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Biomedical subjects

M Vignali

Publications and source records attributed to M Vignali.

At least 91 records · Page 5Linked to original sources
PubMed1994

Immunosuppressive effect of danazol on lymphocyte-mediated cytotoxicity toward human endometrial stromal cells.

The influence of danazol on the expression of lymphocyte-mediated cytotoxicity toward the stromal component of the endometrium was examined. Stromal cells were isolated from endometrial biopsy specimens of nine women without endometrial pathology and 12 patients with endometriosis. A cytotoxicity assay by 51Cr release was performed to determine the effect of danazol on natural killer lymphocyte lytic activity. A significant suppression of spontaneous cytotoxicity against stromal cell targets of both study subjects was observed when effectors were pretreated for 48 h with danazol at a concentration of 5 x 10(-6) mol/l. The time-course of the danazol effect indicated that drug activity was detectable as early as 24 h after the addition of the drug and increase up to 72 h. These findings confirm that danazol may influence the immune system at many levels. In particular, they demonstrate that the drug is also able to inhibit the natural immunity toward the stromal component of the endometrium.

Adult↗
PubMed1993

Mammalian oocytes exhibit specific recognition of the RGD (Arg-Gly-Asp) tripeptide and express oolemmal integrins.

Integrins are a family of cell adhesion receptors involved in many cell-cell and cell-matrix interactions. Some of these heterodimeric receptors, such as alpha 5 beta 1 and alpha v beta 1, specifically recognize the amino acid sequence Arg-Gly-Asp (RGD) within their ligands. The RGD sequence is found in fibronectin, vitronectin, and other extracellular matrix proteins. Our results demonstrate that the oolemmas of eggs from human and several other mammalian species contain receptors capable of binding to RGD ligands, and that integrin subunits are expressed by oocytes. Four distinct techniques were utilized to identify the presence of functional integrins on mammalian eggs. RGD-binding receptors were detected on the surfaces of zona-free eggs from all species tested. Covaspheres coated with PepTite-2000, which contains RGD, bound to the eggs and formed rosettes. Rosetting was competitively inhibited by PepTite-2000 and by GRGDTP, a soluble RGD peptide, but not by RGES. An ELISA using polyclonal antibodies directed against the cytoplasmic tails of the integrin subunits identified the integrin subunits alpha 5, beta 1, and alpha v, but not beta 3, in detergent extracts of Syrian hamster eggs. A dot blot confirmed the presence of alpha v in hamster egg lysates. Finally, the integrin subunits alpha 2, alpha 5, alpha 6, but not alpha 4, were detected on the surfaces of zona-free eggs from human and Syrian hamster. Immunobeads coated with monoclonal antibodies specific for alpha 2, alpha 5, and alpha 6 bound to the eggs and formed rosettes.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

Basic fibroblast growth factor and its receptor messenger ribonucleic acids are expressed in human ovarian epithelial neoplasms.

OBJECTIVE: Our purpose was to determine whether basic fibroblast growth factor is present in, and synthesized by, human ovarian epithelial neoplasms and to evaluate the expression of gene for the basic fibroblast growth factor receptor. STUDY DESIGN: The synthesis of basic fibroblast growth factor and its receptor was investigated in seven primary human ovarian epithelial neoplasms. Neoplastic tissues were homogenized and the cytoplasmic extracts purified by heparin-sepharose chromatography with a linear salt gradient of 0.6 to 3 mol/L sodium chloride in Tris-hydrochloric acid. The in situ synthesis of basic fibroblast growth factor and its receptor was demonstrated by polymerase chain reaction. Total ribonucleic acid was reverse transcribed and then amplified with two oligonucleotide primers specific for the bovine and human basic fibroblast growth factor gene and its human receptor gene. RESULTS: As assessed by both bioassay and radioimmunoassay a peak of basic fibroblast growth factor-like activity was present in all tumors in the chromatographic fractions eluted with 3 mol/L sodium chloride. The mitogenic effect on bovine adrenocortical endothelial cell proliferation varied from 35% to 153% above control cultures. Levels of basic fibroblast growth factor-like immunoreactivity were between 4 and 33 ng/ml. Qualitatively similar results were obtained after purifying the cytoplasmic extract of dispersed human ovarian tumor cells. The mitogenic effect was completely abolished by a specific neutralizing anti-basic fibroblast growth factor antibody. Single major deoxyribonucleic acid bands of the expected size (354 and 661 bp) were detected in all tumors studied. The identify of this material with the human basic fibroblast growth factor sequence was confirmed by restriction enzyme analysis. CONCLUSION: These data demonstrate that both basic fibroblast growth factor and its receptor are present in and synthesized by human ovarian tumor cells. Thus basic fibroblast growth factor might stimulate their abnormal proliferation through an autocrine mechanism.

Carcinoma↗
PubMed1993

Culture of human endometrial cells: a new simple technique to completely separate epithelial glands.

In vitro studies on dispersed human endometrial cells are still difficult to perform as the techniques employed do not allow an optimal separation of the cells. In particular, epithelial glands tend to maintain their tubular structure even after enzymatic dispersion. This could be a disadvantage when monolayers of single well dispersed cells are needed. In this paper we describe a new technique to establish monolayer cultures of isolated endometrial stromal and epithelial cell populations. After a first collagenase digestion, stromal and epithelial cells were separated by differential sedimentation at unity gravity. The epithelial glands obtained were further dispersed in single cells using a short incubation with low amount of trypsin. Morphologic characterization was performed using immunohistochemistry for vimentin and cytokeratins. Compared to previous described methods, this procedure is shorter and could better preserve cell surface structures. Thus, it could be successfully employed for in vitro studies of human endometrial pathophysiology.

Cell Separation↗
PubMed1992

Evidence for the presence of an integrin cell adhesion receptor on the oolemma of unfertilized human oocytes.

The Arg-Gly-Asp peptide (RGD), contained in several extracellular matrix proteins such as fibronectin, laminin, vitronectin, and collagen, is a tripeptide that plays a role as a recognition sequence in many cell-to-cell and cell-to-matrix adhesion mechanisms, through its interaction with several receptors of the integrin family. We previously described the ability of the oolemma of hamster oocytes to bind GRGDTP coupled to the surface of activated immunobeads and demonstrated that RGD-containing oligopeptides inhibit the adhesion of human and hamster spermatozoa to zona-free hamster oocytes and their subsequent penetration. In the present experiments, we show, utilizing immunobeads coated with an RGD-containing peptide (PepTiteTM 2000), that the oolemma of unfertilized human eggs is also able to recognize this adhesion sequence. The binding of PepTiteTM 2000-coated immunobeads to the oolemma was inhibited by the oligopeptide GRGDTP as well as by fibronectin and laminin. When immunobeads were prepared with a PepTiteTM concentration of 10 micrograms/ml, GRGDTP 150 micrograms/ml, laminin 80 micrograms/ml, and fibronectin 60 micrograms/ml inhibited bead rosetting on the egg surface. These data suggest that a specific binding moiety for RGD is present on the human egg surface. The binding of fibronectin to the oolemma was also demonstrated by the rosetting of immunobeads coupled with antifibronectin antibody to human oocytes after their exposure to 1 mg/ml free fibronectin. Such binding of fibronectin to the oolemma could be inhibited by coincubation with a monoclonal antibody directed against the cell adhesion fragment of fibronectin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1992

Two-step tumour targetting in ovarian cancer patients using biotinylated monoclonal antibodies and radioactive streptavidin.

A new method for intraperitoneal tumour targetting in ovarian cancer using biotinylated monoclonal antibodies (MoAb) and radioactive streptavidin is described. Fifteen patients with histologically documented ovarian carcinoma were injected intraperitoneally with 2 mg of biotinylated MoAb MOv18, followed 3-5 days later by 100-150 micrograms of indium-111 streptavidin, at the specific activity of 280-370 MBq/mg in 500 ml of normal saline. No toxicity was observed. Tumours were imaged from 2 to 48 h after radioactivity injection by recording both planar and single photon emission tomography (SPET) data. All patients underwent surgery 1-8 days later (mean 3 days) after scanning. The resected tumour and normal tissue radioactivity were measured. On the day of surgery, the tumour to normal tissue ratio was 9:1 (range 3:1-30:1) and 45:1 (range 12:1-120:1) for intra- and extraperitoneal samples, respectively. The mean tumor to blood ratio was 14:1 (range 4:1-30:1). The injected dose (i.d.) per gram of tumour was 0.112 (range 0.01-0.3) for recurrences and 0.05 for primary tumour (range 0.005-0.2). Over 24-48 h 14% i.d. (range 8-18% i.d.) was found in the urine, 14% i.d. (range 6-29% i.d.) in the blood and 63% i.d. (range 56-70% i.d.) was still in the peritoneal cavity. These preliminary clinical data suggest that this two-step strategy may be superior to the conventional approach (radiolabelled antibodies) for intraperitoneal radioimmunolocalization and radioimmunotherapy of ovarian cancer.

Bacterial Proteins↗
PubMed1992

Danazol suppresses both spontaneous and activated human lymphocyte-mediated cytotoxicity.

Danazol is an effective drug in the treatment of endometriosis. Previous reports suggest that it can act by influencing the immune system. Thus, in this study we evaluated whether danazol is able to affect the expression of natural and activated cytotoxicity by human peripheral blood lymphocytes (PBL). A cytotoxicity assay by 51Cr release was performed to determine the effect of danazol on lymphocyte cytotoxic response toward an erythroleukemic cell line (K562). A significant suppression of spontaneous cytotoxicity was observed when lymphocytes were pretreated for 18 h with danazol at the concentration of 5 x 10(-6) M. In addition, danazol could significantly inhibit both IL-2- and INF alpha-activated cytotoxicity at the concentration of 10(-5) M. Following 18 h exposure to danazol, PBL were able to completely recover their spontaneous but not activated cytotoxic potential upon further in vitro incubation (18 h) in absence of the drug. The biological significance of danazol as an inhibitor of immune function under experimentally defined conditions is discussed in relation to its possible role in vivo.

Cytotoxicity, Immunologic↗
PubMed1992

Leuprorelin acetate depot vs danazol in the treatment of endometriosis: results of an open multicentre trial.

A national multicentre trial was organized in order to compare the efficacy and safety of leuprorelin acetate depot and danazol in the treatment of endometriosis. Sixty-seven patients with pelvic endometriosis of different severity at laparoscopy were included in the study and followed during the 24 weeks of treatment. Leuprorelin acetate depot 3.75 mg was injected every 24 days, while the daily dose of danazol was 600-800 mg. At the end of the study objective improvements induced by the two drugs were observed by a second laparoscopic examination. In addition, at regular intervals during the study semiquantitative evaluation of subjective symptoms were monitored. Scoring the final objective changes in the two patient groups revealed no significant difference, however the women treated with leuprorelin acetate depot registered significantly better control of pelvic pain. Due to its efficacy, tolerability and ease of use, leuprorelin acetate appears to be an excellent drug for the treatment of endometriosis.

Adult↗
PubMed1991

Deficient antiendometrium lymphocyte-mediated cytotoxicity in patients with endometriosis.

OBJECTIVE: To study the possible role of the immune system in the pathogenesis of endometriosis. DESIGN: A cytotoxicity assay by 51Cr release was performed to determine the lymphocyte cytotoxic response toward endometrial targets and an erytroleukemic cell line (K562). SETTING: The assays were performed in an academic research environment. PATIENTS, PARTICIPANTS: Twenty-five control women and 25 patients with endometriosis were selected on the basis of laparoscopic examination. INTERVENTIONS: The lymphocyte cytotoxic activity was evaluated separately on endometrial stromal and epithelial cells after 4 hours' incubation. MAIN OUTCOME MEASURE: The study was designed to determine, in controls and endometriosis patients, the lymphocyte-mediated cytotoxicity toward stromal and epithelial cells of endometrium. RESULTS: The lymphocyte response in the presence of stromal cell antigens was significantly lower (P less than 0.02) in disease-affected women when compared with that obtained in controls (2.89 +/- 0.87 and 7.64 +/- 1.66, respectively). In contrast, when the same assay was performed on K562 cells, no difference was observed between endometriosis patients and controls. CONCLUSIONS: These data suggest that an altered immune recognition might be one of the pathogenic mechanisms of endometriosis. Moreover, they indicate that this is not a general phenomenon but is specific for the endometrial target.

Adult↗
PubMed1990

Magnetic resonance imaging in endometrial carcinoma staging.

Correct evaluation of myometrial infiltration is essential in patients with stage I and II endometrial cancer who are candidates for hysterectomy without lymphadenectomy, if extensive infiltration of the myometrium is not present. The aim of this study was to evaluate the use of magnetic resonance imaging (MRI) to improve staging of patients with endometrial cancer. Thirty patients with histological diagnosis of endometrial cancer were studied with MRI at 1.5 T and subsequently underwent abdominal hysterectomy. The MRI results were compared with those of the histological tests. MRI was performed with a 1.5-T magnet and spin-echo (SE) technique [repetition time/echo time (msec) = 2.000/35-90]. Contiguous 4-mm sections of were obtained from the sagittal plane. Clinical staging was not confirmed in two patients who presented with cervical extension of the tumor. The overall accuracy of MRI in determining the grade of myometrial and cervical invasion was 86 and 90%, respectively.

Aged↗
PubMed1989

Antenatal prevention of respiratory distress syndrome; a multicentre survey.

Obstetric and neonatal data were collected on 934 preterm deliveries in 11 Italian centres in 1980, 1985 and 1986. Therapeutic regimens for prevention of respiratory distress syndrome (RDS) were applied in 42% of the cases in 1980, 32% in 1985 and 42% in 1986. Prevention was made in most cases with corticosteroids, although their use fell progressively from 94% in 1980 to 74% in 1986. A combination of two substances was used in a percentage of cases varying from 5 to 10% in all three years. In 903 non-malformed infants, the overall incidence of RDS was not significantly different in cases in which pharmacological prevention was attempted compared with cases without prevention. The only factors significantly affecting the incidence of RDS were gestational age, birth weight and Apgar score.

Data Collection↗
PubMed1989

[Assessment of the myometrial infiltration of endometrial carcinoma (FIGO stage I-II). The accuracy of magnetic resonance (1.5 T)].

Thirty-three patients with histological diagnosis of endometrial carcinoma were studied with MR imaging at 1.5 T. All patients were clinically diagnosed as having stage I or II disease (FIGO) and underwent abdominal hysterectomy. MR tissue characteristics and morphological features were compared with morphological features were compared with microscopic pathological findings, in all patients. The following initials were adopted for statistical comparison: M0 = tumor confined to endometrium; M1 = infiltration of the inner third of myometrium; M2 = invasion of the central third of myometrium; M3 = infiltration of the outer third of myometrium. T1-weighted (SE 600/30) and T2-weighted (SE 2000/35.90) (SE 1500/28.60) images were obtained in sagittal plane (section thickness = mm 4). Overall MR accuracy in determining the extent of myometrial invasion was 78%. Correct evaluation of myometrial invasion is essential in patients with stage I or II endometrial carcinoma who are to undergo transvaginal hysterectomy without lymphadenectomy (M0, M1).

Aged↗
PubMed1986

Effect of cigarette smoking on plasmatic regulation of vascular prostacyclin in pregnancy and puerperium.

Prostaglandins, particularly prostacyclin, participate in the control of fetal and maternal circulations. In the present study the effect of cigarette smoking on plasma prostacyclin-stimulating activity during late pregnancy and the puerperium (four to six months) and in the newborns was assessed. Plasma samples from 22 apparently healthy nonsmokers and 17 smokers (more than 15 cigarettes per day) were obtained twice during pregnancy and once after delivery. Plasma samples from nine infants born to smokers and seven infants born to nonsmokers were obtained 72 to 96 hours after birth. Plasma activity was evaluated by incubating the plasma samples with cultured rat aortic smooth muscle cells and measuring the prostacyclin released in the culture medium by specific radioimmunoassay of its stable metabolite, 6-keto-prostaglandin F1 alpha. In all of the women, plasma activity did not change significantly during pregnancy or after delivery. In smokers, plasma activity was always lower than in nonsmokers, but not significantly so. The plasma activity of infants born to smokers was significantly lower than that of infants born to nonsmokers.

Animals↗
PubMed1984

Metergoline in the management of normoprolactinemic secondary amenorrhea. A multicentric controlled trial.

In a multicentric study, the effect of the antiserotoninergic agent metergoline was evaluated in the management of patients with idiopathic normoprolactinemic secondary amenorrhea (NSA). The awareness that psychological factors might lead to a spontaneous reappearance of menses was also taken into account, and all the patients, after physical, gynecological and laboratory examinations, and the performance of the progesterone withdrawal bleeding test (100 mg i.m.) and the clomiphene citrate test (100 mg p.o./day for 5 days), were treated for 60 days with placebo; only patients showing no menses during placebo administration were later treated with metergoline. 108 patients entered the trial: of these, 48 experienced menses on admission or during placebo administration, and were withdrawn. Of the 60 patients not responding to placebo, 50 were treated for 90 days with metergoline (4 mg t.i.d.), and 23 had menses, ovulatory in 68.4% of cases. A new placebo treatment was accompanied, in the majority of cases, by recurrence of amenorrhea. These results indicate that many patients with NSA may experience a spontaneous disappearance of the disease: in cases more seriously affected metergoline might be a useful therapeutic agent.

Amenorrhea↗
PubMed1982

Three-day therapy of vaginal candidiasis with clotrimazole vaginal tablets and econazole ovules: a multicenter comparative study.

The activity and tolerance of 200 mg clotrimazole vaginal tablets and 150 mg econazole vaginal ovules were compared in the treatment of vaginal candidiasis in a controlled multicenter (3 centers) study. Treatment was allocated according to a randomized list and each investigator treated 20 patients per group. Duration of therapy was 3 days. Follow-up investigations were carried out approximately 1 and 4 weeks after commencement of treatment. The results were analyzed for 54 patients treated with clotrimazole and 57 with econazole; 9 patients were excluded from the analysis because of insufficient data. At the first follow-up examination, 1 week after starting therapy, 93.7% of patients on clotrimazole and 83.7% of patients on econazole had negative cultures. The success of therapy, as judged by the clinicians, was 95.8% for clotrimazole and 88.0% for econazole. For the first and fourth week combined reviews, 86.0% of clotrimazole patients and 77.8% of econazole patients had negative cultures and the success of therapy was 88.4% for clotrimazole and 80.0% for econazole. These differences were not statistically significant but there was a clear numerical superiority for clotrimazole. Tolerance was good in all the patients and no side-effects were observed. One can conclude that the treatment of vaginal candidiasis with a 200-mg vaginal tablet of clotrimazole over 3 days is highly effective and comparable with other established treatment regimes.

Adolescent↗