Effects of nafarelin on bone density.

The effects of the gonadotropin-releasing hormone analog nafarelin on bone metabolism during treatment of premenopausal women for endometriosis were evaluated in two studies, both of which involved 6 months of medication followed by 6 months without medication. With low doses of nafarelin (200 micrograms/day), bone mineral measurements remained constant, whereas with high doses of the drug (400 micrograms/day) bone mineral levels decreased significantly. Within 6 months after treatment was stopped, however, bone mineral measurements returned to normal levels. Both dosages of nafarelin resulted in significant increases in bone resorption, but these determinations also returned to pretreatment values after drug therapy was stopped. In the second study, patients treated with nafarelin, 400 micrograms/day, and norethindrone, 1.2 mg/day, did not lose significant amounts of bone mineral during the treatment period.

Thymic lymphocytes. III. Cooperative phenomenon in the proliferation of thymocytes under Con A stimulation.

In the present paper, the response of thymocytes to Con A is analyzed in terms of a cooperative phenomenon between medullary thymocytes, cortical thymocytes, thymic accessory cells, and interleukin 2. Medullary thymocytes respond spontaneously to Con A and produce IL-2. The addition of exogenously produced IL-2 enhances their proliferation. Small numbers of cortical (PNA+) thymocytes do not respond to Con A, even in the presence of IL-2-containing supernatant. By increasing the number of PNA+ cells per well, sensitivity to Con A and IL-2 appears. This response may be linked either to the increase in a minor PNA+-responding population and/or to the enhanced contamination by medullary thymocytes and macrophages in non-responding PNA+ thymocyte population. In this hypothesis, either the contaminating cells respond by themselves and/or cooperate with PNA+ cells to induce their proliferation. Coculture of non-responding low numbers of PNA+ thymocytes with Con A- and IL-2-containing supernatant in the presence of PNA- cells containing thymic medullary thymocytes and macrophages always produces a higher response than that of each individual population. These results show that a cooperative phenomenon occurs in the cocultures of PNA+ and PNA- thymic cells. We can show using PNA+ and PNA- thymocytes with different Thy 1 alleles, that indeed both PNA+ and populations participate PNA-thymocytes with different Thy 1 alleles, that indeed both PNA+ and PNA- populations participate in the generation of proliferating cells. We can demonstrate, by lysis experiments with monoclonal antibodies and complement that at the end of coculture, most of the proliferating cells are Lyt 1+, and part are Lyt 2+ or L3T4+. We discuss the fact that the phenotype of the cells after activation does not allow us to deduce the phenotype of their precursors. Lysis of Ia+ cells prior to coculture, reduces the level of the proliferative response but does not modify the percentage of cooperation produced by the coculture. Cooperation with medullary mature thymocytes or the presence of active Ia- accessory cells possibly able to convert to Ia expression during coculture experiments may account for these results.

A new vaginal antifungal agent--butoconazole nitrate.

The continuously increasing incidence of vulvovaginal candidiasis necessitated a search for novel therapeutic modalities. Butoconazole nitrate (BN) a new imidazole, has been singled out for clinical studies since, in experimental vaginal candidiasis, it proved more effective than either miconazole nitrate (MN) or clotrimazole. One hundred and thirty volunteers with vaginal candidiasis, verified by wet mount and positive fungal culture, were randomly assigned to receive daily, for 6 days, either 1% BN (44 patients), or 2% BN (45 patients) vaginal cream or 2% MN (41 patients) vaginal cream. The patients were comparable regarding age (85% were 18-39 years of age), gravidity and parity. Twenty-five per cent had a recent history of unsuccessfully treated fungal vaginitis. Eight days after completion of treatment, negative fungal cultures were found in 98% of the patients using 2% BN, in 91% of patients using 1% BN and in 83% using 2% MN. The recurrence rate of the disease was low; about 80% of patients using 1% and 2% BN and 68% of those using 2% MN were culture-negative 30 days after conclusion of treatment. Rapid relief of clinical symptoms was experienced by patients in all three treatment groups. No significant side effects of the treatment were observed in any of the treatment groups.

Management of the fetus with a urinary tract malformation.

Obstetric sonography revealed urinary tract malformations in 13 fetuses. Six had severe dysplastic lesions incompatible with postnatal life; in all six, oligohydramnios and inability to detect normal kidney or bladder allowed appropriate counseling and management. Four fetuses had unilateral lesions (three hydronephrotic, one multicystic); all had evidence of adequate contralateral function and were successfully treated after delivery near term. Three fetuses had bilateral hydronephrosis secondary to urethral obstruction. The two who were born near term died of hypoplastic lungs, end-stage hydronephrosis, and facial and skeletal deformities. The other, electively delivered at 32 weeks, had none of the stigmata of Potter's syndrome, and early decompression salvaged sufficient renal function for survival. Prenatal sonographic assessment of urinary tract anatomy and function can improve perinatal management. The fetus with hydronephrosis may benefit from early decompression.

The transverse common duct.

The sonographic appearance and incidence of transverse common ducts were studied. A review of 118 intraoperative cholangiograms showed a transverse segment longer than 35 mm in 18% of the dilated extrahepatic ducts and in 6% of the normal caliber ducts. The causes of obstruction varied. When dilated, the transverse segment may be confused with the portal and splenic veins on a sonogram. An awareness of this anatomic variation can be helpful in locating an obstructing lesion of the extrahepatic bile duct.

Antigenic profile of murine natural killer cells.

We have determined the antigenic profile of natural killer (NK) cells from C57BL/6 mice that have lytic activity to RLmale1 tumor cells, in a 5-hr 51Cr release assay. To perform the antigenic typing more effectively, we enriched the NK populations by using discontinous BSA density gradient and established a correlation between NK activity and NK-1 antigen in each layer. As NK activity and Nk-1 antigen are both criteria for measuring NK cells, we determined the antigenic profile of NK cells by monitoring both the change in NK activity and NK-1 cells, after elimination of cells with various antisera to lymphocytic alloantigens. We have found that all NK cells expressed Nk-1, Qa-4, and Qa-5 antigens; about 50% of NK cells expressed Thy-1 and Qa-2 antigens, and 25% of NK cells also expressed Lyt-1 antigen. NK cells did not express significant amounts of Qa-1, Qa-3, Lyt-2, and Lyb-2 antigens. Therefore, spontaneous NK cells are heterogeneous with regard to their surface antigens, some of which are T cell antigens. However, the present antigenic profile does not allow us to classify NK cells in the conventional T cells lineage.

Antigenic modulation in vitro. III. Failure to modulate H-2 antigens on several mouse tumors.

The capacity of various malignant and normal mouse cells to acquire resistance to lysis by guinea pig complement during exposure to H-2 antisera in vitro at 37 degrees C (antigenic modulation) was examined. All tumors tested, including cell lines of the TL+ leukemias RADA1, ASL1, and RLmale1, the TL- leukemia EL 4, myelomas MOPC-70A and S194, and the sarcoma Meth A, failed to modulate when incubated with multispecific or monospecific H-2 antisera up to 24 hours, even though under comparable conditions thymus-leukemia (TL) antigens and surface IgG molecules modulated within several hours. Indirect sensitization of RADA1 leukemia cells with H-2 antisera followed by antiserum against mouse IgG also failed to induce H-2 antigen modulation. Normal peritoneal cells from certain mouse strains were partially modulated with H-2D-specific or H-2K-specific and monospecific antisera within several hours, but normal thymus and lymph node cells did not modulate. Modulation of peritoneal cells occurred without a complete loss of sensitizing H-2 antibody from the cell surface and required a cobra venom factor-sensitive activity that could be restored by human complement component C3. Modulation of TL antigens in vitro had previously been shown to have similar characteristics.

In vivo modulation of thymus-leukemia antigens on mouse leukemia cells and thymocytes: retention of modulating antibody on the cell surface.

Inoculation of RADA1, ASL1, and ERLD murine leukemia cells into the peritoneal cavities of (C57BL/6J x A/TL--)F1 mice hyperimmunized against thymus-leukemia (TL) cell-surface antigens rendered most cells insensitive to lysis in vitro by guinea pig complement even in the presence of TL antiserum. Thymocytes of A/J mice were similarly modulated by passive injection of TL antiserum. In all cases, retention of some modulating antibody on the surfaces of most cells modulated in vivo for 1--27 days was indicated by: 1) acquisition of sensitivity of modulated cells to lysis by absorbed rabbit complement; 2) positive immunofluorescence reactions for mouse IgG on the surfaces of modulated cells; and 3) release of cytolytically active TL antibody from cells into the circulation of unimmunized mice following transfer of modulated cells. Reversal of modulation of RADA1 cells was complete in some experiments within 24 hours after transfer to unimmunized mice, by which time all indications of cell-bound TL antibody were lost. These results indicate that even long-term modulation of TL antigenicity in vivo does not result in a complete loss of modulating antibody (presumably attached to TL antigens) from the cell surface.

Antigenic modulation in vitro. I. Fate of thymus-leukemia (TL) antigen-antibody complexes following modulation of TL antigenicity from the surfaces of mouse leukemia cells and thymocytes.

The modulation or loss of thymus-leukemia (TL) antigenicity from the surfaces of mouse RADA1 leukemia cells and normal thymocytes during incubation with TL antibody in vitro at 37 degrees C was investigated by cytotoxicity, immunofluorescence, and immunoelectron microscopy. The fate of bivalent and monovalent antibody during modulation was visualized by fluorescence microscopy. Considerable antibody remained bound to the cell surface after modulation, bivalent antibody being displaced topographically into "patches" and "caps" while monovalent antibody was only slightly aggregated on the cell surface. Some antibody was internalized, presumably by pinocytosis, and was sequestered into the Golgi region of the cell. Capping usually occurred over the pole of the cell opposite from the Golgi region, which may explain the lack of extensive pinocytosis of modulating bivalent antibody. Since modulation with monovalent antibody occurs without patch or cap formation, gross topographical redistribution of TL antigen-antibody complexes is not required for modulation, although more subtle displacement of these complexes may be involved. Modulation was demonstrable by cytotoxicity with guinea pig C' but not with absorbed rabbit C', indicating that modulated TL antigens remain bound to the cell surface. A heat-labile factor in TL antiserum and in mouse serum in general is responsible for "blocking" the cytolytic interaction of guinea pig C' with modulated TL antigen-antibody complexes.

Hybrid antibody-induced topographical redistribution of surface immunoglobulins, alloantigens, and concanavalin A receptors on mouse lymphoid cells.

Redistribution of surface immunoglobulins, H-2(b), Thy-1.2, and TL.1,2,3 alloantigens, and concanavalin A receptors on mouse lymphoid cells induced by hybrid rabbit F(ab')(2) antibody (anti-mouse immunoglobulin/anti-visual marker or anti-concanavalin A/anti-visual marker) was studied by immunofluorescence. When used directly to label surface immunoglobulin, and indirectly to label alloantigens and concanavalin A receptors, hybrid antibodies induced similar displacement of all surface components from a uniform distribution into "patches" and "caps" at 37 degrees . One hybrid antibody preparation, antimouse immunoglobulin/anti-ferritin, contained negligible amounts of bivalent anti-mouse immunoglobulin antibody, and was therefore "monovalent" for the antimouse immunoglobulin specificity. This observation suggests that factors other than multivalent crosslinking are responsible for hybrid antibody-induced redistribution of cell-surface components. Cap formation induced by hybrid antibody was enhanced markedly by attachment of the visual marker, either ferritin or southern bean mosaic virus, at 37 degrees . At -5 degrees , hybrid antibody does not displace uniformly distributed H-2(b) alloantigen-alloantibody complexes, but patches of label develop when ferritin attaches to the hybrid antibody. These results explain the patchy distribution of cell-surface components, which is a temperature-independent characteristic of labeling with hybrid antibodies and visual markers for electron microscopy.