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See detailValidation of a new pregnancy-associated glycoprotein radioimmunoassay method for the detection of early pregnancy in ewes.
Barbato, Olimpia; Melo de Sousa, Noelita ULg; Debenedetti, A. et al

in Theriogenology (2009), 72

The aim of the current study was to describe the use of a pool of different antisera raised against pregnancy-associated glycoproteins (PAGs; purified from both ovine and caprine placentas) for early ... [more ▼]

The aim of the current study was to describe the use of a pool of different antisera raised against pregnancy-associated glycoproteins (PAGs; purified from both ovine and caprine placentas) for early pregnancy diagnosis in ovine species. Sixty-three pluriparous Sarda ewes (Ovis aries) were synchronized. Blood samples were withdrawn on Days 18, 24, 26, 28, 30, and 50 after mating. These samples were assayed for progesterone (radioimmunoassay [RIA] including an extraction step) and for pregnancy-associated glycoproteins (RIA-706 and RIA-srPool). Progesterone concentrations were under 1.0ng/mL in all nonpregnant Sarda ewes. In pregnant ewes, mean progesterone concentrations ranged from 2.4ng/mL (Day 24, single pregnancies) to 4.4ng/mL (Day 28, multiple pregnancies). During all periods of examination, PAGs remained lower than 0.8ng/mL in nonpregnant ewes. On Day 18 of pregnancy, PAG concentrations could be detected in 26 of 43 (60.5%) and in 41 of 43 (95.3%) pregnant ewes using the RIA-706 and RIA-srPool methods, respectively. From Day 24 to Day 50, using both RIA methods, PAGs could be detected in all pregnant ewes. On Day 24, the best threshold for pregnancy diagnosis was obtained by use of RIA-srPool, maximal concentration in nonpregnant ewes being 0.3ng/mL and minimal concentration in pregnant ewes being 4.8ng/mL. In general, progesterone and PAG concentrations were higher in multiple pregnancies than in single pregnancies. However, because of large individual variations, single pregnancies could not be differentiated from multiple pregnancies [less ▲]

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See detailConcentrations of pregnancy-associated glycoproteins in Water buffaloes females (Bubalus bubalis) during pregnancy and postpartum periods
Barbato, O.; Melo de Sousa, Noelita ULg; Malfatti, A. et al

in O’Leary, M.; Arnett, J. (Eds.) Pregnancy Protein Research (2009)

The concentrations of pregnancy-associated glycoproteins (PAG) were determined in buffalo cows (Bubalus bubalis) using three different radioimmunoassay (RIA) systems (RIA-497, RIA-706, and RIA-708 ... [more ▼]

The concentrations of pregnancy-associated glycoproteins (PAG) were determined in buffalo cows (Bubalus bubalis) using three different radioimmunoassay (RIA) systems (RIA-497, RIA-706, and RIA-708). Samples (10 mL) were collected by jugular venipuncture from Week 0 until Week 28 of pregnancy (9 females), and from parturition until Week 10 postpartum (13 females). During pregnancy, PAG molecules were detectable at Week 6 using the three aforementioned RIA systems (3.9 ± 1.3 ng/mL, 9.7 ± 1.3 ng/mL, and 9.9 ± 0.7 ng/mL for RIA-497, RIA-706, and RIA-708, respectively). These concentrations increased gradually until Week 28, reaching values of 39.6 ± 4.0 ng/mL (RIA-497), 50.5 ± 11.9 ng/mL (RIA-706), and 68.2 ± 20.8 ng/mL (RIA-708). PAG concentrations determined by RIA-497, RIA-706, and RIA-708 were strongly correlated throughout the entire gestation period, RIA-708 giving the higher concentrations. At parturition, the mean concentrations ranged from 34.9 ± 4.0 (RIA-497) to 84.7 ± 10.6 ng/mL (RIA-708). Thereafter, the concentrations decreased steadily, reaching very low levels (< 1.0 ng/mL) at Week 8 postpartum. In conclusion, PAG concentrations measured by the above three RIA systems showed a profile similar to those previously described in bovine species, with higher concentrations being detected by RIA-706 and RIA-708. However, the patterns of PAG concentrations, studied using the three aforementioned PAG-RIA systems, differed around parturition, with very low levels being observed in the female buffaloes [less ▲]

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See detailIsolation of new pregnancy-associated glycoproteins from water buffalo (Bubalus bubalis) placenta by Vicia villosa affinity chromatography.
Barbato, O.; Melo de Sousa, Noelita ULg; Klisch, K. et al

in Research in Veterinary Science (2008), 85(3), 457-66

The present study describes the isolation and characterization of new pregnancy-associated glycoprotein molecules (PAG) from midpregnancy and late-pregnancy placentas in the water buffalo (Bubalus bubalis ... [more ▼]

The present study describes the isolation and characterization of new pregnancy-associated glycoprotein molecules (PAG) from midpregnancy and late-pregnancy placentas in the water buffalo (Bubalus bubalis). After extraction, the homogenates are subjected to acid and ammonium sulfate precipitations followed by DEAE chromatography. Subsequently, the water buffalo PAG (wbPAG) from these solutions are enriched by Vicia villosa agarose (VVA) affinity chromatography. As determined by western blotting with anti-PAG sera, the apparent molecular masses of the immunoreactive bands from the VVA peaks range from 59.5 to 75.8kDa and from 57.8 to 73.3kDa in the midpregnancy and late-pregnancy placentas, respectively. Amino-terminal microsequencing of the immunoreactive proteins has allowed the identification of three distinct wbPAG sequences, which have been deposited in the SwissProt database: RGSXLTIHPLRNIRDFFYVG (acc. no. P85048), RGSXLTILPLRNIID (acc. no. P85049), and RGSXLTHLPLRNI (acc. no. P85050). Their comparison to previously identified proteins has shown that two of them are new because they have not been described before. Our results confirm the suitability of VVA chromatography for the enrichment of the multiple PAG molecules expressed in buffalo placenta. [less ▲]

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See detailPregnancy-Associated Glycoprotein 62 [Fragment] from Bubalus bubalis placenta - Access number P85050
Barbato, O.; Melo de Sousa, Noelita ULg; Debenedetti, A. et al

E-print/Working paper (2007)

N-terminal microsequence obtained after purification and characterization of placental proteins in Bubalus bubalis. Proteins were submitted to SwissProt databank.

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See detailPregnancy-Associated Glycoprotein 70 [Fragment] from Bubalus bubalis placenta - Access number P85049
Barbato, O.; Melo de Sousa, Noelita ULg; Debenedetti, A. et al

E-print/Working paper (2007)

N-terminal microsequence obtained after purification and characterization of placental proteins in Bubalus bubalis. Proteins were submitted to SwissProt databank.

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See detailPregnancy-Associated Glycoprotein 75 [Fragment] from Bubalus bubalis placenta - Access number P85048
Barbato, O.; Melo de Sousa, Noelita ULg; Debenedetti, A. et al

E-print/Working paper (2007)

N-terminal microsequence obtained after purification and characterization of placental proteins in Bubalus bubalis. Proteins were submitted to SwissProt databank.

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See detailThe Pregnancy-Associated Glycoproteins: Biochemical Aspects and Clinical Application for Pregnancy Follow-up
Melo de Sousa, Noelita ULg; Barbato, O.; Bella, Amina ULg et al

Conference (2007)

Pregnancy diagnosis is an important part in reproduction management of ruminants. In the last years, a large polymorphic family of placenta-expressed proteins has been discovered in ruminant species and ... [more ▼]

Pregnancy diagnosis is an important part in reproduction management of ruminants. In the last years, a large polymorphic family of placenta-expressed proteins has been discovered in ruminant species and used for pregnancy diagnosis. Members of this family are named pregnancy-associated glycoproteins (PAG), being synthesized in the mono- and binucleate cells of the ruminant’s trophectoderm. Part of them are released in the maternal blood circulation where they can be assayed by different laboratory techniques. Due to large variety of expressed molecules and to large variations in the post-translational processing of the PAG, different immuno-systems present different ability to quantify the PAG released in blood. The sensitivity (92 to 100%) and specificity of PAG radioimmunoassay when used for pregnancy diagnosis are very high. The assay of PAG can also bring very interesting information for researchers working in programs focused on the study of embryonic and fetal mortalities, as well as on embryo biotechnology (ET, FIV, clonage), animal nutrition, or infections diseases resulting in pathologies affecting the pregnancy. [less ▲]

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See detailIsolation of PAG from buffalo (Bubalus bubalis) placenta
Barbato, O.; Melo de Sousa, Noelita ULg; Klisch, K. et al

in Reproduction (Cambridge, England). Abstract Series (2006), Suppl

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See detailPurificazione delle proteine associate alla gestazione (PAG) nella bufala (Bubalus bubalis): risultati preliminari.
Barbato, O.; Melo de Sousa, Noelita ULg; Malfatti, A. et al

in Atti 2o Congresso Nazionale sull’Allevamento del Bufalo (2003)

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See detailPregnancy-associated glycoprotein detection by RIA in pregnant buffalo cows (Bubalus bubalis)
Debenedetti, A.; Melo de Sousa, Noelita ULg; Sulon, Joseph ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2001), 5(suppl), 62

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