scholarly journals Detection of pregnancy in sheep using an ELISA for pregnancy-specific protein B

2018 ◽  
Vol 66 (2) ◽  
pp. 329-336
Author(s):  
Tímea Milisits-Németh ◽  
Orsolya Gabriella Balogh ◽  
István Egerszegi ◽  
László Kern ◽  
R. Garth Sasser ◽  
...  
Keyword(s):  
Accurate Method ◽  
Specific Protein ◽  
Economic Benefits ◽  
Seasonal Breeding ◽  
Serum Samples ◽  
Mating Period ◽  
Blood Samples ◽  
Sheep Production ◽  
Protein B

The early detection of pregnancy and the determination of fetal numbers have economic benefits in sheep production because of the seasonal breeding patterns where missing a breeding opportunity means the loss of one productive year. The purpose of this study was to evaluate the efficacy of the B6-HRP ELISA for ovine pregnancy-specific protein B (oPSPB) measurement in the detection of pregnancy and estimation of fetal numbers in different sheep breeds. BioPRYN® ELISA assay kit was used for the detection of pregnancy in the experimental animals. Ninety-three ewes of three breeds (British Milksheep – BM, Lacaune – L and Transylvanian Racka – TR), each from three farms in Hungary, were included in the study. BM and L ewes were artificially inseminated (AI). Thirty-five days after AI, all ewes were examined by transabdominal ultrasound. The TR flock was mated naturally over a six-week period. At the end of the mating period, the ewes were similarly examined by ultrasound. Blood samples were taken from all pregnant ewes twice (35 and 65 days after AI), and serum samples were assayed by the BioPRYN test. It can be concluded that the detection of serum PSPB by ELISA is a much easier, safer, less expensive and highly accurate method for the detection of ovine pregnancy. Although some breed-related differences were detectable at 35 and 65 days post breeding, no differences in oPSPB levels were found in pregnant ewes carrying different numbers of fetuses.

scholarly journals Effects of Delayed Sample Processing on Determination of Total and High Molecular Weight (HMW) Adiponectin in Serum and Plasma: A Pilot Study

2016 ◽  
Vol 8 (3) ◽  
pp. 19
Author(s):  
Choaping Ng ◽  
Felicity J Rose ◽  
Sahar Keshvari ◽  
Marina M Reeves ◽  
Goce Dimeski ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Pilot Study ◽  
High Molecular Weight ◽  
Accurate Determination ◽  
Mean Difference ◽  
Serum Samples ◽  
Blood Samples ◽  
Hmw Adiponectin ◽  
Total Adiponectin

<p>Adiponectin is a beneficial adipocyte-secreted hormone, which circulates in a variety of multimeric forms termed low and high molecular weight (LMW/HMW). Effectiveness of clinical therapeutic trials which target adiponectin rely on accurate determination of circulating total and HMW adiponectin levels but the accuracy may be influenced by variations in sample handling processes. The aim of this pilot study was to investigate the effects of delayed processing of blood samples on the concentration of total and HMW adiponectin.</p><p>Materials and Methods: Fasting blood samples were collected for analysis of total and HMW adiponectin concentrations in EDTA plasma and serum from eight healthy participants.  Samples were centrifuged post 15 min storage at 4<sup>o</sup>C as the comparative ‘ideal’ method or after up to 72 h of refrigerated storage or 6 h at room temperature. Total and HMW adiponectin concentrations were measured by ELISA.</p><p>Results: Under ideal handling conditions measurements of total and HMW adiponectin concentrations were significantly higher in serum than in plasma (mean difference: -1.3 µg/mL [95% CI: -1.6, -1.0], p&lt;0.001; and, -0.6 µg/mL [95% CI: -0.7, -0.5], p&lt;0.001, respectively).  Storage of blood samples at 4<sup>o</sup>C for 72 h resulted in significant reductions in concentration of total adiponectin in serum (mean difference: -1.4 µg/mL [95% CI: -2.0, -0.8], p=0.001) and HMW adiponectin in plasma (mean difference: -0.6 µg/mL [95%CI: -0.9, -0.2], p=0.007), compared with ideal conditions.  Further analysis of serum samples showed a significant decrease in total adiponectin concentration after 6 h storage at 4<sup>o</sup>C (mean difference: -1.4 µg/mL [95% CI: -2.0, -0.8], p=0.001) compared with ideal conditions.</p><p>Conclusions: Delayed processing of samples may have differential effects on the concentration of total and HMW adiponectin in serum or plasma. Larger studies are warranted for clinical intervention trials.</p>

PSV-4 Determination of Deoxynivalenol (DON) Content in Biological Samples as an Indicator of DON Intake in Grower-finisher Pigs

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 206-207
Author(s):  
Michael O Wellington ◽  
Michael A Bosompem ◽  
Veronika Nagl ◽  
Daniel A Columbus
Keyword(s):  
Mass Spectrometry ◽  
Biological Samples ◽  
High Performance ◽  
Tandem Mass ◽  
Serum Samples ◽  
Blood Samples ◽  
Swine Diets ◽  
Direct Quantification ◽  
Serum And Urine

Abstract Due to difficulties in obtaining consistent and/or reliable measures of deoxynivalenol (DON) in complete swine diets, we investigated whether measuring DON in biological samples could be used as an indicator of DON ingestion in pigs. In this study, graded levels of DON (1, 3, or 5 ppm) were fed to grower-finisher pigs for a period of 77-d. On d 35 and 77 of the study, urine samples were quantitatively collected over a 24-h period and blood samples were collected between 3 – 4 h after the morning meal on each of those days for serum DON analysis. For direct quantification of DON in urine, high-performance liquid chromatography with tandem mass spectrometry was performed. For serum samples, indirect quantification of DON was performed via enzymatic hydrolysis. We observed that DON content in urine increased linearly as intake of DON increased (Fig.1A; P &lt; 0.05). Analysis of DON in serum follow a similar trend, where serum DON content was increased as DON intake increased (Fig.1B; P &lt; 0.05). An average of 30% of DON ingested was recovered as DON in urine over a 24-h period. In summary, there was a linear relationship between DON intake and DON content in both urine and blood serum, therefore, analyzing DON concentration in serum and urine could be used as a tool to estimate for DON exposure in pigs under controlled conditions.

scholarly journals Pregnancy-Specific Protein B in Yankasa Ewes During Pregnancy and Postpartum Periods

2021 ◽  
Vol 44 (1) ◽  
pp. 55-62
Author(s):  
Adewale Ayodeji Adeyeye ◽  
Yusha'u Usman Abubakar ◽  
Olufisayo Oluwadamilare Leigh ◽  
Iyorhembe Utim Ate ◽  
Jashilagari Stephen ◽  
...  
Keyword(s):  
Specific Protein ◽  
Trophoblast Cells ◽  
Blood Samples ◽  
Pregnancy And Postpartum ◽  
Group A ◽  
Group B ◽  
Protein B

Abstract Pregnancy-specific protein B (PSPB) is produced by mono and binucleate trophoblast cells in the placenta of ruminants during pregnancy. This study was designed to determine the pattern of serum PSPB in Yankasa ewes during pregnancy and postpartum periods. Mature cycling Yankasa ewes were synchronized and divided into two groups A (n=11) and B (n=13). Group A was bred, while group B was unbred. Blood samples for PSPB assessment were collected from the ewes starting from the day of breeding until 4 weeks post-lambing. All pregnant Yankasa ewes lambed with singleton lambs after an average of 151.18 days. There was a significant (p<0.05) increase in PSPB in pregnant compared with the non-pregnant ewes in the period between 3 weeks post-breeding and 3 weeks post-lambing. Peaks were detected in the first (100.60 ng/ml), second (133.90 ng/ml), and third (114.82 ng/ml) trimesters at 5, 10 and 21 weeks of gestation, respectively, but steadily decreased within 4 weeks (2.38 ng/ml) postpartum. In conclusion, PSPB detected pregnancy in Yankasa ewes from 3 weeks post-breeding with peak levels at 5, 10 and 21 weeks post-breeding in the first, second, and third trimesters, respectively. PSPB decreased gradually after lambing until 4 weeks postpartum.

scholarly journals Determination of diazepam and its metabolites in serum by the use of liquid chromatography: Mass spectrometry method

2007 ◽  
Vol 64 (10) ◽  
pp. 659-662 ◽  
Author(s):  
Snezana Djordjevic ◽  
Vesna Kilibarda
Keyword(s):  
Biological Fluids ◽  
Accurate Method ◽  
Reversed Phase ◽  
Active Metabolites ◽  
Serum Samples ◽  
Standard Curve ◽  
Routine Identification ◽  
Selective Ion Monitoring ◽  
Pharmacologically Active

Background/Aim. Diazepam is a benzodiazepine anxyolitic. Metabolism of diazepam takes place in liver which generates pharmacologically active metabolites N-desmethyldiazepam, temazepam and oxazepam. The aim of this study was to develop and validate the method of liquid chromatographymass spectrometry (LC-MS) for separation and determination of diazepam and its active metabolites in the serum of rats samples after i.p. application of diazepam in a dose of 10 mg/kg. Methods. The serum samples taken from Wistar rats, were used in LC-MS analysis after the application of 10 mg/kg of diazepam i.p. Results. After alkaline extraction from the serum samples with diethylether and separation on a C18 reversed-phase column by using mobile phase methanolglacial acetic acid-water (50:1:49 v/v), diazepam and its metabolites were quantified. Determination was performed in a selective ion monitoring (SIM) mode, thereby the other exogenous and endogenous compounds did not interfere with this assay. Diazepam, N-desmethyldiazepam, oxazepam and temazepam were eluted in 14 minutes. The standard curve was linear in the range from 10-2 000 ng/ml. The limits of detection for diazepam, N-desmethyldiazepam, oxazepam and temazepam were 4.37, 3.13, 4.38 and 7.31 ng/ml, respectively. The limits of quantitation for diazepam, Ndesmethyldiazepam, oxazepam and temazepam were 14.58, 10.41, 14.59 and 24.36 ng/ml, respectively. Conclusion. The described LC-MS is a simple, sensitive, specific and accurate method and could be used for routine identification and quantification of small concentrations of diazepam and its metabolites in biological fluids.

Excess serum osmolality gap after ingestion of methanol: a methodology-associated phenomenon?

1994 ◽  
Vol 40 (8) ◽  
pp. 1587-1590 ◽  
Author(s):  
P Demedts ◽  
L Theunis ◽  
A Wauters ◽  
F Franck ◽  
R Daelemans ◽  
...  
Keyword(s):  
Gas Chromatography ◽  
Serum Osmolality ◽  
Ethanol Administration ◽  
Serum Samples ◽  
Blood Samples ◽  
Split Mode ◽  
Head Space Gas Chromatography ◽  
Osmolal Gap ◽  
Gap Values

Abstract A patient intentionally ingested an unknown amount of methanol and was admitted to the hospital 6 h later. On admission, the methanol concentration in blood was estimated as approximately of 134 mmol/L, based on the calculation of the osmolal gap. Intravenous ethanol administration and hemodialysis were promptly started. During hemodialysis, several blood samples were collected for determination of methanol and ethanol concentrations. Initially, we used gas chromatography with split-mode injection of pretreated serum samples; however, methanol concentrations turned out to be significantly lower than expected, based on calculated osmolal gap values. Because no explanation for the excess serum osmolal gap was apparent, we reanalyzed samples, using head-space gas chromatography. The methanol concentrations measured were significantly higher and osmolal gap values were no longer excessive.

Use of pregnancy-specific protein-B and estrone sulfate for determination of pregnancy on Day 49 in fallow deer ()

1993 ◽  
Vol 40 (2) ◽  
pp. 307-312 ◽  
Author(s):  
C. Wilker ◽  
B. Ball ◽  
T. Reimers ◽  
G. Sasser ◽  
M. Brunner ◽  
...  
Keyword(s):  
Fallow Deer ◽  
Specific Protein ◽  
Estrone Sulfate ◽  
Protein B

scholarly journals Discovery of Specific Antigens That Can Predict Microfilarial Intensity inLoa loaInfection

2017 ◽  
Vol 55 (9) ◽  
pp. 2671-2678 ◽  
Author(s):  
Papa M. Drame ◽  
Sasisekhar Bennuru ◽  
Thomas B. Nutman
Keyword(s):  
Point Of Care ◽  
Bioinformatic Analysis ◽  
Potential Utility ◽  
Circulating Biomarkers ◽  
Loa Loa ◽  
Serum Samples ◽  
Blood Samples ◽  
Antigen Capture ◽  
Specific Antigens

ABSTRACTAntigen-based immunoassays are currently needed for point-of-care quantification ofLoa loamicrofilariae (mf). Coupling transcriptomic approaches with bioinformatic analysis, we have identified 11 specific putative proteins (coding mRNAs) with potential utility as biomarkers of patent (mf+)L. loainfection. We successfully developed antigen capture immunoassays to quantify 2 (LOAG_14221 and LOAG_15846) of these proteins in individual plasma/serum samples. Of the 2 quantifiable circulating biomarkers, LOAG_14221 showed the highest degree of specificity, particularly with a monoclonal antibody-based immunoassay. Moreover, the levels of LOAG_14221 inL. loamf+patients were positively correlated to the mf densities in the corresponding blood samples (r= 0.53 andP= 0.008 for polyclonal assay;r= 0.54 andP= 0.004 for monoclonal assay). Thus, LOAG_14221 is a very promising biomarker that will be exploited in a quantitative point-of-care immunoassay for determination ofL. loamf densities.

The Dtermination of Paracetamol by HPLC Validation of the Method and Application on Serum Samples

2018 ◽  
Vol 69 (3) ◽  
pp. 627-631 ◽  
Author(s):  
Viorica Ohriac (Popa) ◽  
Diana Cimpoesu ◽  
Adrian Florin Spac ◽  
Paul Nedelea ◽  
Voichita Lazureanu ◽  
...  
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Hplc Analysis ◽  
Emotional Experience ◽  
Optimum Conditions ◽  
Serum Samples ◽  
Liquid Chromatograph ◽  
Mobile Phase Flow Rate ◽  
Quantification Limit

Pain is defined as a disagreeable sensory and emotional experience related to a tissue or potential lesion. Paracetamol (Acetaminophen) is the most used non-morphine analgesic. For the determination of paracetamol we developed and validated the high performance liquid chromatography (HPLC) analysis using a Dionex Ultimate 3000 liquid chromatograph equipped with a multidimensional detector. After determining the optimum conditions of analysis (80/20 water / acetonitrile mobile phase, flow rate 1.0 mL / min, detection wavelength 245 nm) we validated the method following the following parameters: linearity of response function, linearity of results, limit (LD = 0.66 mg / mL) and quantification limit (LQ = 2.00 mg / mL), and precision. The method of determining paracetamol by HPLC was applied to 30 samples of serum collected from patients who had pain and were treated with paracetamol.

Development of an HPLC-UV Method for Quantification of Stattic

2019 ◽  
Vol 15 (6) ◽  
pp. 568-573
Author(s):  
Soheil Sedaghat ◽  
Ommoleila Molavi ◽  
Akram Faridi ◽  
Ali Shayanfar ◽  
Mohammad Reza Rashidi
Keyword(s):  
High Performance ◽  
Accurate Method ◽  
Plasma Samples ◽  
Promising Target ◽  
Relative Standard ◽  
Dimerization Inhibitor ◽  
Oncogenic Protein ◽  
First Time ◽  
Transcription 3

Background: Signal transducer and activator of transcription 3 (STAT3), an oncogenic protein found constitutively active in many types of human malignancies, is considered to be a promising target for cancer therapy. Objective: In this study for the first time, a simple and accurate method has been developed for the determination of a STAT3 dimerization inhibitor called stattic in aqueous and plasma samples. Methods: A reverse-phase high-performance liquid chromatography (RP-HPLC) composed of C18 column as stationary phase, and the mixture of acetonitrile (60%) and water (40%) as mobile phase with a UV detection at 215 nm were applied for quantification of stattic. The developed method was validated by Food and Drug Administration (FDA) guideline. Results: The method provided a linear range between 1-40 and 2.5-40 µg mL-1 for aqueous and plasma samples, respectively, with a correlation coefficient of 0.999. The accuracy (as recovery) of the developed method was found to be between 95-105% for aqueous medium and 85-115% for plasma samples. The precision (as relative standard deviation) for aqueous and plasma samples was less than 6% and 15%, respectively. The sensitivity of the developed method based on FDA guideline was 1 µg mL-1 for aqueous and 2.5 µg mL-1 for plasma samples. Conclusion: These results show that the established method is a fast and accurate quantification for stattic in aqueous and plasma samples.

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