Daidzein-rich isoflavone aglycones inhibit cell growth and inflammation in endometriosis.

Endometriosis is an estrogen-dependent disease, and isoflavones interact with estrogen receptors. The purposes of this study are to investigate the in vitro and in vivo effects of daidzein-rich isoflavone aglycones (DRIAs), dietary supplements, on cellular proliferation in endometriosis. Stromal cells isolated from ovarian endometrioma (OESCs) and normal endometrium (NESCs) were cultured with DRIAs, i.e., each of the DRIA components (daidzein, genistein, or glycitein), or isoflavone glycosides (IG; DRIA precursors). A mouse model of endometriosis was established by transplanting donor-mouse uterine fragments into recipient mice. Our results showed that DRIAs (0.2-20 µM) inhibited the proliferation of OESCs (P < 0.05 for 0.2 µM; P < 0.01 for 2 and 20 µM) but not of NESCs. However, daidzein, genistein, glycitein, and IG did not inhibit their proliferation. DRIA-induced suppression was reversed by inhibition of the estrogen receptor (ER)ß by an antagonist, PHTPP, or by ERß siRNA (P < 0.05), but not by MPP, an ERα antagonist. In OESCs, DRIAs led to reduced expression of IL-6, IL-8, COX-2, and aromatase, as well as reduced aromatase activity, serum glucocorticoid-regulated kinase levels, and PGE2 levels (P < 0.05). Western blot and immunofluorescence assays revealed that DRIAs inhibited TNF-α-induced IκB phosphorylation and p65 uptake into the nuclei of OESCs. In the mouse model, a DRIA-containing feed significantly decreased the number, weight, and Ki-67 proliferative activity of endometriosis-like lesions compared to in mice fed with an IG-containing feed and the control feed (P < 0.01). In conclusion, DRIAs inhibit cellular proliferation in endometriosis, thus representing a potential therapeutic option for the management of endometriosis.

Identification of porcine follipsin as plasma kallikrein, and its possible involvement in the production of bradykinin within the follicles of porcine ovaries.

To determine the identity of porcine follipsin, a plasma kallikrein cDNA clone was isolated from a porcine liver cDNA library. The clone encoded a protein of 643 amino acids, exhibiting identities 79.7, 72. 9, and 74.4% homologous to human, rat, and mouse plasma prekallikrein, respectively. The amino acid sequences of four internal peptides isolated from the tryptic digest of follipsin were all found in the deduced sequence. Authentic plasma kallikrein was purified from porcine plasma and compared directly with follipsin. Actions on synthetic substrates and behaviors with proteinase inhibitors were indistinguishable between these two enzymes. The cDNA was expressed in COS-7 cells and the recombinant protein was prepared from the culture medium of these cells. No active enzyme could be obtained, but the expressed protein was reacted with anti-porcine plasma kallikrein antibody. The mRNA was detected only in the liver in northern blot analysis. RT-PCR analysis of RNAs revealed that porcine testis, in addition to the liver, expressed the corresponding mRNA. In the ovary, plasma kallikrein was detected as a main band of the active form (Mr = 85,000) and the band of the minor inactive precursor form (Mr = 80,000), respectively. In contrast, the liver extract contained only the precursor form. Incubation of high molecular weight kininogen with follicular fluid plasma kallikrein resulted in an increased production of bradykinin. Further, the fresh fluid of large-sized follicles of porcine ovaries was found to contain this peptide hormone at a detectable level. These results indicate that porcine follipsin is plasma kallikrein, and that the enzyme may be involved in the production of bradykinin within ovarian follicles.

Localization of prolyl endopeptidase mRNA in small growing follicles of porcine ovary.

Prolyl endopeptidase (EC3.4.21.26) has been considered a unique intracellular enzyme catalyzing internal peptide bond hydrolysis of Pro-X. In this study, the distribution of prolyl endopeptidase activity and its mRNA was investigated in the follicles of porcine ovary. Both follicular fluid and granulosa cell fractions from small follicles showed higher activity than those from large follicles. Molecular cloning and Northern blot analysis suggested that only one species of prolyl endopeptidase gene was expressed in the ovary. In addition, in situ hybridization study revealed that the prolyl endopeptidase mRNA expression was more noticeable in the granulosa cell layers of small ovarian follicles than in those of large follicles, suggesting its importance in the early stage of follicular development.

A case of herpes zoster associated with colitis.

A 58-year-old Japanese woman who had herpes zoster in association with colitis was successfully treated with intravenously administrated acyclovir. Vesicular lesions with red haloes ranged from the left side of her buttock to the left extremity, corresponding to the L4 to S2 dermatomes. Her colitis was considered to have been induced by varicella-zoster virus, based on the facts that the clinical courses were correlated and that the innervation of the affected site of the colon corresponded to an infected dermatome (S2).

Necrotizing fasciitis due to Pasteurella multocida infection.

Necrotizing fasciitis is a potentially fatal clinical disease caused by infection with various bacteria in addition to streptococci, which are common causative agents. We report on a rare case of this disease in association with Pasteurella multocida infection. A 58-year-old man had systemic features of shock after a 15-hour history of a painful swelling on the right lower leg. The swelling led to skin blistering and necrosis from which P. multocida was isolated. Those lesions progressed rapidly. The patient also had a history of chronic liver injury as described in previous reports.

Purification, characterization, and localization of follipsin, a novel serine proteinase from the fluid of porcine ovarian follicles.

Follipsin, an enzyme that accumulates in the follicular fluid of porcine ovaries during follicular maturation, was purified to apparent homogeneity. The purified enzyme consists of two different polypeptide chains having M(r) = 45,000 and 32,000 each, associated covalently. The enzyme activity was strongly inhibited by diisopropyl fluorophosphate, benzamidine, leupeptin, and antipain, indicating that follipsin is a serine proteinase. Using synthetic peptide substrates containing 4-methylcoumaryl-7-amide, follipsin was shown to preferentially hydrolyze Arg-X bonds but not Lys-X bonds. The NH2-terminal amino acid sequences of the 45- and 32-kDa polypeptides were highly homologous with those of the heavy and light chains, respectively, of human plasma kallikrein and human factor XIa. Immunological analyses and substrate specificity studies, together with other existing evidence, indicated that follipsin is distinct from kallikrein and factor XIa, thus being a novel type of serine proteinase. Follipsin is immunohistochemically localized in follicular fluid as well as in stroma cells of porcine ovaries. The results strongly suggest that follipsin originates from interstitial cells of the ovarian stroma.