The CD15 carbohydrate epitope is expressed in mature human neutrophils, monocytes, and promyelocytes. We aimed to determine the ␣1,3-fucosyltransferase responsible for the expression of CD15 in each subpopulation of leukocytes. Three ␣1,3-fucosyltransferases, FUT4, FUT7, and FUT9, are expressed in human leukocytes. We demonstrated that FUT9 exhibits 20-fold stronger activity for CD15 synthesis than FUT4, whereas FUT4 exhibits 4.5-fold stronger activity for CDw65 synthesis than FUT9. By competitive reverse transcriptase-polymerase chain reaction, FUT9 was found to be strongly expressed in mature granulocytes and peripheral blood mononuclear cell, but not in monocytes. CD34؉ and CD15 ؉ cells in cord blood and myeloid cell lines (HL-60 and U937) did not express FUT9 at all. FUT4 transcripts were ubiquitously expressed in all blood cells and all cultured cell lines, with HL-60 and U937 cells in particular expressing a number of FUT4 transcripts. Transfection of the FUT9 gene into Jurkat and U937 cells demonstrated that FUT9 has the potential to express CD15 in myeloid and lymphoid cells. These findings suggest that the expression of CD15 in mature granulocytes is directed by FUT9, whereas it is determined in promyelocytes and monocytes by FUT4. Measurement of CD15 synthesizing activity in cell homogenates of each cell population using the polylactosamine acceptor further supported these conclusions.There are three CD 1 markers of human leukocytes comprising fucosylated carbohydrate epitopes. As listed in Fig. 1 below, the distal lactosamine unit (LN; type 2 chain), Gal␤1,4GlcNAc, of the polylactosamine chain is fucosylated through ␣1,3-fucosyltransferase (␣1,3FUT) activity to form the CD15 (Lewis x; Le X ) epitope (1, 2). The CD15s (sialylated CD15; sialyl Le X (sLe X )) and CDw65 (VIM-2) epitopes are also fucosylated structures related to CD15, i.e. CD15s is formed by ␣2,3-sialylation prior to the fucosylation of the distal LN unit of polylactosamine by ␣1,3FUT, and CDw65 is formed by fucosylation of the inner LN unit of ␣2,3-sialylated polylactosamine by ␣1,3FUT (2, 3).The CD15 epitope is expressed in some tissues, such as epithelial cells of intestinal tissues (4 -6), certain neurons and glial cells in the central nervous system (7,8). In human leukocytes, CD15 is expressed preferentially in monocytes, mature neutrophils, and all myeloid cells from the promyelocyte stage onwards, making it a useful cell surface marker (9 -11). CD15 is considered to be involved in neutrophil functions, that is, cell-cell interactions, phagocytosis, stimulation of degranulation, and respiratory burst, although the function of CD15 is not clear (12)(13)(14)(15)(16).Six human ␣1,3FUT genes have been cloned to date, which are FUT3 (Fuc-TIII), FUT4 (Fuc-TIV), FUT5 (Fuc-TV), FUT6 (Fuc-TVI), FUT7 (Fuc-TVII), and FUT9 (Fuc-TIX) (1, 17-23). FUT9, a new member of the human ␣1,3FUT family, which we have recently cloned, is expressed in human leukocytes, glandular compartments of the stomach, and forebrain (23). The FUT9 gene was mapped on ch...

The present study was undertaken to investigate the role of angiotensin II (Ang II) in ovulation and ovarian steroidogenesis and prostaglandin (PG) production via the Ang II receptors in rabbit ovaries. In in vitro perfused rabbit ovaries, PD123319, a selective nonpeptide antagonist for AT2 receptors, reduced the Ang II-induced ovulation in a dose-dependent manner, whereas CV-11974, a selective nonpeptide antagonist for AT1 receptor, did not affect the Ang II-induced ovulation. Ang II also significantly stimulated the meiotic maturation of ovulated ova and follicular oocytes in the absence of gonadotropin. The addition of PD123319 at 10 (-6) M to the perfusate significantly inhibited the Ang II-induced oocyte maturation. Ang II did not stimulate the production of progesterone by perfused rabbit ovaries but significantly stimulated the production of estradiol (E2) and PGs. When PD123319 at 10(-6) M was added to the perfusate 30 min before the onset of Ang II administration, the Ang II-stimulated production of E2 and PGs was significantly blocked. Saralasin, a peptide analog of Ang II, inhibited the specific binding of [125I] iodo-[Sar1, Ile8] Ang II to rabbit ovarian membranes in a concentration-dependent manner, yielding an inhibitory constant (IC50) value of 1.58 x 10(-9) M. PD123319 and CV-11974 also inhibited the binding of [125I]iodo-[Sar1, Ile8] Ang II; however, PD123319 and CV-11974 were 15 and 40 times less potent than saralasin, respectively. Autoradiographic study revealed that an intense localization of Ang II receptors in the rabbit ovaries was present in the granulosa cell layers and the stroma of the preovulatory follicles. AT2 receptors were predominantly located in granulosa cells, whereas AT1 receptors were more concentrated in the stroma and thecal cell layers. In summary, Ang II induced ovulation and oocyte maturation and stimulated the production of E2 and PG by perfused rabbit ovary in vitro via the AT2 receptor. Thus, locally produced Ang II may be part of a novel intraovarian paracrine or autocrine control mechanism during the ovulatory process.

To investigate the possible direct involvement ofangiotensin II (Ang II) in ovulation and oocyte maturation, Ang II ill lOOor IOyg was administered at 2-h intervals in the in-vitro perfused rabbit ovaries. The addition of Ang II in the pcrfusate induced ovulation in vhro in the absence of gonadotropin, while ovulation did not occur in any contralaleral control ovaries. I-Iowcver, the ovulatory eflicicncy in the Ang II-trcatcd ovaries was signiiicantly lower than in hCG-treated ovaries. Ang II signilicantly stimulated the meiolic maturation of ovulated ova and follicular oocytes. Concomitant addition of lhe specific receplor antagonist of Ang II. saralasin, 30 min before the onset of Ang II administrdlion blocked Ang ILinduced ovulation in a complete manner. Although suralasin did not inhibit completely hCG-induced ovulation and oocytc muturalion, these rcsulls suggest that Ang II produced in the ovary may act locally in the process of ovulation.

The present study was undertaken to investigate the effects of GH on follicular growth, oocyte maturation, ovulation, and production of insulin-like growth factor-I (IGF-I) in the in vitro perfused rabbit ovaries. Ovulation did not occur in any ovaries perfused with GH at a concentration of 1, 10, 100, or 200 ng/ml, but the addition of GH to the perfusate increased the follicle diameter in a dose-dependent manner. The production of IGF-I by ovaries perfused with medium alone was very low throughout the perfusion period. The addition of 100 ng/ml GH to the perfusate significantly increased ovarian production of IGF-I at 4, 6, 8, and 12 h compared with the contralateral control ovaries. Changes in the tissue concentrations of IGF-I in ovaries perfused with 100 ng/ml GH paralleled those triggered by exposure to 50 IU human CG (hCG). When the effect of GH on the tissue concentration of IGF-I was determined at 4 h, GH stimulated the tissue concentration of IGF-I in perfused rabbit ovaries in a dose-dependent manner. The percent increase in follicle diameter in ovaries treated with GH was significantly correlated with the intraovarian IGF-I content. The mean number of ovulations per ovary and the ovulatory efficiency were significantly reduced in ovaries perfused with 5 IU hCG, compared with those in ovaries perfused with 50 IU hCG. The addition of 100 ng/ml GH to the perfusate significantly increased the ovulatory efficiency and follicle diameter in the 5 IU hCG-treated ovaries. Exposure to GH significantly stimulated the resumption of meiosis in the follicular oocytes compared with that in ovaries perfused with medium alone. Furthermore, GH significantly stimulated the resumption of meiosis in ovulated ova and follicular oocytes in ovaries treated with 5 IU hCG. Thus, exposure to GH-stimulated follicular growth, oocyte maturation, and production of IGF-I in the in vitro perfused rabbit ovaries, which indicates that the ovary is in fact a site of GH reception and action. Additionally, GH enhanced the effects of gonadotropins, acting synergistically to promote the ovulatory process. These observations suggest that GH may amplify gonadotropin actions in the process of follicular development and ovulation, at least in part, by stimulating ovarian IGF-I production.

We have reported the outcome of a dose escalation study of single-fraction carbon ion radiotherapy for stage I NSCLC, showing the feasibility of obtaining excellent results comparable to those with previous fractionated regimens.