platform that shows potential in biomarker discovery, especially in areas such as disease diagnosis, assessment of drug efficacy or toxicity. Blood is collected in various establishments in conditions that are not standardized. Next, the samples are prepared and analyzed using different methodologies or tools. When targeted analysis of key molecules (e.g., a drug or its metabolite[s]) is the aim, enforcement of certain measures or additional analyses may correct and harmonize these discrepancies. In omics fields such as those performed by holistic analytical approaches, no such rules or tools are available. As a result, comparison or correlation of results or data fusion becomes impractical. However, it becomes evident that such obstacles should be overcome in the near future to allow for large-scale studies that involve the assaying of samples from hundreds of individuals. In this case the effect of sample handling and preparation becomes very serious, in order to avoid wasting months of work from experts and expensive instrument time. The present review aims to cover the different methodologies applied to the pretreatment of blood prior to LC-MS metabolomic/metabonomic studies. The article tries to critically compare the methods and highlight issues that need to be addressed.

Ehrlichia canis morulae and DNA detection in whole blood and spleen aspiration samples.

Science.gov (United States)

Faria, Joice Lara Maia; Dagnone, Ana Sílvia; Munhoz, Thiago Demarchi; João, Carolina Franchi; Pereira, Wanderson Adriano Biscola; Machado, Rosângela Zacarias; Tinucci-Costa, Mirela

2010-01-01

The aim of this study was to compare the detection of Ehrlichia canis morulae and DNA by nPCR in whole blood and spleen aspiration. The sample included 40 dogs showing thrombocytopenia associated to clinical signs suggestive of canine ehrlichiosis. Morulae detection showed that in 35 of the dogs studied, 17 had morulae in spleen tissue, and two in buffy coat smears. E. canis DNA was detected in 29/40 blood samples. We verified that morulae detection is more efficient in cytological preparations from spleen aspiration. On the other hand, nPCR on spleen and blood samples were equally efficient for disease diagnosis.

A comprehensive compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects

DEFF Research Database (Denmark)

Vahidi, O; Kwok, K E; Gopaluni, R B

2016-01-01

We have expanded a former compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects. The former model was a detailed physiological model which considered the interactions of three substances, glucose, insulin and glucagon on regulating the blood sugar. The main...... variations of blood glucose concentrations following an oral glucose intake. Another model representing the incretins production in the gastrointestinal tract along with their hormonal effects on boosting pancreatic insulin production is also added to the former model. We have used two sets of clinical data...... obtained during oral glucose tolerance test and isoglycemic intravenous glucose infusion test from both type 2 diabetic and healthy subjects to estimate the model parameters and to validate the model results. The estimation of model parameters is accomplished through solving a nonlinear optimization...

Suitability of small diagnostic peripheral-blood samples for cell-therapy studies.

Science.gov (United States)

Stephanou, Coralea; Papasavva, Panayiota; Zachariou, Myria; Patsali, Petros; Epitropou, Marilena; Ladas, Petros; Al-Abdulla, Ruba; Christou, Soteroulla; Antoniou, Michael N; Lederer, Carsten W; Kleanthous, Marina

2017-02-01

Primary hematopoietic stem and progenitor cells (HSPCs) are key components of cell-based therapies for blood disorders and are thus the authentic substrate for related research. We propose that ubiquitous small-volume diagnostic samples represent a readily available and as yet untapped resource of primary patient-derived cells for cell- and gene-therapy studies. In the present study we compare isolation and storage methods for HSPCs from normal and thalassemic small-volume blood samples, considering genotype, density-gradient versus lysis-based cell isolation and cryostorage media with different serum contents. Downstream analyses include viability, recovery, differentiation in semi-solid media and performance in liquid cultures and viral transductions. We demonstrate that HSPCs isolated either by ammonium-chloride potassium (ACK)-based lysis or by gradient isolation are suitable for functional analyses in clonogenic assays, high-level HSPC expansion and efficient lentiviral transduction. For cryostorage of cells, gradient isolation is superior to ACK lysis, and cryostorage in freezing media containing 50% fetal bovine serum demonstrated good results across all tested criteria. For assays on freshly isolated cells, ACK lysis performed similar to, and for thalassemic samples better than, gradient isolation, at a fraction of the cost and hands-on time. All isolation and storage methods show considerable variation within sample groups, but this is particularly acute for density gradient isolation of thalassemic samples. This study demonstrates the suitability of small-volume blood samples for storage and preclinical studies, opening up the research field of HSPC and gene therapy to any blood diagnostic laboratory with corresponding bioethics approval for experimental use of surplus material. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

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

Science.gov (United States)

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

2015-11-01

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

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

Science.gov (United States)

Tobias, Karen M; Serrano, Leslie; Sun, Xiaocun; Flatland, Bente

2016-01-01

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. 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. 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. Canine EDTA whole blood samples cool most

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.

Science.gov (United States)

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 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

Relations between a novel, reliable, and rapid index of arterial compliance (PP-HDI) and well-established inidices of arterial blood pressure (ABP) in a sample of hypertensive elderly subjects.

Science.gov (United States)

Bergamini, L; Finelli, M E; Bendini, C; Ferrari, E; Veschi, M; Neviani, F; Manni, B; Pelosi, A; Rioli, G; Neri, M

2009-01-01

Hypertension is a risk factor for a long-lasting arterial wall-remodelling leading to stiffness. The rapid method measuring the pulse pressure (PP) by means of the tool of Hypertension Diagnostic Instruments (HDI) called PP-HDI, overcomes some of the problems arising with more-time consuming methods, like ambulatory blood pressure monitoring (ABPM), and give information about the elasticity of the arterial walls. We studied the relationship between the PP-HDI, the large artery compliance (LA-C) and small artery compliance (SA-C) and few well-established indices of arterial blood pressure (ABP) in a sample of 75 hypertensive subjects, aged 65 years and over. Significant correlations between LA-C and heart rate (HR), PP-ABPM and PP-HDI were found. SA-C relates with HR and systolic blood pressure (SBP) measured in lying and standing positions. Applying a stepwise regression analysis, we found that LA-C variance stems from PP-HDI and HR, while SA-C variance stems from SBP in lying position. Receiver operator characteristic (ROC) curves for thresholds of PP showed that PP-HDI reached levels of sensitivity/specificity similar to PP-ABPM. In conclusion, surveillance of ABP through hemo-dynamic indices, in particular of SBP, is essential, nevertheless the advantage of this control is not known in an elderly population where the organ damage is already evident. PP needs necessarily an instrumental measurement. The PP-HDI result is similar in reliability with respect to PPABPM, but is more rapid and well applicable in an elderly population.

Molecular identification and phylogenetic analysis of Wuchereria bancrofti from human blood samples in Egypt.

Science.gov (United States)

Abdel-Shafi, Iman R; Shoieb, Eman Y; Attia, Samar S; Rubio, José M; Ta-Tang, Thuy-Huong; El-Badry, Ayman A

2017-03-01

Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.

Measurement of glucose area under the curve using minimally invasive interstitial fluid extraction technology: evaluation of glucose monitoring concepts without blood sampling.

Science.gov (United States)

Sato, Toshiyuki; Okada, Seiki; Hagino, Kei; Asakura, Yoshihiro; Kikkawa, Yasuo; Kojima, Junko; Watanabe, Toshihiro; Maekawa, Yasunori; Isobe, Kazuki; Koike, Reona; Nakajima, Hiromu; Asano, Kaoru

2011-12-01

Monitoring postprandial hyperglycemia is crucial in treating diabetes, although its dynamics make accurate monitoring difficult. We developed a new technology for monitoring postprandial hyperglycemia using interstitial fluid (ISF) extraction technology without blood sampling. The glucose area under the curve (AUC) using this system was measured as accumulated ISF glucose (IG) with simultaneous calibration with sodium ions. The objective of this study was to evaluate this technological concept in healthy individuals. Minimally invasive ISF extraction technology (MIET) comprises two steps: pretreatment with microneedles and ISF accumulation over a specific time by contact with a solvent. The correlation between glucose and sodium ion levels using MIET was evaluated in 12 subjects with stable blood glucose (BG) levels during fasting. BG and IG time courses were evaluated in three subjects to confirm their relationship while BG was fluctuating. Furthermore, the accuracy of glucose AUC measurements by MIET was evaluated several hours after a meal in 30 subjects. A high correlation was observed between glucose and sodium ion levels when BG levels were stable (R=0.87), indicating that sodium ion is a good internal standard for calibration. The variation in IG and BG with MIET was similar, indicating that IG is an adequate substitute for BG. Finally, we showed a strong correlation (R=0.92) between IG-AUC and BG-AUC after a meal. These findings validate the adequacy of glucose AUC measurements using MIET. Monitoring glucose using MIET without blood sampling may be beneficial to patients with diabetes.

Parametrically defined cerebral blood vessels as non-invasive blood input functions for brain PET studies

International Nuclear Information System (INIS)

Asselin, Marie-Claude; Cunningham, Vincent J; Amano, Shigeko; Gunn, Roger N; Nahmias, Claude

2004-01-01

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-[ 18 F]fluoro-L-DOPA and one subject injected with [ 11 C]CO-labelled red blood cells. In the phantom study, the diameter of syringes filled with an 11 C 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

The Effect of Pneumatic Tube Systems on the Hemolysis of Biochemistry Blood Samples.

Science.gov (United States)

Cakirca, Gokhan; Erdal, Huseyin

2017-05-01

Pneumatic tube systems (PTSs) are widely used in many hospitals because they lead to reduced turnaround times and cost efficiency. However, PTSs may affect the quality of the blood samples transported to the laboratory. The aim of this study was to investigate the effect of the PTS used in our hospital on the hemolysis of the biochemical blood samples transported to the laboratory. A total of 148 samples were manually transported to the laboratory by hospital staff, 148 samples were transported with the PTS, and 113 were transported with the PTS without use of sponge-rubber inserts (PTSws). Hemolysis rates and the levels of biochemical analytes for the different transportation methods were compared. No significant difference was found between the samples transported manually and with the PTS with regard to hemolysis rate and the levels of biochemical analytes. However, the samples transported with the PTSws showed a significant difference compared with the samples transported manually and with the PTS with regard to hemolysis rate and potassium and lactate dehydrogenase levels. The percentages of the samples that exceeded the permissible threshold for the hemolysis among the samples transported manually, with the PTS, and with the PTSws were 10%, 8%, and 47%, respectively. A PTS can be used safely for transporting biochemistry blood samples to the laboratory. However, a sponge-rubber insert that holds sample tubes must be used with the PTS to prevent the hemolysis of blood samples. Copyright © 2016 Emergency Nurses Association. Published by Elsevier Inc. All rights reserved.

Large-scale prospective T cell function assays in shipped, unfrozen blood samples: experiences from the multicenter TRIGR trial.

Science.gov (United States)

Hadley, David; Cheung, Roy K; Becker, Dorothy J; Girgis, Rose; Palmer, Jerry P; Cuthbertson, David; Krischer, Jeffrey P; Dosch, Hans-Michael

2014-02-01

Broad consensus assigns T lymphocytes fundamental roles in inflammatory, infectious, and autoimmune diseases. However, clinical investigations have lacked fully characterized and validated procedures, equivalent to those of widely practiced biochemical tests with established clinical roles, for measuring core T cell functions. The Trial to Reduce Insulin-dependent diabetes mellitus in the Genetically at Risk (TRIGR) type 1 diabetes prevention trial used consecutive measurements of T cell proliferative responses in prospectively collected fresh heparinized blood samples shipped by courier within North America. In this article, we report on the quality control implications of this simple and pragmatic shipping practice and the interpretation of positive- and negative-control analytes in our assay. We used polyclonal and postvaccination responses in 4,919 samples to analyze the development of T cell immunocompetence. We have found that the vast majority of the samples were viable up to 3 days from the blood draw, yet meaningful responses were found in a proportion of those with longer travel times. Furthermore, the shipping time of uncooled samples significantly decreased both the viabilities of the samples and the unstimulated cell counts in the viable samples. Also, subject age was significantly associated with the number of unstimulated cells and T cell proliferation to positive activators. Finally, we observed a pattern of statistically significant increases in T cell responses to tetanus toxin around the timing of infant vaccinations. This assay platform and shipping protocol satisfy the criteria for robust and reproducible long-term measurements of human T cell function, comparable to those of established blood biochemical tests. We present a stable technology for prospective disease-relevant T cell analysis in immunological diseases, vaccination medicine, and measurement of herd immunity.

Determination of cadmium in whole blood and scalp hair samples of Pakistani male lung cancer patients by electrothermal atomic absorption spectrometer

International Nuclear Information System (INIS)

Kazi, T.G.; Memon, A.R.; Afridi, H.I.; Jamali, M.K.; Arain, M.B.; Jalbani, N.; Sarfraz, R.A.

2008-01-01

A large number of epidemiologic studies have been undertaken to identify potential risk factors for cancer, amongst which the association with cadmium has received considerable attention. There is compelling evidence in support of positive associations between cadmium and risk of lung cancer. In present study we measured the concentration of Cd in whole blood and scalp hair samples of 120 male lung cancer patients (smokers) and 150 controls or referents (smokers and nonsmokers) from different cities of Pakistan. Both referents and patients were of same age group (ranged 40-70 years), socio-economic status, localities and dietary habits. The scalp hair and whole blood samples were oxidized by 65% nitric acid: 30% hydrogen peroxide (2:1) ratio in microwave oven. To check the validity of the proposed method, a conventional wet acid digestion method was used to obtain total Cd concentration in certified samples of human hair BCR 397 and Clincheck control-lyophilized human whole blood. All digests were analyzed for Cd concentration by electrothermal atomic absorption spectrometer (ETAAS). The results of this study showed that the average Cd concentration was higher in the blood and scalp hair of lung cancer patients at different stages as compared to controls (p < 001). The smoker referents have high level of Cd in both biological samples as compared to nonsmoker subjects. These results illustrate that the patients who continued smoking after confirmed diagnosis of lung cancer have 34.2-67.26 and 22.4-57.3% more Cd in blood samples and scalp hair than lung cancer patients who cease smoking. This study is compelling evidence in support of positive associations between cadmium, cigarette smoking and lung cancer risk

Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper.

Science.gov (United States)

Jiang, Weidong; Mao, Ying Qing; Huang, Ruochun; Duan, Chaohui; Xi, Yun; Yang, Kai; Huang, Ruo-Pan

2014-01-31

Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of some toxic metals in blood samples of smokers in ...

African Journals Online (AJOL)

Purpose: To determine some toxic elements in the blood of cigarette and tobacco pipe smokers in Riyadh, Saudi Arabia. Methods: The study setting was Riyadh, the capital city of Saudi Arabia, Riyadh City. Male volunteers, aged 20 - 58 year, whose blood samples were collected, were classified into three groups of ...

Association of ABO and Rh Blood Groups to Blood-Borne Infections among Blood Donors in Tehran-Iran.

Science.gov (United States)

Mohammadali, Fatemeh; Pourfathollah, Aliakbar

2014-07-01

The aim of this study was to investigate the prevalence of hepatitis B, hepatitis C, HIV and syphilis infections in blood donors referred to Tehran Blood Transfusion Center (TBTC), and determine any association between blood groups and blood- borne infections between the years of 2005 and 2011. This was a retrospective study conducted at TBTC. All of the donor serum samples were screened for HBV, HCV, HIV and syphilis by using third generation ELISA kits and RPR test. Initial reactive samples were tested in duplicate. Confirmatory tests were performed on all repeatedly reactive donations. Blood group was determined by forward and reverse blood grouping. The results were subjected to chi square analysis for determination of statistical difference between the values among different categories according to SPSS program. Overall, 2031451 donor serum samples were collected in 2005-2011. Totally, 10451 were positive test for HBV, HCV, HIV and syphilis. The overall seroprevalence of HBV, HCV, HIV, and syphilis was 0.39%, 0.11%, 0.005%, and 0.010%, respectively. Hepatitis B and HIV infections were significantly associated with blood group of donors (P blood group "A" and percentage of HBs Ag was lower in donors who had blood group O. There was no significant association between Hepatitis C and syphilis infections with ABO and Rh blood groups (P>0.05). Compared with neighboring countries and the international standards, prevalence of blood-borne infections is relatively low.

Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum.

Science.gov (United States)

Rosypal, Alexa C; Pick, Leanne D; Hernandez, Jaime O Esquivel; Lindsay, David S

2014-09-15

Collection of blood samples from veterinary and wildlife patients is often challenging because the samples have to be collected on farm or in the wild under various environmental conditions. This poses many technical problems associated with venipuncture materials, their safe use and disposal, transportation and processing of collected samples. Dried blood spot (DBS) sample collection techniques offer a simple and practical alternative to traditional blood collection methods to obtain blood samples from animals for parasite antibody evaluation. The DBS collection devices are compact, simple to use, and are particularly useful for large number of samples. Additionally, DBS samples take up less space and they are easier to transport than traditional venipuncture-collected blood samples. Visceral leishmaniasis (VL) is a potentially fatal parasitic disease of dogs and humans and it is frequently diagnosed by antibody tests. Immunochromatographic tests (ICT) for antibodies to Leishmania infantum are commercially available for dogs and they produce qualitative results in minutes. Measurement of canine antibodies to L. infantum with the ICT using traditional venipuncture has been validated previously, but the use of DBS samples has not been evaluated using this method. The purpose of the present study was to determine the ability of DBS samples to detect antibodies to L. infantum in dogs using a commercial ICT assay. One hundred plasma samples from dogs experimentally infected with the LIVT-1 strain of L. infantum were collected by venipuncture and frozen. Individual samples were thawed, and then 80 μl plasma (2 drops) was aliquotted onto the 8-spoked disk pad on individual DBS sample collection devices (HemaSpot™, Spot-On Sciences, Austin, TX), dried, and stored in the dark at room temperature. After one month and six months, respectively, 2 spokes of the 8 spokes of the disk pad of each DBS sample were removed and eluted in 200 μl PBS. The eluate was used to test

Subjective State, Blood Pressure, and Behavioral Control Changes Produced by an "Energy Shot"

Science.gov (United States)

Marczinski, Cecile A; Stamates, Amy L; Ossege, Julianne; Maloney, Sarah F; Bardgett, Mark E; Brown, Clifford J

2014-06-01

Background: Energy drinks and energy shots are popular consumer beverages that are advertised to increase feelings of alertness. Typically, these products include high levels of caffeine, a mild psychostimulant drug. The scientific evidence demonstrating the specific benefits of energy products to users in terms of subjective state and objective performance is surprisingly lacking. Moreover, there are rising health concerns associated with the use of these products. Therefore, the purpose of this study was to investigate the acute effects of a popular energy shot (5-Hour Energy ® ) on subjective and objective measures that were assessed hourly for 6 hours following consumption. Methods: Participants ( n =14) completed a three-session study where they received the energy shot, a placebo control, and no drink. Following dose administration, participants completed subjective Profile of Mood States ratings hourly for 6 hours. Participants also repeatedly completed a behavioral control task (the cued go/no-go task) and provided blood pressure and pulse rate readings at each hour. Results: Consumption of the energy shot did improve subjective state, as measured by increased ratings of vigor and decreased ratings of fatigue. However, the energy shot did not alter objective performance, which worsened over time. Importantly, the energy shot elevated both systolic and diastolic blood pressure. Conclusions: Consumption of one energy shot may only result in modest benefits to subjective state. Individuals with preexisting hypertension or other medical conditions should be cautious about using these new consumer products.

A cell transportation solution that preserves live circulating tumor cells in patient blood samples

International Nuclear Information System (INIS)

Stefansson, Steingrimur; Adams, Daniel L.; Ershler, William B.; Le, Huyen; Ho, David H.

2016-01-01

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90 % viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs

A cell transportation solution that preserves live circulating tumor cells in patient blood samples.

Science.gov (United States)

Stefansson, Steingrimur; Adams, Daniel L; Ershler, William B; Le, Huyen; Ho, David H

2016-05-06

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90% viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs after

Comparative evaluation of blood and serum samples in rapid immunochromatographic tests for visceral leishmaniasis.

Science.gov (United States)

Kumar, Dinesh; Khanal, Basudha; Tiwary, Puja; Mudavath, Shyam Lal; Tiwary, Narendra K; Singh, Rupa; Koirala, Kanika; Boelaert, Marleen; Rijal, Suman; Sundar, Shyam

2013-12-01

Rapid diagnostic tests (RDTs) based on the detection of specific antibodies in serum are commonly used for the diagnosis of visceral leishmaniasis (VL). Several commercial kits are available, and some of them allow the use of whole-blood samples instead of serum. An RDT is much more user-friendly for blood samples than for serum samples. In this study, we examined the sensitivities and specificities of six different commercially available immunochromatographic tests for their accuracy in detecting Leishmania infection in whole blood and serum of parasitologically confirmed VL cases. This study was performed in areas of India and Nepal where VL is endemic. A total of 177 confirmed VL cases, 208 healthy controls from areas of endemicity (EHCs), 26 malaria patients (MP), and 37 tuberculosis (TB) patients were enrolled. The reproducibilities of the blood and serum results and between-reader and between-laboratory results were tested. In India, the sensitivities of all the RDTs ranged between 94.7 and 100.0%, with no significant differences between whole blood and serum. The specificities ranged between 92.4 and 100.0%, except for the specificity of the Onsite Leishmania Ab RevB kit, which was lower (33.6 to 42.0%). No differences in specificities were observed for blood and serum. In Nepal, the sensitivities of all the test kits, for whole-blood as well as serum samples, ranged between 96.3 and 100.0%, and the specificities ranged between 90.1 and 96.1%, again with the exception of that of the Onsite Leishmania Ab RevB test, which was markedly lower (48.7 to 49.3%). The diagnostic accuracies of all the tests, except for one brand, were excellent for the whole-blood and serum samples. We conclude that whole blood is an adequate alternative for serum in RDTs for VL, with sensitivities and specificities comparable to those obtained in serum samples, provided that the test kit is of overall good quality.

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

Energy Technology Data Exchange (ETDEWEB)

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

1984-01-01

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

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

International Nuclear Information System (INIS)

Rodak, L.; Smid, B.; Valicek, L.; Jurak, E.

1984-01-01

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

Beneficial Effects of Long-Term CPAP Treatment on Sleep Quality and Blood Pressure in Adherent Subjects With Obstructive Sleep Apnea.

Science.gov (United States)

Yang, Mei-Chen; Huang, Yi-Chih; Lan, Chou-Chin; Wu, Yao-Kuang; Huang, Kuo-Feng

2015-12-01

Obstructive sleep apnea (OSA) is associated with increased risk of cardiovascular diseases. Although CPAP is the first treatment choice for moderate-to-severe OSA, acceptance of and adherence to CPAP remain problematic. High CPAP adherence is generally defined as ≥4 h of use/night for ≥70% of the nights monitored. We investigated the long-term beneficial effects of CPAP on sleep quality and blood pressure in subjects with moderate-to-severe OSA according to high or low CPAP adherence. We retrospectively analyzed 121 subjects with moderate-to-severe OSA from August 2008 to July 2012. These subjects were divided into 3 groups: (1) no CPAP treatment (n = 29), (2) low CPAP adherence (n = 28), and (3) high CPAP adherence (n = 64). All subjects were followed up for at least 1 y. The 3 groups were compared regarding anthropometric and polysomnographic variables, presence of cardiovascular comorbidities, and blood pressure at baseline and at the last follow-up. The no-treatment group showed significant increases in oxygen desaturation index and blood pressure. The high-adherence group showed significant improvement in daytime sleepiness, apnea-hypopnea index (AHI), oxygen desaturation index, and blood pressure. Although the AHI was also significantly decreased after CPAP treatment in the low-adherence group, blood pressure remained unchanged. CPAP treatment had beneficial effects on both sleep quality and blood pressure only in subjects with OSA and high CPAP adherence who used CPAP for ≥4 h/night for ≥70% of nights monitored. Subjects with low CPAP adherence received beneficial effects on AHI, but not blood pressure. Copyright © 2015 by Daedalus Enterprises.

Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.

Science.gov (United States)

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

2017-08-23

Dried blood spots are commonly used for sample collection in clinical and non-clinical settings. This method is simple, and biomolecules in the samples remain stable for months at room temperature. In the field, blood samples for the study and diagnosis of malaria are often collected on dried blood spot cards, so development of a biomarker extraction and analysis method is needed. A simple extraction procedure for the malarial biomarker Plasmodium falciparum histidine-rich protein 2 (HRP2) from dried blood spots was optimized to achieve maximum extraction efficiency. This method was used to assess the stability of HRP2 in dried blood spots. Furthermore, 328 patient samples made available from rural Zambia were analysed for HRP2 using the developed method. These samples were collected at the initial administration of artemisinin-based combination therapy and at several points following treatment. An average extraction efficiency of 70% HRP2 with a low picomolar detection limit was achieved. In specific storage conditions HRP2 was found to be stable in dried blood spots for at least 6 months. Analysis of patient samples showed the method to have a sensitivity of 94% and a specificity of 89% when compared with microscopy, and trends in HRP2 clearance after treatment were observed. The dried blood spot ELISA for HRP2 was found to be sensitive, specific and accurate. The method was effectively used to assess biomarker clearance characteristics in patient samples, which prove it to be ideal for gaining further insight into the disease and epidemiological applications.

Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia.

Science.gov (United States)

Krleza, Jasna Lenicek

2014-01-01

Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about the laboratory's parent institution, patient population, types and frequencies of laboratory tests based on capillary blood samples, choice of reference intervals, and policies and procedures specifically related to capillary sampling. Sampling practices were compared with guidelines from the Clinical and Laboratory Standards Institute (CLSI) and the World Health Organization (WHO). Of the 204 laboratories surveyed, 174 (85%) responded with complete questionnaires. Among the 174 respondents, 155 (89%) reported that they routinely perform capillary sampling, which is carried out by laboratory staff in 118 laboratories (76%). Nearly half of respondent laboratories (48%) do not have a written protocol including order of draw for multiple sampling. A single puncture site is used to provide capillary blood for up to two samples at 43% of laboratories that occasionally or regularly perform such sampling. Most respondents (88%) never perform arterialisation prior to capillary blood sampling. Capillary blood sampling is highly prevalent in Croatia across different types of clinical facilities and patient populations. Capillary sampling procedures are not standardised in the country, and the rate of laboratory compliance with CLSI and WHO guidelines is low.

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

Science.gov (United States)

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

Autonomic neuropathy in nondiabetic offspring of type 2 diabetic subjects is associated with urinary albumin excretion rate and 24-h ambulatory blood pressure: the Fredericia Study

DEFF Research Database (Denmark)

Foss, Anne-Catherine; Vestbo, Else; Frøland, Anders

2001-01-01

The aim of this study was to examine the impact of parental type 2 diabetes on the autonomic nervous system and to determine whether autonomic neuropathy is present and associated with changes in 24-h ambulatory blood pressure (AMBP) and urinary albumin excretion rate (UAER) in nondiabetic subjects......, Redmond, WA), and UAER was determined through three overnight urine samples. The subjects with parental type 2 diabetes had significantly lower heart rate variation in all three bedside tests (P

Gene expression patterns in CD4+ peripheral blood cells in healthy subjects and stage IV melanoma patients.

Science.gov (United States)

Felts, Sara J; Van Keulen, Virginia P; Scheid, Adam D; Allen, Kathleen S; Bradshaw, Renee K; Jen, Jin; Peikert, Tobias; Middha, Sumit; Zhang, Yuji; Block, Matthew S; Markovic, Svetomir N; Pease, Larry R

2015-11-01

Melanoma patients exhibit changes in immune responsiveness in the local tumor environment, draining lymph nodes, and peripheral blood. Immune-targeting therapies are revolutionizing melanoma patient care increasingly, and studies show that patients derive clinical benefit from these newer agents. Nonetheless, predicting which patients will benefit from these costly therapies remains a challenge. In an effort to capture individual differences in immune responsiveness, we are analyzing patterns of gene expression in human peripheral blood cells using RNAseq. Focusing on CD4+ peripheral blood cells, we describe multiple categories of immune regulating genes, which are expressed in highly ordered patterns shared by cohorts of healthy subjects and stage IV melanoma patients. Despite displaying conservation in overall transcriptome structure, CD4+ peripheral blood cells from melanoma patients differ quantitatively from healthy subjects in the expression of more than 2000 genes. Moreover, 1300 differentially expressed genes are found in transcript response patterns following activation of CD4+ cells ex vivo, suggesting that widespread functional discrepancies differentiate the immune systems of healthy subjects and melanoma patients. While our analysis reveals that the transcriptome architecture characteristic of healthy subjects is maintained in cancer patients, the genes expressed differentially among individuals and across cohorts provide opportunities for understanding variable immune states as well as response potentials, thus establishing a foundation for predicting individual responses to stimuli such as immunotherapeutic agents.

Trace-element measurement in human blood samples

International Nuclear Information System (INIS)

Hamidian, M.R.; Ebrahimi-Fakhar, F.

1992-01-01

It is conceivable that some essential elements such as zinc, iron, calcium, copper, phosphorus, selenium, etc., have a major impact on biological and metabolical functions in the human body. The concentration of these elements is normally very minute and changes within a naturally set tolerance. The accurate measurement of these elements in biological samples, such as in blood, is one of the objectives of medical physics in diagnosis. There are many sophisticated methods to measure the accurate amount of each element in biological samples. The methods used in this project are a combination of proton-induced X-ray emission (PIXE) and neutron activation analysis (NAA). The PIXE and NAA are fast and reliable techniques for multielement analysis at the level of parts per million and less

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

Science.gov (United States)

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

2015-12-01

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

Automated dried blood spots standard and QC sample preparation using a robotic liquid handler.

Science.gov (United States)

Yuan, Long; Zhang, Duxi; Aubry, Anne-Francoise; Arnold, Mark E

2012-12-01

A dried blood spot (DBS) bioanalysis assay involves many steps, such as the preparation of standard (STD) and QC samples in blood, the spotting onto DBS cards, and the cutting-out of the spots. These steps are labor intensive and time consuming if done manually, which, therefore, makes automation very desirable in DBS bioanalysis. A robotic liquid handler was successfully applied to the preparation of STD and QC samples in blood and to spot the blood samples onto DBS cards using buspirone as the model compound. This automated preparation was demonstrated to be accurate and consistent. However the accuracy and precision of automated preparation were similar to those from manual preparation. The effect of spotting volume on accuracy was evaluated and a trend of increasing concentrations of buspirone with increasing spotting volumes was observed. The automated STD and QC sample preparation process significantly improved the efficiency, robustness and safety of DBS bioanalysis.

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

Science.gov (United States)

Yuan, Yuan; Kitamura-Muramatsu, Yuri; Saito, Susumu; Ishizaki, Hiroshi; Nakano, Miwa; Haga, Satoshi; Matoba, Kazuhiro; Ohno, Ayumu; Murakami, Hironobu; Takeshima, Shin-Nosuke; Aida, Yoko

2015-12-02

Bovine leukemia virus (BLV) induces enzootic bovine leukosis, which is the most common neoplastic disease in cattle. Sero-epidemiological studies show that BLV infection occurs worldwide. Direct contact between infected and uninfected cattle is thought to be one of the risk factors for BLV transmission. Contact transmission occurs via a mixture of natural sources, blood, and exudates. To confirm that BLV provirus is detectable in these samples, matched blood, nasal secretion, and saliva samples were collected from 50 cattle, and genomic DNA was extracted. BLV-CoCoMo-qPCR-2, an assay developed for the highly sensitive detection of BLV, was then used to measure the proviral load in blood (n=50), nasal secretions (n=48), and saliva (n=47) samples. The results showed that 35 blood samples, 14 nasal secretion samples, and 6 saliva samples were positive for the BLV provirus. Matched blood samples from cattle that were positive for the BLV provirus (either in nasal secretion or saliva samples) were also positive in their blood. The proviral load in the positive blood samples was >14,000 (copies/1×10(5) cells). Thus, even though the proviral load in the nasal secretion and saliva samples was much lower (blood, prolonged direct contact between infected and healthy cattle may be considered as a risk factor for BLV transmission. Copyright © 2015 Elsevier B.V. All rights reserved.

Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia

Science.gov (United States)

Krleza, Jasna Lenicek

2014-01-01

Introduction: Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. Materials and methods: All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about the laboratory’s parent institution, patient population, types and frequencies of laboratory tests based on capillary blood samples, choice of reference intervals, and policies and procedures specifically related to capillary sampling. Sampling practices were compared with guidelines from the Clinical and Laboratory Standards Institute (CLSI) and the World Health Organization (WHO). Results: Of the 204 laboratories surveyed, 174 (85%) responded with complete questionnaires. Among the 174 respondents, 155 (89%) reported that they routinely perform capillary sampling, which is carried out by laboratory staff in 118 laboratories (76%). Nearly half of respondent laboratories (48%) do not have a written protocol including order of draw for multiple sampling. A single puncture site is used to provide capillary blood for up to two samples at 43% of laboratories that occasionally or regularly perform such sampling. Most respondents (88%) never perform arterialisation prior to capillary blood sampling. Conclusions: Capillary blood sampling is highly prevalent in Croatia across different types of clinical facilities and patient populations. Capillary sampling procedures are not standardised in the country, and the rate of laboratory compliance with CLSI and WHO guidelines is low. PMID:25351353

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

Science.gov (United States)

Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

2015-04-11

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

Sources of pre-analytical variations in yield of DNA extracted from blood samples: analysis of 50,000 DNA samples in EPIC.

Directory of Open Access Journals (Sweden)

Elodie Caboux

Full Text Available The European Prospective Investigation into Cancer and nutrition (EPIC is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88% performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies.

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

Science.gov (United States)

Tsuchiya, Kazuyoshi; Nakanishi, Naoyuki; Nakamachi, Eiji

2005-02-01

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

Blood gas sample spiking with total parenteral nutrition, lipid emulsion, and concentrated dextrose solutions as a model for predicting sample contamination based on glucose result.

Science.gov (United States)

Jara-Aguirre, Jose C; Smeets, Steven W; Wockenfus, Amy M; Karon, Brad S

2018-05-01

Evaluate the effects of blood gas sample contamination with total parenteral nutrition (TPN)/lipid emulsion and dextrose 50% (D50) solutions on blood gas and electrolyte measurement; and determine whether glucose concentration can predict blood gas sample contamination with TPN/lipid emulsion or D50. Residual lithium heparin arterial blood gas samples were spiked with TPN/lipid emulsion (0 to 15%) and D50 solutions (0 to 2.5%). Blood gas (pH, pCO2, pO2), electrolytes (Na+, K+ ionized calcium) and hemoglobin were measured with a Radiometer ABL90. Glucose concentration was measured in separated plasma by Roche Cobas c501. Chart review of neonatal blood gas results with glucose >300 mg/dL (>16.65 mmol/L) over a seven month period was performed to determine whether repeat (within 4 h) blood gas results suggested pre-analytical errors in blood gas results. Results were used to determine whether a glucose threshold could predict contamination resulting in blood gas and electrolyte results with greater than laboratory-defined allowable error. Samples spiked with 5% or more TPN/lipid emulsion solution or 1% D50 showed glucose concentration >500 mg/dL (>27.75 mmol/L) and produced blood gas (pH, pO 2 , pCO 2 ) results with greater than laboratory-defined allowable error. TPN/lipid emulsion, but not D50, produced greater than allowable error in electrolyte (Na + ,K + ,Ca ++ ,Hb) results at these concentrations. Based on chart review of 144 neonatal blood gas results with glucose >250 mg/dL received over seven months, four of ten neonatal intensive care unit (NICU) patients with glucose results >500 mg/dL and repeat blood gas results within 4 h had results highly suggestive of pre-analytical error. Only 3 of 36 NICU patients with glucose results 300-500 mg/dL and repeat blood gas results within 4 h had clear pre-analytical errors in blood gas results. Glucose concentration can be used as an indicator of significant blood sample contamination with either TPN

Population pharmacokinetic analysis of clopidogrel in healthy Jordanian subjects with emphasis optimal sampling strategy.

Science.gov (United States)

Yousef, A M; Melhem, M; Xue, B; Arafat, T; Reynolds, D K; Van Wart, S A

2013-05-01

Clopidogrel is metabolized primarily into an inactive carboxyl metabolite (clopidogrel-IM) or to a lesser extent an active thiol metabolite. A population pharmacokinetic (PK) model was developed using NONMEM(®) to describe the time course of clopidogrel-IM in plasma and to design a sparse-sampling strategy to predict clopidogrel-IM exposures for use in characterizing anti-platelet activity. Serial blood samples from 76 healthy Jordanian subjects administered a single 75 mg oral dose of clopidogrel were collected and assayed for clopidogrel-IM using reverse phase high performance liquid chromatography. A two-compartment (2-CMT) PK model with first-order absorption and elimination plus an absorption lag-time was evaluated, as well as a variation of this model designed to mimic enterohepatic recycling (EHC). Optimal PK sampling strategies (OSS) were determined using WinPOPT based upon collection of 3-12 post-dose samples. A two-compartment model with EHC provided the best fit and reduced bias in C(max) (median prediction error (PE%) of 9.58% versus 12.2%) relative to the basic two-compartment model, AUC(0-24) was similar for both models (median PE% = 1.39%). The OSS for fitting the two-compartment model with EHC required the collection of seven samples (0.25, 1, 2, 4, 5, 6 and 12 h). Reasonably unbiased and precise exposures were obtained when re-fitting this model to a reduced dataset considering only these sampling times. A two-compartment model considering EHC best characterized the time course of clopidogrel-IM in plasma. Use of the suggested OSS will allow for the collection of fewer PK samples when assessing clopidogrel-IM exposures. Copyright © 2013 John Wiley & Sons, Ltd.

Effect of whey on blood glucose and insulin responses to composite breakfast and lunch meals in type 2 diabetic subjects

DEFF Research Database (Denmark)

Frid, Anders H; Nilsson, Mikael; Holst, Jens Juul

2005-01-01

glycemic index (GI) with whey proteins may increase insulin secretion and improve blood glucose control in type 2 diabetic subjects. DESIGN: Fourteen diet-treated subjects with type 2 diabetes were served a high-GI breakfast (white bread) and subsequent high-GI lunch (mashed potatoes with meatballs...... insulin release and reduces postprandial blood glucose excursion after a lunch meal consisting of mashed potatoes and meatballs in type 2 diabetic subjects....

Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia.

Science.gov (United States)

Nordentoft, Merete; Larsen, Janne Tidselbak; Pedersen, Carsten Bøcker; Sørensen, Holger Jelling; Hollegaard, Mads Villiam; Hougaard, David Michael; Mortensen, Preben Bo; Petersen, Liselotte

2015-03-01

The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have found that early and late blood sampling, compared to sampling at day 5, was associated with increased risk of schizophrenia. As delay in sampling of blood for neonatal screening cannot in itself influence the risk of schizophrenia, it must be seen as a proxy for unknown underlying causes responsible for this association. Therefore, we investigated whether the increased risk can be explained by other risk factors for schizophrenia. A case-control design was applied. A total of 846 cases with schizophrenia were selected from the Danish Psychiatric Case Register. One control was selected for each case, matched on sex and exact date of birth. Both early and late blood sampling was associated with increased risk for schizophrenia. Compared to blood sampling at day 5, sampling at days 0 to 4 after birth was associated with an incidence rate ratio (IRR) of 1.46 (95% CI 1.15-1.87) for development of schizophrenia, and sampling at days 6 to 9 and at days 10 to 53 was associated with an IRR of 1.5 (95% CI 1.13-1.98) and 3.00 (95% CI 1.59-5.67), respectively. After adjusting the estimates for place of birth, both parents' psychiatric illness, maternal and paternal age, parents' country of origin, child admission, and parental education and income, the estimates were slightly different. Thus, blood collection at 0-4days was associated with an IRR of 1.27 (95% CI 0.94-1.71), 6-9days 1.31 (95% CI 0.94-1.84) and 10+days 3.52 (95% CI 1.50 to 8.24). After adjusting risk estimates for well-known risk factors, delay in sampling of blood for neonatal screening was associated with unexplained increased risk of schizophrenia. Thus, a key finding is that age at test is a proxy for unobserved risk factors

Effect of exercise on erythrocyte count and blood activity concentration after /sup 99m/Tc in vivo red blood cell labeling

International Nuclear Information System (INIS)

Konstam, M.A.; Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

1982-01-01

We studied the effect of exercise on blood radiotracer concentration after /sup 99m/Tc in vivo red blood cell labeling. After red blood cell labeling, 13 subjects underwent maximal supine bicycle exercise. Radioactivity, analyzed with a well counter, was measured in heparinized venous blood samples drawn at rest and during peak exercise. Changes in activity were compared with changes in erythrocyte count. Activity and erythrocyte counts increased during exercise in all 13 subjects. Percent increase in activity correlated with percent increase in erythrocyte count (r . -0.78), but did not correlate with either duration of exercise or maximal heart rate. Twenty minutes after termination of exercise, activity and erythrocyte count had decreased from peak exercise values but remained higher than preexercise values. In nine nonexercised control subjects, samples drawn 20 minutes apart showed no change in activity or in erythrocyte count. We conclude that exercise increases blood activity, primarily because of an increase in erythrocyte count. During radionuclide ventriculography, blood activity must be measured before and after any intervention, particularly exercise, before a change in left ventricular activity can be attributed to a change in left ventricular volume

Fabrication and Characterization of a Microfluidic Device to Ultrapurify Blood Samples

KAUST Repository

Tallerico, Marco

2015-05-04

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

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

DEFF Research Database (Denmark)

Losacco, Valentina; Cuttini, Marina; Greisen, Gorm

2011-01-01

Objective To describe the use of heel blood sampling and non-pharmacological analgesia in a large representative sample of neonatal intensive care units (NICUs) in eight European countries, and compare their self-reported practices with evidence-based recommendations. Methods Information on use...... of heel blood sampling and associated procedures (oral sweet solutions, non-nutritive sucking, swaddling or positioning, topical anaesthetics and heel warming) were collected through a structured mail questionnaire. 284 NICUs (78% response rate) participated, but only 175 with >/=50 very low birth weight...... admissions per year were included in this analysis. Results Use of heel blood sampling appeared widespread. Most units in the Netherlands, UK, Denmark, Sweden and France predominantly adopted mechanical devices, while manual lance was still in use in the other countries. The two Scandinavian countries...

Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples

Czech Academy of Sciences Publication Activity Database

Rájecký, Michal; Lojek, Antonín; Číž, Milan

2012-01-01

Roč. 34, č. 2 (2012), s. 136-142 ISSN 1751-5521 R&D Projects: GA MŠk(CZ) OC10044 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : chemiluminescence * isoluminol * whole blood Subject RIV: BO - Biophysics Impact factor: 1.293, year: 2012

Evaluation of a lateral flow-based technology card for blood typing using a simplified protocol in a model of extreme blood sampling conditions.

Science.gov (United States)

Clavier, Benoît; Pouget, Thomas; Sailliol, Anne

2018-02-01

Life-threatening situations requiring blood transfusion under extreme conditions or in remote and austere locations, such as the battlefield or in traffic accidents, would benefit from reliable blood typing practices that are easily understood by a nonscientist or nonlaboratory technician and provide quick results. A simplified protocol was developed for the lateral flow-based device MDmulticard ABO-D-Rh subgroups-K. Its performance was compared to a reference method (PK7300, Beckman Coulter) in native blood samples from donors. The method was tested on blood samples stressed in vitro as a model of hemorrhage cases (through hemodilution using physiologic serum) and dehydration (through hemoconcentration by removing an aliquot of plasma after centrifugation), respectively. A total of 146 tests were performed on 52 samples; 126 in the hemodilution group (42 for each native, diluted 1/2, and diluted 1/4 samples) and 20 in the hemoconcentration group (10 for each native and 10% concentrated samples). Hematocrit in the tested samples ranged from 9.8% to 57.6% while hemoglobin levels ranged from 3.2 to 20.1 g/dL. The phenotype profile detected with the MDmulticard using the simplified protocol resulted in 22 A, seven B, 20 O, and three AB, of which nine were D- and five were Kell positive. No discrepancies were found with respect to the results obtained with the reference method. The simplified protocol for MDmulticard use could be considered a reliable method for blood typing in extreme environment or emergency situations, worsened by red blood cell dilution or concentration. © 2017 AABB.

Assessing the associations of blood metabolites with osteoporosis: a Mendelian randomization study.

Science.gov (United States)

Liu, Li; Wen, Yan; Zhang, Lei; Xu, Peng; Liang, Xiao; Du, Yanan; Li, Ping; He, Awen; Fan, QianRui; Hao, Jingcan; Wang, Wenyu; Guo, Xiong; Shen, Hui; Tian, Qing; Zhang, Feng; Deng, Hong-Wen

2018-03-01

Osteoporosis is a metabolic bone disease. The impact of blood metabolites on the development of osteoporosis remains elusive now. To explore the relationship between blood metabolites and osteoporosis. We used 2,286 unrelated Caucasian subjects as discovery samples and 3,143 unrelated Caucasian subjects from the Framingham heart study (FHS) as replication samples. Bone mineral density (BMD) were measured using dual-energy X-ray absorptiometry. Genome-wide SNP genotyping was performed using Affymetrix Human SNP Array 6.0 (for discovery samples) and Affymetrix SNP 500K and 50K array (for FHS replication samples). The SNP sets significantly associated with blood metabolites were obtained from a published whole-genome sequencing study. For each subject, the genetic risk score (GRS) of metabolite was calculated from the genotype data of metabolite associated SNP sets. Pearson correlation analysis was conducted to evaluate the potential impact of blood metabolites on the variations bone phenotypes. 10,000 permutations were conducted to calculate the empirical P value and false discovery rate (FDR). 481 blood metabolites were analyzed in this study. We identified multiple blood metabolites associated with hip BMD, such as 1,5-anhydroglucitol(1,5-AG) (Pdiscovery metabolites on the variations of BMD, and identified several candidate blood metabolites for osteoporosis.

Consumption of 125I labelled fibrinogen in normal subjects

International Nuclear Information System (INIS)

Langer, B.; Camargo, E.E.; Reis, J.M.M. dos; Carvalho, N.; Leao, L.E.P.

1978-01-01

The metabolism of iodine- 125 labeled human fibrinogen is studied by using three different sets of the radiopharmaceutical (0.9, 1.3 and 1.84 iodine atoms/fibrinogen molecule ratios) in 19 normal subjects. An aliquot of 40 μCi of fibrinogem- 125 I is injected in each subject, on normal dietary conditions and blood samples are withdrawn at 30, 60, 180, 36 and 720 minutes after the injection and, thereafter, one daily sample during 10 days. The compartmental distribution of the tracer is defined by plotting plasma and serum sample counts on a semilogarithmic graph paper. A rapid phase and 3 compartments are obtained. A 'rapid' consumption half-life and a 'real' consumption half-life are defined. The fibrinogen clottability is followed up to the last blood sample by checking the ratios of serum and plasma radioactivities [pt

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

Science.gov (United States)

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

2015-01-01

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

Radioimmunoassay screening and GC/MS confirmation of whole blood samples for drugs of abuse

Energy Technology Data Exchange (ETDEWEB)

Spiehler, V.R.; Sedgwick, P.

From 1981 to 1984, an average of 300 radioimmunoassay screens on whole blood were performed each week in the authors laboratory. Most samples were screened for opiates phencyclidine and its analogs, barbiturates, and cocaine or its metabolite benzoylecgonine. A commercially available radioimmunoassay was used with modifications to facilitate screening of whole blood. Increasing sample size increased the sensitivity of the assay. Changing reagent concentration (1:1 dilution), incubation time, sample matrix (water, urine, or blood), or fraction counted (precipitate or supernatant) did not affect the utility of the standard curve or the sensitivity of the assay. All positive results for phencyclidine, opiates, cocaine, and related compounds were confirmed by GC/MA. Barbiturate positives were confirmed by UV spectrophotometry.

Creatinine measurement on dry blood spot sample for chronic kidney disease screening.

Science.gov (United States)

Silva, Alan Castro Azevedo E; Gómez, Juan Fidel Bencomo; Lugon, Jocemir Ronaldo; Graciano, Miguel Luis

2016-03-01

Chronic kidney disease (CKD) screening is advisable due to its high morbidity and mortality and is usually performed by sampling blood and urine. Here we present an innovative and simpler method, by measuring creatinine on a dry blood spot on filter paper. One-hundred and six individuals at high risk for CKD were enrolled. The creatinine values obtained using both tests and the demographic data of each participant allowed us to determinate the eGFR. The adopted cutoff for CKD was an eGFR creatinine values differences (+ 0.68mg/dl to -0.55mg/dl) inside the ± 1.96 SD, without systematic differences. Measurement of creatinine on dry blood sample is an easily feasible non-invasive diagnostic test with good accuracy that may be useful to screen chronic kidney disease.

Efficacy of the FilmArray blood culture identification panel for direct molecular diagnosis of infectious diseases from samples other than blood.

Science.gov (United States)

Micó, Miquel; Navarro, Ferran; de Miniac, Daniela; González, Yésica; Brell, Albert; López, Cristina; Sánchez-Reus, Ferran; Mirelis, Beatriz; Coll, Pere

2015-12-01

Molecular-based techniques reduce the delay in diagnosing infectious diseases and therefore contribute to better patient outcomes. We assessed the FilmArray blood culture identification (BCID) panel (Biofire Diagnostics/bioMérieux) directly on clinical specimens other than blood: cerebrospinal, joint, pleural and ascitic fluids, bronchoscopy samples and abscesses. We compared the results from 88 samples obtained by culture-based techniques. The percentage of agreement between the two methods was 75 % with a Cohen κ value of 0.51. Global sensitivity and specificity using the FilmArray BCID panel were 71 and 97 %, respectively. Sensitivity was poorer in samples with a low bacterial load, such as ascitic and pleural fluids (25 %), whereas the sensitivity for abscess samples was high (89 %). These findings suggest that the FilmArray BCID panel could be useful to perform microbiological diagnosis directly from samples other than positive blood cultures, as it offers acceptable sensitivity and moderate agreement with conventional microbiological methods. Nevertheless, cost-benefit studies should be performed before introducing this method into algorithms for microbiological diagnostics.

Modification of a two blood sample method used for measurement of GFR with 99mTc-DTPA.

Science.gov (United States)

Surma, Marian J; Płachcińska, Anna; Kuśmierek, Jacek

2018-01-01

Measurements of GFR may be performed with a slope/intercept method (S/I), using only two blood samples taken in strictly defined time points. The aim of the study was to modify this method in order to extend time intervals suitable for blood sampling. Modification was based on a variation of a Russel et al. model parameter, selection of time intervals suitable for blood sampling and assessment of uncertainty of calculated results. Archived values of GFR measurements of 169 patients with different renal function, from 5.5 to 179 mL/min, calculated with a multiple blood sample method were used. Concentrations of a radiopharmaceutical in consecutive minutes, from 60th to 190th after injection, were calculated theoretically, using archived parameters of biexponential functions describing a decrease in 99mTc-DTPA concentration in blood plasma with time. These values, together with injected activities, were treated as measurements and used for S/I clearance calculations. Next, values of S/I clearance were compared with the multiple blood sample method in order to calculate suitable values of exponent present in a Russel's model, for every combination of two blood sampling time points. A model was considered accurately fitted to measured values when SEE ≤ 3.6 mL/min. Assessments of uncertainty of obtained results were based on law of error superposition, taking into account mean square prediction error and also errors introduced by pipetting, time measurement and stochastic radioactive decay. The accepted criteria resulted in extension of time intervals suitable for blood sampling to: between 60 and 90 minutes after injection for the first sample and between 150 and 180 minutes for the second sample. Uncertainty of results was assessed as between 4 mL/min for GFR = 5-10 mL/min and 8 mL/min for GFR = 180 mL/min. Time intervals accepted for blood sampling fully satisfy nuclear medicine staff and ensure proper determination of GFR. Uncertainty of results is entirely

Improved removal of blood contamination from ThinPrep cervical cytology samples for Raman spectroscopic analysis.

Science.gov (United States)

Traynor, Damien; Duraipandian, Shiyamala; Martin, Cara M; O'Leary, John J; Lyng, Fiona M

2018-05-01

There is an unmet need for methods to help in the early detection of cervical precancer. Optical spectroscopy-based techniques, such as Raman spectroscopy, have shown great potential for diagnosis of different cancers, including cervical cancer. However, relatively few studies have been carried out on liquid-based cytology (LBC) pap test specimens and confounding factors, such as blood contamination, have been identified. Previous work reported a method to remove blood contamination before Raman spectroscopy by pretreatment of the slides with hydrogen peroxide. The aim of the present study was to extend this work to excessively bloody samples to see if these could be rendered suitable for Raman spectroscopy. LBC ThinPrep specimens were treated by adding hydrogen peroxide directly to the vial before slide preparation. Good quality Raman spectra were recorded from negative and high grade (HG) cytology samples with no blood contamination and with heavy blood contamination. Good classification between negative and HG cytology could be achieved for samples with no blood contamination (sensitivity 92%, specificity 93%) and heavy blood contamination (sensitivity 89%, specificity 88%) with poorer classification when samples were combined (sensitivity 82%, specificity 87%). This study demonstrates for the first time the improved potential of Raman spectroscopy for analysis of ThinPrep specimens regardless of blood contamination. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Blood transfusion sampling and a greater role for error recovery.

Science.gov (United States)

Oldham, Jane

Patient identification errors in pre-transfusion blood sampling ('wrong blood in tube') are a persistent area of risk. These errors can potentially result in life-threatening complications. Current measures to address root causes of incidents and near misses have not resolved this problem and there is a need to look afresh at this issue. PROJECT PURPOSE: This narrative review of the literature is part of a wider system-improvement project designed to explore and seek a better understanding of the factors that contribute to transfusion sampling error as a prerequisite to examining current and potential approaches to error reduction. A broad search of the literature was undertaken to identify themes relating to this phenomenon. KEY DISCOVERIES: Two key themes emerged from the literature. Firstly, despite multi-faceted causes of error, the consistent element is the ever-present potential for human error. Secondly, current focus on error prevention could potentially be augmented with greater attention to error recovery. Exploring ways in which clinical staff taking samples might learn how to better identify their own errors is proposed to add to current safety initiatives.

Levels of PCBs, DDT, DDE and DDD in Italian human blood samples

Energy Technology Data Exchange (ETDEWEB)

Rocca, C. La; Abate, V.; Alivernini, S.; Iacovella, N.; Mantovani, A.; Turrio-Baldassarri, L. [Ist. Superiore di Sanita, Roma (Italy); Silvestroni, L.; Spera, G. [Dept. of Medical Pathophysiology, Univ. (Italy)

2004-09-15

The environmental contamination from polychlorinated biphenyls (PCBs) is effecting the exposure of the general population in a direct way through air inhalation, ingestion of particulate matter and dermal absorption and, most of all, in an indirect way through diet. Diet represents, in fact, the main way of human exposure to PCBs. PCBs have potential teratogenic, carcinogenic, hormonal and immunological effects. An association between endometriosis and high levels of PCB in plasma has also been reported3. Moreover, some congeners (PCB 105, PCB 118, PCB 153) have effects on thyroid hormones in animal models, although the PCB dose used in these experiments was an order of magnitude higher than the estimated human exposure. Humans are, however, exposed to a complex mixtures of PCB congeners. In this study identification and quantification of 60 PCB congeners and 3 chlorinated pesticides in human whole blood samples are presented. The subjects examined in this pilot study were a small group of patients with possible endocrine-related problems and unknown specific exposure. The aim of this study was to increase the present understanding about the distribution of the PCBs in human whole blood. The levels of DDT and metabolites were measured as well, since these compounds are consistently reported to contribute to the whole body burden of persistent chlorinated compounds, together with PCBs.

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

Directory of Open Access Journals (Sweden)

Williams Adam R

2009-12-01

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

Nocturnal variations in subcutaneous blood flow rate in lower leg of normal human subjects

DEFF Research Database (Denmark)

Sindrup, J H; Kastrup, J; Jørgensen, B

1991-01-01

in central and local postural sympathetic vasoconstrictor activity. During sleep, characteristic variations in subcutaneous blood flow were disclosed. The 133Xe washout curve could be divided into three segments with significantly different slopes. Approximately 90 min after the subject went to sleep...

Hybrid image and blood sampling input function for quantification of small animal dynamic PET data

International Nuclear Information System (INIS)

Shoghi, Kooresh I.; Welch, Michael J.

2007-01-01

We describe and validate a hybrid image and blood sampling (HIBS) method to derive the input function for quantification of microPET mice data. The HIBS algorithm derives the peak of the input function from the image, which is corrected for recovery, while the tail is derived from 5 to 6 optimally placed blood sampling points. A Bezier interpolation algorithm is used to link the rightmost image peak data point to the leftmost blood sampling point. To assess the performance of HIBS, 4 mice underwent 60-min microPET imaging sessions following a 0.40-0.50-mCi bolus administration of 18 FDG. In total, 21 blood samples (blood-sampled plasma time-activity curve, bsPTAC) were obtained throughout the imaging session to compare against the proposed HIBS method. MicroPET images were reconstructed using filtered back projection with a zoom of 2.75 on the heart. Volumetric regions of interest (ROIs) were composed by drawing circular ROIs 3 pixels in diameter on 3-4 transverse planes of the left ventricle. Performance was characterized by kinetic simulations in terms of bias in parameter estimates when bsPTAC and HIBS are used as input functions. The peak of the bsPTAC curve was distorted in comparison to the HIBS-derived curve due to temporal limitations and delay in blood sampling, which affected the rates of bidirectional exchange between plasma and tissue. The results highlight limitations in using bsPTAC. The HIBS method, however, yields consistent results, and thus, is a substitute for bsPTAC

Vaccination and blood sampling acceptability during Ramadan fasting month: A cross-sectional study in Conakry, Guinea.

Science.gov (United States)

Peiffer-Smadja, Nathan; Ouedraogo, Ramatou; D'Ortenzio, Eric; Cissé, Papa Ndiaga; Zeggani, Zahra; Beavogui, Abdoul Habib; Faye, Sylvain Landry; Le Marcis, Frédéric; Yazdanpanah, Yazdan; Nguyen, Vinh-Kim

2017-05-02

There are few data on the acceptability of vaccination or blood sampling during Ramadan fasting month in Muslim countries. This could impact vaccination campaigns, clinical trials or healthcare during Ramadan. Using a semi-structured questionnaire, we conducted a cross-sectional study on 201 practising Muslims and 10 religious leaders in Conakry, Guinea in the wake of the recent epidemic Ebola epidemic. Acceptability of vaccination and blood sampling during Ramadan were investigated as well as reasons for refusal. Vaccination was judged acceptable during Ramadan by 46% (93/201, 95% CI 0.40-0.53) of practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) of religious leaders (p=0.11). Blood sampling was judged acceptable during Ramadan by 54% (108/201, 95% CI 0.47-0.60) of practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) of religious leaders (p=0.19). The percentage of participants that judged both blood sampling and vaccination acceptable during Ramadan was 40% (81/201, 95% CI 0.34-0.47) for practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) for religious leaders (p=0.048). The most common reasons for refusal of vaccination or blood sampling were that nothing should enter or leave the body during Ramadan (43%), that adverse events could lead to breaking the fast (32%), that blood should not be seen during Ramadan (9%) and that the Quran explicitly forbids it (9%). Although most Muslims leaders and scientists consider that injections including immunization and blood sampling should be authorized during Ramadan, many Muslims in our study judged vaccination or blood sampling unacceptable when fasting. Widely available recommendations on healthcare during Ramadan would be useful to inform Muslims. Copyright © 2017. Published by Elsevier Ltd.

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

International Nuclear Information System (INIS)

Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

2015-01-01

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

Large-scale prospective T cell function assays in shipped, unfrozen blood samples

DEFF Research Database (Denmark)

Hadley, David; Cheung, Roy K; Becker, Dorothy J

2014-01-01

, for measuring core T cell functions. The Trial to Reduce Insulin-dependent diabetes mellitus in the Genetically at Risk (TRIGR) type 1 diabetes prevention trial used consecutive measurements of T cell proliferative responses in prospectively collected fresh heparinized blood samples shipped by courier within...... cell immunocompetence. We have found that the vast majority of the samples were viable up to 3 days from the blood draw, yet meaningful responses were found in a proportion of those with longer travel times. Furthermore, the shipping time of uncooled samples significantly decreased both the viabilities...... North America. In this article, we report on the quality control implications of this simple and pragmatic shipping practice and the interpretation of positive- and negative-control analytes in our assay. We used polyclonal and postvaccination responses in 4,919 samples to analyze the development of T...

Upward Shift and Steepening of the Blood Pressure Response to Exercise in Hypertensive Subjects at High Altitude.

Science.gov (United States)

Caravita, Sergio; Faini, Andrea; Baratto, Claudia; Bilo, Grzegorz; Macarlupu, Josè Luis; Lang, Morin; Revera, Miriam; Lombardi, Carolina; Villafuerte, Francisco C; Agostoni, Piergiuseppe; Parati, Gianfranco

2018-06-09

Acute exposure to high-altitude hypobaric hypoxia induces a blood pressure rise in hypertensive humans, both at rest and during exercise. It is unclear whether this phenomenon reflects specific blood pressure hyperreactivity or rather an upward shift of blood pressure levels. We aimed at evaluating the extent and rate of blood pressure rise during exercise in hypertensive subjects acutely exposed to high altitude, and how these alterations can be counterbalanced by antihypertensive treatment. Fifty-five subjects with mild hypertension, double-blindly randomized to placebo or to a fixed-dose combination of an angiotensin-receptor blocker (telmisartan 80 mg) and a calcium-channel blocker (nifedipine slow release 30 mg), performed a cardiopulmonary exercise test at sea level and after the first night's stay at 3260 m altitude. High-altitude exposure caused both an 8 mm Hg upward shift ( P blood pressure/oxygen consumption relationship during exercise, independent of treatment. Telmisartan/nifedipine did not modify blood pressure reactivity to exercise (blood pressure/oxygen consumption slope), but downward shifted ( P blood pressure and oxygen consumption by 26 mm Hg, both at sea level and at altitude. Muscle oxygen delivery was not influenced by altitude exposure but was higher on telmisartan/nifedipine than on placebo ( P blood pressure response to exercise. The effect of the combination of telmisartan/nifedipine slow release outweighed these changes and was associated with better muscle oxygen delivery. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01830530. © 2018 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR.

Science.gov (United States)

Prabagaran, Solai Ramatchandirane; Kalaiselvi, Vellingiri; Chandramouleeswaran, Naganathan; Deepthi, Krishnan Nair Geetha; Brahmadathan, Kootallur Narayanan; Mani, Mariappa

2017-08-01

A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered. Copyright © 2017 Elsevier B.V. All rights reserved.

24-h monitoring of calcineurin phosphatase activity in healthy subjects

DEFF Research Database (Denmark)

Koefoed-Nielsen, P.B.; Karamperis, N.; Jørgensen, Kaj Anker

2005-01-01

The calcineurin inhibitors cyclosporine and tacrolimus are the cornerstone immunosuppressants used in solid organ transplantation. Studies investigating calcineurin (CaN) activity in renal transplanted patients have been published, but basic properties of the enzyme activity in healthy subjects...... remain to be described. The aim of this study was to investigate whether CaN displays circadian variation or sex difference is present in healthy subjects. Twenty subjects had blood samples drawn every 4 h for a 24-h period. CaN activity was determined in whole blood as the release of 32P from...

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

Science.gov (United States)

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

Proposing an Empirically Justified Reference Threshold for Blood Culture Sampling Rates in Intensive Care Units

Science.gov (United States)

Castell, Stefanie; Schwab, Frank; Geffers, Christine; Bongartz, Hannah; Brunkhorst, Frank M.; Gastmeier, Petra; Mikolajczyk, Rafael T.

2014-01-01

Early and appropriate blood culture sampling is recommended as a standard of care for patients with suspected bloodstream infections (BSI) but is rarely taken into account when quality indicators for BSI are evaluated. To date, sampling of about 100 to 200 blood culture sets per 1,000 patient-days is recommended as the target range for blood culture rates. However, the empirical basis of this recommendation is not clear. The aim of the current study was to analyze the association between blood culture rates and observed BSI rates and to derive a reference threshold for blood culture rates in intensive care units (ICUs). This study is based on data from 223 ICUs taking part in the German hospital infection surveillance system. We applied locally weighted regression and segmented Poisson regression to assess the association between blood culture rates and BSI rates. Below 80 to 90 blood culture sets per 1,000 patient-days, observed BSI rates increased with increasing blood culture rates, while there was no further increase above this threshold. Segmented Poisson regression located the threshold at 87 (95% confidence interval, 54 to 120) blood culture sets per 1,000 patient-days. Only one-third of the investigated ICUs displayed blood culture rates above this threshold. We provided empirical justification for a blood culture target threshold in ICUs. In the majority of the studied ICUs, blood culture sampling rates were below this threshold. This suggests that a substantial fraction of BSI cases might remain undetected; reporting observed BSI rates as a quality indicator without sufficiently high blood culture rates might be misleading. PMID:25520442

Hanford Environmental Information System (HEIS). Volume 7: Sample and Data Tracking subject area

International Nuclear Information System (INIS)

1994-06-01

The Hanford Environmental Information System (HEIS) Sample and Data Tracking subject area allows insertion of tracking information into a central repository where the data is immediately available for viewing. For example, a technical coordinator is able to view the current status of a particular sampling effort, from sample collection to data package validation dates. Four major types of data comprise the Sample and Data Tracking subject area: data about the mechanisms that groups a set of samples for a particular sampling effort; data about how constituents are grouped and assigned to a sample; data about when, where, and how samples are sent to a laboratory for analysis; and data bout the status of a sample's constituent analysis requirements, i.e., whether the analysis results have been returned from the laboratory

Determination of Ethanol in Blood Samples Using Partial Least Square Regression Applied to Surface Enhanced Raman Spectroscopy.

Science.gov (United States)

Açikgöz, Güneş; Hamamci, Berna; Yildiz, Abdulkadir

2018-04-01

Alcohol consumption triggers toxic effect to organs and tissues in the human body. The risks are essentially thought to be related to ethanol content in alcoholic beverages. The identification of ethanol in blood samples requires rapid, minimal sample handling, and non-destructive analysis, such as Raman Spectroscopy. This study aims to apply Raman Spectroscopy for identification of ethanol in blood samples. Silver nanoparticles were synthesized to obtain Surface Enhanced Raman Spectroscopy (SERS) spectra of blood samples. The SERS spectra were used for Partial Least Square (PLS) for determining ethanol quantitatively. To apply PLS method, 920~820 cm -1 band interval was chosen and the spectral changes of the observed concentrations statistically associated with each other. The blood samples were examined according to this model and the quantity of ethanol was determined as that: first a calibration method was established. A strong relationship was observed between known concentration values and the values obtained by PLS method (R 2 = 1). Second instead of then, quantities of ethanol in 40 blood samples were predicted according to the calibration method. Quantitative analysis of the ethanol in the blood was done by analyzing the data obtained by Raman spectroscopy and the PLS method.

Effect of exercise on erythrocyte count and blood activity concentration after technetium-99m in vivo red blood cell labeling

International Nuclear Information System (INIS)

Konstom, M.A.; Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

1982-01-01

The effects of exercise on blood radiotracer concentration after technetium-99m in vivo red blood cell labeling was studied. After red blood cell labeling, 13 subjects underwent maximal supine bicycle exercise. Radioactivity, analyzed with a well counter, was measured in heparinized venous blood samples drawn at rest and during peak exercise. Changes in activity were compared with changes in erythrocyte count. Activity and erythrocyte counts increased in erythrocyte count (r=0.78), but did not correlate with either duration of exercise or maximal heart rate. Twenty minutes after termination of exercise, activity and erythrocyte count had decreased from peak exercise values but remained higher than preexercise values. In nine nonexercised control subjects, samples drawn 20 minutes apart showed no change in activity or in erythrocyte count. It was concluded that exercise increases blood activity, primarily because of an increase in erythrocyte count. During radionuclide ventriculography, blood activity must be measured before and after any intervention, particularly exercise, before a change in left ventricular activity can be attributed to a change in left ventricular volume

EVALUATION OF ZEBU NELLORE CATTLE BLOOD SAMPLES USING THE CELL-DYN 3500 HEMATOLOGY ANALYZER

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Alexandre Secorun Borges

2014-12-01

Full Text Available The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Celldyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510 and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson’s correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semiautomated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

Detection of Merkel Cell Polyomavirus DNA in Serum Samples of Healthy Blood Donors

Science.gov (United States)

Mazzoni, Elisa; Rotondo, John C.; Marracino, Luisa; Selvatici, Rita; Bononi, Ilaria; Torreggiani, Elena; Touzé, Antoine; Martini, Fernanda; Tognon, Mauro G.

2017-01-01

Merkel cell polyomavirus (MCPyV) has been detected in 80% of Merkel cell carcinomas (MCC). In the host, the MCPyV reservoir remains elusive. MCPyV DNA sequences were revealed in blood donor buffy coats. In this study, MCPyV DNA sequences were investigated in the sera (n = 190) of healthy blood donors. Two MCPyV DNA sequences, coding for the viral oncoprotein large T antigen (LT), were investigated using polymerase chain reaction (PCR) methods and DNA sequencing. Circulating MCPyV sequences were detected in sera with a prevalence of 2.6% (5/190), at low-DNA viral load, which is in the range of 1–4 and 1–5 copies/μl by real-time PCR and droplet digital PCR, respectively. DNA sequencing carried out in the five MCPyV-positive samples indicated that the two MCPyV LT sequences which were analyzed belong to the MKL-1 strain. Circulating MCPyV LT sequences are present in blood donor sera. MCPyV-positive samples from blood donors could represent a potential vehicle for MCPyV infection in receivers, whereas an increase in viral load may occur with multiple blood transfusions. In certain patient conditions, such as immune-depression/suppression, additional disease or old age, transfusion of MCPyV-positive samples could be an additional risk factor for MCC onset. PMID:29238698

Transcutaneous bilirubinometry reduces the need for blood sampling in neonates with visible jaundice.

Science.gov (United States)

Mishra, S; Chawla, D; Agarwal, R; Deorari, A K; Paul, V K; Bhutani, V K

2009-12-01

We determined usefulness of transcutaneous bilirubinometry to decrease the need for blood sampling to assay serum total bilirubin (STB) in the management of jaundiced healthy Indian neonates. Newborns, > or =35 weeks' gestation, with clinical evidence of jaundice were enrolled in an institutional approved randomized clinical trial. The severity of hyperbilirubinaemia was determined by two non-invasive methods: i) protocol-based visual assessment of bilirubin (VaB) and ii) transcutaneous bilirubin (TcB) determination (BiliCheck). By a random allocation, either method was used to decide the need for blood sampling, which was defined to be present if assessed STB by allocated method exceeded 80% of hour-specific threshold values for phototherapy (2004 AAP Guidelines). A total of 617 neonates were randomized to either TcB (n = 314) or VaB (n = 303) groups with comparable gestation, birth weight and postnatal age. Need for blood sampling to assay STB was 34% lower (95% CI: 10% to 51%) in the TcB group compared with VaB group (17.5% vs 26.4% assessments; risk difference: -8.9%, 95% CI: -2.4% to -15.4%; p = 0.008). Routine use of transcutaneous bilirubinometry compared with systematic visual assessment of bilirubin significantly reduced the need for blood sampling to assay STB in jaundiced term and late-preterm neonates. (ClinicalTrials.gov number, NCT00653874).

Trace element analysis of whole blood and urine samples of diabetic patients

Energy Technology Data Exchange (ETDEWEB)

Lodhi, A S; Rashiduzzaman Khan, M [Atomic Energy Organization of Iran, Teheran. Nuclear Research Centre

1979-01-01

A number of samples of whole blood, and urine from diabetic and non-diabetic persons have been analyzed for their trace elemental contents using the proton-induced X-ray emission. The elemental contents of the diabetic and non-diabetic samples are compared.

Quantitative Assessment of the Impact of Blood Pulsation on Intraocular Pressure Measurement Results in Healthy Subjects

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Robert Koprowski

2017-01-01

Full Text Available Background. Blood pulsation affects the results obtained using various medical devices in many different ways. Method. The paper proves the effect of blood pulsation on intraocular pressure measurements. Six measurements for each of the 10 healthy subjects were performed in various phases of blood pulsation. A total of 8400 corneal deformation images were recorded. The results of intraocular pressure measurements were related to the results of heartbeat phases measured with a pulse oximeter placed on the index finger of the subject’s left hand. Results. The correlation between the heartbeat phase measured with a pulse oximeter and intraocular pressure is 0.69±0.26 (p<0.05. The phase shift calculated for the maximum correlation is equal to 60±40° (p<0.05. When the moment of measuring intraocular pressure with an air-puff tonometer is not synchronized, the changes in IOP for the analysed group of subjects can vary in the range of ±2.31 mmHg (p<0.3. Conclusions. Blood pulsation has a statistically significant effect on the results of intraocular pressure measurement. For this reason, in modern ophthalmic devices, the measurement should be synchronized with the heartbeat phases. The paper proposes an additional method for synchronizing the time of pressure measurement with the blood pulsation phase.

A policy of routine umbilical cord blood gas analysis decreased missing samples from high-risk births.

Science.gov (United States)

Ahlberg, M; Elvander, C; Johansson, S; Cnattingius, S; Stephansson, O

2017-01-01

This study compared obstetric units practicing routine or selective umbilical cord blood gas analysis, with respect to the risk of missing samples in high-risk deliveries and in infants with birth asphyxia. This was a Swedish population-based cohort study that used register data for 155 235 deliveries of live singleton infants between 2008 and 2014. Risk ratios and 95% confidence intervals were calculated to estimate the association between routine and selective umbilical cord blood gas sampling strategies and the risk of missing samples. Selective sampling increased the risk ratios when routine sampling was used as the reference, with a value of 1.0, and these were significant in high-risk deliveries and birth asphyxia. The risk ratios for selective sampling were large-for-gestational age (9.07), preterm delivery at up to 36 weeks of gestation (8.24), small-for-gestational age (7.94), two or more foetal scalp blood samples (5.96), an Apgar score of less than seven at one minute (2.36), emergency Caesarean section (1.67) and instrumental vaginal delivery (1.24). Compared with routine sampling, selective umbilical cord blood gas sampling significantly increased the risks of missing samples in high-risk deliveries and in infants with birth asphyxia. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

Improved sample capsule for determination of oxygen in hemolyzed blood

Science.gov (United States)

Malik, W. M.

1967-01-01

Sample capsule for determination of oxygen in hemolyzed blood consists of a measured section of polytetrafluoroethylene tubing equipped at each end with a connector and a stopcock valve. This method eliminates errors from air entrainment or from the use of mercury or syringe lubricant.

Effects of Blood Sample Collection Pre- and Post- Slaughter, Edta ...

African Journals Online (AJOL)

The samples were immediately subjected to Wet mount (WM), Haemotocrit centrifugation test (HCT) and Thin smear (TS) tests. The results revealed that, of the 100 samples examined, 19 (19%) were positive for the presence of Microfilaria spp while 6(6%) yielded Trypanosome spp. Of the 19 samples detected having ...

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

DEFF Research Database (Denmark)

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

time interval from blood sampling to culture. The objective of the study was to assess options for use of day-old blood samples for early-stage diagnosis of MAP infections. Bovine interleukin 12 (IL-12) can induce, and IL-10 reduce, IFN-γ production. Therefore, addition of IL-12 and anti-IL-10 could...... result in production of IFN-γ in samples previously exposed to MAP antigens. Whole blood samples were collected from heifers in a Danish dairy herd known to be infected with MAP. The samples were collected on three sample dates, and on each date the blood samples were stimulated with PPDj and recombinant......The interferon gamma (IFN-γ) test measuring specific cell-mediated immune responses in whole blood can be used for diagnosis at an early stage of Mycobacterium avium subsp. paratuberculosis (MAP) infection. A major obstacle for the practical use of IFN-γ testing is the recommended maximum 8 hour...

Sex identification of polar bears from blood and tissue samples

Science.gov (United States)

Amstrup, Steven C.; Garner, G.W.; Cronin, M.A.; Patton, J.C.

1993-01-01

Polar bears (Ursus maritimus) can be adversely affected by hunting and other human perturbations because of low population densities and low reproduction rates. The sustainable take of adult females may be as low as 1.5% of the population. Females and accompanying young are most vulnerable to hunting, and hunters have not consistently reported the sex composition of the harvest, therefore a method to confirm the sexes of polar bears harvested in Alaska is needed. Evidence of the sex of harvested animals is often not available, but blood or other tissue samples often are. We extracted DNA from tissue and blood samples, and amplified segments of zinc finger (ZFX and ZFY) genes from both X and Y chromosomes with the polymerase chain reaction. Digestion of amplified portions of the X chromosome with the restriction enzyme HaeIII resulted in subdivision of the original amplified segment into four smaller fragments. Digestion with HaeIII did not subdivide the original segment amplified from the Y chromosome. The differing fragment sizes produced patterns in gel electrophoresis that distinguished samples from male and female bears 100% of the time. This technique is applicable to the investigation of many wildlife management and research questions.

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

Science.gov (United States)

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

1985-08-05

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

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

Science.gov (United States)

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

1988-01-01

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

Standardised Resting Time Prior to Blood Sampling and Diurnal Variation Associated with Risk of Patient Misclassification

DEFF Research Database (Denmark)

Bøgh Andersen, Ida; Brasen, Claus L.; Christensen, Henry

2015-01-01

.9×10-7) and sodium (p = 8.7×10-16). Only TSH and albumin were clinically significantly influenced by diurnal variation. Resting time had no clinically significant effect. CONCLUSIONS: We found no need for resting 15 minutes prior to blood sampling. However, diurnal variation was found to have a significant......BACKGROUND: According to current recommendations, blood samples should be taken in the morning after 15 minutes' resting time. Some components exhibit diurnal variation and in response to pressures to expand opening hours and reduce waiting time, the aims of this study were to investigate...... the impact of resting time prior to blood sampling and diurnal variation on biochemical components, including albumin, thyrotropin (TSH), total calcium and sodium in plasma. METHODS: All patients referred to an outpatient clinic for blood sampling were included in the period Nov 2011 until June 2014 (opening...

Quantitative detection of epstein-barr virus DNA in cerebrospinal fluid and blood samples of patients with relapsing-remitting multiple sclerosis.

Directory of Open Access Journals (Sweden)

Clementina E Cocuzza

Full Text Available The presence of Epstein-Barr Virus (EBV DNA in cerebrospinal fluid (CSF and peripheral blood (PB samples collected from 55 patients with clinical and radiologically-active relapsing-remitting MS (RRMS and 51 subjects with other neurological diseases was determined using standardized commercially available kits for viral nucleic acid extraction and quantitative EBV DNA detection. Both cell-free and cell-associated CSF and PB fractions were analyzed, to distinguish latent from lytic EBV infection. EBV DNA was detected in 5.5% and 18.2% of cell-free and cell-associated CSF fractions of patients with RRMS as compared to 7.8% and 7.8% of controls; plasma and peripheral blood mononuclear cells (PBMC positivity rates were 7.3% and 47.3% versus 5.8% and 31.4%, respectively. No significant difference in median EBV viral loads of positive samples was found between RRMS and control patients in all tested samples. Absence of statistically significant differences in EBV positivity rates between RRMS and control patients, despite the use of highly sensitive standardized methods, points to the lack of association between EBV and MS disease activity.

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

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Andreas Lamkowski

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

Elements concentrations and relationship of whole blood and urine in 40 identical adult men in China

International Nuclear Information System (INIS)

Zhu, H.D.; Wu, Q; Fan, T.J.; Liu, Q.F; Wang, J.Y; Wang, N.F; Liu, H.S; Wang, X.Y; Ou-Yang, L.; Liu, Y.Q.; Xie, Q.

2008-01-01

Objective: To determine elemental concentrations in whole blood and 24 hr. urine of identical adult men, relative daily urinary excretion and verify relationship between both of concentrations in the blood and urine. Methods: During the same time as sampling organ or tissue samples from autopsy, whole blood and 24 hr. urine samples of identical subjects were obtained from each of 10 healthy adult male volunteers, living in 4 areas with different dietary types in China. The concentrations of 56 elements in both the two kinds of samples were analyzed by using ICP-MS as the principal, assisted with ICP-AES as well GFAAS techniques and necessary QC measures. The concentrations of urinary creatinine in the urine samples were determined by using spectrophotometric method. Results: Concentrations of both the 56 elements in these whole blood and urine samples of identical subjects and urinary creatinine and related daily urinary excretions were obtained. Conclusion: This research obtained the new data on both concentrations of these elements in whole blood and urine samples of identical subjects and their daily urinary excretions for the first time in China. These results have provided preliminary basis for understanding concentrations of these elements in the whole blood, daily urinary excretions of identical subjects as well their differences for different areas, and developing relative background values and parameters for Chinese Reference Man. Furthermore, the obtained results have been compared with both internal and external literature data and discussed. (author)

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

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

1988-02-01

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

Dried blood spots on carboxymethyl cellulose sheets: Rapid sample preparation based on dissolution and precipitation

DEFF Research Database (Denmark)

Skoglund Ask, Kristine; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

2016-01-01

This short communication describes the use of carboxymethyl cellulose sheets as sampling material for dried blood spots. Whole blood, spiked with quetiapine, a hydrophobic and basic small molecule drug substance, was spotted on the sheet and subsequently dried. The dried spot was then almost...... completely dissolved in acidified aqueous solution. It was shown that the dissolved polymer, together with major blood components can easily be precipitated and removed with acetonitrile. The presented sampling on a water-soluble biopolymer derivative followed by precipitation resulted in a simple protocol...

Evaluation of dried blood spot samples for hepatitis C virus detection and quantification.

Science.gov (United States)

Marques, Brunna Lemos Crespo; do Espírito-Santo, Márcia Paschoal; Marques, Vanessa Alves; Miguel, Juliana Custódio; da Silva, Elisangela Ferreira; Villela-Nogueira, Cristiane Alves; Lewis-Ximenez, Lia Laura; Lampe, Elisabeth; Villar, Livia Melo

2016-09-01

Dried blood spots (DBS) could be an excellent alternative for HCV diagnosis, since it is less invasive and can be stored and transported without refrigeration. The aim of this study was to optimize quantitative and qualitative methods for HCV detection in DBS. DBS and serum samples were collected from 99 subjects (59 anti-HCV/HCV RNA positive and 40 seronegative samples). Seven extraction methods and different PCR parameters were evaluated in DBS samples in the quantitative RT-PCR (qRT-PCR) developed to amplify the 5' noncoding region of HCV. A qualitative PCR for amplification of NS5B region of HCV was also valued and the nested-PCR sequenced. The qRT-PCR showed good correlation to commercial assay for HCV viral measurement in serum. To quantify HCV RNA in DBS, it was necessary to increase reverse transcriptase and cDNA concentration. HCV RNA quantification in DBS demonstrated sensitivity of 65.9%, 100% of specificity and kappa statistic of 0.65. The median viral load of DBS samples was 5.38 log10 copies/ml (minimum value=1.76 and maximum value=10.48 log10 copies/ml). HCV RNA was detected in NS5B regions and nucleotide sequences obtained in 43 serum and 11 DBS samples. The presence of the same subtype was observed in paired serum and DBS samples. In this study, it was possible to demonstrate that, despite the low sensitivity, the optimized protocol was able to determine the viral load, as well as, the infecting HCV genotype, validating the usefulness of DBS for viral load determination and molecular epidemiology studies of HCV. Copyright © 2016 Elsevier B.V. All rights reserved.

Acute effects of traditional Japanese alcohol beverages on blood glucose and polysomnography levels in healthy subjects

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Megumi Kido

2016-04-01

Full Text Available Background. Alcohol consumption is a lifestyle factor associated with type 2 diabetes. This relationship is reportedly different depending on the type of alcohol beverage. The purpose of this study was to examine the acute effects of traditional Japanese alcohol beverages on biochemical parameters, physical and emotional state, and sleep patterns. Methods. Six healthy subjects (three men and three women; age, 28.8 ± 9.5 years; body mass index, 21.4 ± 1.6 kg/m2 consumed three different types of alcohol beverages (beer, shochu, and sake, each with 40 g ethanol or mineral water with dinner on different days in the hospital. Blood samples were collected before and 1, 2, and 12 h after drinking each beverage, and assessments of physical and emotional state were administered at the same time. In addition, sleep patterns and brain waves were examined using polysomnography. Results. Blood glucose levels at 1 h and the 12-h area under the curve (AUC value after drinking shochu were significantly lower than that with water and beer. The 12-h blood insulin AUC value after drinking shochu was significantly lower than that with beer. Blood glucose × insulin level at 1 h and the 2-h blood glucose × insulin AUC value with shochu were significantly lower than that with beer. The insulinogenic indexes at 2 h with beer and sake, but not shochu, were significantly higher than that with water. The visual analogue scale scores of physical and emotional state showed that the tipsiness levels with beer, shochu, and sake at 1 h were significantly higher than that with water. These tipsiness levels were maintained at 2 h. The polysomnography showed that the rapid eye movement (REM sleep latency with shochu and sake were shorter than that with water and beer. Conclusions. Acute consumption of alcohol beverages with a meal resulted in different responses in postprandial glucose and insulin levels as well as REM sleep latency. Alcohol beverage type should be taken into

Acute effects of traditional Japanese alcohol beverages on blood glucose and polysomnography levels in healthy subjects.

Science.gov (United States)

Kido, Megumi; Asakawa, Akihiro; Koyama, Ken-Ichiro K; Takaoka, Toshio; Tajima, Aya; Takaoka, Shigeru; Yoshizaki, Yumiko; Okutsu, Kayu; Takamine, Kazunori T; Sameshima, Yoshihiro; Inui, Akio

2016-01-01

Background. Alcohol consumption is a lifestyle factor associated with type 2 diabetes. This relationship is reportedly different depending on the type of alcohol beverage. The purpose of this study was to examine the acute effects of traditional Japanese alcohol beverages on biochemical parameters, physical and emotional state, and sleep patterns. Methods. Six healthy subjects (three men and three women; age, 28.8 ± 9.5 years; body mass index, 21.4 ± 1.6 kg/m(2)) consumed three different types of alcohol beverages (beer, shochu, and sake, each with 40 g ethanol) or mineral water with dinner on different days in the hospital. Blood samples were collected before and 1, 2, and 12 h after drinking each beverage, and assessments of physical and emotional state were administered at the same time. In addition, sleep patterns and brain waves were examined using polysomnography. Results. Blood glucose levels at 1 h and the 12-h area under the curve (AUC) value after drinking shochu were significantly lower than that with water and beer. The 12-h blood insulin AUC value after drinking shochu was significantly lower than that with beer. Blood glucose × insulin level at 1 h and the 2-h blood glucose × insulin AUC value with shochu were significantly lower than that with beer. The insulinogenic indexes at 2 h with beer and sake, but not shochu, were significantly higher than that with water. The visual analogue scale scores of physical and emotional state showed that the tipsiness levels with beer, shochu, and sake at 1 h were significantly higher than that with water. These tipsiness levels were maintained at 2 h. The polysomnography showed that the rapid eye movement (REM) sleep latency with shochu and sake were shorter than that with water and beer. Conclusions. Acute consumption of alcohol beverages with a meal resulted in different responses in postprandial glucose and insulin levels as well as REM sleep latency. Alcohol beverage type should be taken into consideration

Clinical evaluation of Statstrip(R) Lactate for use in fetal scalp blood sampling

NARCIS (Netherlands)

Heinis, A.M.F.; Dillen, J. van; Oosting, J.D.; Rhose, S.; Vandenbussche, F.P.; Drongelen, J. van

2017-01-01

INTRODUCTION: Point-of-care testing of fetal scalp blood lactate is used as an alternative to pH analysis in fetal scalp blood sampling (FBS) during labor. Lactate measurements are not standardized and values vary with each device used. The aim of this study was to evaluate StatStrip(R) Lactate

Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia

DEFF Research Database (Denmark)

Nordentoft, Merete; Tidselbak Larsen, Janne; Pedersen, Carsten Bøcker

2015-01-01

for this association. Therefore, we investigated whether the increased risk can be explained by other risk factors for schizophrenia. METHODS: A case-control design was applied. A total of 846 cases with schizophrenia were selected from the Danish Psychiatric Case Register. One control was selected for each case......BACKGROUND: The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have...... found that early and late blood sampling, compared to sampling at day 5, was associated with increased risk of schizophrenia. As delay in sampling of blood for neonatal screening cannot in itself influence the risk of schizophrenia, it must be seen as a proxy for unknown underlying causes responsible...

Hypomethylation of serum blood clot DNA, but not plasma EDTA-blood cell pellet DNA, from vitamin B12-deficient subjects.

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Eoin P Quinlivan

Full Text Available Vitamin B12, a co-factor in methyl-group transfer, is important in maintaining DNA (deoxycytidine methylation. Using two independent assays we examined the effect of vitamin B12-deficiency (plasma vitamin B12<148 pmol/L on DNA methylation in women of childbearing age. Coagulated blood clot DNA from vitamin B12-deficient women had significantly (p<0.001 lower percentage deoxycytidine methylation (3.23±0.66%; n = 248 and greater [3 H]methyl-acceptance (42,859±9,699 cpm; n = 17 than DNA from B12-replete women (4.44±0.18%; n = 128 and 26,049±2,814 cpm; n = 11 [correlation between assays: r = -0.8538; p<0.001; n = 28]. In contrast, uncoagulated EDTA-blood cell pellet DNA from vitamin B12-deficient and B12-replete women exhibited similar percentage methylation (4.45±0.15%; n = 77 vs. 4.47±0.15%; n = 47 and [3 H]methyl-acceptance (27,378±4,094 cpm; n = 17 vs. 26,610±2,292 cpm; n = 11. Therefore, in simultaneously collected paired blood samples, vitamin B12-deficiency was associated with decreased DNA methylation only in coagulated samples. These findings highlight the importance of sample collection methods in epigenetic studies, and the potential impact biological processes can have on DNA methylation during collection.

Selection of blood sampling times for determination of 51Cr-EDTA clearance in a screeening procedure

International Nuclear Information System (INIS)

Gullquist, R.; Askergren, A.; Brandt, R.; Silk, B.; Strandell, T.; Huddinge University Hospital

1983-01-01

In a group of 44 construction workers various blood sampling protocols were compared with regard to variability of the 51 Cr-EDTA clearance on repeated determinations. A comparison was also made among the different blood sampling protocols with a reference method using Simpson's formula in the area calculation. A double slope method lasting for two and a half hours was finally choosen and suggested as a screening procedure in industrial environment with blood sampling at 5, 15, 90, 120, 135 and 150 minutes after injection and with the patient resting in a semirecumbent position. (orig.) [de

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

Science.gov (United States)

Huszank, Robert; Csedreki, László; Török, Zsófia

2017-02-07

There are various liquid materials whose elemental composition is of interest in various fields of science and technology. In many cases, sample preparation or the extraction can be complicated, or it would destroy the original environment before the analysis (for example, in the case of biological samples). However, multielement direct analysis of liquid samples can be realized by an external PIXE-PIGE measurement system. Particle-induced X-ray and gamma-ray emission spectroscopy (PIXE, PIGE) techniques were applied in external (in-air) microbeam configuration for the trace and main element determination of liquid samples. The direct analysis of standard solutions of several metal salts and human blood samples (whole blood, blood serum, blood plasma, and formed elements) was realized. From the blood samples, Na, P, S, Cl, K, Ca, Fe, Cu, Zn, and Br elemental concentrations were determined. The focused and scanned ion beam creates an opportunity to analyze very small volume samples (∼10 μL). As the sample matrix consists of light elements, the analysis is possible at ppm level. Using this external beam setup, it was found that it is possible to determine elemental composition of small-volume liquid samples routinely, while the liquid samples do not require any preparation processes, and thus, they can be analyzed directly. In the case of lower concentrations, the method is also suitable for the analysis (down to even ∼1 ppm level) but with less accuracy and longer measurement times.

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

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

2017-03-01

ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol. The AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods. ABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do.

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

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Rima Abdul Razzak

2013-01-01

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

Detection of Salmonella typhi by nested polymerase chain reaction in blood, urine, and stool samples

NARCIS (Netherlands)

Hatta, Mochammad; Smits, Henk L.

2007-01-01

A nested polymerase chain reaction (PCR) specific for Salmonella enterica serovar Typhi was used for the detection of the pathogen in blood, urine, and stool samples from 131 patients with clinical suspicion of typhoid fever. The sensitivity of blood culture, the PCRs with blood, urine, and feces,

Plasma concentrations of endothelial vasoactive substances in clinically healthy subjects. associations with urinary albumin excretion and ambulatory blood pressure

DEFF Research Database (Denmark)

Clausen, P; Jensen, J S; Jensent, G

2000-01-01

controls with normoalbuminuria. Plasma concentration of ET-1 was measured using an ELISA method and plasma concentration of nitrate/nitrite using a photometric method. Twenty-four-hour blood pressure was measured using a portable recorder (TM-2421). No significant differences in the concentrations...... of nitrate/nitrite and ET-1 were found between the groups, e.g. 21 (10-105) vs. 18 (11 -152) (p=0.33) and 0.98 (0.58 1.95) vs. 1.10 (0.54 -1.50) (p = 0.27), respectively. However, plasma nitrate/nitrite was significantly positively correlated to systolic and diastolic blood pressure in subjects...... in clinically healthy subjects. However, nitrate/nitrite is positively correlated to BP only in subjects with normoalbuminuria, and ET-1 is positively correlated to BP only in subjects with elevated UAE....

The effect of local heat on term neonates pain intensity during heel-blood sampling

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R. GHobadi Mohebi

2017-04-01

Full Text Available Aims: Newborns are more sensitive to pain than adults and are more susceptible to the long-term complications of pain. So, it is necessary to use procedures for reducing pain in newborns. The aim of this study was to determine the effect of local heat on the pain intensity of heel-blood sampling in the term newborns. Material & Methods: In this randomized controlled clinical trial study, in 2012, 63 healthy 3 to 5-day newborns who were referred to Shahid Delkhah Health Center in Ferdows were selected by random sampling method and randomly divided into 3 groups (21 people in each group: test (heat, placebo (sound and control. The pain intensity of newborns before, during and after heel-blood sampling was evaluated. The data collection tools were demographic questionnaire and Neonatal Infant Pain Scale (NIPS. Data were analyzed by SPSS 14.5 software and chi-square test, one-way ANOVA, Tukey's post hoc test, and ANOVA with repeated observations. Finding: The mean pain intensity in the three groups was not significantly different before intervention (p=0.86, but the mean pain intensity was lower in the test group than in the other two groups (p=0.006. After heel-blood sampling, the mean pain intensity was the least in the test group and was the most in the control group (p<0.001. Conclusion: Local heat during and after heel blood sampling decreases pain intensity in the term newborns.

Influence of partial pressure of oxygen in blood samples on measurement performance in glucose-oxidase-based systems for self-monitoring of blood glucose.

Science.gov (United States)

Baumstark, Annette; Schmid, Christina; Pleus, Stefan; Haug, Cornelia; Freckmann, Guido

2013-11-01

Partial pressure of oxygen (pO2) in blood samples can affect blood glucose (BG) measurements, particularly in systems that employ the glucose oxidase (GOx) enzyme reaction on test strips. In this study, we assessed the impact of different pO2 values on the performance of five GOx systems and one glucose dehydrogenase (GDH) system. Two of the GOx systems are labeled by the manufacturers to be sensitive to increased blood oxygen content, while the other three GOx systems are not. Aliquots of 20 venous samples were adjusted to the following pO2 values: oxygen sensitive. © 2013 Diabetes Technology Society.

Fetal scalp blood sampling in labor - a review

DEFF Research Database (Denmark)

Jørgensen, Jan Stener; Weber, Tom

2014-01-01

During the 1970s and 1980s, electronic fetal monitoring and fetal scalp blood sampling (FBS) were introduced without robust evidence. With a methodical review of the published literature, and using one randomized controlled trial, seven controlled studies, nine randomized studies of various...... surveillance methods and data from the Danish National Birth Registry, we have assessed the usefulness of FBS as a complementary tool to improve the specificity and sensitivity of electronic cardiotocography (CTG). Based on heterogeneous studies of modest quality with somewhat inconsistent results, we conclude...

Comparison of three methods for recovery of Brucella canis DNA from canine blood samples.

Science.gov (United States)

Batinga, Maria Cryskely A; Dos Santos, Jaíne C; Lima, Julia T R; Bigotto, Maria Fernanda D; Muner, Kerstin; Faita, Thalita; Soares, Rodrigo M; da Silva, David A V; Oliveira, Trícia M F S; Ferreira, Helena L; Diniz, Jaqueline A; Keid, Lara B

2017-12-01

Brucella canis, a gram-negative, facultative intracellular and zoonotic bacterium causes canine brucellosis. Direct methods are the most appropriate for the detection of canine brucellosis and bacterial isolation from blood samples has been employed as gold-standard method. However, due to the delay in obtaining results and the biological risk of the bacterial culturing, the polymerase chain reaction (PCR) has been successfully used as an alternative method for the diagnosis of the infection. Sample preparation is a key step for successful PCR and protocols that provide high DNA yield and purity are recommended to ensure high diagnostic sensitivity. The objective of this study was to evaluate the performance of PCR for the diagnosis of B. canis infection in 36 dogs by testing DNA of whole blood obtained through different extraction and purification protocols. Methods 1 and 2 were based on a commercial kit, using protocols recommended for DNA purification of whole blood and tissue samples, respectively. Method 3 was an in-house method based on enzymatic lysis and purification using organic solvents. The results of the PCR on samples obtained through three different DNA extraction protocols were compared to the blood culture. Of the 36 dogs, 13 (36.1%) were positive by blood culturing, while nine (25.0%), 14 (38.8%), and 15 (41.6%) were positive by PCR after DNA extraction using methods 1, 2 and 3, respectively. PCR performed on DNA purified by Method 2 was as efficient as blood culturing and PCR performed on DNA purified with in-house method, but had the advantage of being less laborious and, therefore, a suitable alternative for the direct B. canis detection in dogs. Copyright © 2017. Published by Elsevier B.V.

Preanalytical blood sample workup for cell-free DNA analysis using Droplet Digital PCR for future molecular cancer diagnostics.

Science.gov (United States)

van Ginkel, Joost H; van den Broek, Daan A; van Kuik, Joyce; Linders, Dorothé; de Weger, Roel; Willems, Stefan M; Huibers, Manon M H

2017-10-01

In current molecular cancer diagnostics, using blood samples of cancer patients for the detection of genetic alterations in plasma (cell-free) circulating tumor DNA (ctDNA) is an emerging practice. Since ctDNA levels in blood are low, highly sensitive Droplet Digital PCR (ddPCR) can be used for detecting rare mutational targets. In order to perform ddPCR on blood samples, a standardized procedure for processing and analyzing blood samples is necessary to facilitate implementation into clinical practice. Therefore, we assessed the technical sample workup procedure for ddPCR on blood plasma samples. Blood samples from healthy individuals, as well as lung cancer patients were analyzed. We compared different methods and protocols for sample collection, storage, centrifugation, isolation, and quantification. Cell-free DNA (cfDNA) concentrations of several wild-type targets and BRAF and EGFR-mutant ctDNA concentrations quantified by ddPCR were primary outcome measurements. Highest cfDNA concentrations were measured in blood collected in serum tubes. No significant differences in cfDNA concentrations were detected between various time points of up to 24 h until centrifugation. Highest cfDNA concentrations were detected after DNA isolation with the Quick cfDNA Serum & Plasma Kit, while plasma isolation using the QIAamp Circulating Nucleic Acid Kit yielded the most consistent results. DdPCR results on cfDNA are highly dependent on multiple factors during preanalytical sample workup, which need to be addressed during the development of this diagnostic tool for cancer diagnostics in the future. © 2017 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

Impaired myocardial blood flow reserve in subjects with metabolic syndrome analyzed using positron emission tomography and N-13 labeled ammonia

Energy Technology Data Exchange (ETDEWEB)

Teragawa, Hiroki; Kihara, Yasuki [Hiroshima University Graduate School of Biomedical Sciences, Department of Cardiovascular Medicine, Hiroshima (Japan); Morita, Koichi; Tamaki, Nagara [Hokkaido University Graduate School of Medicine, Department of Nuclear Medicine, Sapporo (Japan); Shishido, Hiroki; Otsuka, Nobuaki; Hirokawa, Yutaka [Hiroshima Heiwa Clinic, Hiroshima (Japan); Chayama, Kazuaki [Hiroshima University Graduate School of Biomedical Sciences, Department of Molecular Science and Medicine, Hiroshima (Japan)

2010-02-15

Coronary vasomotor response might be impaired in metabolic syndrome (MS); however, the precise abnormality has not been elucidated. The aim of this study was to assess coronary-vasomotor response in MS subjects using N-13 labeled ammonia and positron emission tomography. Myocardial blood flow (MBF) was measured at rest and during adenosine infusion in MS subjects (n = 13, MS group) with no definite evidence of heart disease and in subjects without MS (n = 14, non-MS group). Coronary vascular resistance (CVR) was calculated by dividing the mean aortic blood pressure by MBF. Myocardial blood flow reserve (MFR) was calculated as the ratio of the MBF during adenosine infusion to that during rest. Blood chemical parameters were measured to evaluate their relationship with MFR. During adenosine infusion, MBF was lower (p = 0.0085) and CVR higher (p = 0.0128) in the MS group than in the non-MS group and MFR was significantly lower in the MS group than in the non-MS group (2.13 {+-} 0.99 vs. 3.38 {+-} 0.95, p = 0.0027). Multivariate analysis demonstrated that the homeostasis model assessment-insulin resistance (p < 0.05) and the presence of hypertension (p < 0.05) were independent determinants of MFR. The results indicate that MFR was impaired in MS subjects, suggesting that an abnormal coronary microvascular response occurred in these subjects. This abnormality may have been partially due to insulin resistance and hypertension. (orig.)

Impaired myocardial blood flow reserve in subjects with metabolic syndrome analyzed using positron emission tomography and N-13 labeled ammonia

International Nuclear Information System (INIS)

Teragawa, Hiroki; Kihara, Yasuki; Morita, Koichi; Tamaki, Nagara; Shishido, Hiroki; Otsuka, Nobuaki; Hirokawa, Yutaka; Chayama, Kazuaki

2010-01-01

Coronary vasomotor response might be impaired in metabolic syndrome (MS); however, the precise abnormality has not been elucidated. The aim of this study was to assess coronary-vasomotor response in MS subjects using N-13 labeled ammonia and positron emission tomography. Myocardial blood flow (MBF) was measured at rest and during adenosine infusion in MS subjects (n = 13, MS group) with no definite evidence of heart disease and in subjects without MS (n = 14, non-MS group). Coronary vascular resistance (CVR) was calculated by dividing the mean aortic blood pressure by MBF. Myocardial blood flow reserve (MFR) was calculated as the ratio of the MBF during adenosine infusion to that during rest. Blood chemical parameters were measured to evaluate their relationship with MFR. During adenosine infusion, MBF was lower (p = 0.0085) and CVR higher (p = 0.0128) in the MS group than in the non-MS group and MFR was significantly lower in the MS group than in the non-MS group (2.13 ± 0.99 vs. 3.38 ± 0.95, p = 0.0027). Multivariate analysis demonstrated that the homeostasis model assessment-insulin resistance (p < 0.05) and the presence of hypertension (p < 0.05) were independent determinants of MFR. The results indicate that MFR was impaired in MS subjects, suggesting that an abnormal coronary microvascular response occurred in these subjects. This abnormality may have been partially due to insulin resistance and hypertension. (orig.)

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

Science.gov (United States)

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 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.

Isolation of endothelial colony-forming cells from blood samples collected from the jugular and cephalic veins of healthy adult horses.

Science.gov (United States)

Sharpe, Ashley N; Seeto, Wen J; Winter, Randolph L; Zhong, Qiao; Lipke, Elizabeth A; Wooldridge, Anne A

2016-10-01

OBJECTIVE To evaluate optimal isolation of endothelial colony-forming cells (ECFCs) from peripheral blood of horses. SAMPLE Jugular and cephalic venous blood samples from 17 adult horses. PROCEDURES Each blood sample was divided; isolation was performed with whole blood adherence (WBA) and density gradient centrifugation (DGC). Isolated cells were characterized by uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-Ac-LDL), vascular tubule formation, and expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor-2, and von Willebrand factor) and hematopoietic (CD14) cell markers by use of indirect immunofluorescence assay (IFA) and flow cytometry. RESULTS Colonies with cobblestone morphology were isolated from 15 of 17 horses. Blood collected from the cephalic vein yielded colonies significantly more often (14/17 horses) than did blood collected from the jugular vein (8/17 horses). Of 14 cephalic blood samples with colonies, 13 were obtained with DGC and 8 with WBA. Of 8 jugular blood samples with colonies, 8 were obtained with DGC and 4 with WBA. Colony frequency (colonies per milliliter of blood) was significantly higher for cephalic blood samples and samples isolated with DGC. Cells formed vascular tubules, had uptake of DiI-Ac-LDL, and expressed endothelial markers by use of IFA and flow cytometry, which confirmed their identity as ECFCs. CONCLUSIONS AND CLINICAL RELEVANCE Maximum yield of ECFCs was obtained for blood samples collected from both the jugular and cephalic veins and use of DGC to isolate cells. Consistent yield of ECFCs from peripheral blood of horses will enable studies to evaluate diagnostic and therapeutic uses.

Analytical and between-subject variation of thrombin generation measured by calibrated automated thrombography on plasma samples.

Science.gov (United States)

Kristensen, Anne F; Kristensen, Søren R; Falkmer, Ursula; Münster, Anna-Marie B; Pedersen, Shona

2018-05-01

The Calibrated Automated Thrombography (CAT) is an in vitro thrombin generation (TG) assay that holds promise as a valuable tool within clinical diagnostics. However, the technique has a considerable analytical variation, and we therefore, investigated the analytical and between-subject variation of CAT systematically. Moreover, we assess the application of an internal standard for normalization to diminish variation. 20 healthy volunteers donated one blood sample which was subsequently centrifuged, aliquoted and stored at -80 °C prior to analysis. The analytical variation was determined on eight runs, where plasma from the same seven volunteers was processed in triplicates, and for the between-subject variation, TG analysis was performed on plasma from all 20 volunteers. The trigger reagents used for the TG assays included both PPP reagent containing 5 pM tissue factor (TF) and PPPlow with 1 pM TF. Plasma, drawn from a single donor, was applied to all plates as an internal standard for each TG analysis, which subsequently was used for normalization. The total analytical variation for TG analysis performed with PPPlow reagent is 3-14% and 9-13% for PPP reagent. This variation can be minimally reduced by using an internal standard but mainly for ETP (endogenous thrombin potential). The between-subject variation is higher when using PPPlow than PPP and this variation is considerable higher than the analytical variation. TG has a rather high inherent analytical variation but considerable lower than the between-subject variation when using PPPlow as reagent.

A comprehensive compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects.

Science.gov (United States)

Vahidi, O; Kwok, K E; Gopaluni, R B; Knop, F K

2016-09-01

We have expanded a former compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects. The former model was a detailed physiological model which considered the interactions of three substances, glucose, insulin and glucagon on regulating the blood sugar. The main drawback of the former model was its restriction on the route of glucose entrance to the body which was limited to the intravenous glucose injection. To handle the oral glucose intake, we have added a model of glucose absorption in the gastrointestinal tract to the former model to address the resultant variations of blood glucose concentrations following an oral glucose intake. Another model representing the incretins production in the gastrointestinal tract along with their hormonal effects on boosting pancreatic insulin production is also added to the former model. We have used two sets of clinical data obtained during oral glucose tolerance test and isoglycemic intravenous glucose infusion test from both type 2 diabetic and healthy subjects to estimate the model parameters and to validate the model results. The estimation of model parameters is accomplished through solving a nonlinear optimization problem. The results show acceptable precision of the estimated model parameters and demonstrate the capability of the model in accurate prediction of the body response during the clinical studies.

Sensitivity of nested-PCR for plasmodium detection in pooled whole blood samples and its usefulness to blood donor screening in endemic areas.

Science.gov (United States)

de Freitas, Daniel Roberto Coradi; Gomes, Luciano Teixeira; Fontes, Cor Jesus F; Tauil, Pedro Luiz; Pang, Lorrin W; Duarte, Elisabeth Carmen

2014-04-01

Transfusion-transmitted malaria is a severe disease with high fatality rate. Most Brazilian blood banks in the Amazon region perform malaria screening using microscopic examination (thick smears). Since low parasite concentrations are expected in asymptomatic blood donors a high sensitivity test should be used for donor screening. This study determined the sensitivity of a nested-PCR for plasmodium detection in pooled samples. We performed a one-stage criterion validation study with 21 positive samples pooled with samples from ten negative volunteer until three different concentrations were reached (0.33; 0.25; 0.20 parasites/μL - p/μL). Nested PCR was performed as described by Snounou et al. (1993). Sensitivities (and confidence intervals) were determined by stratum of final parasite concentration on the pooled samples. All samples with parasitemia values of 0.33 and 0.25 p/μL had 100% sensitivity (95%CI=86.3-100). One negative result was obtained from a sample with 0.20 p/μL sensitivity=95.2% (95%CI=76.2-99.9). Compared to parasitemia detectable under ideal conditions of thick smear, this nested-PCR in pooled sample was able to detect 40 times more parasites per microliter. Nested-PCR in pooled samples should be considered as a high sensitive alternative to thick smear for donor screening in blood banks at endemic regions. Local authorities need to assess cost:benefit advantages of this method compared to alternatives. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

Science.gov (United States)

Stein, Claudia; Makarewicz, Oliwia; Pfeifer, Yvonne; Brandt, Christian; Pletz, Mathias W

2015-05-01

Bloodstream infections with ESBL-producers are associated with increased mortality, which is due to delayed appropriate treatment resulting in clinical failure. Current routine diagnostics for detection of bloodstream infections consists of blood culture followed by species identification and susceptibility testing. In attempts to improve and accelerate diagnostic procedures, PCR-based methods have been developed. These methods focus on species identification covering only a limited number of ESBL coding genes. Therefore, they fail to cover the steadily further evolving genetic diversity of clinically relevant β-lactamases. We have recently designed a fast and novel RNA targeting method to detect and specify CTX-M alleles from bacterial cultures, based on an amplification-pyrosequencing approach. We further developed this assay towards a diagnostic tool for clinical use and evaluated its sensitivity and specificity when applied directly to human blood samples. An optimized protocol for mRNA isolation allows detection of specific CTX-M groups from as little as 100 CFU/mL blood via reverse transcription, amplification, and pyrosequencing directly from human EDTA blood samples as well as from pre-incubated human blood cultures with a turnaround time for test results of <7 h. Copyright © 2015 Elsevier GmbH. All rights reserved.

Gene methylation parallelisms between peripheral blood cells and oral mucosa samples in relation to overweight.

Science.gov (United States)

San-Cristobal, Rodrigo; Navas-Carretero, Santiago; Milagro, Fermín I; Riezu-Boj, J Ignacio; Guruceaga, Elizabeth; Celis-Morales, Carlos; Livingstone, Katherine M; Brennan, Lorraine; Lovegrove, Julie A; Daniel, Hannelore; Saris, Wim H; Traczyk, Iwonna; Manios, Yannis; Gibney, Eileen R; Gibney, Michael J; Mathers, John C; Martinez, J Alfredo

2016-08-01

Epigenetics has an important role in the regulation of metabolic adaptation to environmental modifications. In this sense, the determination of epigenetic changes in non-invasive samples during the development of metabolic diseases could play an important role in the procedures in primary healthcare practice. To help translate the knowledge of epigenetics to public health practice, the present study aims to explore the parallelism of methylation levels between white blood cells and buccal samples in relation to obesity and associated disorders. Blood and buccal swap samples were collected from a subsample of the Spanish cohort of the Food4Me study. Infinium HumanMethylation450 DNA Analysis was carried out for the determination of methylation levels. Standard deviation for β values method and concordance correlation analysis were used to select those CpG which showed best parallelism between samples. A total of 277 CpGs met the criteria and were selected for an enrichment analysis and a correlation analysis with anthropometrical and clinical parameters. From those selected CpGs, four presented high associations with BMI (cg01055691 in GAP43; r = -0.92 and rho = -0.84 for blood; r = -0.89 and rho = -0.83 for buccal sample), HOMA-IR (cg00095677 in ATP2A3; r = 0.82 and rho = -0.84 for blood; r = -0.8 and rho = -0.83 for buccal sample) and leptin (cg14464133 in ADARB2; r = -0.9182 and rho = -0.94 for blood; r = -0.893 and rho = -0.79 for buccal sample). These findings demonstrate the potential application of non-invasive buccal samples in the identification of surrogate epigenetic biomarkers and identify methylation sites in GAP43, ATP2A3 and ADARB2 genes as potential targets in relation to overweight management and insulin sensibility.

The gingival vein as a minimally traumatic site for multiple blood sampling in guinea pigs and hamsters.

Science.gov (United States)

Rodrigues, Mariana Valotta; de Castro, Simone Oliveira; de Albuquerque, Cynthia Zaccanini; Mattaraia, Vânia Gomes de Moura; Santoro, Marcelo Larami

2017-01-01

Laboratory animals are still necessary in scientific investigation and vaccine testing, but while novel methodological approaches are not available for their replacement, the search for new, humane, easy, and painless methods is necessary to diminish their stress and pain. When multiple blood samples are to be collected from hamsters and guinea pigs, the number of available venipuncture sites-which are greatly diminished in these species in comparison with other rodents due to the absence of a long tail-, harasses animal caregivers and researchers. Thus, this study aimed to evaluate if gingival vein puncture could be used as an additional route to obtain multiple blood samples from anesthetized hamsters and guinea pigs in such a way that animal behavior, well-being or hematological parameters would not be altered. Thus, twelve anesthetized Syrian golden hamsters and English guinea pigs were randomly allocated in two groups: a control group, whose blood samples were not collected, and an experimental group in which blood samples (200 microliters) were collected by gingival vein puncture at weekly intervals over six weeks. Clinical assessment, body weight gain and complete blood cell count were evaluated weekly, and control and experimental animals were euthanized at week seven, when the mentolabial region was processed to histological analyses. Multiple blood sampling from the gingival vein evoked no clinical manifestations or alteration in the behavioral repertoire, nor a statistically significant difference in weight gain in both species. Guinea pigs showed no alteration in red blood cell, leukocyte or platelet parameters over time. Hamsters developed a characteristic pattern of age-related physiological changes, which were considered normal. Histological analyses showed no difference in morphological structures in the interdental gingiva, confirming that multiple blood sampling is barely traumatic. Thus, these results evidence that blood collection from multiple

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

DEFF Research Database (Denmark)

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

2014-01-01

eosinophilia in routine blood samples as a potential biomarker of solid tumor development in a prospective design. MATERIAL AND METHODS: From the Copenhagen Primary Care Differential Count (CopDiff) Database, we identified 356 196 individuals with at least one differential cell count (DIFF) encompassing...... was increased with mild eosinophilia [OR 1.93 (CI 1.29-2.89), p = 0.0013]. No associations with eosinophilia were observed for the remaining solid cancers. CONCLUSION: We demonstrate that eosinophilia in routine blood samples associates with an increased risk of bladder cancer. Our data emphasize...... that additional preclinical studies are needed in order to shed further light on the role of eosinophils in carcinogenesis, where it is still unknown whether the cells contribute to tumor immune surveillance or neoplastic evolution....

Local heat stress and skin blood flowmotion in subjects with familial predisposition or newly diagnosed hypertension.

Science.gov (United States)

Gryglewska, Barbara; Nęcki, Mirosław; Cwynar, Marcin; Baron, Tomasz; Grodzicki, Tomasz

2010-12-01

The aim of the study was to investigate the skin microcirculation blood flow and flowmotion response to heat stress in normotensive subjects with familial predisposition to hypertension and in hypertensive patients. Normotensives without [NT(-)] or with [NT(+)] familial predisposition and subjects with newly diagnosed hypertension (HT) were studied. Clinic blood pressure (BP) measurements and ambulatory BP monitoring as well as laboratory assessments were performed. Resting (RF), heat (HF) and maximal heat (MHF) blood flows were measured using PeriFlux laser Doppler flowmetry (LDF) and expressed as absolute units (AU) and as index of cutaneous vascular conductance (CVC). Spectral analysis of the skin LDF signal was performed by means of the Perisoft dedicated software. Kruskall-Wallis analysis of variance, χ(2) statistic and multivariate reverse regression analysis were used for calculation. The studied population consisted of 70 persons (mean age 36.1 ± 10.3 years, 44.3% women): 17 NT(-), 22 NT(+) and 31 HT, age and gender matched. Higher values of body mass index (BMI), and insulin, glucose and triglyceride levels were observed in HT than in NT groups. RF, HF and MHF were similar in all study groups, but CVC of maximal heat flow differed (p=0.02); in particular, lower values were observed in the HT than in NT(-) group (p=0.01). The study groups differed with regard to total power (p=0.01) and myogenic (p=0.03) origin flowmotion with the lowest values in the NT(+) group. BMI and night BP characteristics were strong predictors of reduction of CVC, MHF and myogenic origin flowmotion. Skin microcirculation response to local heat stress is altered in hypertensive patients with decrease in maximal heat CVC values. Moreover, normotensive subjects with familial predisposition to hypertension are characterized by diminished myogenic origin of skin blood flowmotion.

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

DEFF Research Database (Denmark)

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

2008-01-01

BACKGROUND: In many countries and especially in Scandinavia, blood samples drawn in primary healthcare are sent to a hospital laboratory for analysis. The samples are exposed to various conditions regarding storage time, storage temperature and transport form. As these factors can have a severe...... impact on the quality of results, we wanted to study which combination of transport conditions could fulfil our pre-defined goals for maximum allowable error. METHODS: Samples from 406 patients from nine general practitioners (GPs) in two Danish counties were sent to two hospitals for analyses, during......, centrifuged and separated at the doctor's office within 45-60 min. This sample was considered as the best estimate of a comparison value. RESULTS: The pre-set quality goals were fulfilled for all the investigated components for samples transported to hospital by courier either as whole blood or as "on gel...

The Acute Effect of Resistance Exercise with Blood Flow Restriction with Hemodynamic Variables on Hypertensive Subjects

Directory of Open Access Journals (Sweden)

Araújo Joamira P.

2014-12-01

Full Text Available The purpose of this study was to analyze systolic blood pressure (SBP, diastolic blood pressure (DBP and the heart rate (HR before, during and after training at moderate intensity (MI, 50%-1RM and at low intensity with blood flow restriction (LIBFR. In a randomized controlled trial study, 14 subjects (average age 45±9,9 years performed one of the exercise protocols during two separate visits to the laboratory. SBP, DBP and HR measurements were collected prior to the start of the set and 15, 30, 45 and 60 minutes after knee extension exercises. Repeated measures of analysis of variance (ANOVA were used to identify significant variables (2 x 5; group x time. The results demonstrated a significant reduction in SBP in the LIBFR group. These results provide evidence that strength training performed acutely alters hemodynamic variables. However, training with blood flow restriction is more efficient in reducing blood pressure in hypertensive individuals than training with moderate intensity.

Gram-negative and -positive bacteria differentiation in blood culture samples by headspace volatile compound analysis.

Science.gov (United States)

Dolch, Michael E; Janitza, Silke; Boulesteix, Anne-Laure; Graßmann-Lichtenauer, Carola; Praun, Siegfried; Denzer, Wolfgang; Schelling, Gustav; Schubert, Sören

2016-12-01

Identification of microorganisms in positive blood cultures still relies on standard techniques such as Gram staining followed by culturing with definite microorganism identification. Alternatively, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry or the analysis of headspace volatile compound (VC) composition produced by cultures can help to differentiate between microorganisms under experimental conditions. This study assessed the efficacy of volatile compound based microorganism differentiation into Gram-negatives and -positives in unselected positive blood culture samples from patients. Headspace gas samples of positive blood culture samples were transferred to sterilized, sealed, and evacuated 20 ml glass vials and stored at -30 °C until batch analysis. Headspace gas VC content analysis was carried out via an auto sampler connected to an ion-molecule reaction mass spectrometer (IMR-MS). Measurements covered a mass range from 16 to 135 u including CO2, H2, N2, and O2. Prediction rules for microorganism identification based on VC composition were derived using a training data set and evaluated using a validation data set within a random split validation procedure. One-hundred-fifty-two aerobic samples growing 27 Gram-negatives, 106 Gram-positives, and 19 fungi and 130 anaerobic samples growing 37 Gram-negatives, 91 Gram-positives, and two fungi were analysed. In anaerobic samples, ten discriminators were identified by the random forest method allowing for bacteria differentiation into Gram-negative and -positive (error rate: 16.7 % in validation data set). For aerobic samples the error rate was not better than random. In anaerobic blood culture samples of patients IMR-MS based headspace VC composition analysis facilitates bacteria differentiation into Gram-negative and -positive.

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

DEFF Research Database (Denmark)

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

2001-01-01

in humans. The overall recovery of radioiron from blood is more than 90%, and the coefficient of variation, as judged by the variation in the ratio Fe-55/Fe-59 is in the order of 4%. Combined with whole-body counting of 59Fe and direct gamma -counting of Fe-59 on blood samples, this method represents......For studies on iron absorption in man radioisotopes represent an easy and simple tool. However, measurement of the orbital electron emitting radioiron, Fe-55, in blood is difficult and insufficiently described in the literature. The present study describes a relatively simple method...... for simultaneous determination of Fe-55 and Fe-59 in blood, using a dry-ashing procedure and recrystallization of the remaining iron. The detection Limit of the method permits measurements of 0.1 Bq/ml blood thus allowing detection of Less than 1% absorption from a 40 kBq dose, which is ethically acceptable...

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

Science.gov (United States)

Ifegwu, Clinton; Igwo-Ezikpe, Miriam N.; Anyakora, Chimezie; Osuntoki, Akinniyi; Oseni, Kafayat A.; Alao, Eragbae O.

2013-01-01

Polynuclear Aromatic Hydrocarbons (PAHs) are a major component of fuel generator fumes. Carcinogenicity of these compounds has long been established. In this study, 37 Swiss albino rats were exposed to generator fumes at varied distances for 8 hours per day for a period of 42 days and the level of 1-hydroxypyrene in their blood was evaluated. This study also tried to correlate the level of blood 1-hyroxypyrene with the distance from the source of pollution. Plasma was collected by centrifuging the whole blood sample followed by complete hydrolysis of the conjugated 1-hydroxypyrene glucuronide to yield the analyte of interest, 1-hydroxypyrene, which was achieved using beta glucuronidase. High performance liquid chromatography (HPLC) with UV detector was used to determine the 1-hydroxypyrene concentrations in the blood samples. The mobile phase was water:methanol (12:88 v/v) isocratic run at the flow rate of 1.2 mL/min with CI8 stationary phase at 250 nm. After 42 days of exposure, blood concentration level of 1-hydroxypyrene ranged from 34 μg/mL to 26.29 μg/mL depending on the distance from source of exposure. The control group had no 1-hydroxypyrene in their blood. After the period of exposure, percentage of death correlated with the distance from the source of exposure. Percentage of death ranged from 56% to zero depending on the proximity to source of pollution. PMID:24179393

On-chip acoustophoretic isolation of microflora including S. typhimurium from raw chicken, beef and blood samples.

Science.gov (United States)

Ngamsom, Bongkot; Lopez-Martinez, Maria J; Raymond, Jean-Claude; Broyer, Patrick; Patel, Pradip; Pamme, Nicole

2016-04-01

Pathogen analysis in food samples routinely involves lengthy growth-based pre-enrichment and selective enrichment of food matrices to increase the ratio of pathogen to background flora. Similarly, for blood culture analysis, pathogens must be isolated and enriched from a large excess of blood cells to allow further analysis. Conventional techniques of centrifugation and filtration are cumbersome, suffer from low sample throughput, are not readily amenable to automation and carry a risk of damaging biological samples. We report on-chip acoustophoresis as a pre-analytical technique for the resolution of total microbial flora from food and blood samples. The resulting 'clarified' sample is expected to increase the performance of downstream systems for the specific detection of the pathogens. A microfluidic chip with three inlets, a central separation channel and three outlets was utilized. Samples were introduced through the side inlets, and buffer solution through the central inlet. Upon ultrasound actuation, large debris particles (10-100 μm) from meat samples were continuously partitioned into the central buffer channel, leaving the 'clarified' outer sample streams containing both, the pathogenic cells and the background flora (ca. 1 μm) to be collected over a 30 min operation cycle before further analysis. The system was successfully tested with Salmonella typhimurium-spiked (ca. 10(3)CFU mL(-1)) samples of chicken and minced beef, demonstrating a high level of the pathogen recovery (60-90%). When applied to S. typhimurium contaminated blood samples (10(7)CFU mL(-1)), acoustophoresis resulted in a high depletion (99.8%) of the red blood cells (RBC) which partitioned in the buffer stream, whilst sufficient numbers of the viable S. typhimurium remained in the outer channels for further analysis. These results indicate that the technology may provide a generic approach for pre-analytical sample preparation prior to integrated and automated downstream detection of

Rapid Fractionation and Isolation of Whole Blood Components in Samples Obtained from a Community-based Setting.

Science.gov (United States)

Weckle, Amy; Aiello, Allison E; Uddin, Monica; Galea, Sandro; Coulborn, Rebecca M; Soliven, Richelo; Meier, Helen; Wildman, Derek E

2015-11-30

Collection and processing of whole blood samples in a non-clinical setting offers a unique opportunity to evaluate community-dwelling individuals both with and without preexisting conditions. Rapid processing of these samples is essential to avoid degradation of key cellular components. Included here are methods for simultaneous peripheral blood mononuclear cell (PBMC), DNA, RNA and serum isolation from a single blood draw performed in the homes of consenting participants across a metropolitan area, with processing initiated within 2 hr of collection. We have used these techniques to process over 1,600 blood specimens yielding consistent, high quality material, which has subsequently been used in successful DNA methylation, genotyping, gene expression and flow cytometry analyses. Some of the methods employed are standard; however, when combined in the described manner, they enable efficient processing of samples from participants of population- and/or community-based studies who would not normally be evaluated in a clinical setting. Therefore, this protocol has the potential to obtain samples (and subsequently data) that are more representative of the general population.

Regional cerebral blood flow measurements by a noninvasive microsphere method using 123I-IMP. Comparison with the modified fractional uptake method and the continuous arterial blood sampling method

International Nuclear Information System (INIS)

Nakano, Seigo; Matsuda, Hiroshi; Tanizaki, Hiroshi; Ogawa, Masafumi; Miyazaki, Yoshiharu; Yonekura, Yoshiharu

1998-01-01

A noninvasive microsphere method using N-isopropyl-p-( 123 I)iodoamphetamine ( 123 I-IMP), developed by Yonekura et al., was performed in 10 patients with neurological diseases to quantify regional cerebral blood flow (rCBF). Regional CBF values by this method were compared with rCBF values simultaneously estimated from both the modified fractional uptake (FU) method using cardiac output developed by Miyazaki et al. and the conventional method with continuous arterial blood sampling. In comparison, we designated the factor which converted raw SPECT voxel counts to rCBF values as a CBF factor. A highly significant correlation (r=0.962, p<0.001) was obtained in the CBF factors between the present method and the continuous arterial blood sampling method. The CBF factors by the present method were only 2.7% higher on the average than those by the continuous arterial blood sampling method. There were significant correlation (r=0.811 and r=O.798, p<0.001) in the CBF factor between modified FU method (threshold for estimating total brain SPECT counts; 10% and 30% respectively) and the continuous arterial blood sampling method. However, the CBF factors of the modified FU method showed 31.4% and 62.3% higher on the average (threshold; 10% and 30% respectively) than those by the continuous arterial blood sampling method. In conclusion, this newly developed method for rCBF measurements was considered to be useful for routine clinical studies without any blood sampling. (author)

Quantification of human polyomavirus JC virus load in urine and blood samples of healthy tribal populations of North-Eastern part of West Bengal, India.

Science.gov (United States)

Chattaraj, S; Bera, N K; Dutta, C; Bhattacharjee, S

2015-01-01

Human polyomavirus JC (JCV) is a widespread human virus with profound pathogenic potential. A study was undertaken to quantify JCV load in urine and peripheral blood samples of immunocompetent, apparently healthy tribal individuals of North-Eastern part of West Bengal, India for the first time. One hundred and thirteen samples of urine or blood were collected from different tribal groups of this region. For the quantitative estimation of the viral load in each sample, real-time polymerase chain reaction method using the SYBR Green dye was employed. The viral load estimated was found in the range between 3.5 × 102 and 2.12 × 106 copies/ml of samples having a mean and median viral copy numbers of 8.67 × 105 and 9.19 × 105 copies/ml of sample respectively. The mean viral DNA load in urine samples of the studied immunocompetent population was found to be higher than that found in a study conducted in the USA, but lower than similar groups of Italy and healthy adult women in the USA. However when compared with median values of viral DNA loads in urine samples of immunocompetent human subjects of Kuwait, Portugal, and Switzerland the observed viral DNA load was found to be substantially higher.

Nonsmoker Exposure to Secondhand Cannabis Smoke. III. Oral Fluid and Blood Drug Concentrations and Corresponding Subjective Effects.

Science.gov (United States)

Cone, Edward J; Bigelow, George E; Herrmann, Evan S; Mitchell, John M; LoDico, Charles; Flegel, Ronald; Vandrey, Ryan

2015-09-01

The increasing use of highly potent strains of cannabis prompted this new evaluation of human toxicology and subjective effects following passive exposure to cannabis smoke. The study was designed to produce extreme cannabis smoke exposure conditions tolerable to drug-free nonsmokers. Six experienced cannabis users smoked cannabis cigarettes [5.3% Δ(9)-tetrahydrocannabinol (THC) in Session 1 and 11.3% THC in Sessions 2 and 3] in a closed chamber. Six nonsmokers were seated alternately with smokers during exposure sessions of 1 h duration. Sessions 1 and 2 were conducted with no ventilation and ventilation was employed in Session 3. Oral fluid, whole blood and subjective effect measures were obtained before and at multiple time points after each session. Oral fluid was analyzed by ELISA (4 ng/mL cutoff concentration) and by LC-MS-MS (limit of quantitation) for THC (1 ng/mL) and total THCCOOH (0.02 ng/mL). Blood was analyzed by LC-MS-MS (0.5 ng/mL) for THC, 11-OH-THC and free THCCOOH. Positive tests for THC in oral fluid and blood were obtained for nonsmokers up to 3 h following exposure. Ratings of subjective effects correlated with the degree of exposure. Subjective effect measures and amounts of THC absorbed by nonsmokers (relative to smokers) indicated that extreme secondhand cannabis smoke exposure mimicked, though to a lesser extent, active cannabis smoking. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Blood sample collection and patient identification demand improvement: a questionnaire study of preanalytical practices in hospital wards and laboratories.

Science.gov (United States)

Wallin, Olof; Söderberg, Johan; Van Guelpen, Bethany; Stenlund, Hans; Grankvist, Kjell; Brulin, Christine

2010-09-01

Scand J Caring Sci; 2010; 24; 581-591 
 Blood sample collection and patient identification demand improvement: a questionnaire study of preanalytical practices in hospital wards and laboratories   Most errors in venous blood testing result from human mistakes occurring before the sample reach the laboratory.   To survey venous blood sampling (VBS) practices in hospital wards and to compare practices with hospital laboratories.   Staff in two hospitals (all wards) and two hospital laboratories (314 respondents, response rate 94%), completed a questionnaire addressing issues relevant to the collection of venous blood samples for clinical chemistry testing.   The findings suggest that instructions for patient identification and the collection of venous blood samples were not always followed. For example, 79% of the respondents reported the undesirable practice (UDP) of not always using wristbands for patient identification. Similarly, 87% of the respondents noted the UDP of removing venous stasis after the sampling is finished. Compared with the ward staff, a significantly higher proportion of the laboratory staff reported desirable practices regarding the collection of venous blood samples. Neither education nor the existence of established sampling routines was clearly associated with VBS practices among the ward staff.   The results of this study, the first of its kind, suggest that a clinically important risk of error is associated with VBS in the surveyed wards. Most important is the risk of misidentification of patients. Quality improvement of blood sample collection is clearly needed, particularly in hospital wards. © 2009 The Authors. Journal compilation © 2009 Nordic College of Caring Science.

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

Science.gov (United States)

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.

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

Science.gov (United States)

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

Molecular genotyping of HCV infection in seropositive blood donor

Science.gov (United States)

Zarin, Siti Noraziah Abu; Ibrahim, Nazlina

2013-11-01

This study is to investigate the prevalence of hepatitis C virus infection in seropositive blood donor. RNA was extracted from 32 positive samples in National Blood Centre and Melaka Hospital. The core and NS5B sequences were obtained from 23 samples. Genotype 3a is most prevalent in this study followed by genotype 1a. Evidence of mixed-genotypes (3a and 1b) infections was found in 5 subjects.

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

OpenAIRE

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

2016-01-01

Background ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. Methods In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to...

Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study.

Science.gov (United States)

De Kesel, Pieter M M; Lambert, Willy E; Stove, Christophe P

2015-06-30

Volumetric absorptive microsampling (VAMS) is a novel sampling technique that allows the straightforward collection of an accurate volume of blood (approximately 10μL) from a drop or pool of blood by dipping an absorbent polymeric tip into it. The resulting blood microsample is dried and analyzed as a whole. The aim of this study was to evaluate the potential of VAMS to overcome the hematocrit bias, an important issue in the analysis of dried blood microsamples. An LC-MS/MS method for analysis of the model compounds caffeine and paraxanthine in VAMS samples was fully validated and fulfilled all pre-established criteria. In conjunction with previously validated procedures for dried blood spots (DBS) and blood, this allowed us to set up a meticulous comparative study in which both compounds were determined in over 80 corresponding VAMS, DBS and liquid whole blood samples. These originated from authentic human patient samples, covering a wide hematocrit range (0.21-0.50). By calculating the differences with reference whole blood concentrations, we found that analyte concentrations in VAMS samples were not affected by a bias that changed over the evaluated hematocrit range, in contrast to DBS results. However, VAMS concentrations tend to overestimate whole blood concentrations, as a consistent positive bias was observed. A different behavior of VAMS samples prepared from incurred and spiked blood, combined with a somewhat reduced recovery of caffeine and paraxanthine from VAMS tips at high hematocrit values, an effect that was not observed for DBS using a very similar extraction procedure, was found to be at the basis of the observed VAMS-whole blood deviations. Based on this study, being the first in which the validity and robustness of VAMS is evaluated by analyzing incurred human samples, it can be concluded that VAMS effectively assists in eliminating the effect of hematocrit. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

The subtyping of primary aldosteronism by adrenal vein sampling: sequential blood sampling causes factitious lateralization.

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Rossitto, Giacomo; Battistel, Michele; Barbiero, Giulio; Bisogni, Valeria; Maiolino, Giuseppe; Diego, Miotto; Seccia, Teresa M; Rossi, Gian Paolo

2018-02-01

The pulsatile secretion of adrenocortical hormones and a stress reaction occurring when starting adrenal vein sampling (AVS) can affect the selectivity and also the assessment of lateralization when sequential blood sampling is used. We therefore tested the hypothesis that a simulated sequential blood sampling could decrease the diagnostic accuracy of lateralization index for identification of aldosterone-producing adenoma (APA), as compared with bilaterally simultaneous AVS. In 138 consecutive patients who underwent subtyping of primary aldosteronism, we compared the results obtained simultaneously bilaterally when starting AVS (t-15) and 15 min after (t0), with those gained with a simulated sequential right-to-left AVS technique (R ⇒ L) created by combining hormonal values obtained at t-15 and at t0. The concordance between simultaneously obtained values at t-15 and t0, and between simultaneously obtained values and values gained with a sequential R ⇒ L technique, was also assessed. We found a marked interindividual variability of lateralization index values in the patients with bilaterally selective AVS at both time point. However, overall the lateralization index simultaneously determined at t0 provided a more accurate identification of APA than the simulated sequential lateralization indexR ⇒ L (P = 0.001). Moreover, regardless of which side was sampled first, the sequential AVS technique induced a sequence-dependent overestimation of lateralization index. While in APA patients the concordance between simultaneous AVS at t0 and t-15 and between simultaneous t0 and sequential technique was moderate-to-good (K = 0.55 and 0.66, respectively), in non-APA patients, it was poor (K = 0.12 and 0.13, respectively). Sequential AVS generates factitious between-sides gradients, which lower its diagnostic accuracy, likely because of the stress reaction arising upon starting AVS.

Staphylococcus aureus detection in blood samples by silica nanoparticle-oligonucleotides conjugates.

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Borsa, Baris A; Tuna, Bilge G; Hernandez, Frank J; Hernandez, Luiza I; Bayramoglu, Gulay; Arica, M Yakup; Ozalp, V Cengiz

2016-12-15

A fast, specific and sensitive homogeneous assay for Staphylococcus aureus detection was developed by measuring the activity of secreted nuclease from the bacteria via a modified DNA oligonucleotide. As biosensor format, an effective system, Nanokeepers as previously reported, were used for triggered release of confined fluorophores, and hence specific detection of S. aureus on nuclease activity was obtained. The interference from blood components for fluorescent quantification was eliminated by a pre-purification by aptamer-functionalized silica magnetic nanoparticles. The reported assay system was exclusively formed by nucleic acid oligos and magnetic or mesoporous silica nanoparticles, that can be used on blood samples in a stepwise manner. The assay was successfully used as a sensing platform for the specific detection of S. aureus cells as low as 682 CFU in whole blood. Copyright © 2016 Elsevier B.V. All rights reserved.

Influence of Partial Pressure of Oxygen in Blood Samples on Measurement Performance in Glucose-Oxidase-Based Systems for Self-Monitoring of Blood Glucose

Science.gov (United States)

Baumstark, Annette; Schmid, Christina; Pleus, Stefan; Haug, Cornelia; Freckmann, Guido

2013-01-01

Background Partial pressure of oxygen (pO2) in blood samples can affect blood glucose (BG) measurements, particularly in systems that employ the glucose oxidase (GOx) enzyme reaction on test strips. In this study, we assessed the impact of different pO2 values on the performance of five GOx systems and one glucose dehydrogenase (GDH) system. Two of the GOx systems are labeled by the manufacturers to be sensitive to increased blood oxygen content, while the other three GOx systems are not. Methods Aliquots of 20 venous samples were adjusted to the following pO2 values: pO2 ~70 mmHg, which is considered to be similar to pO2 in capillary blood samples, and the mean BG result at pO2 pO2 pO2 ≥150 mmHg. For both pO2 levels, relative differences of all tested GOx systems were significant (p pO2 values pO2 variations lead to clinically relevant BG measurement deviations in GOx systems, even in GOx systems that are not labeled as being oxygen sensitive. PMID:24351177

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

Directory of Open Access Journals (Sweden)

Ironside James W

2007-08-01

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

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

Science.gov (United States)

Fagge, Timothy J; Barclay, G Robin; Stove, G Colin; Stove, Gordon; Robinson, Michael J; Head, Mark W; Ironside, James W; Turner, Marc L

2007-01-01

Background Sub-clinical variant Creutzfeldt-Jakob disease (vCJD) infection and reports of vCJD transmission through blood transfusion emphasise the need for blood screening assays to ensure the safety of blood and transplanted tissues. Most assays aim to detect abnormal prion protein (PrPSc), although achieving required sensitivity is a challenge. Methods We have used innovative Atomic Dielectric Resonance Spectroscopy (ADRS), which determines dielectric properties of materials which are established by reflectivity and penetration of radio/micro waves, to analyse blood samples from patients and controls to identify characteristic ADR signatures unique to blood from vCJD and to sCJD patients. Initial sets of blood samples from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) were screened as training samples to determine group-specific ADR characteristics, and provided a basis for classification of blinded sets of samples. Results Blood sample groups from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) screened by ADRS were classified with 100% specificity and sensitivity, discriminating these by a co-variance expert analysis system. Conclusion ADRS appears capable of recognising and discriminating serum samples from vCJD, sCJD, non-CJD neurological diseases, and normal healthy adults, and might be developed to provide a system for primary screening or confirmatory assay complementary to other screening systems. PMID:17760958

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

Science.gov (United States)

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

2017-01-01

This study focuses on the detection and identification of DNA and antibodies to Ehrlichia spp. in samples of blood bank donors in Costa Rica using molecular and serological techniques. Presence of Ehrlichia canis was determined in 10 (3.6%) out of 280 blood samples using polymerase chain reaction (PCR) targeting the ehrlichial dsb conserved gene. Analysis of the ehrlichial trp36 polymorphic gene in these 10 samples revealed substantial polymorphism among the E. canis genotypes, including divergent tandem repeat sequences. Nucleotide sequences of dsb and trp36 amplicons revealed a novel genotype of E. canis in blood bank donors from Costa Rica. Indirect immunofluorescence assay (IFA) detected antibodies in 35 (35%) of 100 serum samples evaluated. Thirty samples showed low endpoint titers (64-256) to E. canis, whereas five sera yielded high endpoint titers (1024-8192); these five samples were also E. canis-PCR positive. These findings represent the first report of the presence of E. canis in humans in Central America. Copyright © 2016 Elsevier GmbH. All rights reserved.

Quantifying regional cerebral blood flow with N-isopropyl-p-[123I]iodoamphetamine and SPECT by one-point sampling method

International Nuclear Information System (INIS)

Odano, Ikuo; Takahashi, Naoya; Noguchi, Eikichi; Ohtaki, Hiro; Hatano, Masayoshi; Yamazaki, Yoshihiro; Higuchi, Takeshi; Ohkubo, Masaki.

1994-01-01

We developed a new non-invasive technique; one-point sampling method, for quantitative measurement of regional cerebral blood flow (rCBF) with N-isopropyl-p-[ 123 I]iodoamphetamine and SPECT. Although the continuous withdrawal of arterial blood and octanol treatment of the blood are required in the conventional microsphere method, the new technique dose not require these two procedures. The total activity of 123 I-IMP obtained by the continuous withdrawal of arterial blood is inferred by the activity of 133 I-IMP obtained by the one point arterial sample using a regression line. To determine when one point sampling time was optimum for inferring integral input function of the continuous withdrawal and whether the treatment of sampled blood for octanol fraction was required, we examined a correlation between the total activity of arterial blood withdrawn from 0 to 5 min after the injection and the activity of one point sample obtained at time t, and calculated a regression line. As a result, the minimum % error for the inference using the regression line was obtained at 6 min after the 123 I-IMP injection, moreover, the octanol treatment was not required. Then examining an effect on the values of rCBF when the sampling time was deviated from 6 min, we could correct the values in approximately 3% error when the sample was obtained at 6±1 min after the injection. The one-point sampling method provides accurate and relatively non-invasive measurement of rCBF without octanol extraction of arterial blood. (author)

Blood meal analysis of tabanid fly after it biting the rare Sumatran rhinoceros.

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Rovie-Ryan, Jeffrine Japning; Zainuddin, Zainal Zahari; Marni, Wahap; Ahmad, Abdul Hamid; Ambu, Laurentius N; Payne, Junaidi

2013-02-01

To demonstrate a noninvasive large mammalian genetic sampling method using blood meal obtained from a tabanid fly. Blood meal was recovered from the abdomen of an engorged tabanid fly (Haematopota sp.) which was captured immediately after biting a Sumatran rhino in captivity. The blood was applied on to a Whatman FTA(®) blood card. Subsequent laboratory work was conducted to extract, amplify and sequence the DNA from the sample. Validation was done by sampling the hair follicles and blood samples from the rhinoceros and subjecting it to the same laboratory process. BLAST search and constructed phylogenetic trees confirmed the blood meal samples were indeed from the rhino. This method could be used in the field application to noninvasively collect genetic samples. Collection of tabanids and other haematophagous arthropods (e.g. mosquitoes and ticks) and other blood-sucking parasites (e.g. leeches and worms) could also provide information on vector-borne diseases.

Effect on blood lipids of two daily servings of Camembert cheese. An intervention trial in mildly hypercholesterolemic subjects.

Science.gov (United States)

Schlienger, Jean-Louis; Paillard, Francois; Lecerf, Jean-Michel; Romon, Monique; Bonhomme, Cécile; Schmitt, Bernard; Donazzolo, Yves; Defoort, Catherine; Mallmann, Cécilia; Le Ruyet, Pascale; Bresson, Jean-Louis

2014-12-01

As a concentrated source of saturated fat, cheese consumption is considered to be associated with increased cholesterolemia and generally forbidden in dietary guidelines for adults with hypercholesterolemia. The aim of this study was to evaluate the impact of saturated fatty acids on lipid parameters and blood pressure with regards to different types of dairy products: Camembert and full-fat yoghurt. One-hundred and fifty-nine moderate hypercholesterolemic subjects without treatment were instructed to consume two full-fat yoghurts (2 × 125 g) per day for 3 weeks (run-in period) and then for a further period of 5 weeks, either two full-fat yoghurts or two 30 g servings of Camembert cheese per day. We observed that over the 5-week daily consumption of two servings of Camembert cheese, blood pressure and serum lipids did not change in moderate hypercholesterolemic subjects. These results suggest that fermented cheese such as Camembert could be consumed daily without affecting serum lipids or blood pressure.

Progressive sensorineural hearing loss, subjective tinnitus and vertigo caused by elevated blood lipids.

Science.gov (United States)

Pulec, J L; Pulec, M B; Mendoza, I

1997-10-01

The otologist frequently sees patients with progressive sensorineural hearing loss, subjective aural tinnitus and vertigo with no apparent cause. Elevated blood lipids may be a cause of inner ear malfunction on a biochemical basis. To establish the true incidence of this condition, all new patients (4,251) seen during an eight-year period were evaluated; of these, 2,332 patients had complaints of inner ear disease. All had a complete neurotologic examination, appropriate audiometric and vestibular studies and imaging, and blood tests including lipid phenotype studies. Hyperlipoproteinemia was found in 120 patients (5.1%). Most patients were found to be overweight and had additional coexisting conditions such as diabetes mellitus. Treatment with vasodilators and a 500-calorie, high-protein, low-carbohydrate diet yielded improvement of symptoms in 83% of patients within five months of initiation of treatment.

Gingival crevicular blood for screening of blood glucose level in patients with & without diabetes: a chair-side test.

Science.gov (United States)

Bhavsar, M V; Brahmbhatt, N A; Sahayata, V; Bhavsar, N V

2016-05-01

Diabetes is a pandemic disease with increasing prevalence and serious complications. Periodontitis being one of its presentation and is its sixth recognized complication. This study compares blood glucose levels in gingival crevicular blood of patients with and without diabetes elicited during routine periodontal probing and venous blood sample. Seventy patients with moderate gingivitis and periodontitis positive for bleeding on probing were chosen. All the subjects were divided in two groups, group I consisted of 35 diabetic and group II of 35 non-diabetic subjects. Blood from the gingiva of the most inflamed site was collected with the test strip of a glucose self-monitoring device, and the blood glucose levels were measured. At the same time, intravenous blood was collected for measurement in a laboratory glucose analyzer. Gingival index and probing pocket depth were evaluated for each subject at same time. The mean GCB levels and VB derived from all samples were 156.07 ± 49.23 mg dl(-1) and 156 ± 49.89 mg dl(-1) , respectively, for diabetic group and 90.80 ± 11.07 and 93.41 ± 9.30 for non-diabetic group. In both the groups, the difference between GCB and VB glucose levels was non-significant (P > 0.005). Highly significant correlation between GCB and VB (r = 0.972 for diabetic and r = 0.721 for non-diabetic) in both the groups was found. The data from this study show that GCB collected during diagnostic periodontal examination can be an excellent source for estimation of blood sugar or glucometric analysis. This technique is also suitable for routine screening of diabetic and early diagnosis of unknown diabetic cases. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Immunosuppressant therapeutic drug monitoring by LC-MS/MS: workflow optimization through automated processing of whole blood samples.

Science.gov (United States)

Marinova, Mariela; Artusi, Carlo; Brugnolo, Laura; Antonelli, Giorgia; Zaninotto, Martina; Plebani, Mario

2013-11-01

Although, due to its high specificity and sensitivity, LC-MS/MS is an efficient technique for the routine determination of immunosuppressants in whole blood, it involves time-consuming manual sample preparation. The aim of the present study was therefore to develop an automated sample-preparation protocol for the quantification of sirolimus, everolimus and tacrolimus by LC-MS/MS using a liquid handling platform. Six-level commercially available blood calibrators were used for assay development, while four quality control materials and three blood samples from patients under immunosuppressant treatment were employed for the evaluation of imprecision. Barcode reading, sample re-suspension, transfer of whole blood samples into 96-well plates, addition of internal standard solution, mixing, and protein precipitation were performed with a liquid handling platform. After plate filtration, the deproteinised supernatants were submitted for SPE on-line. The only manual steps in the entire process were de-capping of the tubes, and transfer of the well plates to the HPLC autosampler. Calibration curves were linear throughout the selected ranges. The imprecision and accuracy data for all analytes were highly satisfactory. The agreement between the results obtained with manual and those obtained with automated sample preparation was optimal (n=390, r=0.96). In daily routine (100 patient samples) the typical overall total turnaround time was less than 6h. Our findings indicate that the proposed analytical system is suitable for routine analysis, since it is straightforward and precise. Furthermore, it incurs less manual workload and less risk of error in the quantification of whole blood immunosuppressant concentrations than conventional methods. © 2013.

Reproducibility of ambulatory blood pressure values and circadian blood pressure patterns in untreated subjects in a 1-11 month interval.

Science.gov (United States)

Monte, Miguel; Cambão, Mariana; Mesquita Bastos, José; Polónia, Jorge

2015-11-01

To evaluate in untreated subjects the reproducibility of mean values and four circadian patterns between two ambulatory blood pressure monitoring (ABPM) recordings separated by 1-11 months. We performed a retrospective analysis of 481 individuals (59% women) evaluated by ABPM on two occasions, visit 1 (V1) and 2 (V2), separated by 5.5+0.2 months. Four circadian patterns were defined by night/day systolic blood pressure (SBP) ratios: reverse dippers (RD), ratio >1.0; non-dippers (ND), ratio 0.9-1.0; dippers (D), ratio 0.8-patterns were calculated by the percentage of the same profile from V1 to V2. Mean 24-h blood pressure (BP) at V1 and V2 was 126.8/75.9±0.5/0.5 vs. 126.5/75.7±0.5/0.4 mmHg (NS). Nighttime SBP fall was 9.8±0.4 (V1) and 9.6±0.3% (V2) (NS). The correlation coefficient of ABPM data at V1 vs. at V2 was 0.41-0.69 (ppatterns. Copyright © 2015 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

XRF and TXRF techniques for multi-element determination of trace elements in whole blood and human hair samples

International Nuclear Information System (INIS)

Khuder, A.; Karjou, J.; Sawan, M.Kh.; Bakir, M.A.

2007-01-01

XRF and TXRF were established as useful techniques for multi-element analysis of whole blood and human head hair samples. Direct-XRF with different collimation units and different X-ray excitation modes was successfully used for the determination of S, P, K, Ca, Fe, and Br elements in blood samples and K, Ca, Mn, Fe elements in human hair samples. Direct analysis by TXRF was used for the determination of Rb and Sr in digested blood and human hair samples, respectively, while, the co-precipitation method using APDC for TXRF analysis was used for the determination of Ni, Cu, Zn, and Pb elements in both matrices. As a result, the improved XRF and TXRF methods were applied for multi-element determination of elements in whole blood and human hair samples in non-occupational exposed population living in Damascus city. The mean concentrations of analyzed elements in both matrices were on the reported range values for non-occupational population in other countries. (author)

XRF and TXRF techniques for multi-element determination of trace elements in whole blood and human hair samples

International Nuclear Information System (INIS)

Khuder, A.; Karjou, J.; Sawan, M.Kh.; Bakir, M.A.

2008-01-01

XRF and TXRF were established as useful techniques for multi-element analysis of whole blood and human head hair samples. Direct-XRF with different collimation units and different X-ray excitation modes was successfully used for the determination of S, P, K, Ca, Fe, and Br elements in blood samples and K, Ca, Mn, Fe elements in human hair samples. Direct analysis by TXRF was used for the determination of Rb and Sr in digested blood and human hair samples, respectively, while, the co-precipitation method using APDC for TXRF analysis was used for the determination of Ni, Cu, Zn, and Pb elements in both matrices. As a result, the improved XRF and TXRF methods were applied for multi-element determination of elements in whole blood and human hair samples in non-occupational exposed population living in Damascus city. The mean concentrations of analyzed elements in both matrices were on the reported range values for non-occupational population in other countries. (author)

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

Science.gov (United States)

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.

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

Science.gov (United States)

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

2016-05-01

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

Association between subjective actual sleep duration, subjective sleep need, age, body mass index, and gender in a large sample of young adults.

Science.gov (United States)

Kalak, Nadeem; Brand, Serge; Beck, Johannes; Holsboer-Trachsler, Edith; Wollmer, M Axel

2015-01-01

Poor sleep is a major health concern, and there is evidence that young adults are at increased risk of suffering from poor sleep. There is also evidence that sleep duration can vary as a function of gender and body mass index (BMI). We sought to replicate these findings in a large sample of young adults, and also tested the hypothesis that a smaller gap between subjective sleep duration and subjective sleep need is associated with a greater feeling of being restored. A total of 2,929 university students (mean age 23.24±3.13 years, 69.1% female) took part in an Internet-based survey. They answered questions related to demographics and subjective sleep patterns. We found no gender differences in subjective sleep duration, subjective sleep need, BMI, age, or feeling of being restored. Nonlinear associations were observed between subjective sleep duration, BMI, and feeling of being restored. Moreover, a larger discrepancy between subjective actual sleep duration and subjective sleep need was associated with a lower feeling of being restored. The present pattern of results from a large sample of young adults suggests that males and females do not differ with respect to subjective sleep duration, BMI, or feeling of being restored. Moreover, nonlinear correlations seemed to provide a more accurate reflection of the relationship between subjective sleep and demographic variables.

Assessment of lead in blood samples of children residing in the vicinity of industries

International Nuclear Information System (INIS)

Shah, F.; Kazi, T.G.; Afridi, H.I.; Brahaman, K.D.; Arain, S.S.; Panhwar, A.H.

2013-01-01

The aim of present study was to determine the lead (Pb) distributions in blood and prevalence of elevated Pb exposure among children, age ranged (5-10 years), residing near industrialized region of Hyderabad city, Pakistan. For comparison, biological samples of children of same age group from non-industrial area were also analyzed. The Pb concentration in blood samples was determined by electrothermal atomic absorption spectrometry, prior to microwave assisted acid digestion. The results showed that significantly higher proportion of children living in the vicinity of industrial area, had blood Pb levels (BLL) in the range of 15.4-35.6 micro g/dL, and 8.51-16.7 micro g/dL for those of non-industrial area. The blood Pb level was higher in boys of both groups as compared to girls of same age group, but the difference was not significant (p=0.178). Negative correlation was observed between BLL and hemoglobin levels (p<0.001), while positive correlation was observed between BLL and age. (author)

Hemoglobin in samples with leukocytosis can be measured on ABL 700 series blood gas analyzers

NARCIS (Netherlands)

Scharnhorst, V.; Laar, van der P.D.; Vader, H.

2003-01-01

To compare lactate, bilirubin and Hemoglobin F concentrations obtained on ABL 700 series blood gas analyzers with those from laboratory methods. Pooled neonatal plasma, cord blood and adult plasma samples were used for comparison of bilirubin, hemoglobin F and lactate concentrations respectively.

Evaluation of a nested-PCR for mycobacterium tuberculosis detection in blood and urine samples.

Science.gov (United States)

da Cruz, Heidi Lacerda Alves; de Albuquerque Montenegro, Rosana; de Araújo Lima, Juliana Falcão; da Rocha Poroca, Diogo; da Costa Lima, Juliana Figueirêdo; Maria Lapa Montenegro, Lílian; Crovella, Sergio; Charifker Schindler, Haiana

2011-01-01

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

Blood pathway analyses reveal differences between prediabetic subjects with or without dyslipidaemia. The Cardiovascular Risk in Young Finns Study.

Science.gov (United States)

Laaksonen, Jaakko; Taipale, Tuukka; Seppälä, Ilkka; Raitoharju, Emma; Mononen, Nina; Lyytikäinen, Leo-Pekka; Waldenberger, Melanie; Illig, Thomas; Hutri-Kähönen, Nina; Rönnemaa, Tapani; Juonala, Markus; Viikari, Jorma; Kähönen, Mika; Raitakari, Olli; Lehtimäki, Terho

2017-10-01

Prediabetes often occurs together with dyslipidaemia, which is paradoxically treated with statins predisposing to type 2 diabetes mellitus. We examined peripheral blood pathway profiles in prediabetic subjects with (PR D ) and without dyslipidaemia (PR 0 ) and compared these to nonprediabetic controls without dyslipidaemia (C 0 ). The participants were from the Cardiovascular Risk in Young Finns Study, including 1240 subjects aged 34 to 49 years. Genome-wide expression data of peripheral blood and gene set enrichment analysis were used to investigate the differentially expressed genes and enriched pathways between different subtypes of prediabetes. Pathways for cholesterol synthesis, interleukin-12-mediated signalling events, and downstream signalling in naïve CD8+ T-cells were upregulated in the PR 0 group in comparison with controls (C 0 ). The upregulation of these pathways was independent of waist circumference, blood pressure, smoking status, and insulin. Adjustment for CRP left the CD8+ T-cell signalling and interleukin-12-mediated signalling event pathway upregulated. The cholesterol synthesis pathway was also upregulated when all prediabetic subjects (PR 0 and PR D ) were compared with the nonprediabetic control group. No pathways were upregulated or downregulated when the PR D group was compared with the C 0 group. Five genes in the PR 0 group and 1 in the PR D group were significantly differentially expressed in comparison with the C 0 group. Blood cell gene expression profiles differ significantly between prediabetic subjects with and without dyslipidaemia. Whether this classification may be used in detection of prediabetic individuals at a high risk of cardiovascular complications remains to be examined. Copyright © 2017 John Wiley & Sons, Ltd.

General guidelines for safe and expeditious international transport of samples subjected to biological dosimetry assessment.

Science.gov (United States)

Di Giorgio, Marina; Radl, Analía; Taja, María R; Bubniak, Ruth; Deminge, Mayra; Sapienza, Carla; Vázquez, Marina; Baciu, Florian; Kenny, Pat

2014-06-01

It has been observed that victims of accidental overexposures show better chance of survival if they receive medical treatment early. The increased risk of scenarios involving mass casualties has stimulated the scientific community to develop tools that would help the medical doctors to treat victims. The biological dosimetry has become a routine test to estimate the dose, supplementing physical and clinical dosimetry. In case of radiation emergencies, in order to provide timely and effectively biological dosimetry assistance it is essential to guarantee an adequate transport of blood samples in principal, for providing support to countries that do not have biodosimetry laboratories. The objective of the present paper is to provide general guidelines, summarised in 10 points, for timely and proper receiving and sending of blood samples under National and International regulations, for safe and expeditious international transport. These guidelines cover the classification, packaging, marking, labelling, refrigeration and documentation requirements for the international shipping of blood samples and pellets, to provide assistance missions with a tool that would contribute with the preparedness for an effective biodosimetric response in cases of radiological or nuclear emergencies. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Standardization and application of indirect ELISA for diagnosis of Mycoplasma bovis in bovine blood serum samples

Directory of Open Access Journals (Sweden)

Samira Moraes Cunha de Mesquita

2015-06-01

Full Text Available ABSTRACT. Mesquita S.M.C., Mansur F.J., Nascimento E.R., Barreto M.L. & Kimura L.M.S. [Standardization and application of indirect ELISA for diagnosis of Mycoplasma bovis in bovine blood serum samples.] Padroniza- ção e aplicação de ELISA indireto para diagnóstico de Mycoplasma bovis em amostras de soro sanguíneo bovino. Revista Brasileira de Medicina Veterinária, 37(2:101-107, 2015. Universidade Federal Fluminense, Faculdade de Veteriná- ria, Rua Vital Brazil Filho, 64, Vital Brazil, Niterói, RJ 24230-340, Brasil. E-mail: samira.veterinaria@gmail.com International researchers presented results indicating frequent involvement of Mycoplasma spp. as a causative agent of mastitis in cattle, associating its presence with significant economic losses to farmers. Mycoplasma bovis is the species most reported and relevant, because it causes more severe disease. The level of antibodies against M. bovis remains high for several months and can be detected by ELISA. The aim of this work was to develop an indirect ELISA with whole cell antigen of M. bovis (strain Donetta PG 45 with subsequent application in bovine blood serum samples for detection of antibodies against M. bovis. The immunization of cows A and B by inoculating an immunogen against M. bovis to obtain hyperimmune blood serum was the first stage of this work, then the stage of standardization of ELISA was proceeded. The concentration of 2 mg of antigen/mL for coating the microtiter plates was decided by statistical analyses. The optical density value 0,2 was determined as the limit of reactivity discrimination of samples (the cut-off point. The hyperimmune blood serum sample of the cow A (collected 30 days after immunization was chosen as the positive control and, the fetal calf serum was chosen as negative control of the assay. In addition, the ideal optimal dilutions found for blood serum samples was 1:400 and for conjugate was 1:10.000 and the substrate used was the ortho

Comparison of Hematologic and Biochemical Test Results in Blood Samples Obtained by Jugular Venipuncture Versus Nail Clip in Moluccan Cockatoos (Cacatua moluccensis).

Science.gov (United States)

Bennett, Tracy D; Lejnieks, Daniel V; Koepke, Hoyt; Grimson, Fiona; Szucs, Jennifer; Omaits, Kerri; Lane, Rosalie

2015-12-01

In birds, blood samples are often collected from the jugular, medial metatarsal, and basilic vein. Samples are sometimes collected by toe nail clip, but concerns to avoid drawing blood from the nail include pain after nail clips for blood collection, potential differences in complete blood count (CBC) results, and potential contamination with uric acid values. To compare differences in biochemical and hematologic values in blood samples obtained by jugular venipuncture versus toenail clip, blood samples were collected from Moluccan cockatoos (Cacatua moluccensis) (N = 23) and sent to a commercial laboratory for routine CBCs and serum biochemical analysis. Results showed good agreement between venipuncture and nail clip blood samples in red blood cell count, packed cell volume, heterophil count and percentage, lymphocyte count and percentage, aspartate aminotransferase, chloride, creatine phosphokinase, glucose, lactate dehydrogenase, total protein, and uric acid values. Constant bias was found in values of bile acids, cholesterol, and hemoglobin. Proportional bias toward higher values in the jugular sample were found in total white blood cell (WBC) count and inorganic phosphorus. Serum calcium plots revealed a proportional bias toward higher values in the toe nail blood when values were increased. Results suggest some differences in WBC count, bile acids, calcium, cholesterol, hemoglobin, and phosphorus values between blood samples collected by jugular venipuncture and samples collected by toe nail clip, but the differences are mostly minor and, with the possible exception of inorganic phosphorus and marginally elevated or very low WBC counts, are unlikely to affect the use or interpretation of the avian blood panel.

Wuchereria bancrofti in Tanzania: microfilarial periodicity and effect of blood sampling time on microfilarial intensities

DEFF Research Database (Denmark)

Simonsen, Poul Erik; Niemann, L.; Meyrowitsch, Dan Wolf

1997-01-01

The circadian periodicity of Wuchereria bancrofti microfilarial (mf) intensities in peripheral blood was analysed in a group of infected individuals from an endemic community in north-eastern Tanzania. The mf density was quantified at two-hourly intervals for 24 hours. A clear nocturnal periodic...... of blood sampling before peak time is discussed, and the importance of taking sampling time into consideration when analysing data from epidemiological studies is emphasized. A simple method is devised which can be used to adjust for the influence of time on mf intensities, in studies where accurate...... information on mf intensities is necessary, and where it is impossible to obtain all samples at peak time....

Focal physiological uncoupling of cerebral blood flow and oxidative metabolism during somatosensory stimulation in human subjects

International Nuclear Information System (INIS)

Fox, P.T.; Raichle, M.E.

1986-01-01

Coupling between cerebral blood flow (CBF) and cerebral metabolic rate of oxygen (CMRO 2 ) was studied using multiple sequential administrations of 15 O-labeled radiotracers and positron emission tomography. In the resting state an excellent correlation between CBF and CMRO 2 was found when paired measurements of CBF and CMRO 2 from multiple (30-48) brain regions were tested in each of 33 normal subjects. Regional uncoupling of CBF and CMRO 2 was found, however, during neuronal activation induced by somatosensory stimulation. Stimulus-induced focal augmentation of cerebral blood flow (29% mean) far exceeded the concomitant local increase in tissue metabolic rate (mean, 5%), when resting-state and stimulated-state measurements were obtained in each of 9 subjects. Stimulus duration had no significant effect on response magnitude or on the degree of CBF-CMRO 2 uncoupling observed. Dynamic, physiological regulation of CBF by a mechanism (neuronal or biochemical) dependent on neuronal firing per se, but independent of the cerebral metabolic rate of oxygen, is hypothesized

Basal blood parameters of horses subjected to aerobic activity fed with lipidic concentrated

Directory of Open Access Journals (Sweden)

Kátia de Oliveira

2012-02-01

Full Text Available The feeding diets were evaluated containing low and high levels of soybean oil for horses athletes subjected to two protocols of aerobic training on the response of basal blood biochemical parameters. Four horses were used in latin square design with treatments in a 2 x 2 factorial arrangement. Treatments consisted levels of 5 and 15% oil concentrates and two aerobic training, 40' and 60' minutes. Plasmatic parameters were monitored, triglyceride (TG, total cholesterol (TC, glucose (GLU and lactate (LAC, during basal metabolism. The TG, TC, GLU and LAC from horses at rest were not affected (P> 0.05 neither of diet and physical activity, 0.21, 3.79, 4.18, 0.93 mmol L-1, respectively. It can be concluded that offer concentrate with high content of soybean oil to athletic horses in aerobic activities can be performed without altering the blood biochemical profile of basal metabolism.

Cerebral blood flow and metabolism during isoflurane-induced hypotension in patients subjected to surgery for cerebral aneurysms

DEFF Research Database (Denmark)

Madsen, J B; Cold, G E; Hansen, E S

1987-01-01

Cerebral blood flow and cerebral metabolic rate for oxygen were measured during isoflurane-induced hypotension in 10 patients subjected to craniotomy for clipping of a cerebral aneurysm. Flow and metabolism were measured 5-13 days after the subarachnoid haemorrhage by a modification of the classi......Cerebral blood flow and cerebral metabolic rate for oxygen were measured during isoflurane-induced hypotension in 10 patients subjected to craniotomy for clipping of a cerebral aneurysm. Flow and metabolism were measured 5-13 days after the subarachnoid haemorrhage by a modification......). Controlled hypotension to an average MAP of 50-55 mm Hg was induced by increasing the dose of isoflurane, and maintained at an inspired concentration of 2.2 +/- 0.2%. This resulted in a significant decrease in CMRO2 (to 1.73 +/- 0.16 ml/100 g min-1), while CBF was unchanged. After the clipping...

Maternal red blood cell alloantibodies identified in blood samples obtained from Iranian pregnant women: the first population study in Iran.

Science.gov (United States)

Shahverdi, Ehsan; Moghaddam, Mostafa; Gorzin, Fateme

2017-01-01

The objective was to determine the frequency of occurrence of alloantibodies among pregnant women in Iran. This was a prospective cross-sectional study, which was carried out in the immunohematology reference laboratory of the Iranian Blood Transfusion Organization in Tehran, Iran, in 2008 to 2015. Screening and identification of red blood cell (RBC) alloantibodies was done on the sera of 7340 pregnant females using the standard tube method and gel column agglutination technique. Alloantibodies were identified in the serum of 332 of the 7340 (4.5%) pregnant women. A total of 410 antibodies were detected in 332 positive maternal serum samples with no previous history of blood transfusion. Anti-D was the most common antibody accounting for 70.5% of all the antibodies formed in D- women. The incidence of specific alloimmunization other than Rh group was 14.4%. We concluded that the alloimmunization rate was high in comparison with wide pattern in previous studies. In Iran, like other developing countries, alloimmunization screening tests are performed only to detect anti-D in pregnant D- women. This high rate of alloimmunization, quite possibly, is due to the fact that the majority of blood samples came from pregnant women known to have previous obstetric problems. However, we suggest that RBC antibody screening tests should be extended to all D+ women. © 2016 AABB.

Pneumatic tube-transported blood samples in lithium heparinate gel separator tubes may be more susceptible to haemolysis than blood samples in serum tubes.

Science.gov (United States)

Böckel-Frohnhöfer, Nicole; Hübner, Ulrich; Hummel, Björn; Geisel, Jürgen

2014-10-01

Pneumatic tube systems are widely used in hospitals. Advantages are high speed and rapid availability of the samples. However, the transportation by pneumatic tube promotes haemolysis. Haemolysis interferes with many spectrophotometric assays and is a common problem in clinical laboratories. The haemolysis index (HI) as a semi-quantitative representation of the level of haemolysis was compared in unpaired tube-transported and hand-delivered routine lithium heparinate plasma samples (n = 1368 and n = 837, respectively). Additionally, the HI distribution was measured in lithium heparinate plasma samples with a HI above the threshold value of 20 and in paired serum samples after transportation by pneumatic tube system. HI values above 20 can interfere with the selected assays: Creatine kinase (CK), creatine kinase-MB (CK-MB) and alanine aminotransferase (ALT) activities. These parameters were determined to demonstrate how haemolysis affects the results. 17.5% of the tube-transported plasma samples and 2.6% of the hand-delivered plasma samples had a HI above 20. The median HI in pneumatic tube-transported lithium heparinate plasma was 85 and 33 in the paired serum samples. The median HI difference between paired plasma and serum was 46. Blood samples in lithium heparinate tubes may be substantially more susceptible to haemolysis by pneumatic tube transportation than serum tube samples. Although our results cannot be universally applied to laboratories with different pneumatic tube systems, it is recommended that each laboratory evaluate carefully the degree of haemolysis after the transportation by the own pneumatic tube system and in terms of the sample type.

Effect of commercial rye whole-meal bread on postprandial blood glucose and gastric emptying in healthy subjects

Directory of Open Access Journals (Sweden)

Darwich Gassan

2009-06-01

Full Text Available Abstract Background The intake of dietary fibre has been shown to reduce the risk of developing diabetes mellitus. The aim of this study was to compare the effects of commercial rye whole-meal bread containing whole kernels and white wheat bread on the rate of gastric emptying and postprandial glucose response in healthy subjects. Methods Ten healthy subjects took part in a blinded crossover trial. Blood glucose level and gastric emptying rate (GER were determined after the ingestion of 150 g white wheat bread or 150 g whole-meal rye bread on two different occasions after fasting overnight. The GER was measured using real-time ultrasonography, and was calculated as the percentage change in antral cross-sectional area 15 and 90 minutes after completing the meal. Results No statistically significant difference was found between the GER values or the blood glucose levels following the two meals when evaluated with the Wilcoxon signed rank sum test. Conclusion The present study revealed no difference in postprandial blood glucose response or gastric emptying after the ingestion of rye whole-meal bread compared with white wheat bread. Trial registration NCT00779298

High-Quality Exome Sequencing of Whole-Genome Amplified Neonatal Dried Blood Spot DNA

DEFF Research Database (Denmark)

Poulsen, Jesper Buchhave; Lescai, Francesco; Grove, Jakob

2016-01-01

Stored neonatal dried blood spot (DBS) samples from neonatal screening programmes are a valuable diagnostic and research resource. Combined with information from national health registries they can be used in population-based studies of genetic diseases. DNA extracted from neonatal DBSs can...... be amplified to obtain micrograms of an otherwise limited resource, referred to as whole-genome amplified DNA (wgaDNA). Here we investigate the robustness of exome sequencing of wgaDNA of neonatal DBS samples. We conducted three pilot studies of seven, eight and seven subjects, respectively. For each subject...... we analysed a neonatal DBS sample and corresponding adult whole-blood (WB) reference sample. Different DNA sample types were prepared for each of the subjects. Pilot 1: wgaDNA of 2x3.2mm neonatal DBSs (DBS_2x3.2) and raw DNA extract of the WB reference sample (WB_ref). Pilot 2: DBS_2x3.2, WB...

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

CERN Document Server

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

2002-01-01

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

Discrimination of lymphoma using laser-induced breakdown spectroscopy conducted on whole blood samples

Science.gov (United States)

Chen, Xue; Li, Xiaohui; Yang, Sibo; Yu, Xin; Liu, Aichun

2018-01-01

Lymphoma is a significant cancer that affects the human lymphatic and hematopoietic systems. In this work, discrimination of lymphoma using laser-induced breakdown spectroscopy (LIBS) conducted on whole blood samples is presented. The whole blood samples collected from lymphoma patients and healthy controls are deposited onto standard quantitative filter papers and ablated with a 1064 nm Q-switched Nd:YAG laser. 16 atomic and ionic emission lines of calcium (Ca), iron (Fe), magnesium (Mg), potassium (K) and sodium (Na) are selected to discriminate the cancer disease. Chemometric methods, including principal component analysis (PCA), linear discriminant analysis (LDA) classification, and k nearest neighbor (kNN) classification are used to build the discrimination models. Both LDA and kNN models have achieved very good discrimination performances for lymphoma, with an accuracy of over 99.7%, a sensitivity of over 0.996, and a specificity of over 0.997. These results demonstrate that the whole-blood-based LIBS technique in combination with chemometric methods can serve as a fast, less invasive, and accurate method for detection and discrimination of human malignancies. PMID:29541503

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

Czech Academy of Sciences Publication Activity Database

Zhang, H.; Korenková, Vlasta; Sjoback, R.; Švec, David; Bjorkman, J.; Kruhoffer, M.; Verderio, P.; Pizzamiglio, S.; Ciniselli, Ch.M.; Wyrich, R.; Oelmueller, U.; Kubista, Mikael; Lindahl, T.; Lonneborg, A.; Rian, E.

2014-01-01

Roč. 9, č. 11/e111644 (2014) E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional research plan: CEZ:AV0Z50520701 Institutional support: RVO:86652036 Keywords : GENE-EXPRESSION PROFILES * HUMAN WHOLE-BLOOD * TIME RT-PCR Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2014

Levels of metals in blood samples from Mallards (Anas platyrhynchos) from urban areas in Poland

International Nuclear Information System (INIS)

Binkowski, Łukasz J.; Meissner, Włodzimierz

2013-01-01

In this paper we present the studies conducted on blood samples taken from Mallards (Anas platyrhynchos). Birds were captured for ringing purposes (n = 43) in two small and two big towns (including highly urbanized areas). For comparison samples of blood from birds shot on fish ponds were used (n = 26). Based on the body mass all sampled individuals can be assessed as being in good condition. Levels of cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), nickel (Ni), lead (Pb) and zinc (Zn) in blood samples were measured with AAS. Concentrations of metals did not differ statistically between sexes and made up a following order: Fe > Zn > Cu > Cr ≈ Ni > Pb > Cd. Mallards from towns revealed lower concentrations of Zn and Cu but higher concentration of Fe. There was no difference in exposition to Pb between birds from towns and fish ponds. -- Highlights: •Concentrations of seven metals were measured in blood of Mallards. •Birds represent different kind of areas: urban (towns) and non-urban ones (fish ponds). •Concentrations of studied metals made up a following order: Fe > Zn > Cu > Cr ≈ Ni > Pb > Cd. •Birds from towns revealed higher amount of Fe and lower concentrations of Zn and Cu. •There was no difference in exposition to Pb between birds from towns and fish ponds. -- Exposition to iron is higher on industrialized areas in towns, but exposition to zinc and copper on such areas is lower in comparison to country fish ponds

Venepuncture versus heel lance for blood sampling in term neonates.

Science.gov (United States)

Shah, Vibhuti S; Ohlsson, Arne

2011-10-05

Heel lance has been the conventional method of blood sampling in neonates for screening tests. Neonates undergoing heel lance experience pain which cannot be completely alleviated. To determine whether venepuncture or heel lance is less painful and more effective for blood sampling in term neonates. Randomized or quasi-randomised controlled trials comparing pain response to venepuncture versus heel lance were identified by searching the Cochrane Central Regsiter of Controlled Trials (CENTRAL, The Cochrane Library), MEDLINE, EMBASE, CINAHL, and clinical trials registries in May 2011. Trials comparing pain response to venepuncture versus heel lance with or with out the use of a sweet tasting solution as a co-intervention in term neonates. Outcomes included pain response to venepuncture versus heel lance with or without the use of a sweet tasting solution using validated pain measures, the need of repeat sampling and cry characteristics. Analyses included typical relative risk (RR), risk difference (RD), number needed to treat (NNT), weighted mean difference (WMD) and standardized mean difference (SMD) with their 95% confidence intervals (CI). Between study heterogeneity was reported including the I squared (I(2)) test. Six studies (n = 478) of variable quality were included. A composite outcome of Infant Pain Scale (NIPS), Neonatal Facial Action Coding System (NFCS) and/or Premature Infant Pain Profile (PIPP) score was reported in 288 infants, who did not receive a sweet tasting solution. Meta-analysis showed a significant reduction in the venepuncture versus the heel lance group (SMD -0.76, 95% CI -1.00 to -0.52; I(2) = 0%). When a sweet tasting solution was provided the SMD remained significant favouring the venepuncture group (SMD - 0.38, 95% CI -0.69 to -0.07). The typical RD for requiring more than one skin puncture for venepuncture versus heel lance (reported in 4 studies; n = 254) was -0.34 (95% CI -0.43 to -0.25; I(2) = 97%). The NNT to avoid one repeat skin

Detection of cytomegalovirus in blood donors by PCR using the digene SHARP signal system assay: effects of sample preparation and detection methodology.

OpenAIRE

Krajden, M; Shankaran, P; Bourke, C; Lau, W

1996-01-01

Cytomegalovirus (CMV) is an important cause of transfusion-associated morbidity and mortality; however, only 0.4 to 12% of the blood products obtained from seropositive blood donors transmit infection. The effects of three commercially available whole-blood sample preparation kits on the detection of CMV PCR products by a semiquantitative adaptation of the Digene SHARP Signal System Assay (DSSSA) in samples from volunteer blood donors was assessed. Of 101 samples from seropositive blood donor...

Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia

OpenAIRE

Lenicek Krleza, Jasna

2014-01-01

Introduction: Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. Materials and methods: All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about t...

Determination of the technetium-99m mercaptoacetyltriglycine plasma clearance in children by means of a single blood sample: a multicentre study

International Nuclear Information System (INIS)

Piepsz, A.; Gordon, I.; Hahn, K.; Kolinska, J.; Kotzerke, J.; Sixt, R.

1993-01-01

A multicentree European study was undertaken in order to determine a reasonable algorithm allowing the determination of overall technetium-99m mercaptoacetyltriglycine clearance using a single blood sample. Employing multiple blood sample clearance as a reference method, it was shown that an acceptable estimation of the MAG3 renal clearance could be obtained using a blood sample taken at any time between 30 and 40 min after tracer injection. After correction for body surface area comparison of clearance determined using (a) the single blood sample and (b) the multiple blood samples provided a coefficient of correlation of 0.949 and an SEE of 27 ml/min. This algorithm is valid for clearance values higher than 100 ml/min/1.73 m 2 and for children older than 1 year of age. (orig.)

Evaluation of dry blood spot technique for quantification of an Anti-CD20 monoclonal antibody drug in human blood samples.

Science.gov (United States)

Lin, Yong-Qing; Zhang, Yilu; Li, Connie; Li, Louis; Zhang, Kelley; Li, Shawn

2012-01-01

To evaluate the dried blood spot (DBS) technique in ELISA quantification of larger biomolecular drugs, an anti-CD20 monoclonal antibody drug was used as an example. A method for the quantification of the anti-CD20 drug in human DBS was developed and validated. The drug standard and quality control samples prepared in fresh human blood were spotted on DBS cards and then extracted. A luminescent ELISA was used for quantification of the drug from DBS samples. The assay range of the anti-CD20 drug standards in DBS was 100-2500ng/mL. The intra-assay precision (%CV) ranged from 0.4% to 10.1%, and the accuracy (%Recovery) ranged from 77.9% to 113.9%. The inter assay precision (%CV) ranged from 5.9% to 17.4%, and the accuracy ranged from 81.5% to 110.5%. The DBS samples diluted 500 and 50-fold yielded recovery of 88.7% and 90.7%, respectively. The preparation of DBS in higher and lower hematocrit (53% and 35%) conditions did not affect the recovery of the drug. Furthermore, the storage stability of the anti-CD20 drug on DBS cards was tested at various conditions. It was found that the anti-CD20 drug was stable for one week in DBS stored at room temperature. However, it was determined that the stability was compro]mised in DBS stored at high humidity, high temperature (55°C), and exposed to direct daylight for a week, as well as for samples stored at room temperature and high humidity conditions for a month. Stability did not change significantly in samples that underwent 3 freeze/thaw cycles. Our results demonstrated a successful use of DBS technique in ELISA quantification of an anti-CD20 monoclonal antibody drug in human blood. The stability data provides information regarding sample storage and shipping for future clinical studies. It is, therefore, concluded that the DBS technique is applicable in the quantification of other large biomolecule drugs or biomarkers. Copyright © 2011 Elsevier Inc. All rights reserved.

Sex based levels of C-reactive protein and white blood cell count in subjects with metabolic syndrome: Isfahan Healthy Heart Program.

Science.gov (United States)

Gharipour, Mojgan; Ramezani, Mohammad Arash; Sadeghi, Masuomeh; Khosravi, Alireza; Masjedi, Mohsen; Khosravi-Boroujeni, Hossein; Rafieian-Kopaei, Mahmoud; Sarrafzadegan, Nizal

2013-06-01

C-reactive protein (CRP) and white blood cell (WBC) are proinflammatory markers. They are major pathophysiological for the development of metabolic syndrome (MetS). This study aimed to address the independent associations between MetS and WBC counts and serum CRP levels and evaluation of their magnitude in relation to the MetS, based on the sex in the Iranian adults. In this cross-sectional study, subjects who met the MetS criteria, based on the Adult Treatment Panel III were selected from the Isfahan Healthy Heart Program database. A questionnaire containing the demographic data, weight, height, waist, and hip circumference of the respondents was completed for each person. Blood pressure was measured and the anthropometric measurements were done, and fasting blood samples were taken for 2 h postload plasma glucose (2 hpp). Serum [total, high-density lipoprotein (HDL), and low-density lipoprotein] levels of cholesterol, triglyceride, and CRP as well as WBC counts were determined. The univariate analyses were carried out to assess the relation between the CRP levels, WBC counts with the MetS in both sexes the. In men with the abdominal obesity, the higher levels of WBC count, high serum triglyceride and blood glucose levels, a low serum HDL level, and raised systolic and diastolic blood pressure were observed. However, the higher serum CRP levels were only observed in those with the low serum HDL-cholesterol levels. The mean values of the WBC counts were statistically different between the men with and without MetS, but the mean values of the CRP levels were similar between the two groups. In women, the mean values of WBC count and CRP levels were statistically different in the subjects with and without a MetS components (except for the low serum HDL levels and high diastolic blood pressure for the WBC measures and abdominal obesity for the CRP measures) and for those with and without MetS. The age and smoking adjusted changes in the CRP levels and WBC counts

Post-mortem detection of gasoline residues in lung tissue and heart blood of fire victims.

Science.gov (United States)

Pahor, Kevin; Olson, Greg; Forbes, Shari L

2013-09-01

The purpose of this study was to determine whether gasoline residues could be detected post-mortem in lung tissue and heart blood of fire victims. The lungs and heart blood were investigated to determine whether they were suitable samples for collection and could be collected without contamination during an autopsy. Three sets of test subjects (pig carcasses) were investigated under two different fire scenarios. Test subjects 1 were anaesthetized following animal ethics approval, inhaled gasoline vapours for a short period and then euthanized. The carcasses were clothed and placed in a house where additional gasoline was poured onto the carcass post-mortem in one fire, but not in the other. Test subjects 2 did not inhale gasoline, were clothed and placed in the house and had gasoline poured onto them in both fires. Test subjects 3 were clothed but had no exposure to gasoline either ante- or post-mortem. Following controlled burns and suppression with water, the carcasses were collected, and their lungs and heart blood were excised at a necropsy. The headspace from the samples was analysed using thermal desorption-gas chromatography-mass spectroscopy. Gasoline was identified in the lungs and heart blood from the subjects that were exposed to gasoline vapours prior to death (test subjects 1). All other samples were negative for gasoline residues. These results suggest that it is useful to analyse for volatile ignitable liquids in lung tissue and blood as it may help to determine whether a victim was alive and inhaling gases at the time of a fire.

Determination of degree of RBC agglutination for blood typing using a small quantity of blood sample in a microfluidic system.

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Chang, Yaw-Jen; Ho, Ching-Yuan; Zhou, Xin-Miao; Yen, Hsiu-Rong

2018-04-15

Blood typing assay is a critical test to ensure the serological compatibility of a donor and an intended recipient prior to a blood transfusion. This paper presents a microfluidic blood typing system using a small quantity of blood sample to determine the degree of agglutination of red blood cell (RBC). Two measuring methods were proposed: impedimetric measurement and electroanalytical measurement. The charge transfer resistance in the impedimetric measurement and the power parameter in the electroanalytical measurement were used for the analysis of agglutination level. From the experimental results, both measuring methods provide quantitative results, and the parameters are linearly and monotonically related to the degree of RBC agglutination. However, the electroanalytical measurement is more reliable than the impedimetric technique because the impedimetric measurement may suffer from many influencing factors, such as chip conditions. Five levels from non-agglutination (level 0) to strong agglutination (level 4+) can be discriminated in this study, conforming to the clinical requirement to prevent any risks in transfusion. Copyright © 2017 Elsevier B.V. All rights reserved.

Does Pneumatic Tube System Transport Contribute to Hemolysis in ED Blood Samples?

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Phelan, Michael P; Reineks, Edmunds Z; Hustey, Fredric M; Berriochoa, Jacob P; Podolsky, Seth R; Meldon, Stephen; Schold, Jesse D; Chamberlin, Janelle; Procop, Gary W

2016-09-01

Our goal was to determine if the hemolysis among blood samples obtained in an emergency department and then sent to the laboratory in a pneumatic tube system was different from those in samples that were hand-carried. The hemolysis index is measured on all samples submitted for potassium analysis. We queried our hospital laboratory database system (SunQuest(®)) for potassium results for specimens obtained between January 2014 and July 2014. From facility maintenance records, we identified periods of system downtime, during which specimens were hand-carried to the laboratory. During the study period, 15,851 blood specimens were transported via our pneumatic tube system and 92 samples were hand delivered. The proportions of hemolyzed specimens in the two groups were not significantly different (13.6% vs. 13.1% [p=0.90]). Results were consistent when the criterion was limited to gross (3.3% vs 3.3% [p=0.99]) or mild (10.3% vs 9.8% [p=0.88]) hemolysis. The hemolysis rate showed minimal variation during the study period (12.6%-14.6%). We found no statistical difference in the percentages of hemolyzed specimens transported by a pneumatic tube system or hand delivered to the laboratory. Certain features of pneumatic tube systems might contribute to hemolysis (e.g., speed, distance, packing material). Since each system is unique in design, we encourage medical facilities to consider whether their method of transport might contribute to hemolysis in samples obtained in the emergency department.

PIXE elemental analysis of erythrocyte and blood plasma samples from human pregnancies

International Nuclear Information System (INIS)

Borbely-Kiss, I.; Koltay, E.; Laszlo, S.; Szabo, Gy.

1984-01-01

Elemental concentrations of P, S, Cl, K, Ca, Fe, Ni, Cu, Zn, Br, Rb have been determined in erythrocyte and blood plasma samples from normal and diabetic human pregnancies. Average values, the dependence of the concentrations on the time during gestation period, the correlation coefficients for pairs of elements as well as for the same elements in plasma and erythrocyte samples are given. A marked difference appeared in a number of cases between normal and diabetic pregnancies. (author)

PIXE elemental analysis of erythrocyte and blood plasma samples from human pregnancies

Energy Technology Data Exchange (ETDEWEB)

Borbely-Kiss, I; Koltay, E; Laszlo, S; Szabo, Gy [Magyar Tudomanyos Akademia, Debrecen. Atommag Kutato Intezete; Goedeny, S [Orvostudomanyi Egyetem, Szeged (Hungary). Szueleszeti es Noegyogyaszati Klinika; Seif El-Nasr, S [Teachers' Coll. for Women, Samia (Kuwait)

1984-07-01

Elemental concentrations of P, S, Cl, K, Ca, Fe, Ni, Cu, Zn, Br, Rb have been determined in erythrocyte and blood plasma samples from normal and diabetic human pregnancies. Average values, the dependence of the concentrations on the time during gestation period, the correlation coefficients for pairs of elements as well as for the same elements in plasma and erythrocyte samples are given. A marked difference appeared in a number of cases between normal and diabetic pregnancies. 11 refs.

Suppressive response of confections containing the extractive from leaves of Morus Alba on postprandial blood glucose and insulin in healthy human subjects

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Oku Tsuneyuki

2009-07-01

Full Text Available Abstract Background The first aim of this study was to clarify the effective ratio of extractive from leaves of Morus Alba (ELM to sucrose so as to apply this knowledge to the preparation of confections that could effectively suppress the elevation of postprandial blood glucose and insulin. The second aim was to identify the efficacy of confections prepared with the optimally effective ratio determined from the first study, using healthy human subjects. Methods Ten healthy females (22.3 years, BMI 21.4 kg/m2 participated in this within-subject, repeated measures study. For the first aim of this study, the test solutions containing 30 g of sucrose and 1.2 or 3.0 g of ELM were repeatedly and randomly given to each subject. To identify the practically suppressive effects on postprandial blood glucose and insulin, some confections with added ELM were prepared as follows: Mizu-yokan, 30 g of sucrose with the addition of 1.5 or 3.0 g ELM; Daifuku-mochi, 9.0 g of starch in addition to 30 g of sucrose and 1.5 or 3.0 g ELM; Chiffon-cake, 24 g of sucrose, starch, and 3.0 or 6.0 g of ELM, and were ingested by each subject. Blood and end-expiration were collected at selected periods after test food ingestion. Results When 30 g of sucrose with 1.2 or 3.0 g of ELM were ingested by subjects, the elevations of postprandial blood glucose and insulin were effectively suppressed (p p Conclusion ELM-containing confections for which the ratio of ELM and sucrose is one-tenth effectively suppress the postprandial blood glucose and insulin by inhibiting the intestinal sucrase, thus creating a prebiotic effect. The development of confections with ELM can therefore contribute to the prevention and the quality of life for prediabetic and diabetic patients.

Use of standard laboratory methods to obviate routine dithiothreitol treatment of blood samples with daratumumab interference.

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Lintel, Nicholas J; Brown, Debra K; Schafer, Diane T; Tsimba-Chitsva, Farai M; Koepsell, Scott A; Shunkwiler, Sara M

2017-01-01

Daratumumab is an antibody currently used in the treatment of patients with refractory multiple myeloma. Blood samples from patients being treated with daratumumab may show panreactivity during pre-transfusion testing. To facilitate the provision of blood components for such patients, it is recommended that a baseline phenotype or genotype be established prior to starting treatment with daratumumab. If patient red blood cells (RBCs) require phenotyping after the start of daratumumab treatment, dithiothreitol (DTT) treatment of the patient's RBCs should be performed. The medical charts of four patients treated with daratumumab were reviewed. The individual number of doses ranged from 1 to 14; patient age ranged from 55 to 78 years; two men and two women were included in the review. Type and screen data were obtained from samples collected over 33 encounters with a range of 1 to 13 encounters per patient. All samples were tested initially by automated solid-phase testing. Any reactivity with solid phase led to tube testing with either low-ionic-strength saline, polyethylene glycol, or both. If incubation failed to eliminate the reactivity, the sample was sent to a reference laboratory for DTT treatment and phenotyping. Of the 33 samples tested, 23 (69.7%) samples had reactivity in solid-phase testing. In 8 of the 10 samples that did not react in solid-phase, testing was conducted more than four half-lives after the last dose of daratumumab. Of the 23 that had reactivity in solid-phase, 16 (69.6%) samples demonstrated loss of reactivity using common laboratory methods. For the seven patients whose sample reactivity was not initially eliminated, six were provided with phenotypically matched blood based on prior molecular testing. Only one sample was sent out for DTT treatment. These results suggest that daratumumab interference with pre-transfusion testing can be addressed using common laboratory methods. This finding could save time and money for laboratories that do

A Computational Framework to Optimize Subject-Specific Hemodialysis Blood Flow Rate to Prevent Intimal Hyperplasia

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Mahmoudzadeh, Javid; Wlodarczyk, Marta; Cassel, Kevin

2017-11-01

Development of excessive intimal hyperplasia (IH) in the cephalic vein of renal failure patients who receive chronic hemodialysis treatment results in vascular access failure and multiple treatment complications. Specifically, cephalic arch stenosis (CAS) is known to exacerbate hypertensive blood pressure, thrombosis, and subsequent cardiovascular incidents that would necessitate costly interventional procedures with low success rates. It has been hypothesized that excessive blood flow rate post access maturation which strongly violates the venous homeostasis is the main hemodynamic factor that orchestrates the onset and development of CAS. In this article, a computational framework based on a strong coupling of computational fluid dynamics (CFD) and shape optimization is proposed that aims to identify the effective blood flow rate on a patient-specific basis that avoids the onset of CAS while providing the adequate blood flow rate required to facilitate hemodialysis. This effective flow rate can be achieved through implementation of Miller's surgical banding method after the maturation of the arteriovenous fistula and is rooted in the relaxation of wall stresses back to a homeostatic target value. The results are indicative that this optimized hemodialysis blood flow rate is, in fact, a subject-specific value that can be assessed post vascular access maturation and prior to the initiation of chronic hemodialysis treatment as a mitigative action against CAS-related access failure. This computational technology can be employed for individualized dialysis treatment.

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

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

1990-01-01

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

In vitro immunomodulation of a whole blood IFN-γ release assay enhances T cell responses in subjects with latent tuberculosis infection.

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Rajiv L Gaur

Full Text Available Activation of innate immunity via pathogen recognition receptors (PRR modulates adaptive immune responses. PRR ligands are being exploited as vaccine adjuvants and as therapeutics, but their utility in diagnostics has not been explored. Interferon-gamma (IFN-γ release assays (IGRAs are functional T cell assays used to diagnose latent tuberculosis infection (LTBI; however, novel approaches are needed to improve their sensitivity.In vitro immunomodulation of a whole blood IGRA (QuantiFERON®-TB GOLD In-Tube with Toll-like receptor agonists poly(I:C, LPS, and imiquimod was performed on blood from subjects with LTBI and negative controls.In vitro immunomodulation significantly enhanced the response of T cells stimulated with M. tuberculosis antigens from subjects with LTBI but not from uninfected controls. Immunomodulation of IGRA revealed T cell responses in subjects with LTBI whose T cells otherwise do not respond to in vitro stimulation with antigens alone. Similar to their in vivo functions, addition of poly(I:C and LPS to whole blood induced secretion of inflammatory cytokines and IFN-α and enhanced the surface expression of antigen presenting and costimulatory molecules on antigen presenting cells.In vitro immunomodulation of whole blood IGRA may be an effective strategy for enhancing the sensitivity of T cells for diagnosis of LTBI.

Direct analysis of site-specific N-glycopeptides of serological proteins in dried blood spot samples.

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Choi, Na Young; Hwang, Heeyoun; Ji, Eun Sun; Park, Gun Wook; Lee, Ju Yeon; Lee, Hyun Kyoung; Kim, Jin Young; Yoo, Jong Shin

2017-08-01

Dried blood spot (DBS) samples have a number of advantages, especially with respect to ease of collection, transportation, and storage and to reduce biohazard risk. N-glycosylation is a major post-translational modification of proteins in human blood that is related to a variety of biological functions, including metastasis, cell-cell interactions, inflammation, and immunization. Here, we directly analyzed tryptic N-glycopeptides from glycoproteins in DBS samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS) without centrifugation of blood samples, depletion of major proteins, desalting of tryptic peptides, and enrichment of N-glycopeptides. Using this simple method, we identified a total of 41 site-specific N-glycopeptides from 16 glycoproteins in the DBS samples, from immunoglobulin gamma 1 (IgG-1, 10 mg/mL) down to complement component C7 (50 μg/mL). Of these, 32 N-glycopeptides from 14 glycoproteins were consistently quantified over 180 days stored at room temperature. The major abundant glycoproteins in the DBS samples were IgG-1 and IgG-2, which contain nine asialo-fucosylated complex types of 16 different N-glycopeptide isoforms. Sialo-non-fucosylated complex types were primarily detected in the other glycoproteins such as alpha-1-acid glycoprotein 1, 2, alpha-1-antitypsin, alpha-2-macroglobulin, haptoglobin, hemopexin, Ig alpha 1, 2 chain C region, kininogen-1, prothrombin, and serotransferrin. We first report the characterization of site-specific N-glycoproteins in DBS samples by LC-MS/MS with minimal sample preparation.

Vagal Blocking Improves Glycemic Control and Elevated Blood Pressure in Obese Subjects with Type 2 Diabetes Mellitus

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S. Shikora

2013-01-01

Full Text Available Background. An active device that downregulates abdominal vagal signalling has resulted in significant weight loss in feasibility studies. Objective. To prospectively evaluate the effect of intermittent vagal blocking (VBLOC on weight loss, glycemic control, and blood pressure (BP in obese subjects with DM2. Methods. Twenty-eight subjects were implanted with a VBLOC device (Maestro Rechargeable System at 5 centers in an open-label study. Effects on weight loss, HbA1c, fasting blood glucose, and BP were evaluated at 1 week to 12 months. Results. 26 subjects (17 females/9 males, 51±2 years, BMI 37±1 kg/m2, mean ± SEM completed 12 months followup. One serious adverse event (pain at implant site was easily resolved. At 1 week and 12 months, mean excess weight loss percentages (% EWL were 9±1% and 25±4% (P<0.0001, and HbA1c declined by 0.3±0.1% and 1.0±0.2% (P=0.02, baseline 7.8±0.2%. In DM2 subjects with elevated BP (n=15, mean arterial pressure reduced by 7±3 mmHg and 8±3 mmHg (P=0.04, baseline 100 ± 2 mmHg at 1 week and 12 months. All subjects MAP decreased by 3 ± 2 mmHg (baseline 95 ± 2 mmHg at 12 months. Conclusions. VBLOC was safe in obese DM2 subjects and associated with meaningful weight loss, early and sustained improvements in HbA1c, and reductions in BP in hypertensive DM2 subjects. This trial is registered with ClinicalTrials.gov NCT00555958.

On the improvement of blood sample collection at clinical laboratories.

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Grasas, Alex; Ramalhinho, Helena; Pessoa, Luciana S; Resende, Mauricio G C; Caballé, Imma; Barba, Nuria

2014-01-09

Blood samples are usually collected daily from different collection points, such hospitals and health centers, and transported to a core laboratory for testing. This paper presents a project to improve the collection routes of two of the largest clinical laboratories in Spain. These routes must be designed in a cost-efficient manner while satisfying two important constraints: (i) two-hour time windows between collection and delivery, and (ii) vehicle capacity. A heuristic method based on a genetic algorithm has been designed to solve the problem of blood sample collection. The user enters the following information for each collection point: postal address, average collecting time, and average demand (in thermal containers). After implementing the algorithm using C programming, this is run and, in few seconds, it obtains optimal (or near-optimal) collection routes that specify the collection sequence for each vehicle. Different scenarios using various types of vehicles have been considered. Unless new collection points are added or problem parameters are changed substantially, routes need to be designed only once. The two laboratories in this study previously planned routes manually for 43 and 74 collection points, respectively. These routes were covered by an external carrier company. With the implementation of this algorithm, the number of routes could be reduced from ten to seven in one laboratory and from twelve to nine in the other, which represents significant annual savings in transportation costs. The algorithm presented can be easily implemented in other laboratories that face this type of problem, and it is particularly interesting and useful as the number of collection points increases. The method designs blood collection routes with reduced costs that meet the time and capacity constraints of the problem.

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

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

2016-07-01

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