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Biomedical subjects

G Giacomini

Publications and source records attributed to G Giacomini.

At least 19 recordsLinked to original sources

Measurement of DNA cross-linking in patients on ifosfamide therapy using the single cell gel electrophoresis (comet) assay.

The single cell gel electrophoresis comet assay has become established as a sensitive technique for measuring DNA strand breaks. The technique has been modified to allow the sensitive detection and quantitation of DNA interstrand cross-linking at the single cell level. Cells are irradiated immediately before analysis to deliver a fixed level of random strand breakage. After embedding of cells in agarose and lysis, the presence of cross-links retards the electrophoretic mobility of the alkaline denatured cellular DNA. Cross-links are, therefore, quantitated as the decrease in the comet tail moment compared with irradiated controls. Using this method, a linear response of cross-linking versus dose of chlorambucil over a wide dose range was demonstrated in human lymphocytes after drug treatment ex vivo. The method was also sensitive enough to determine cross-linking in clinical samples after chemotherapy. For example, crosslinking was observed in the lymphocytes of patients receiving ifosfamide (3 g/m2/day) as a continuous infusion for 3-5 days or as a 3-h infusion daily for 3 days. Cross-links were detected in all patients within 3 h, with no evidence of DNA single strand break formation. In patients receiving continuous infusion, a plateau of cross-linking was reached by 24 h. In the patients receiving ifosfamide over 3 h, a clear decrease in the peak level of cross-linking was observed before subsequent infusions.

Antineoplastic Agents, Alkylating↗

Accuracy of cervical/vaginal cytology in the diagnosis of bacterial vaginosis.

BACKGROUND AND OBJECTIVES: The cytological pattern of bacterial vaginosis (BV) in the Papanicolaou smear has a controversial history (coccoid vs. Gardnerella type) that has not allowed an efficient use of cervical/vaginal cytology in the diagnosis of this condition. Our study is an attempt to clarify this topic. GOALS: To evaluate the accuracy of cervical/vaginal cytology in the diagnosis of BV. STUDY DESIGN: 1,896 separate examinations were performed on 1,471 women attending our sexually transmitted diseases center between 1990 and 1993. Amsel's composite clinical criteria were used as a gold standard. The Pap smear was prepared on two slides with the addition of a vaginal specimen. RESULTS: In the 166 cases of BV, sensitivity is 88.7%, and specificity is 98.8%. Positive predictive value is 89.8, and negative predictive value is 98.7. Kendall's correlation coefficient is 0.88 (p < 0.001). CONCLUSIONS: Our findings support the validity of the vaginal Pap smear in diagnosing BV and suggest the screening of such a disease.

Female↗

HIV infection increases the risk of squamous intra-epithelial lesions in women with HPV infection: an analysis of HPV genotypes. DIANAIDS Collaborative Study Group.

We assessed the association between different HPV genotypes, HIV infection, and cervical squamous intra-epithelial lesions (SIL) in 236 women with known HIV serostatus enrolled in a longitudinal multicentric study in Italy. Of these women, 135 were HIV-infected, and were not markedly different from HIV-negative women with regard to demographic characteristics, sexual practices, smoking, or intravenous drug use. We obtained 232 cervical smears suitable for cytological examination and HPV-genotype analysis (134 from HIV-positive women and 98 from HIV-negative women). For 86 HIV-positive and 89 HIV-negative women, the smears appeared normal at cytomorphological analysis. Cytological dysplasia of varying degrees was detected in 48 smears from HIV-positive women and in 9 from HIV-negative women. HPV prevalence, assessed using polymerase-chain-reaction analysis, did not significantly differ between HIV-positive and HIV-negative women. The prevalence of HPV-associated SIL was much greater among HIV-infected women. The most frequently detected genotypes in both groups were HPV 16 and HPV 18. The prevalence of HPV 16 among HIV-positive women was similar to that for HIV-negative women; this was also true for HPV 18. However, in the HIV-positive group, most of these genotypes were associated with SIL. HIV-positive women showed a wider spectrum of genotypes, including non-oncogenic and rare types. An association between SIL and HIV infection was confirmed for all HPV genotype classes.

Adult↗

Determinants of squamous intraepithelial lesions (SIL) on Pap smear: the role of HPV infection and of HIV-1-induced immunosuppression. DIANAIDS Collaborative Study Group.

This study aimed to identify risk factors for squamous intraepithelial lesions (SIL) in women with known HIV status and to explore the association between SIL, HPV subtype, and HIV-induced immunosuppression. The study population consisted of women with known HIV serological status who were attending a network of 16 clinical centres in Italy. Detailed behavioural data, clinical and laboratory parameters, and samples for diagnosis of SIL by Papanicolau smear and HPV infection using a polymerase chain reaction (PCR) were obtained from each study participant. The strength of the association between SIL and possible risk factors was assessed calculating crude and adjusted odds ratios derived from univariate analysis and multivariate models. We enrolled 236 women, of whom 135 (57.2%) were HIV-infected. SIL was diagnosed in 57 women (24.1%); of these, 48 (35.6%) were HIV-infected and 9 (8.9%) were HIV-negative. HPV-DNA was detected in 41 (72%) women with SIL and in 45 (25%) women without SIL. HPV-DNA was more often detected among HIV-infected women than among HIV-negative women (40% vs. 32%), but the difference was not statistically significant. Women infected with high-risk types or with low-risk-uncharacterised types of HPV both had a higher risk of SIL compared with HPV-negative women (respectively, AOR: 17.53 and AOR: 2.89). HIV-infected women with severe or moderate immunosuppression were more likely to have SIL than HIV-negative women (respectively, AOR: 7.29 and AOR: 3.09) also independently from HPV infection. Women reporting use of a contraceptive pill had a 2.5 times higher risk of SIL compared with those who never used hormonal contraceptives. The results confirm that high-risk HPV types are strongly associated with SIL, and that HIV infection may strengthen the effect of HPV at cervical level. The use of oral contraceptives may slightly increase the risk of SIL in women with at-risk behaviour for HIV infection.

Adult↗

Ovarian mesothelial and extramesothelial cells in interactive culture.

The ovarian mesothelium (OM) represents the tissue of origin of ovarian epithelial cancer. To gain insight into the regulation of this tissue, OM organoids and submesothelial ovarian stromal cells (SC) were isolated from New Zealand White rabbits by a stepwise tissue dispersal technique, while granulosa cells (GC) were aspirated from mature follicles (14 +/- 4 groups/animal). OM and SC dispersal were sequentially accomplished by: a) 1-h incubation in collagenase type I (300 U/ml), gentle scraping of the ovarian surface, and 1 g sedimentation of OM organoids (equivalent to 0.93 +/- 0.40 x 10(6) cells/animal) on 5% bovine serum albumin (BSA); b) 2-h incubation in pronase-collagenase (0.5%-300 U/ml) under periodical resuspension and gentle scraping of SC (1.40 +/- 0.25 x 10(6)/animal) from OM-denuded ovaries. After a week-long in vitro expansion, OM cells (OMC) were cultured alone and with SC or GC within monocameral vessels or bicameral transfilter vessels in serumless, fibronectinrich (4 micrograms/ml) HL-1 medium. After 7 d of contact cell-cell interaction, cytokeratin-positive OMC became surrounded by fibroblastoid, vimentin-positive SC or by cytokeratin and vimentin-weakly positive GC. Filter-bound OMC humorally interacting with underlying SC or GC displayed a biphasic, epithelioid and spindle, morphology with universal cytokeratin expression. Bromo-2'-deoxyuridine (BrdU) immunoperoxidase revealed mean cell proliferation indices of 14.88% for OMC cultured alone, 11.21% and 19.39% for OMC cultured with GC or SC in monocameral dishes, and 15.25% or 22.47% for OMC cultured in bicameral vessels over GC or SC, respectively. This model provides an experimental tool for investigating the unexplored role of stromal-mesothelial interaction in OM pathobiology.

Animals↗

Epithelial cells are the major source of biologically active granulocyte macrophage colony-stimulating factor in human endometrium.

Granulocyte macrophage colony-stimulating factor (GM-CSF) has emerged as an important growth factor for trophoblast and other placental cells, leading to improved placental functioning and fetal survival. Recent observations have indicated that GM-CSF is synthesized by epithelial cells in the murine pregnant and non-pregnant uterus. In this study, the production of GM-CSF by cells derived from human endometrium is assessed using a sensitive bioassay and specific neutralization of the cytokine bioactivity with a monoclonal antibody to GM-CSF. Originally, GM-CSF was assayed in the culture supernatants of explant cultures of human endometria. Concentrations of GM-CSF up to 4440 pg/ml were detected. Subsequently, enriched epithelial cell cultures were prepared from glands isolated from human endometrium. The purity of epithelial cultures was demonstrated by the expression of cytokeratin, a weak immunoreactivity for vimentin and a lack of immunoreactivity for leukocyte common antigen, CD68, a macrophage-specific protein and endothelial marker (factor VIII-related antigens). Detected concentrations of GM-CSF were as high as 18,800 pg/ml. Furthermore, pure epithelial cells of a neoplastic endometrial cell line ECC1 secreted GM-CSF, confirming the ability of endometrial epithelial cells to secrete this cytokine. The immunostaining of dated endometria from proliferative and secretory phases showed primarily that epithelial cells, and to a lesser extent stromal cells, exhibited immunoreactivity for GM-CSF. A Western blot analysis, performed to validate the immunohistochemical data, confirmed the presence of an immunoreactive gene product for GM-CSF in human endometrium throughout the menstrual cycle. These findings indicate that human endometrium synthesizes GM-CSF and that epithelial cells are a major contributor to its production.

Adult↗

Cervical intraepithelial neoplasia and human papillomavirus related lesions of the genital tract in HIV positive and negative women.

Two hundred and twenty one women at high risk for HIV (intravenous drug users and/or those with infected partners) were investigated, through a self-filled questionnaire and gynaecological examination, to define the relationship between genital Human Papilloma Virus (HPV) infections, preneoplastic cervical intraepithelial lesions (CIN) and behavioural risk factors. In the 121 HIV positive women, 58 (47%) had HPV lesions at colposcopic and/or cytologic examination and, out of these 58, 23 (40%) had CIN 1, CIN 2 or CIN 3. Six out of the 16 cases with CIN 1 and CIN 2 (37%) followed-up showed a rapid progression of the lesion to CIN 3; in 3 women the interval was 6 months, in the other 3 about 12 months. Only 5 (7%) of the remaining 66 women without HPV lesions had a CIN lesion, with an obviously significant difference on comparison with HPV positive subjects. Sixty two women out of the 121 (52%) had a previous diagnosis of condylomata. In the 100 HIV negative women, 23 (23%) had HPV lesions and, among these 23, 6 (26%) had CIN 1, CIN 2 or CIN 3; 1 of them had rapid progression from CIN 1 to CIN 3 within a year. Only 5 (3%) without HPV infection showed any kind of CIN. 33 women out of 100 (33%) had a previous clinical history of condylomata. Our findings strongly suggest that HIV infection is associated with HPV lesions and that cervical cytological abnormalities develop in this situation. There is a need for short interval cytological and colposcopic follow-up for women at high risk of HIV infection.

Adult↗

The diagnostic cytology of nonspecific vaginitis.

Diagnostic cytology has not kept pace with the general growing interest in nonspecific vaginitis (NSV). This study on 114 patients compares Papanicolaou vaginal smears with bacteriologic cultures, whiff test, gram-stained vaginal smears, and vaginal pH in the evaluation of nonspecific vaginitis. Cytology is a valid practical test for diagnosis for nonspecific vaginitis; however, it must be emphasized that the coccoid morphotype is the same as the Gardnerella morphotype. Perhaps the Gardnerella morphotype is the most typical of all the inflammatory typologies found in vaginal smears, and it was indeed one of the first to be described in the history of diagnostic cytology--albeit with a slight misnomer. The question is of considerable importance since NSV is the most frequent and persistent of all collapses of the vaginal ecosystem. Cases of NSV must be screened to prevent more serious diseases of the female genital tract.

Adolescent↗

Synchronous adenoacanthoma of the ovary and endometrial adenocarcinoma. Report of two cases.

Two cases of synchronous endometrioid adenocarcinoma of the endometrium and ovary are here presented. The clinical aspects and the pathologic findings support an autochthonous origin of these neoplasias. The existence of the extended Müllerian system can explain such a possibility. The recognition of this possible association is really important because it implies a different therapeutic approach.

Adenocarcinoma↗