Biomedical subjects

M Majewska

Publications and source records attributed to M Majewska.

At least 19 recordsLinked to original sources

PubMed2006

Radiocompetition of secretory pregnancy-associated glycoproteins as chorionic ligands with luteal and uterine gonadotrophin receptors of pregnant pigs.

Porcine pregnancy-associated glycoprotein (pPAG) family is very promiscuous and its role(s) remains unknown. The objective of this study was to identify whether secretory placental proteins (including pPAGs), produced in vitro by porcine chorionic explants, may interact with other proteins/targets, i.e. luteal and uterine binding sites of pregnant pigs. Trophoblast (TRF) and trophectoderm (TRD) were harvested during peri-implantation and placentation periods (14-61 dpc-day post coitum). In vitro-produced TRF/TRD proteins were isolated from media by ultrafractionation (>10 kDa MWCO) or precipitation with 20-75% saturation of (NH(4))(2)SO(4) and pPAG proteins were monitored by Western blotting. Secretory TRF/TRD ligands (including PAGs) were serially diluted (0.78-25 microg/ligand) and examined by radioreceptor assay (RRA). Luteal and uterine membrane receptors of pregnant pigs (pRc) were isolated from corpora lutea (pCLRc), myometrium (pMYORc) and endometrium (pENDRc). The three pRc types were harvested during three periods of pregnancy: 14 dpc (14 Rc), 21-26 dpc (21-26 Rc) and 31 dpc (31 Rc). The RRA competitions of individual TRF or TRD ligands were performed with (125)I-hCG as tracer and different pRc types. The RRA results of TRF/TRD were compared to hCG/pLH ligands--as positive controls (0.39-50 ng/ml), and endometrial (END) proteins (0.78-25 microg/ml) produced in vitro by END explants of cyclic, pseudopregnant and pregnant gilts (cEND, PsEND and pEND, respectively)--as negative control ligands. Results indicated that secretory TRF/TRD proteins (+pPAGs) were able to compete with (125)I-hCG for binding with other proteins/targets, i.e. luteal and uterine receptors of pregnant pigs (pCLRc, pMYORc and pENDRc) in a concentration- and pregnancy stage-dependent manner. This study indicated that porcine secretory 14-15 dpc TRF (pPAG; 30-73 kDa) ligands, effectively displaced (125)I-hCG tracer from pCL14Rc (up to P< or =0.01), corresponding to displacement by hCG and porcine LH. During the early stage of pregnancy, some competition tendency (P< or =0.01) was also detected for TRF ligands (14-15 dpc) with pEND14Rc. As pregnancy advanced, significant (125)I-hCG competition (at least P< or =0.05) with secretory semi-purified TRD ligands (30-42 dpc) was determined for all types of examined receptors pCL31Rc, pMYO31Rc and pEND31Rc, mainly with TRD fractions precipitated by 20% saturation of (NH(4))(2)SO(4). It seems that chorionic pPAG family can be involved in luteoprotective mechanism during implantation and placentation, according to the binding-interaction with luteal and uterine gonadotropin receptors of pregnant pigs.

Animals↗

PubMed2006

Pregnancy-associated glycoprotein family (PAG)--as chorionic signaling ligands for gonadotropin receptors of cyclic animals.

The chorionic pregnancy-associated glycoprotein (PAG) family was identified in pigs, cattle and other eutherian mammals. The objective of this study was to examine whether secretory chorionic proteins (including PAGs), produced in vitro by explants of porcine and bovine placental membranes, may interact with other proteins, i.e. gonadal and extragonadal binding sites. Trophoblast (TRF) and trophectoderm (TRD) explants of pigs (n=38; 14-61 dpc-day post coitum) or cotyledons (CT) of cows (n=5; 40-110 dpc) were long-term cultured. Released chorionic proteins were ultra-fractionated from media (>10 kDa) or precipitated [20-75% of (NH(4))(2)SO(4)]. The PAGs were monitored by Western/PAGE (30-73 kDa). Secretory TRF/TRD/CT (+PAG) proteins (0.78-25 microg/ligand) were examined by radioreceptor assay (RRA) with iodinated hCG ((125)I-hCG) for binding-effectiveness by gonadotropin receptors of cyclic pigs and cows (cRc). Gonadal and extragonadal cRc isolated from luteal-phase corpora lutea and uteri (cCLRc, cMYORc and cENDRc) were tested with positive control ligands: porcine LH and hCG (0.39-50 ng/ml). Control proteins produced in vitro by endometrial (END) explants of cyclic (cEND), pseudopregnant (PsEND) and pregnant (pEND) gilts were utilised as negative ligands (0.78-25 microg/ligand). Positive control ligands competed with (125)I-hCG for binding by cCLRc, cMYORc and cENDRc (18-61%/B(0) for hCG and 27-57%/B(0) for LH). Negative ligands (cEND, PsEND and pEND) did not show cRc bindings. This is the first RRA report indicating that in vitro produced porcine TRF/TRD proteins (+PAG) competed (P< or =0.05) with (125)I-hCG for binding by cCLRc, cMYORc and cENDRc in a concentration- and pregnancy stage-dependent manner. The highest competition with (125)I-hCG (up to P< or =0.001) was found for ultra-fractionated TRF/TRD proteins (>10 kDa) during early pregnancy (<22 dpc). The greatest competition (P< or =0.05) of precipitated porcine TRD proteins (>30 dpc) was detected for fractions obtained by saturation with use of 20% of (NH(4))(2)SO(4). Bovine CT proteins revealed lower competition of (125)I-hCG for bovine cCLRc (during 45 dpc only) that was more efficient with CT (up to 71%) than with non-labelled hCG (82%). The PAG proteins may play a role as potential "signal molecules", because they were able to interact with gonadotropin receptors of luteal-phase animals. It seems that the pPAG proteins may be luteoprotective chorionic-origin signals during implantation and placentation, according to binding-effectiveness of the chorionic ligands that was comparable to LH/hCG ligands with gonadal and extragonadal receptors of cyclic animals.

Animals↗

PubMed2005

Chorionic mRNA expression and N-glycodiversity of pregnancy-associated glycoprotein family (PAG) of the European bison (Bison bonasus).

Placental PAG mRNA expression and N-glycodiversity of multiple PAG proteins secreted in vitro by trophectoderm (chorion epithelium) of wild pecoran Bovidae taxons was not examined previously. The study on European bison (Eb) aimed: (1) to determine placental PAG mRNA expression by in situ hybridisation; (2) to identify a profile of pecoran PAG protein family secreted in vitro by cotyledonary (CT) explants; (3) to examine N-glycodiversity of the PAG proteins in this wild taxon. In addition, we compared (4) a profile and N-glycodiversity of the PAG protein family secreted in vitro by CT and interCT-trophectoderm (intCT-TRD) explants of domestic ruminants. Cotyledonary sections of the Eb were used for in situ hybridisation (ISH) with (35)S-labelled probes produced with porcine PAG cDNA as templates. Various CT and intCT-TRD explants were long-term cultured in vitro. Chorionic proteins were isolated from media, ultra-filtrated (>10 kDa MWCO) and analysed by PAGE-Western blotting with various polyclonal anti-PAG sera. Protein samples with or without enzymatic deglycosylation were examined after different times of explant cultures. Released chorionic proteins were deglycosylated by N-glycanase F (PNGase F+) and compared to glycosylated forms (PNGase F-). This is the first paper demonstrating the PAG-like mRNA transcript expression (by ISH) and N-glycodiversity of immuno-reactive PAG-like proteins (produced in vitro by chorionic explants) of European bison. Various PAG proteins of Eb (EbPAG) were secreted by CT explants during long-term in vitro studies. Major approximately 78, approximately 67 and approximately 65 kDa EbPAG-like proteins were reduced by enzymatic deglycosylation (at least by 10 kDa). Considerably smaller amounts of approximately 45 kDa EbPAG-like proteins were also observed. In addition, we have found that various PAG proteins (30-73 kDa) were secreted by bovine CT explants, during long-term in vitro cultures. Corresponding amounts of PAG proteins, similar in M(r), were also secreted by intCT-TRD explants, whose tissues were not utilised for PAG protein extraction during other scientists' previous studies. It seems that the M(r)-heterogeneity and N-glycodiversity of the PAG protein family can play very important role during feto-placental interactions in Bovidae species.

Animals↗

PubMed1991

Comparative studies of serotype-specific Clostridium difficile strains.

The following properties of serotype-specific Clostridium difficile strains were studied: toxigenicity, encapsulation, susceptibility to certain antibiotics, biochemical properties, enzymatic activity. No correlation between toxin titer and frequency of capsule production as well as serogroup affiliation and sensitivity to antibiotics was observed. The strain representative of serogroup C attracts attention because of its distinct properties.

Clostridioides difficile↗

PubMed1978