J. A. Keenan

Interferon‐Gamma (IFN‐γ) and Interleukin‐6 (IL‐6) in Peritoneal Fluid and Macrophage‐Conditioned Media of Women With Endometriosis

PROBLEM: The presence of the various cytokines in human peritoneal fluid has been incompletely evaluated. Changes in cytokine levels may be related to activation of peritoneal macrophages, development of endometriosis, and infertility. This study assesses peritoneal fluid levels of interferon gamma (IFN‐γ) and interleukin‐6 (IL‐6), and peritoneal macrophage production of IL‐6, in women with and without endometriosis.

Postnatal androgenization induces premature aging of rat ovaries.

In the present paper, we report that ovaries of adult rats treated with testosterone propionate (TP) on a critical postnatal Day 5 exhibit histologic and immunohistochemical findings which resemble those of the anovulatory ovaries in middle-aged female rats. The sterile rat model has been long known whereas ovarian failure seems to be a reason for anovulation with normal hypothalamo-pituitary-gonadotropin background. Appropriate function of ovarian steroidogenic cells is also regulated by mesenchymal cells. To characterize the ovarian failure, we studied the histology, luteinizing hormone receptor (LHR) expression, and characterized changes of vascular pericytes, T cells, and dendritic cells in ovarian steroidogenic compartments consisting of interstitial cells (ISC) of ovarian interstitial glands, and granulosa and theca interna cells of ovarian follicles. Normal adult ovaries contained 63% of mature interstitial glands. The mature ISC exhibited moderate cytoplasmic and strong surface LHR expression and fine (<5 micrometer) cytoplasmic vacuoles (ISC of luteal type). They originated from young ISC of thecal type, which exhibited strong cytoplasmic LHR expression. Remaining 37% were aged interstitial glands, which consisted of aged ISC (increased cytoplasmic vacuolization, nuclear pyknosis, and reduced surface LHR expression) and regressing ISC (weak cytoplasmic and no surface LHR expression). However, no mature ISC of luteal type were detected in anovulatory ovaries of adult rats (45- and 60-day-old) injected with TP (100 or 500 microgram) on postnatal Day 5 (TP rats). Their ovaries contained 96% of aged interstitial glands with aged and regressing ISC. Remaining 4% were abnormal interstitial glands with direct transition of young ISC of thecal type into aged ISC (young/aged glands). Lack of mature ISC, and similar amount of aged (96%) and young/aged interstitial glands (4%) was also detected in anovulatory ovaries of untreated persistently estrous middle-aged (10-month-old) females (aging PE rats). The aging process in TP and aging PE rats was accompanied by regression of vascular pericytes, T cells, and dendritic cells within the interstitial glands. In addition, anovulatory ovaries of TP rats and aging PE females contained mature follicles exhibiting LHR overexpression by granulosa cells, and aged (cystic) follicles with reduced layers of granulosa cells lacking LHR expression. In contrast, when the rats were injected with 500 microgram of TP later, on postnatal Day 10, the adult females exhibited estrous cycles and normal ovaries with corpora lutea. These results show that injection of TP during the critical postnatal period causes a lack of mature and preponderance of aged ISC in adult ovaries, accompanied by degeneration of mesenchymal cells. We suggest that mesenchymal cells regulate qualitative aspects of tissue-specific cells, and this function of mesenchymal cells is programmed during the critical period of development.

Is irregular regression of corpora lutea in climacteric women caused by age-induced alterations in the "tissue control system"?

PROBLEM: We have recently observed that the regression of corpora lutea (CL) in women during the reproductive period of life is accompanied by a diminution of Thy‐1 differentiation protein release from vascular pericytes and an accumulation of T lymphocytes and activated macrophages among both degenerating granulosa lutein cells (GLC) and theca lutein cells. These data suggest that the immune system and other stromal factors, representing components of the “tissue control system,” may play a role in regression of the CL. We investigated degenerating CL from climacteric women to address the possibility that the decline of immune functions with advancing age may result in incomplete regression of luteal tissue. This could contribute to the altered hormonal profiles and abnormal uterine bleeding that frequently occur during the climacteric.

Expression of cell cycle regulatory proteins (p53, pRB) in the human female genital tract

PurposeRecent studies have shown that proliferation and differentiation of various cell types is regulated by cell-cycle-related proteins, such as protein p53 and retinoblastoma protein pRb.MethodsThree monoclonal antibodies to p53 (PAb240, PAb421, and PAb1801) and 3H9 monoclonal antibody to pRb were utilized for localization of proteins by peroxidase immunohistochemistry in frozen tissue sections.ResultsNuclear and nucleolar p53 expression was detected in nondividing and relatively stable cells, e.g., oocytes in primordial follicles and granulosa lutein cells. On the other hand, strong cytoplasmic p53 expression was detected in proliferating and low differentiated epithelial cells of the ovarian surface epithelium, amnion, endocervix and ectocervix, indicating enhanced p53 synthesis. Not all three p53 antibodies reacted with each tissue, perhaps due to structural and conformational changes in the p53 molecule, accompanying p53 association with other proteins, e.g., tissue specific transcription factor interactions. pRb expression was usually restricted to the cell nuclei and nucleoli. However, glandular cells of the female reproductive tract showed cytoplasmic pRb expression in juxtaluminal (secretory) segments of cells, a feature not previously described in any cell type. p53 and pRb immunoreactivities declined with advanced differentiation of cells. No p53 or pRb was detected in placental syncytiotrophoblast or terminally differentiated squamous epithelial cells.ConclusionOur data indicate that large quantities of p53 are synthesized in cells leaving the cell cycle and entering differentiation. Except in glandular cells, the pRb expression is confined to the cell nuclei and nucleoli. A unique cytoplasmic expression of pRb in juxtaluminal segments of glandular cells suggests a role for pRb in human female fertility and conception.

Development of the National Embryo Donation Center

The use of Assisted Reproductive Technologies (ARTs) has proliferated rapidly since the birth of Louise Brown in England in 1978 via in vitro fertilization with embryo transfer (IVF–ET, or more commonly, IVF). The ability to cryopreserve embryos followed shortly thereafter, increasing the potential success rates while decreasing the costs, thereby becoming a standard practice among ART clinics. Unfortunately, the proliferation of these technologies led to an unanticipated problem, i.e., the prolonged storage of large numbers of frozen embryos. The reasons that couples fail to use or make a decision on the fate of their frozen embryos has been the subject of a number of studies. Some couples are simply uncomfortable with the idea of anyone else raising their biological child(ren). Others are concerned about whether their embryos will go to a “good home”. Another concern is that the offspring will eventually find them and demand to know why they were not raised by their genetic parents. A less rational, but still common concern, is that their “unknown” children might eventually marry their brother or sister. Finally, other couples simply don’t understand the processes involved, and due to a lack of information or someone to speak to, simply avoid the decision entirely (Elford et al., 2004, pp. 1154–1155; de Lacey, 2005, p. 1668). Whatever the reasons, the result is that over 400,000 embryos are currently being cryopreserved in the USA alone. And, although surveys indicate that 90% or more of couples are keeping them “for their own use” (Hoffman et al., 2003, p. 1063), experience shows that a large percentage, perhaps one-half of them, will never be used by their genetic parents. The National Embryo Donation Center (NEDC) was founded to address this dilemma. The idea of a national center that provided comprehensive embryo adoption and donation services was originally the idea of Dr. David Stevens, who serves as CEO of the Christian Medical and Dental Associations (CMDA). Dr. Stevens presented me with this idea in late 2002. We agreed that the large, and increasing, number of frozen embryos was of concern from medical, legal, and ethical perspectives. We saw the potential to assist both couples who have remaining frozen embryos, as well as infertile couples who desire to experience the