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Association between Breus' mole and partial hydatidiform mole: chance or can hydropic villi precipitate placental massive subchorionic thrombosis?

Breus' mole (massive subchorionic hematoma) is a rare entity most often found in the placentae of macerated stillborn fetuses. Previously considered to represent a postmortem event, recent evidence suggests that it occurs prior to fetal demise. A 23-week gestation male neonate was delivered of a 23-year-old gravida 3, para 2 woman and survived for 49 min. An autopsy with chromosomal studies resulted in a diagnosis of triploidy. Placental examination showed the presence of both Breus' mole and also partial hydatidiform mole. DNA samples extracted from portions of the fresh hematoma and from the fetal spleen were compared using molecular techniques. PCR analysis showed the presence of Y chromosome specific DNA in the placental clot, but a semiquantitative Southern blot demonstrated that roughly 85% of the clot DNA was of maternal origin. These findings suggest that Breus' mole represents primarily maternal thrombosis rather than fetal hemorrhage. We hypothesize that the partial mole could have contributed to the formation of the Breus' mole as some of the hydropic villi may have focally obstructed the maternal venous return from the intervillus space causing sluggish flow and promoting thrombosis. A review of the literature on Breus' mole shows that the majority of reported cases have not included cytogenetic findings. However, several authors have reported an association with triploidy and other chromosomal anomalies characterized by scattered placental hydropic villi. Thus, we suggest that obstruction of maternal venous return by hydropic villi may have played a contributory role in some of these other reported cases.

Adult↗

Measurement of nucleoside kinases in crude tissue extracts.

The measurements of deoxyadenosine kinase, adenosine kinase, and deoxycytidine kinase were examined in human placental cytosol to achieve a valid and reliable assay linear with time and protein. Our studies confirm the need to inhibit deaminase enzymes, since deoxyadenosine and deoxycytidine undergo extensive deamination and phosphorolysis. The use of a uniformly labeled nucleoside substrate introduced an artifact because the chromatographic behavior of the deoxyribose-1-phosphate, formed during the assay, was difficult to distinguish from the deoxynucleoside phosphate product. Accurate product identification was also essential. Finally, the substitution of GTP in place of ATP as the phosphate donor, the addition of a sulfhydryl reducing agent and a monovalent cation need to be considered when an assay is optimized. The use of these methods have lead to valid assays in placental cytosol that are linear with time and protein. Consideration of these important principles are necessary when establishing a valid and reliable nucleoside kinase assay in a crude tissue preparation.

Adenosine Kinase↗

A search for trace expression of placental-like alkaline phosphatase in non-malignant human tissues: demonstration of its occurrence in lung, cervix, testis and thymus.

A search for placental or placental-like human alkaline phosphatase (ALP) was made in human tissues. The tissue extracts were assayed for ALP before and after heating at 65 degrees C for 1 h. Trace amounts of heat-stable ALP activity (greater than 0.01 IU/g) were found in lung, testis, cervix and thymus. The heat-stable ALP in these four tissues gave in Ouchterlony double diffusion plates lines of apparent identity with placental ALP when a rabbit anti-human placental antiserum was used. Inhibition studies with L-phenyl-alanine (Phe), L-homoarginine (Har), L-phenylalanylglycylglycine (Pgg), L-leucine (Leu) and levamisole (Leva), were carried out on the heat-stable ALP and on the total ALP. The heat-stable ALPs from cervix and lung gave [I]50 values with each inhibitor comparable to those of placental ALP. The heat-stable ALPs froM testis and thymus gave [I]50 values for Leu and Pgg which were significantly different from the placental isoenzyme. Electrophoresis of heat-stable lung ALP from different individuals showed polymorphic differences similar to those seen with placental ALPs. Such differences were not seen with heat-stable testis ALP. We conclude that human non-malignant testis, cervix, lung and thymus tissues contain small amounts of placental or placental-like ALPs. The heat-stable ALPs in cervix and lung appear to have the same characteristics as placental ALP and are probably encoded by the same gene locus. The heat stable ALPs in testis and thymus, though immunologically very similar to placental ALP differ from it in inhibition profile and electrophoretically. The significance of the results in relation to the "ectopic' expression of placental and placental-like ALPs in malignancy is discussed.

Alkaline Phosphatase↗

Placental-type alkaline phosphatase in ovarian cancer fluids and tissues.

Placental-type alkaline phosphatase was measured in sera, cyst, and ascites fluids, and from tumor extracts obtained from gynecologic cancer patients, particularly those with cancer of the ovary. A modified assay was used that depended on long incubation (20 to 24 hours) to measure the heat-stable, phenylalanine-sensitive placental isoenzyme. The concentration of enzyme in ascites and cyst fluids was markedly higher than in serum. Cyst fluid values were generally higher than ascites fluids from the same individual. The median enzyme levels for malignant cyst fluids were 50 times greater than for benign cyst fluids. When tumor tissue and fluids were available from the same patients, it was observed that the levels in each were proportional. Determination of this isoenzyme in serum did not give a useful index of tumor burden, as metastatic disease did not consistently result in elevated serum enzyme levels. When ovarian cancer patients were divided into two groups--those in whose sera placental-type phosphatase was elevated, and those in whom it was not--the presence of the enzyme in serum at the time of tumor diagnosis appeared to be a negative prognostic indicator, judged from survival data.

Alkaline Phosphatase↗

Improved purification of the human placental transferrin receptor and a novel immunoradiometric assay for receptor protein.

A simplified method for the purification of human placental transferrin receptor is described. The procedure involves chromatography of a detergent extract of placenta on immobilized iron-loaded transferrin. Knowledge of the physiology of the interaction between transferrin and its receptor is applied to enable bound receptor to be eluted under mild conditions and essentially free of containing transferrin. Purified transferrin receptor and a monoclonal antibody to the receptor were used to develop a novel immunoradiometric assay for the receptor in which the monoclonal antibody is the radiolabelled species. A competition between two populations of receptor, one immobilized on a particulate support and the other in solution, provides the basis for the assay. Using this assay we have measured transferrin receptor levels in placental and hepatic tissue and in three cell lines during both the logarithmic and stationary phases of cell growth.

Antibodies, Monoclonal↗

Quantitative analysis of matrix metalloproteinases-2 and -9, and their tissue inhibitors-1 and -2 in human placenta throughout gestation.

To elucidate the implication of type IV collagenases(MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2) for placental development, we quantified their levels in the conditioned media of placental organ culture and primary culture of the trophoblast as well as in the tissue extracts of placentas from different stages of gestation using specific enzyme-linked immunosorbent assays. First trimester villous tissue secreted about 10 times more pro-MMP-2 than pro-MMP-9, and pro-MMP-2 levels dramatically decreased in the second trimester. On the other hand, pro-MMP-9 levels were more than 10 times higher than those of pro-MMP-2 in the primary culture of the first trimester trophoblast, indicating the involvement of stromal cells for prominent pro-MMP-2 secretion from first trimester villous tissue described above. Levels of TIMPs, especially those of TIMP-2, remained constant throughout gestation both in the culture media and tissue extracts. Gelatin zymography revealed abundant secretion of the active form of MMP-2 as well as pro-MMP-2 from first trimester villous tissue. Western immunoblot analysis confirmed the presence of both TIMP-1 and TIMP-2 in placental tissue. These results suggest that active secretion of MMP-2 from villous tissue in the first trimester and constant production of TIMPs throughout gestation are characteristic of placental development.

Blotting, Western↗

Partial purification, characterization, and assay of a slightly acidic insulin-like peptide (ILAs) from human plasma.

An insulin radioreceptor assay (RRA) using human placental microsomal membranes was used to measure insulin-like activity (ILA) extracted from human plasma concentrates (Cohn fraction IV-4) by acid ethanol. The soluble activity (ILAs), chromatographed on Sephadex G-75 in 1 M acetic acid, migrated as a small molecule (fractional elution volume, 0.56) ahead of insulin (fractional elution volume, 0.70), whereas at neutral pH, ILAs migrated as a large molecular weight species. The ILAs peak from acid gel filtration on Sephadex was further purified by chromatography on carboxymethyl cellulose (CMC). The ILAs peak from both Sephadex and CMC diluted parallel to the porcine insulin standard in the insulin RRA and was totally unreactive in an insulin RIA. The CMC-purified material was iodinated and purified by binding to and elution from human placental membranes. The binding of [125I]ILAs to human placental membranes was inhibited only minimally by insulin and proinsulin and not at all by epidermal growth factor, nerve growth factor, glucagon, or lactogenic hormones, including human growth hormone. Multiplication-stimulating activity (MSA) inhibited in a manner parallel to ILAs. A Scatchard plot of the binding data was nonlinear. Sephadex ILAs was subjected to isoelectric focusing. The fractions assayed in both insulin and ILAs RRAs yielded comparable results. Peaks of ILA were observed at pHs 5.3, 6.6, and 8.4. When CMC-ILA was subjected to isoelectric focusing in polyacrylamide, a single peak of activity migrating between pH 6.2-6.8 was seen. [125I]ILAs focused at exactly the same pH. Electrophoresis of CMC-ILAs in acid-urea revealed a sharp peak of activity migrating with one of the five protein bands seen after staining. Again, [125I]ILAs comigrated with unlabeled ILAs. The molecular weight of ILAs, as determined on a calibrated Sephadex G-150 column at neutral pH, was 9,000-10,000 daltons. CMC-ILAs stimulated [14C]glucose incorporation into triglycerides of rat adipose tissue and augmented [3H]thymidine incorporation into human fibroblasts, chicken embryo fibroblasts, and BALB 3T3 cells as well as [35S]sulfate incorporation into macromolecules of rabbit chondrocyte culture medium. In summary, ILAs isolated on the basis of a RRA for insulin is a slightly acidic peptide with some of the biological activities expected of a somatomedin.

Adipose Tissue↗

Isolation of a major human placental substrate for the epidermal growth factor (urogastrone) receptor kinase: immunological cross-reactivity with transducin and sequence homology with lipocortin.

Using as a starting material either a detergent extract or a protein fraction eluted from membranes with ethylene glycol bis (beta-aminoethyl ether)-N,N'-tetraacetic acid, we have isolated from human placental membranes a major substrate for the epidermal growth factor (urogastrone) receptor kinase (EGF kinase). The substrate was isolated both in an intact form, having a molecular mass of approximately 38-kDa (p38), and in a 35-kDa form (p35) representing a proteolytic cleavage product of p38. Both p38 and p35 cross-reacted with antibodies directed against bovine retinal transducin, but did not cross-react with antibodies directed against the 35-kDa beta subunit of human placental G-protein. Antisera directed against the placental EGF kinase substrate failed to react with either bovine or human placental src kinase substrate, p36. Conversely, antisera directed against p36 reacted only poorly with placental p38 or p35. Although p38 had a blocked amino terminus that precluded sequence analysis, p35 yielded an N-terminal sequence that was identical with residues 13-36 of human lipocortin. Our data clearly distinguish p38 from the previously described intestinal calcium binding protein calpactin I or p36 that is also a tyrosine kinase substrate, and our work points to a close relationship (if not identity) between p35 and a 35-kDa EGF receptor kinase substrate previously characterized in A431 cells. We conclude that p38 and p35, which very likely represent human placental lipocortin, may share only limited epitope homology with transducin alpha subunit; however, the possibility that p38, along with intestinal p36 and with a family of related calcium binding proteins, may, like transducin, play a role in receptor-mediated transmembrane signaling is discussed.

Amino Acid Sequence↗

Simultaneous determination of zidovudine and lamivudine from rat plasma, amniotic fluid and tissues by HPLC.

A sensitive HPLC method has been developed and validated for the simultaneous quantification of zidovudine (AZT) and lamivudine (3TC) in rat plasma, amniotic fluid and placental and fetal tissues. Samples were processed by solid-phase extraction using C2 cartridges. Chromatography was performed using a phenyl column (5 microm, 150 x 2 mm i.d.) under a flow rate of 0.2 mL/min. The mobile phase consisted of 8% acetonitrile in 5 mM 1-heptane sulfonic acid dissolved in 30 mM ammonium formate buffer (pH 3.3). The method was validated in the range 0.25-50 microg/mL for both 3TC and AZT in the four biological matrices. Finally, the method was applied to a study involving fetal transport of co-administration of these compounds in a pregnant rat.

Amniotic Fluid↗

New placental and pregnancy proteins as possible markers in oncology.

A number of placental and pregnancy proteins has been detected by immunochemical methods in extracts from human term placentae. Several of these proteins already have been isolated and characterized and are now investigated for their usefulness as markers in oncology. Several of these proteins so long tested appear to be useful as markers in monitoring tumour patients, i.e. in the early detection of tumour recurrence or metastases and in the control of therapy. In addition, we hope that a combination of several of these new markers with already established tumour markers will enable to better diagnose tumours. Also will the use of solitary tissue proteins aid in the differential diagnosis of cancers.

Diagnosis, Differential↗

Isolation and characterization of a flavin-containing placental protein (PP3).

Placental protein 3 (PP3) has been isolated to purity from saline extracts of human term placentas. PP3 turned out to be a flavin-containing protein and thus appears to be an enzyme. The prosthetic group was identified as flavin-adenine dinucleotide (FAD) which was found to be noncovalently bound to the protein. The physical characterization of PP3 showed that the molecules are composed of two identical subunits having molecular weights of 55000 daltons which are noncovalently linked. Each subunit appears to contain one FAD group. In addition PP3 was found to have an electrophoretic mobility in between the alpha 1 and alpha 2 globulins, an isoelectric point in the range of 5.3-5.7, a sedimentation coefficient of 6.3 S and an extinction coefficient of 15.7 (E1%(1cm) 280 nm). Immunochemical methods were used to detect and quantitate PP3 in extracts of placentas and other human tissues. From one human term placenta an average of around 4 mg PP3 could be extracted; PP3 was also found to occur in extracts of adult human stomach. In concentrated extracts of other human tissues and in human body fluids this protein could not be detected, at least not in concentrations higher than 2 mg/dl.

Adult↗

Binding of diamine oxidase activity to rat and guinea pig microvascular endothelial cells. Comparisons with lipoprotein lipase binding.

Microvascular endothelial cells from rat and guinea pig fat pads were shown to bind diamine oxidase (DAO) activity when incubated with soluble extracts of placenta (33 DAO U/mg of placenta) and a purified placental enzyme preparation (94 U/micrograms of protein). The extent of binding was dependent on the concentration of enzyme activity and tissue. Saturation of binding sites with 5,000 U of DAO/ml resulted in levels of bound activity (up to 11-13 U/mg of endothelial cells) in excess of that observed in all tissues except placenta. Scatchard plots suggested that there were at least two DAO binding sites (apparent Km 92 and 2,450 U/ml). Although the same cell preparations bound 125I-labeled lipoprotein lipase (LPL), the presence of LPL on the endothelial cell surface did not interfere with the binding of DAO activity except when cells were exposed to high concentrations of LPL. Alternatively, bound DAO activity was partially displaced (up to 33%) only with high concentrations (30 micrograms/ml) of LPL. DAO activity may thus be bound to at least two populations of sites, one of which may bind LPL. Both enzymes, however, were displaced by heparin (0.05-5 U/ml) and DAO binding was impaired by prior treatment of cells with proteolytic and glycosaminoglycandegrading enzymes. The demonstration of DAO binding to vascular endothelial cells provides a further example of the ability of these cells to bind enzymes at their surface and thereby act on biologically active substances in the circulation.

Adipose Tissue↗

Presence of human epidermal growth factor in human cerebrospinal fluid.

Human epidermal growth factor (gEGF), a potent stimulator of the growth of many tissues in culture, has been isolated from human urine and subsequently identified in many human biological fluids. We have partially purified and characterized hEGF-like substance(s) from human cerebrospinal fluid (CSF) and determined the concentrations of immunoreactive (IR) hEGF in CSF obtained from patients with a variety of neurological diseases. Competitive binding curves generated by the CSF samples appeared to be parallel to standard hEGF both in RIA and radioreceptor assays using human placental membrane. Sephadex G-50 gel exclusion chromatography of the CSF extract revealed a single peak of IR-hEGF which coeluted with standard hEGF. The apparent mol wt of the CSF hEGF-like substances(s) was estimated to be 8000 by sodium dodecyl sulfate poplyacrylamide gel electrophoresis, and its approximate pI was 4.5 as determined by isoelectric focusing. The concentrations of IR-hEGF in CSF from patients with pituitary and brain tumors and radiculomyelopathy were significantly higher than those from control subjects, while neither patients with hydrocephalus nor encephalomeningitis had CSF IR-hEGF levels statistically different from the control subjects. The presence of hEGF-like substance(s) in human CSF suggests that it may play an important physiological role in the function of the human nervous tissues but does not provide any evidence of its source.

Adolescent↗

Molecular cloning and characterization of the cyclic AMP-responsive ovine CYP11A1 (cholesterol side-chain cleavage) gene promoter: DNase 1 protection of conserved consensus elements.

DNA elements governing transcription of the ovine cytochrome P-450 side-chain cleavage (CYP11A1) gene were investigated. Three overlapping genomic clones for the ovine CYP11A1 gene were isolated and characterized. The transcriptional start site was located 51 nucleotides upstream from the initiating methionine. Gene transfer experiments were conducted in murine adrenocortical Y1 cells and human choriocarcinoma JEG-3 cells using chloramphenicol acetyltransferase reporter gene constructs containing promoter fragments from -2700 to -177 bp. The results demonstrate that DNA elements sufficient to convey a basal level of expression and cyclic AMP (cAMP) responsiveness lie within 177 bp of the transcriptional start, although the possibility that additional regulatory elements reside outside this 177 bp has not been excluded. The ovine 5' flanking sequence demonstrated 92% homology with the bovine sequence, extending over the entire fragment. In contrast, only four significant regions of conservation between the ovine, murine, rat and human CYP11A1 promoters were found. These regions are positioned within 200 bp upstream of the transcriptional start site. DNase 1 footprinting was performed to identify DNA elements able to bind nuclear proteins. Primary adrenocortical and placental tissues from sheep were used as the source of nuclear extracts to detect DNA-protein interactions relevant to CYP11A1 gene expression in vivo. Five regions of protection were detected in the first -634 bp of the ovine CYP11A1 promoter. Three of these elements corresponded to the regions which are well-conserved between species. The other two elements resembled activating protein-1 (AP-1) and AP-4 sites and overlapping AP-2/Sp1 sites, and are conserved in the bovine gene but not in other species. Nuclear protein extracts from adrenals of sheep with different serum ACTH levels (i.e. ACTH-treated, dexamethasone-treated and untreated sheep) protected similar regions of the ovine CYP11A1 promoter fragment. Similarly, the regions protected did not differ when nuclear protein from JEG-3 cells treated with cAMP was compared with that of untreated JEG-3 cells. These results suggest that induction of CYP11A1 gene transcription by ACTH in the ovine adrenal and by cAMP in JEG-3 cells in culture is not mediated by changes in binding of the proteins that interact directly with these footprinted elements. The elements footprinted by extracts from primary ovine tissue lie within the 177 bp sufficient for cAMP-regulated expression. The correspondence of these elements either to regions conserved between species or to known consensus binding sites suggests that these sequences are cis elements involved in regulating transcription of the ovine CYP11A1 gene in vivo.

Amino Acid Sequence↗

Aerobic exercise during pregnancy. Special considerations.

Alterations in maternal physiology during pregnancy affect the physiological respect to aerobic exercise. Maternal resting oxygen consumption (VO2) and cardiac output increase during pregnancy. Heart rate (HR) becomes progressively elevated through gestation, whereas stroke volume (SV) increases until the third trimester and then declines until term, probably because of diminished venous return. Plasma volume increases earlier and to a greater magnitude than red cell volume, resulting in the 'haemodilutional anaemia' of pregnancy and a decline in the oxygen-carrying capacity. Ventilation is greater during pregnancy because of elevated tidal volume and unchanged rate of breathing. The acute and chronic (training) responses to aerobic exercise during pregnancy have not been thoroughly investigated. Specifically, the effect of gestational age, maternal activity status, and type, duration and intensity of exercise on maternal cardiovascular response have only recently begun to be explored. During pregnancy cardiac output during submaximal exertion increases above values in non-pregnant women, except perhaps late in gestation. Both heart rate and stroke volume contribute to the elevated cardiac output. Changes in submaximal exercise VO2 during pregnancy are dependent on the mode of exercise. At the same workload, VO2 increases during weight-bearing exercise, but usually does not differ from postpartum values during weight-supported exercise. One study found no change in VO2max during pregnancy compared to postpartum values. Some recent evidence indicates that the cardiac output vs VO2 relationship for pregnant women is within the range of average values reported for non-pregnant individuals. Exercise arterial-venous oxygen difference is lower during pregnancy, suggesting that the higher cardiac output is distributed to non-exercising vascular beds. The data are limited but suggest that the perfusion of exercising muscle is unchanged during pregnancy and that the major haemodynamic change is an augmented cardiac output so that blood flow to the uterus and fetus is not compromised. Only one study has measured blood flow during exercise in pregnant women. The reported 25% decrease in uterine blood flow during supine cycle exercise in women late in gestation must be interpreted cautiously because the uterus may obstruct the vena cava in the supine position. Studies of exercising pregnant animals usually indicate a decreased uterine blood flow but an enhanced oxygen extraction; the lower blood flow may be limited to non-placental areas. The applicability of these results to humans is unknown.(ABSTRACT TRUNCATED AT 400 WORDS)

Exercise↗

Determination of placental estriol in urine by gas-liquid chromatography, with equilenin as internal standard.

Placental estriol in urine can be determined by rapid enzymatic hydrolysis, extraction with neutral XAD-2 resin, and quantitation by gas-liquid chromatography, with equilenin as the internal standard. Glucose, methenamine mandelate, hydrochlorothiazide, phenolphthalein, or salicylate do not interfere. Analytical recovery of estriol added to urine was 92-108%. Day-to-day precison (CV), established by 15 repilicate analyses at a mean concentration of 21.3 mg/liter, was 6.6%.

17-Ketosteroids↗

Placental abruption is more frequent in women with the angiotensinogen Thr235 mutation.

OBJECTIVE: Obstetrical complications such as preeclampsia, fetal growth restriction, and placental abruption are associated with inadequate placental perfusion. Previous studies have shown that the angiotensinogen (AGT) Thr235 mutation is associated with abnormal remodeling of the uterine spiral arteries and occurs at higher frequencies in preeclampsia. This study was done to evaluate whether the AGT Thr235 mutation increases the risk of placental abruption. MATERIALS AND METHODS: We compared 62 placentas from women who had placental abruption with 240 control patients of similar age and ethnicity. DNA was extracted from paraffin blocks from placentas. AGT Met235Thr mutation status was determined by single fluoresceine labeled probe real-time PCR using a LightCycler system. RESULT: AGT genotypes were divided into three groups: MM (homozygous wild), TT (homozygous mutant), and MT (heterozygous). The constituent ratio of AGT genotype in abrupted placentas (MM 14.5%, MT 43.5%, TT 41.9%) was significantly different from in control group (MM42.5%, MT 39.6%, TT 17.9%) (p<0.001). AGT mutant allele frequency in placental abruption (0.637) was significantly higher than in the control group (0.377) (p<0.001). CONCLUSION: The AGT Thr235 mutation was observed more frequently in placental abruption. AGT Thr235 mutation may be considered a risk factor for placental abruption.

Abruptio Placentae↗

Evaluation of murine placental degradation and transfer of [125I]iodo-epidermal growth factor.

Since epidermal growth factor (EGF) has been postulated to play a role in embryonic and fetal growth, a study was undertaken to assess the placental degradation and transfer of maternally administered EGF. Before iodination, mouse EGF was purified to homogeneity by reversed-phase HPLC. Approximately 5 ng [125I]iodo-EGF (approximately 10(6) cpm) were injected iv into day 10, day 13, or day 17 pregnant CD-1 mice; radioactivity in plasma, placentas, and conceptuses was measured up to 2 h after injection. The time course analysis revealed an initial rapid decline in total plasma radioactivity followed by an increase that was maximal by approximately 30 min. Gel filtration (G-15, G-50) chromatography of plasma revealed that by 5 min, radioactivity was associated with free 125I and with material much larger than EGF. No apparent degradation of [125I]iodo-EGF occurred after direct incubation with maternal plasma. Placental radioactivity had an initial phase of decay between 1 and 5 min followed by an increase that became maximal between 30 and 60 min. Extracts of placentas made with 4 M urea in 0.2 M Tris-HCl, pH 8.0, and taken 1-30 min after injection revealed radioactivity coeluting predominantly with [125I]iodo-EGF at 1 min but shifting to mostly free 125I by 30 min. Uptake of radioactivity by conceptuses was not evident until about 15 min, and only free 125I was detected in extracts; the same results were obtained when 5 micrograms unlabeled EGF were injected simultaneously with [125I]iodo-EGF. Incubation of placental mince with [125I]iodo-EGF yielded [125I]MIT as the apparent major radioactive degradation product. Formation of [125I]MIT in vitro was both time- and temperature-dependent. At 37 C, marked formation of [125I]MIT was observed; at 22 C, only a negligible amount was formed after incubation of mince with [125I]iodo-EGF for 60 min. Incubation of [125I]iodo-EGF with kidney mince yielded predominantly free 125I. When tissues were incubated directly with [125I]MIT, kidney tissue deiodinated the [125I]MIT, but placental tissue did not. When [125I]iodo-EGF was incubated for up to 2 h with kidney tissue in the presence of excess MIT (unlabeled), the major degradation product eluted as [125I]MIT, instead of free 125I. These findings suggest that the mouse placenta can readily bind and then degrade [125I]iodo-EGF to constituent amino acids.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗