Term Placenta Tissue Research Articles

Linkage of two human pregnancy-specific beta 1-glycoprotein genes: one is associated with hydatidiform mole.

A genomic clone containing two linked human pregnancy-specific beta 1-glycoprotein (PS beta G) genes has been isolated and characterized. The two genes are arranged in the same 5'----3' orientation; the 3' region (including the A2 and B-C exons) of the upstream gene, PSGGA, is linked to the 5' region (including the 5'/L and L/N exons) of PSGGB, the downstream gene. Depending upon the domains compared, PSGGA and PSGGB share 92-98% nucleotide and 86-95% amino acid sequence identity with PSG93, the most abundant PS beta G transcript. The 3' exon (B-C) of PSGGA contains four alternative splice sites and three polyadenylylation sites, which account for the 3' heterogeneity previously reported in the PS beta G family. Each of the predicted PSGGA-encoded proteins would have a different carboxyl terminus. PSGGB corresponds to the previously identified cDNA PSG6, which encodes proteins containing a 34-amino acid leader peptide and a 108-amino acid N domain, which is one amino acid shorter than the majority of PS beta G N domains. Additionally, the PSGGB-encoded proteins contain the cell-surface recognition tripeptide Arg-Gly-Asp, shared by several previously reported PS beta Gs as deduced from cDNA sequences. Northern blot hybridization performed with a PSGGB-specific oligonucleotide probe to the N domain revealed that PSGGB or a PSGGB-like gene encodes a major 1.7-kilobase mRNA in hydatidiform mole tissues and a major 2.0-kilobase mRNA in term placenta tissues. Moreover, the PSGGB-specific probe hybridized most strongly with mRNA from molar trophoblastic tissue, suggesting that the PSGGB-like species may be the gene preferentially expressed in gestational trophoblastic disease. Additionally, the sequence of a 2315-base-pair PS beta G cDNA (PSG95) that contains an N-A1-A2-B2-C domain arrangement is reported. The coding region of PSG95 is identical to the previously reported cDNA clones PSG1d and FL-NCA, but PSG95 contains an additional 518 and 523 base pairs in the 3' end as compared with PSG1d and FL-NCA, respectively.

Thiamphenicol during the first trimester of human pregnancy: Placental transfer in vivo, placental uptake in vitro, and inhibition of mitochondrial function

Thiamphenicol (TAP) was administered as a single oral dose of 500 mg (8–9 mg/kg) to pregnant women about to undergo therapeutic abortions. The length of gestations varied from 7 to 12 weeks. The concentrations of TAP were determined by electron capture gas chromatography in maternal serum, placenta, amniotic fluid, and various embryonal/fetal tissues when abortion occurred 1.5 to 20 hr after drug administration. TAP was rapidly transferred across the placental membranes and was fairly evenly distributed in the tissues of the embryonal/fetal compartment. The placental concentrations of TAP were usually higher than the corresponding maternal serum levels (1.1 to 7-fold), suggesting some active role of the placenta in TAP uptake. This was confirmed by in vitro studies with immature and term placenta tissue. From a bathing medium containing 10 μg/ml (comparable to therapeutic blood levels) TAP was accumulated in the intracellular compartment to concentrations about 2.6 times those in the medium. This process was time and temperature dependent. Inspite of the low content of mitochondrial particles in human fetal liver the relatively few mitochondria present exhibited high protein synthesis in vitro which decreased with increasing gestational age. Mitochondrial protein synthesis was inhibited by TAP, and a 50% reduction of amino acid incorporation was observed at 10–15 μg TAP/ml. Such levels of the drug are required for successful chemotherapy. To ascertain such therapeutic drug levels, TAP is usually given at 3 g daily (divided, e.g., into three doses 1 g each). The data obtained for a single application of 0.5 g TAP suggest inhibition of mitochondrial protein synthesis during therapy. While the pharmacokinetic characteristics of TAP suggested that this compound might be well suited to treat intrauterine bacterial infections sensitive to TAP, the drug concentrations reached in human embryonal tissues in vivo are known to produce embryolethality in pregnant rats at comparable developmental stages. The findings reported here raise serious concerns about the suitability of TAP for repeated administration to pregnant women during the first trimester of gestation.

THE SUBCELLULAR DISTRIBUTION OF 17β-HYDROXYSTEROID DEHYDROGENASE IN THE HUMAN TERM PLACENTA

ABSTRACT Human term placenta tissue homogenates were subjected to differential centrifugation procedures. The composition of the subcellular fractions was monitored with a number of marker enzymes and the effectiveness of these enzyme systems was evaluated. The subcellular fractions were tested for their 17β-hydroxy dehydrogenase activity on testosterone, oestradiol and the synthetic substrate retrotestosterone. Buffer medium composition showed a direct influence upon enzyme distribution patterns of all fractions during the same differential centrifugation procedure. All enzyme activities tested became less sedimentable when glycerol was present in the fractionation buffer. Glycerol stabilized soluble 17β-hydroxy dehydrogenase activity during fractionation. The activity of steroid-converting enzymes was inhibited by the presence of glycerol in the medium. Subcellular distribution of marker enzymes did not sustain the presence of mitochondrial 17β-hydroxy dehydrogenase but related it to microsomal contamination. In general the proportion of 17β-hydroxy dehydrogenase activity in the particulate fractions showed a decrease in the substrate order retrotestosterone > testosterone > oestradiol which was independent from the buffer medium used. Specific activities for both particle-bound and soluble 17β-hydroxy dehydrogenase increased in the substrate order retrotestosterone < testosterone < oestradiol. The particulate enzyme activity was maximal with NAD+ for the three substrates tested, but in the cytosol fraction NADP+ was the preferential co-enzyme only when oestradiol was used as the substrate.