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Biomedical subjects

P G Groothuis

Publications and source records attributed to P G Groothuis.

25 records · Page 2Linked to original sources

Adhesion of human endometrial fragments to peritoneum in vitro.

OBJECTIVE: To evaluate the adhesion of endometrial fragments obtained during the proliferative phase of the menstrual cycle to fresh human peritoneum obtained during abdominal surgery. DESIGN: A prospective, descriptive, morphologic and cell biologic study. SETTING: Tertiary care university medical center. PATIENT(S): Six female volunteers. INTERVENTION(S): After endometrial biopsies performed during diagnostic laparoscopy, endometrial fragments were generated by enzymatic digestion and mechanical separation. Peritoneum was obtained during abdominal operations for benign indications. MAIN OUTCOME MEASURE(S): Adhesion of endometrial fragments was studied by histologic examination and scanning and transmission electron microscopy. RESULT(S): After incubation, the mesothelium was intact in some areas, whereas in other areas mesothelial cells were damaged or absent. Adhesion of endometrial fragments was observed only at locations where the basement membrane was exposed. In areas largely denuded of mesothelial cells, endometrial fragments spread over the basement membrane to form monolayers. CONCLUSION(S): Human peritoneum is suitable for studying the adhesion of endometrial fragments. Intact mesothelium prevents the adhesion of endometrial fragments, suggesting that trauma to the mesothelial lining is a prerequisite for endometrial cell adhesion.

Basement Membrane↗

Adhesion of shed menstrual tissue in an in-vitro model using amnion and peritoneum: a light and electron microscopic study.

We have investigated the adhesion of endometrial tissue isolated from antegradely shed menstrual effluent to amnion and peritoneum. This endometrial tissue was cultured overnight on either side of intact and stripped amnion and on the mesothelial side of peritoneum. Light and electron microscopy were applied to evaluate adhesion. With light microscopy adhesion of endometrial fragments to stripped membranes was observed in nine out of 12 specimens and in 12 out of 13 specimens when layered on the extracellular matrix side of amnion. Adhesion when layered on the epithelial side was seen in only four out of 13 specimens. However, when using scanning electron microscopy adhesion of menstrual endometrial tissue could be visualized in all samples. Numerous adhering fragments were seen when layered on the extracellular matrix side of untreated amnion. On several occasions not only adhesion but also spreading of cells was observed. When layered on the epithelial side of untreated amnion or peritoneum, adhesion was exclusively seen at locations where the epithelium was damaged or absent. These findings were confirmed by transmission electron microscopy. These observations indicate that endometrial tissue isolated from antegradely shed menstrual effluent preferentially adheres to subepithelial structures of amnion and peritoneum. The lack of adhesion to epithelial cells suggests that an intact mesothelial lining prevents adhesion of menstrual endometrial tissue.

Amnion↗

Adhesion of human endometrium to the epithelial lining and extracellular matrix of amnion in vitro: an electron microscopic study.

One of the first steps in the pathogenesis of endometriosis is the attachment of the endometrium to the peritoneal lining. Since the peritoneum is extremely fragile and hard to obtain, amnion has been used as an in-vitro model to study adhesion. Scanning and transmission electron microscopy was applied to evaluate the adhesion of endometrial cells isolated in the proliferative and secretory phases of the menstrual cycle. Endometrial fragments obtained in either phase of the cycle were able to adhere to the extracellular matrix of the amnion. Fragments from proliferative phase endometrium showed active spreading and growth over the matrix surface, whereas fragments from secretory phase endometrium did not. Fragments from proliferative as well as secretory phase endometrium were able to adhere to the epithelial side of the amnion, but only at locations where the amniotic epithelium was damaged or partly absent. These observations indicate that the basement membrane and extracellular matrix provide a suitable substrate for endometrial cell attachment and growth and that endometrial cell adhesion occurs preferentially to subepithelial structures, whereas an intact epithelium prevents the adhesion of endometrial fragments to the amnion.

Amnion↗

Uterine response to progesterone in prepubertal gilts.

During early pregnancy, progesterone stimulates the secretion of proteins and other molecules that support the developing conceptus. Some gilts are able to support conceptus development as early as 110 days of age. The objective of this study was to evaluate the onset of responsiveness of the prepubertal uterus to progesterone. Thirty gilts were assigned to receive 2.2 mg progesterone kg-1 body mass per day or corn oil daily for 14 days starting at 6, 46, 76, 106, and 136 days of age. Hysterectomies were performed the day after the last treatment of progesterone, and the uterine horns were weighed and flushed with sterile saline (0.85% NaCl). Recovered flushings were analysed for total luminal protein, retinol binding protein, uteroferrin, prostaglandin E and prostaglandin F. An interaction between age and progesterone occurred for uterine wet mass (P < 0.001). Progesterone did not affect the uterine mass of gilts that underwent hysterectomy at 20 days of age, but did increase the uterine mass (P < 0.05) in other age groups. Progesterone increased (P < 0.01) the amount of total luminal protein in all but the youngest gilts. An increase in the amounts of retinol binding protein and uteroferrin (P < 0.001) by progesterone was first observed in 90-day-old gilts. Prostaglandins exhibited a different age-related pattern. The amount of prostaglandin E was increased (P < 0.001) by progesterone treatment in gilts aged 90-150 days, with a greater (P < 0.05) response at 120 days than at 90 days old. The response at 150 days old decreased (P < 0.05) to that observed at day 90. The response of prostaglandin F to progesterone followed a similar age-related pattern. Therefore, uterine responsiveness to progesterone develops between 20 and 90 days after birth, and uterine mass responds earlier than the secretory responses measured in our study.

Acid Phosphatase↗

Porcine conceptus proteins: antiviral activity and effect on 2',5'-oligoadenylate synthetase.

Interferon-like proteins synthesized by conceptuses of domestic ruminants inhibit luteolysis during early pregnancy. Although pig conceptuses secrete trophoblast interferons during the period of CL maintenance, estrogen is involved with maintenance of the CL. The principal purposes of this work were to confirm production of trophoblast interferons by porcine conceptuses and to compare the effect of trophoblast interferons on endometrium of pigs and cattle. When measured using Madin-Darby bovine kidney (MDBK) cells challenged with vesicular stomatitis virus, antiviral activity in uterine flushings from cyclic gilts was not detectable throughout the estrous cycle; however, in pregnant gilts, antiviral activity increased from undetectable amounts to 4-11 x 10(3) U on Days 14, 16, and 18. Porcine embryos in culture produced 1,100 U/embryo/ml/24 h. Porcine conceptus secretory proteins induced 2',5'-oligo(A) synthetase in MDBK cells and in endometrial explants of cows but had no measurable effect on 2',5'-oligo(A) synthetase activity of endometrial explants of pigs. Similarly, endometrial 2',5'-oligo(A) synthetase of pregnant pigs was unaffected in vivo during the period of maximal synthesis of conceptus secretory proteins. Porcine conceptus secretory proteins produced no detectable increase in serum antiviral activity or 2',5'-oligo(A) synthetase activity of blood mononuclear leukocytes in utero-ovarian venous blood. These results suggest that conceptus interferons of pigs play different roles in the establishment of pregnancy compared to their roles in ruminants.

2',5'-Oligoadenylate Synthetase↗

APC, beta-catenin, and E-cadherin and the development of recurrent endometrial carcinoma.

Endometrial carcinoma, generally, has a good prognosis. However, in some patients, the tumor appears to behave very aggressively, a course that cannot be explained with histopathological characteristics. More insight into the molecular background can be valuable to clarify these differences in tumor behavior. The three components associated with the Wnt pathway--i.e., adenomatous polyposis coli (APC), beta-catenin, and E-cadherin--were evaluated in a case-control study of 28 patients with stage-I endometrial carcinomas to determine their involvement in the development of recurrent disease. Mutation analysis of the mutation cluster region of the APC gene, determination of gene promoter methylation status of the APC-1A and E-cadherin genes, and immunohistochemical analysis of APC, E-cadherin, and beta-catenin were performed using paraffin-embedded tumor tissue. Twenty-one APC gene mutations were detected in 12 of 28 (43%) patients. Only three mutations would result in a stopcodon in the APC gene. APC gene promoter methylation was assessed in 12 of 28 (43%) patients. APC immunostaining was absent in two of 24 (8.3%) patients. The occurrence of APC mutations, APC gene promoter methylation, and APC immunostaining were not predictive for recurrence. No E-cadherin expression was observed in four of 24 patients (17%). E-cadherin gene promoter methylation could not be detected in any of the patients. The absence of E-cadherin expression was predictive for distant metastases, but not for local recurrence. Nuclear localization of beta-catenin was present in nine of 24 (38%) patients and was not predictive for recurrent disease. Involvement of epigenetic and genetic aberrations in APC and beta-catenin genes seems to be of minor importance for the development of local recurrences and distant metastases. Although the number of patients is limited, E-cadherin expression appears to be predictive for the development of distant metastases in endometrial carcinoma.

Adenomatous Polyposis Coli↗