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PubMed1996

Tissue phospholipids during human pregnancy by 31P NMR: myometrium, decidua, placenta and fetal membranes.

The biophysical environment formed by phospholipids, rather than the amount of functional proteins, can be rate limiting for factors controlling myometrial contractility and pregnancy maintenance. We therefore studied myometrial, decidual, placental and fetal membrane phospholipids using the 31P NMR spectrum. This enabled us to identify bulk phospholipids over 0.05 mmol/kg. The method was checked for reliability for the reproductive tissues studied. The chemical shift of phospholipid standards was slightly different according to whether a single compound or a mixture was analyzed. The bulk phospholipids found were phosphatidylcholine (PC), phosphatidylethanolamine, sphingomyelin (SM) and phosphatidylinositol. The ratio PC/SM decreased during pregnancy in the decidua, placenta and fetal membranes, but not in the myometrium. Pregnancy did not induce significant changes in the total myometrial phospholipids. Their composition was stable even during clinical labor. The fetal tissues, placenta and fetal membranes contained about twice as much phospholipid as the maternal tissues, myometrium and decidua. There was no sign of lysocompounds, cardiolipin or phosphatidic acid. This supports the view that the extraction and analyzing techniques used earlier probably created artefacts. The increased fluidity of the myometrial and placental phospholipids during pregnancy may depend on factors other than the composition of phospholipids.

Decidua↗
PubMed1989

High performance liquid chromatographic measurement of lignocaine in tissue samples following transabdominal placental biopsy.

A method is described for the measurement of lignocaine in small samples of fetal and placental tissue. Tissue samples (ca 100 mg) are digested using a preteolytic enzyme. Lignocaine and an internal standard are extracted into methyl tert-butyl ether and analysed by high-performance liquid chromatography with electrochemical detection (+1.0 V vs Ag/AgCl). The limit of accurate measurement is better than 0.1 mg/kg wet weight for a 100 mg sample. This method has been used to assess fetal exposure to the drug when used as a local anaesthetic during transabdominal placental biopsy (chorionic villus sampling). The range of lignocaine concentrations found in the tissue samples was large (from less than 0.04 mg/kg wet weight to 15.4 mg/kg wet weight) although most samples contained less than 1.0 mg lignocaine/kg wet weight.

Animals↗
PubMed1995

Effect of histamine on endothelial permeability and structure and adhesion molecules of the paracellular junctions of perfused human placental microvessels.

The microvessels of the human placenta resemble those of skeletal muscle, both in endothelial cell junctional organization and in the single passage extraction of radiolabeled tracers. Addition of histamine (100 microM) to the fetal perfusate in an isolated term lobule resulted in a rapid and sustained rise (40-80%) over a 30-min perfusion period in the single circulation extraction values of 57Co-labeled cyanocobalamin and of 51Cr-labeled EDTA, but not of 22Na. Extraction values for 125I-albumin were not increased in histamine-perfused vessels nor was any focal leakage of label observed in serial cryostat sections of lobules perfused with rhodamine-conjugated albumin. There was no electron microscopic evidence of transendothelial channels in the microvascular bed; no gaps were seen at the paracellular cleft regions or in neighboring cytoplasm in any microvessels. Tilting of sections on a goniometric stage showed a significant increase in the separation between adjoining endothelial membrane leaflets at tight junctional regions (from 4.1 to 6.1 nm) although the dimensions of the wide zones remained unchanged. Placental microvessels contain the endothelial adhesion molecules PECAM-1 and VE-cadherin in the wide regions of paracellular clefts: PECAM-1 is also localized on the luminal membrane. Histamine-stimulated microvessels showed an altered staining pattern for both of these molecules, at the expense of the cleft regions. These changes in adhesion molecule distribution and tight junctional membrane separation may be parts of a series of events which leads to increased permeability during inflammatory events.

Antigens, CD↗
PubMed1996

Bioproduction of human enzymes in transgenic tobacco.

Transgenic plants have significant potential in the bioproduction of complex human therapeutic proteins due to ease of genetic manipulation, lack of potential contamination with human pathogens, conservation of eukaryotic cell machinery mediating protein modification, and low cost of biomass production. Tobacco has been used as our initial transgenic system because Agrobacterium-mediated transformation is highly efficient, prolific seed production greatly facilitates biomass scale-up, and development of new "health-positive" uses for tobacco has significant regional support. We have targeted bioproduction of complex recombinant human proteins with commercial potential as human pharmaceuticals. Human protein C (hPC), a highly processed serum protease of the coagulation/anticoagulation cascade, was produced at low levels in transgenic tobacco leaves. Analogous to its processing in mammalian systems, tobacco-synthesized hPC appears to undergo multiple proteolytic cleavages, disulfide bond formation, and N-linked glycosylation. Although tobacco-derived hPC has not yet been tested for all posttranslational modifications or for enzymatic (anticlotting) activity, these results are promising and suggest considerable conservation of protein processing machinery between plants and animals. CropTech researchers have also produced the human lysosomal enzyme glucocerebrosidase (hGC) in transgenic tobacco. This glycoprotein has significant commercial potential as replacement therapy in patients with Gaucher's disease. Regular intravenous administration of modified glucocerebrosidase, derived from human placentae or CHO cells, has proven highly effective in reducing disease manifestations in patients with Gaucher's disease. However, the enzyme is expensive (dubbed the "world's most expensive drug" by the media), making it a dramatic model for evaluating the potential of plants to provide a safe, low-cost source of bioactive human enzymes. Transgenic tobacco plants were generated that contained the human glucocerebrosidase cDNA under the control of an inducible plant promoter. hGC expression was demonstrated in plant extracts by enzyme activity assay and immunologic cross-reactivity with anti-hGC antibodies. Tobacco-synthesized hGC comigrates with human placental-derived hGC during electrophoretic separations, is glycosylated, and, most significantly, is enzymatically active. Although expression levels vary depending on transformant and induction protocol, hGC production of > 1 mg/g fresh weight of leaf tissue has been attained in crude extracts. Our studies provide strong support for the utilization of tobacco for high-level production of active hGC for purification and eventual therapeutic use at potentially much reduced costs. Furthermore, this technology should be directly adaptable to the production of a variety of other complex human proteins of biologic and pharmaceutical interest.

Enzymes↗
PubMed2003

Agaricus blazei (Himematsutake) does not alter the development of rat diethylnitrosamine-initiated hepatic preneoplastic foci.

The modifying potential of crude extracts of the mushroom Agaricus blazei Murrill (Himematsutake) on the development and growth of glutathione S-transferase placental form (GST-P)-positive liver foci (liver preneoplastic lesion) was investigated in adult male Wistar rats. Six groups of animals were used. Groups 2 to 5 were given a single i.p. injection of 200 mg/kg b.w. of diethylnitrosamine (DEN) and groups 1 and 6 were treated with saline at the beginning of the experiment. After 2 weeks, animals of groups 3 to 6 were orally treated with three dose levels of aqueous extracts of the mushroom A. blazei (1.2, 5.6, 11.5, and 11.5 mg/ml of dry weight of solids) for 6 weeks. All animals were subjected to two-thirds partial hepatectomy at week 3 and sacrificed at week 8. Two hours before sacrifice, ten animals of each group were administered a single i.p injection of 100 mg/kg of bromodeoxyuridine (BrdU). Apoptotic bodies and BrdU-positive hepatocyte nuclei were quantified in liver sections stained for hematoxylin and eosin (H&E) (eosinophilic foci) and simultaneously stained for GST-P expression (GST-P-positive foci), respectively. The 6-week treatment with A. blazei did not alter the development (number and size) of GST-P-positive foci and did not affect the growth kinetics of liver normal parenchyma or foci in DEN-initiated animals. Our results indicate that the treatment with aqueous extracts of the mushroom A. blazei during the post-initiation stage of rat liver carcinogenesis does not exert any protective effect against the development of GST-P-positive foci induced by DEN.

Agaricus↗
PubMed2006

Expression profiling of mouse placental lactogen II and its correlative genes using a cDNA microarray analysis in the developmental mouse placenta.

The placenta is a highly differentiated organ essential for embryonic growth and development. In order to search for key molecules that are associated with mouse placental lactogen II (mPL-II) gene expression, we applied mouse cDNA microarray analysis to RNAs extracted from placentae on days 10, 12, 14, 16 and 18 of pregnancy. Changes in gene expression were categorized between days 10 and 12, 12 and 14, 14 and 16 and 16 and 18 of pregnancy. After microarray analysis, which had a minimum detectable fold change for differential expression of 2, we selected 10 genes, Apoa2, Apoc2, Ceacam14, Creg1, Fmo1, Igf2, Slc2a1, Spink3, Spi1-1 and Tpbpa, exhibiting a expression pattern similar to the mPL-II gene. Furthermore, we performed real-time PCR analysis and in situ hybridization (ISH) to find correlative expression genes for the mPL-II gene. From these results, we identified a resemblance in gene expression between mPL-II and Igf2 and selected these genes for performance of double-fluorescence immunohistochemical staining. We colocalized these proteins in labyrinthine trophoblast cells. These results strongly suggest that the expression of mPL-II and Igf2 is highly related to placental development in mice. This large-scale identification of genes regulated during placentogenesis assists in further elucidation of the molecular basis of extraembryonic development and function.

Animals↗
PubMed1980

Unusual patterns of benzo[a]pyrene metabolites and DNA-benzo[a]pyrene adducts produced by human placental microsomes in vitro.

Human placental microsomes were incubated with [3H]benzo[a]pyrene (BP) and Salmon sperm DNA and the resulting metabolite-nucleoside complexes resolved by Sephadex LH-20 chromatography. The metabolite pattern was analyzed by high-pressure liquid chromatography (HPLC). The incubates were also co-chromatographed with extracts obtained from incubates with rat liver microsomes and [14C] BP. Phenols, quinones and 7,8-dihydrodiol were detected in the placental incubates. Both 9,10- and 4,5-dihydrodiols were very low as compared with control rat liver samples. Placental microsomes catalyzed the binding of BP metabolites to DNA in vitro, giving rise to two main complexes which co-chromatographed with rat liver-produced peaks attributable to 7,8-diol-9,10-epoxide and 7,8-oxide and/or quinones when metabolized further. The nucleoside metabolite peaks attributable to 4,5-oxide and 9-phenol-4,5-oxide were lacking when compared with the binding pattern catalyzed by rat liver. Both the total binding and specific metabolite-nucleoside adducts in the placenta correlated with fluorometrically measured aryl hydrocarbon hydroxylase (AHH) activity and with the amount of dihydrodiol formed. The results demonstrate that both the metabolite pattern and the nucleoside-metabolite complexes formed by the placental microsomes in vitro differed greatly from thos produced by rat liver microsomes. These sstudies also suggest that it is not possible to predict specific patterns of DNA binding from AHH measurements or even from BP metabolite patterns, especially when comparing different tissues and species.

Aryl Hydrocarbon Hydroxylases↗
PubMed1987

Nuclear acceptor sites for progesterone-receptor complexes in rat placenta.

A cell-free system was used to characterize the binding reaction between the progesterone receptor and nuclear acceptor sites prepared from rat placenta. Two forms of receptor-acceptor complex were examined. One was extracted from nuclei by exposure to 0.6 M KCl; the other type was resistant to salt extraction. Kinetic analysis indicated that the binding reactions were saturable (3-4 pmol binding sites/mg DNA) and of high affinity (Kd = 3-6 nM). Acceptor binding was specific for placental nuclei and did not occur with nuclei prepared from spleen or with denatured nuclei from placenta. Acceptor sites were further characterized by their sensitivity to RNase, DNase I, and protease. RNase treatment had no influence on receptor-acceptor binding. However, DNase I reduced the number of KCl-resistant acceptor sites by 41%, but only a 19% reduction occurred in KCl-extractable acceptor sites (P less than 0.05). Protease removed 34% and 48% of the KCl-resistant and -extractable acceptor sites, respectively, and combined treatment with DNase and protease eliminated 76% of acceptor-binding activity. The endogenous inhibitor previously described from rat placental cytosol blocked acceptor-binding sites in a concentration-dependent manner, a decrease of 1.15 pmol sites/mg inhibitor protein for resistant sites and 0.76 pmol/mg inhibitor protein for extractable sites. However, receptor-acceptor binding was not altered by treating nuclei with actinomycin D or chloroquine. Mercurial reagents reduced receptor-acceptor interaction by 80% and 94% in KCl-resistant and -extractable sites, respectively, whereas sulfhydryl alkylating agents reduced binding 35% and 76%. Pyridoxal phosphate destroyed 88-93% of acceptor binding. The results of these studies suggest that the progesterone receptor acceptor sites are composed of a complex of chromatin protein and DNA in rat placenta. Furthermore, the binding reaction requires the participation of sulfhydryl and terminal amino groups.

Animals↗
PubMed1978

The placental transfer and materno-fetal disposition of methadone in monkeys.

A method is described for the quantitation of levo-methadone[3H] in biological samples which involves sample extraction and thin-layer chromatographic separation. Four pregnant Macaca mulata monkeys, two in early gestation and two in late gestation, were given single i.m. injections of levo-methadone[3H]. Twenty-nine fetal and maternal tissues and fluids were assayed to provide quantities of unchanged methadone and methadone plus metabolites. Little placental transfer of methadone or its metabolites occurred during early gestation, but equivalent concentrations of unchanged methadone were found in maternal and fetal tissues during late gestation (maternal brain, 172 ng/g; fetal brain, 123 ng/g). With few exceptions, tissues or fluids from the late gestation mothers showed higher levels of unchanged methadone than those from early gestation mothers at both 1 hour (P less than .001) and 6 hours (P less than .010) after administration. The late gestation mother had a 40.2% greater concentration of unchanged methadone at 6 hours and a 50.1% greater concentration at 1 hour than the early gestation mothers. These data suggest a slowing of metabolism during advanced pregnancy. At 6 hours after administration the eyes of both early and late gestation mothers and late gestation fetuses showed the highest concentrations of unchanged methadone of any maternal or fetal tissue. This localization of methadone appears to be associated with pigmented epithelium.

Animals↗
PubMed1987

Subunit and functional size of human placental DNA methyltransferase involved in de novo and maintenance methylation.

The subunit molecular size of human DNA methyltransferase isolated from nuclear extracts of placenta was determined on the electroblotted polypeptides after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and compared with the functional size by high performance size exclusion chromatography on Superose 12 and gamma radiation inactivation analysis. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis results indicated a subunit mass of 120 +/- 10 kDa, while the functional size data indicates that the enzyme operates both in de novo and maintenance modes as a dimer of molecular mass 220 +/- 15 kDa with no evidence of monomers in solution of ionic strength between 0.1 and 0.8 M NaCl. The 220-kDa activity carried out the transmethylation of both hemi- and unmethylated DNA substrates. There was no evidence for separate functional catalytic sites on each monomer subunit acting independently when engaged in methylation of hemimethylated or single-stranded DNA from the invariance of radiation inactivation target size with these substrates. The radiation inactivation target size was 230 +/- 15 kDa.

DNA (Cytosine-5-)-Methyltransferases↗
PubMed2000

Familial cryptic translocation with del 4q34-->qter and dup 12pter-->p13 in sibs with tracheal stenosis: clinical, classical and molecular cytogenetic studies and CGH analyses from archival placental tissues evidencing tertiary trisomy 4 in one abortion specimen.

We report on two retarded half-sibs of different sex and seemingly normal karyotype who had the same syndrome of minor anomalies, heart defect and a distal tracheal stenosis, and who shared a healthy mother. These findings raised suspicions of a cryptic chromosome translocation. A translocation t(4;12)(q34;p13), balanced in the mother and unbalanced in the sibs with loss of terminal 4q and gain of terminal 12p regions, was verified by FISH using whole chromosome painting, subtelomeric and YAC probes. Clinical features could be explained by partial monosomy 4q and partial trisomy 12p. Tracheal stenosis was interpreted as a consequence of the same developmental disturbance leading to esophageal atresia and tracheo-esophageal fistula. It was attributed to the 4q deletion in which esophageal atresia as also respiratory difficulties and airway obstructions had been described. Paraffin-embedded placental tissues were available from three of the five abortions of the mother allowing DNA extraction and comparative genome hybridization (CGH). Two of the abortion specimens had the same der(4)t(4;12)(q34;p13) unbalanced translocation as identified in the sibs. In the third abortion specimen, suspicious of triploidy because of partial hydatidiform mole, CGH uncovered a tertiary trisomy 4 resulting from a 3:1 segregation of the translocation chromosomes and their homologs during maternal meiosis I. Differences in CGH results using DNA generated directly or after DOP-PCR were explained by DNA fragmentation in paraffin-embedded tissues and unequal amplification. Am. J. Med. Genet. 94:271-280, 2000.

Adult↗
PubMed2006

An introduction to mass spectrometry based proteomics-detection and characterization of gonadotropins and related molecules.

This review introduces fundamental aspects of mass spectrometry (MS) based proteomics and illustrates how MS is an effective tool for the analysis of glycoprotein hormones. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) and electrospray ionization (ESI) MS are complementary approaches that have been applied for the analysis of gonadotropins, e.g. to characterize differences in the oligosaccharide distribution of commercial human chorionic gonadotropin preparations, for isolated nicked beta-subunit, and identification of a metabolite of placental transforming growth factor in pharmaceutical hCG preparations. Immunoaffinity trapping and concentration of digested sample extract prior to MS analysis confers analytical sensitivity akin to immunoassay. A desirable objective would be to develop for clinical purposes a rapid procedure for MS detection and characterization of gonadotropins. Refinement of on-target immobilization and digestion for subsequent ionization by MALDI may eventually help to provide this capability. The advent of hybrid mass spectrometers will further advance the characterization of these complex molecules.

Animals↗
PubMed1982

Effect of maternal serum insulin on umbilical extraction of glucose and lactate in fed and fasted sheep.

In 18 chronically instrumented pregnant ewes in late gestation, umbilical extractions of glucose and lactate were determined before and during the continuous infusion of ovine insulin, 0.25 mU/kg . min, into one uterine artery. Studies were conducted in both well-nourished and fasting ewes. Exogenous insulin increased the umbilical extraction of glucose without altering the umbilical extraction of lactate over the range of concentrations of maternal blood glucose encountered in the fed and fasted states. Moreover, the efflux of lactate into the uterine venous circulation in two additional ewes was not altered by the administration of insulin. These studies support the hypothesis that maternal circulating insulin binds to insulin receptors on the microvillous brush border of the placenta to effect an increase in the carrier-mediated transfer of glucose rather than cause an intraplacental decrease in the catabolism of glucose to lactate. The results suggest that, as maternal levels of blood glucose and serum insulin spontaneously rise in concert over the physiologic range, both factors may contribute to the increasing umbilical extraction of glucose. Furthermore, these observations raise the possibility that decreased binding of insulin by placental insulin receptors, which is reported to occur in placentas from diabetic women, may be accompanied by a relatively decreased umbilical uptake of glucose for a given maternal concentration of glucose, but not of lactate.

Animals↗
PubMed1999

Globoside expression within the human placenta.

This report demonstrates the presence of the neutral glycosphingolipid, globoside, on the villous trophoblast layer of human placenta. Immunoreactivity for globoside which is the receptor used by human parvovirus B19 was strongest in villous trophoblast cells of first trimester placentae, with diminished reactivity in second trimester placentae, and a near lack of staining for the antigen in those of third trimester. This relative reduction in globoside-specific immunoreactivity in placentae of increasing gestational ages was confirmed using thin-layer chromatographic analyses of extracted neutral glycolipids from the syncytiotrophoblast layer and cytotrophoblast cells of first and third trimester placental villi. The presence of globoside on the protective trophoblast layer of the villi provides a potential pathway whereby B19 may be transmitted from an infected mother to the fetus. The virus once across the placental barrier, may gain access to its erythroid precursor target cells within fetal villus capillaries. The observed change found in globoside immunoreactivity correlates well with the observation that fetal outcome is worse when maternal infection occurs during first or second trimester as compared to an infection occurring near term. The reason for this observed difference in fetal outcome may be due not only to the presence of more target cells potentially to infect during the first and second trimesters, but also to the greater number of viral receptors present on the villous trophoblast layer.

Chromatography, Thin Layer↗
PubMed2005

Individual identification by DNA polymorphism using formalin-fixed placenta with whole genome amplification.

Polymerase chain reaction (PCR) amplification using formalin-fixed material is very limited. In the present study the use of 6 week formalin-fixed placenta for individual identification was examined based on DNA analyses. The objective of the examination was to prove whether the placenta was from a woman who had just given birth. DNA extraction was carried out from the maternal blood sample and from the formalin-fixed placental samples composed of three parts: maternal side, infant side and umbilical cord. One minisatellite (D1S80), 12 short tandem repeat (STR) polymorphisms and amelogenin X, Y were investigated. All the polymorphic systems were detected in the maternal blood sample. The majority of the DNA isolated from the placental tissues had molecular weights of approximately 500 bp, and only two to four STR loci were amplified using the DNA. In order to amplify more DNA polymorphic markers from the formalin-fixed tissues, whole genome amplification was performed. After amplification by degenerate oligonucleotide-primed PCR (DOP-PCR), the products contained DNA with increased molecular weight up to >10 kbp. More DNA loci were typed using the DOP-PCR products. Furthermore, large molecular size fragments were purified from the DOP-PCR products by agarose electrophoresis, and then the D1S80 locus and 12 STR loci were successfully amplified using these fragments.

Adult↗
PubMed1984

Placental protein 12 (PP12) in the human endometrium: tissue concentration in relation to histology and serum levels of PP12, progesterone and oestradiol.

Placental protein 12 (PP12) was measured by radio-immuno-assay in serum and endometrial extracts of 25 women at various phases of the menstrual cycle, and in the decidua from six women undergoing legal abortion in early pregnancy. The results were analysed with respect to endometrial histology and serum oestradiol, progesterone and PP12 concentrations. PP12 was detected in the secretory phase endometrium and decidua, but not in the proliferative phase endometrium. The PP12 concentration in tissue increased from early (1.3-9.4 micrograms/g protein) to late secretory phase (5.3-155 micrograms/g), and considerably higher levels were found in decidua (282-3846 micrograms/g). There was a correlation (P less than 0.001) between endometrial PP12 and serum progesterone levels in non-pregnant women, and there also was a slight correlation (P less than 0.05) between endometrial and serum PP12 concentrations. These results show that PP12 is not pregnancy specific and suggest that the occurrence of PP12 is dependent on progesterone.

Adult↗
PubMed1992

Interleukin-1 is induced in the human placenta by endotoxin and isolation procedures for trophoblasts.

Although freshly obtained placenta contains little or no interleukin-1 (IL-1) mRNA, placenta and isolated trophoblast have been reported to produce significant quantities of bioactive IL-1 in vitro. The present study was designed to determine if endotoxin, a common contaminant of culture medium, and trophoblast isolation procedures could induce IL-1 expression in the placenta. Tissue-extractable IL-1 alpha and IL-1 beta immunoreactive proteins were readily detected in fresh placental membranes, but not placental villi. As little as 10 ng/mL endotoxin were found to induce the expression of IL-1 alpha and IL-1 beta mRNA in intact placental villi cultured in vitro. Intact placenta cultured in the presence of 1.0 microgram/mL endotoxin demonstrated expression of IL-1 alpha and IL-1 beta mRNA and cumulative production and release of immunoreactive IL-1 alpha and IL-1 beta into the medium during 24 h of culture. Placenta incubated in endotoxin-free medium, however, exhibited no detectable IL-1 alpha or IL-1 beta mRNA expression and little or no release of IL-1 alpha or IL-1 beta immunoreactive protein into the medium. When trophoblast cells were freshly isolated by enzymatic digestion, followed by Percoll separation at reduced temperatures to inhibit cell activation, no IL-1 alpha or IL-1 beta mRNA expression was initially detectable. However, IL-1 alpha and IL-1 beta mRNA in isolated trophoblast cells were induced after 30 min of culture in endotoxin-free medium, with maximal induction at 4 h. These results suggest that normally the placenta produces very little, if any, IL-1, and that endotoxin and trophoblast isolation procedures induce IL-1 expression in placental tissues cultured in vitro.

Cell Separation↗
PubMed1982

[Monodeiodination of thyroxine to 3,3',5'-triiodothyronine in the human placenta].

We investigated the characteristics of the monodeiodination of thyroxine to T3 and rT3 in human placentas which were obtained at normal delivery. The placentas were homogenized in a cold sucrose Tris-HCl buffer, pH 7.5. The microsomal fraction was incubated at 37 degrees C in air for 1 hr with 2 micrograms of T4 in the presence of 50mM DTT. The T3 and rT3 generated in the reaction mixture were extracted into cold ethanol and measured by RIA. Among the usal subcellular fractions of the placental homogenate, microsomes were the most potent in deiodinating T4 to rT3. In microsomes, production of rT3 increased with protein concentration, incubation temperature up to 37 degrees C, incubation time up to 120 min and T4 concentration up to 16 micrograms/tube. The production of rT3 from T4 was lost by prior heating of the microsomal fraction to 56 degrees C for 30 min. The net production rate of T4 to rT3 in the microsomal fraction was 17.9 ng/mg protein/micrograms T4/60 min at pH 7.5. RT3 production from T4 was maximal at pH 7.0. The production of T3 from T4 was negligible in the present system. Degradation of T3 in the placentas was rapid. Although the addition of anti-T3 antibody to the reaction mixture suppressed the degradation of T3, it had no effect on the net production of T3, suggesting that the obtained net T3 production rate had not been influenced by its degradation. Degradation of rT3 was negligible. These results indicate that the human placenta actively deiodinates T4 to rT3 enzymatically. This enzyme system might have some influence on the transplacental passage of the thyroid hormone from the mother to the fetus.

Female↗