scholarly journals Analysis of Circulating microRNA During Early Gestation in Japanese Black Cattle

2021 ◽  
Author(s):  
Kasumi Ono ◽  
Sakura Okamoto ◽  
Chiaki Ninomiya ◽  
Noriyuki Toji ◽  
Tomomi Kanazawa ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Early Pregnancy ◽  
Circulating Microrna ◽  
Circulating Mirnas ◽  
Japanese Black Cattle ◽  
Circulating Micrornas ◽  
Physiological Conditions ◽  
Pregnancy Diagnosis ◽  
Bovine Plasma ◽  
Differentially Expressed Mirnas

Abstract Background Circulating microRNAs (miRNAs) are biomarkers for various diseases and physiological conditions in humans and mice; studies in domestic animals, particularly cattle, are limited. The importance of early pregnancy diagnosis (especially within the 21 day cow estrous cycle) in the livestock industry is extremely high. This study compared the circulating miRNAs in non-pregnant and pregnant Japanese Black cows, explored miRNAs as biomarkers for early pregnancy diagnosis, and established a measurement system that includes reference miRNA selection and the effect of hemolysis. Methods miRNA was extracted from the plasma of Japanese Black cows on day 21 after artificial insemination and subjected to a customized bovine oligonucleotide microarray for expression analysis. Differentially expressed miRNAs and reference miRNA candidates were selected and validated by real-time quantification PCR (RT-qPCR). Their stability was evaluated using NormFinder software. Hemolyzed samples were prepared using plasma from five cows in estrous cycle and subjected to RT-qPCR. Results A total of 124 miRNAs were detected in bovine plasma by microarray analysis in non-pregnant and pregnant cows. The levels of five circulating miRNAs were significantly higher in pregnant cows than in non-pregnant cows, and 24 miRNAs were detected only in the pregnant group. NormFinder analysis and RT-qPCR validation showed that miR-2455 was an appropriate reference miRNA in the plasma of non-pregnant and pregnant Japanese Black cows and miR-19b, miR-25, miR-29a, and miR-148a were significantly higher in the pregnant group. These four circulating miRNAs did not change during the estrous cycle and were less affected by hemolysis. Conclusions In the current study, we found high levels of four miRNAs in the plasma of pregnant Japanese Black cows. Since these miRNAs are less affected by hemolysis, they may potentially be used as markers for early pregnancy diagnosis in cattle.

scholarly journals Analysis of circulating miRNA levels in coronary heart disease patients with varying degrees of cardiovascular complications risk. correlations with the MSCT-CA data

2020 ◽  
Vol 75 (4) ◽  
pp. 283-291
Author(s):  
Andrey N. Rozhkov ◽  
Dmitriy Yu. Shchekochikhin ◽  
Natalia М. Baulina ◽  
Natalia A. Matveeva ◽  
Olga O. Favorova ◽  
...  
Keyword(s):  
Coronary Heart Disease ◽  
Heart Disease ◽  
Cardiovascular Complications ◽  
Demographic Characteristics ◽  
Atherosclerotic Plaques ◽  
Circulating Microrna ◽  
Circulating Mirnas ◽  
Circulating Micrornas ◽  
Estimated Risk

Rationale. Cardiovascular diseases remain the leading cause of human death in the world. Studying the role of regulatory non-coding RNAs, which include short single-stranded miRNA molecules, allows a more detailed understanding of the pathological processes underlying the progression of atherosclerosis. Objective to compare the levels of circulating miRNAs in patients with coronary heart disease, confirmed by multislice computed tomography-coronarography (MSCT-CA), with risks of cardiovascular complications and clinical and demographic characteristics. To compare the profiles of circulating miRNAs in groups of patients with stable and unstable atherosclerotic plaques. Methods. MicroRNA levels in the plasma of peripheral blood of patients with coronary heart disease were determined using the miScript miRNA PCR Array MIHS-105Z kit (Qiagen). The significance of differences in miRNA levels between the compared groups was determined using the MannWhitney U-test. The correlations of the levels of circulating miRNAs with clinical and demographic parameters were evaluated using the Spearman correlation coefficient. Risk assessment of cardiovascular complications in these patients was carried out using validated scales (ACC/AHA, Framinghm, SCORE, MESA). Atherosclerotic plaque stability was evaluated using MSCT-CA. Results. The study showed a significant (p 0.05) decrease in miR-16, miR-211, miR-195 miRNA levels in the plasma of patients with coronary heart disease, which correlated with an increase in cardiac vascular risk (CVR) according to ACC/AHA, Framingham and MESA. When comparing groups of patients with stable and unstable atherosclerotic plaques, the latter revealed an increase in the level of let-7b-5p circulating microRNA (p 0.05). Conclusion. Significant associations of the three studied microRNAs with the estimated risk of CVR were identified. It is important to find circulating let-7b-5p in a group of patients with unstable atherosclerotic plaques. Correlations were established between the levels of circulating microRNAs and clinical and demographic characteristics of patients. The study shows the involvement of some microRNAs in the regulation of atherosclerosis.

Validation of Buffalo PAG Antisera for Early Pregnancy Diagnosis in Bovine

2020 ◽  
Author(s):  
Nguyen Van Hanh ◽  
Son Hoang Nghia ◽  
Noelita Melo De Sousa ◽  
Olimpia Barbato ◽  
Jean François Beckers
Keyword(s):  
Early Pregnancy ◽  
Dairy Cow ◽  
The Other ◽  
Plasma Samples ◽  
Vicia Villosa ◽  
Blood Samples ◽  
Pregnancy Diagnosis ◽  
Bovine Plasma ◽  
Bovine Pregnancy ◽  
High Enrichment

This study was designed ability using antibody against buffalo PAG to detect concentration of pregnancy-associated glycoprotein (PAG) in bovine plasma samples. Successful purification of buffalo PAG molecules with high enrichment degree by using Vicia villosa aûnity chromatography was recently reported. In this research, the RIA system using novel anti buffalo-PAG was used for bovine pregnancy diagnosis. In a total 437 blood sampling of dairy cow in Belgium were used. Three PAG-RIA systems were undertaken to measure PAG concentration in bovine routine blood samples: AS#497 (or RIA-1) using rabbit antisera against purified bovine PAG67kDa, AS#706 (or RIA-2) using rabbit antisera against caprine PAG55+62kDa and new AS#859 (or RIA-3) using rabbit antisera against buffalo PAG59.5-75.8kDa). The measured PAG concentrations were similar for the systems As#497 and As#859; although the dilution was shown parallel, the measured PAG concentrations were significantly different between systems As#706 and As#859. On the other hand, the coefficients of correlations between concentrations obtained by the use of As#859 and As#497 were highly comparable to those between As#859 and As#706 or As#497 and As#706. In conclusion, the present study clearly shows that antisera against buffalo PAG can be used to develop new PAG-RIA system for bovine pregnancy diagnosis with high comparable results.

scholarly journals Ultrasonographic Analysis of the Number of Large, Middle and Small Follicles of Japanese Black Cattle during the Estrous Cycle and Early Pregnancy

1997 ◽  
Vol 43 (5) ◽  
pp. j1-j7 ◽  
Author(s):  
Tomoji MANABE ◽  
Kazufumi GOTO ◽  
Atsushi IDETA ◽  
Shinichiro TANIGUCHI ◽  
Shoji OOKUTSU ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Early Pregnancy ◽  
Japanese Black Cattle

75 COMPARISON OF THE DG29TM ENZYME-LINKED IMMUNOSORBENT ASSAY KIT COMPARED WITH TRANSRECTAL ULTRASONOGRAPHY FOR EARLY PREGNANCY DIAGNOSIS FOLLOWING TRANSFER OF JAPANESE BLACK CATTLE EMBRYOS

2012 ◽  
Vol 24 (1) ◽  
pp. 150 ◽  
Author(s):  
Y. Nakamura ◽  
M. Urakawa ◽  
A. Ideta ◽  
A. Shirasawa ◽  
Y. Oono ◽  
...  
Keyword(s):  
Negative Predictive Value ◽  
Clinical Application ◽  
Immunosorbent Assay ◽  
Early Pregnancy ◽  
Predictive Value ◽  
Transrectal Ultrasonography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Japanese Black Cattle ◽  
Elisa Kit ◽  
Pregnancy Diagnosis

In commercial embryo transfer industry, accurate early detection of pregnant and nonpregnant cows plays a key role in achieving an optimum calving-to-conception interval. The DG29TM enzyme-linked immunosorbent assay (ELISA) kit (Conception, Animal Reproduction Technologies, Beaumont, Canada) measures the level of pregnancy-related glycoproteins in blood that are linked to pregnancy. Here, we compared the DG29 kit with transrectal ultrasonography (TU) to evaluate the possibility of the clinical application of the ELISA kit for early pregnancy diagnosis. Embryos recovered from superovulated Japanese Black cows were transferred to 110 recipient Holstein heifers on Day 6 to 8 of the oestrous cycle (oestrus = Day 0). Pregnancy was diagnosed between Day 29 and Day 40 by TU with a 5.0/7.5-MHz linear array transducer (Tringa, Pie Medical Equipment B.V., Maastricht, the Netherlands). Blood samples were collected from the tail vein or artery into vacuum serum tubes after TU and serum was separated and stored at –30°C until the ELISA was performed. The ELISA results were interpreted as positive (pregnant, >1000 pg mL–1) or negative (nonpregnant, <300 pg mL–1). Ninety-nine of the 110 heifers were judged as pregnant or nonpregnant by TU. Seventy-six of the 99 heifers were judged as pregnant, in which fetuses were visualised clearly in the uterine horn. The following measures, sensitivity, specificity, predictive value and accuracy of pregnancy outcomes based on the ELISA results, were assessed by comparing with those based on the definite TU results. The values for sensitivity and specificity were 100% (76/76) and 91.3% (21/23), the positive and negative predictive value were 97.4% (76/78) and 100% (21/21), respectively and accuracy was 98.0% (97/99). On the other hand, chorioallantoic fluids in 11 of the 110 heifers were detected by TU around Day 30 of gestation, but fetuses were not identified or were unclearly visualised, which suggests that the embryos died during the peri-implantation period after transfer. Ten of the 11 heifers were classified as pregnant by the ELISA, but only 3 heifers were identified as pregnant with reexamination by later TU, which indicates that the pregnancy-related glycoproteins residue from embryo mortality was detected by the DG29 kit. However, the negative predictive value of the DG29 kit was 100% in this study. In conclusion, except for early embryonic death, the DG29 kit was highly accurate and suitable for clinical application in early pregnancy determination following transfer of Japanese Black cattle embryos.

scholarly journals Plasma level and expression of visfatin in the porcine hypothalamus during the estrous cycle and early pregnancy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tadeusz Kaminski ◽  
Marta Kiezun ◽  
Ewa Zaobidna ◽  
Kamil Dobrzyn ◽  
Barbara Wasilewska ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Estrous Cycle ◽  
Early Pregnancy ◽  
Energy Homeostasis ◽  
Plasma Concentrations ◽  
Tissue Samples ◽  
Hormone Synthesis ◽  
Energy Sensor ◽  
Gene And Protein Expression ◽  
Hormonal Milieu

AbstractVisfatin appears to be an energy sensor involved in the regulation of female fertility, which creates a hormonal link integrating the control of energy homeostasis and reproduction. This study evaluates the expression levels of visfatin gene and protein in selected areas of the porcine hypothalamus responsible for gonadotropin-releasing hormone synthesis: the mediobasal hypothalamus (MBH) and preoptic area (POA), and visfatin concentrations in the blood plasma. The tissue samples were harvested from gilts on days 2–3, 10–12, 14–16, and 17–19 of the estrous cycle, and on days 10–11, 12–13, 15–16, 27–28 of pregnancy. Visfatin was localized in the cytoplasm and nucleus of cells creating both studied hypothalamic structures. The study demonstrated that visfatin gene and protein expression in MBH and POA depends on hormonal status related to the phase of the estrous cycle or early pregnancy. Blood plasma concentrations of visfatin during the estrous cycle were higher on days 2–3 in relation to other studied phases of the cycle, while during early pregnancy, the highest visfatin contents were observed on days 12–13. This study demonstrated visfatin expression in the porcine hypothalamus and its dependence on the hormonal milieu related to the estrous cycle and early pregnancy.

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