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Biomedical subjects

A Ruggiero

Publications and source records attributed to A Ruggiero.

At least 19 recordsLinked to original sources
PubMed1995

[The knowledge, attitudes and practice in regard to the risk for occupational HIV infection in a group of gynecologists from 3 Italian regions].

The risk of occupational infection by blood-borne pathogens has been well evaluated. However, a low perception of this risk among health care workers (HCWs) and a low compliance to infection control measures has been documented by several studies. During a meeting of gynecologists of Southern Italy, a survey of knowledge, attitudes and practices (K.A.P.) on HIV and other blood-borne viral infections was conducted among 143 obstetricians-gynecologists. 133 (93%) HCWs answered the questionnaire. 81% of them recalled almost one occupational exposure during their career and 54% almost two in the past year. Our data suggest a low knowledge about HIV and the other blood-borne viral infections and Universal Precautions (UP): only 16% of HCWs knew the rate of vertical transmission of HIV, less than 15% knew the rate of seroconversion after occupational exposure to HBV and HCV, and finally only 33% knew to which body fluids apply UP. Moreover, 93.7% of HCWs believed that HIV antibody screening of all patients is an effective strategy to reduce the risk of occupation HIV infection. More than 90% of gynecologists used to request HIV-antibodies to pregnant women, 21% after the first trimester. Although HCWs interviewed can not be considered representative of Italian gynecologists, our data suggest the need of an intensive training to increase gynecologists' knowledge about HIV, other blood-borne pathogens and the risk of occupational infections also in order to modify attitudes and practices.

Adult↗
PubMed1994

Differential effects of protein kinase A sub-units on Chinese-hamster-ovary cell cycle and proliferation.

It has been shown that a marked increase in the levels of RI alpha sub-units and a decrease in RII beta sub-unit levels correlate with neoplastic transformation or with the mitogenic response of normal cells to hormones and growth factors. The selective down-regulation of RI alpha and the following induction of RII beta determine cell-growth arrest and differentiation of several cancer cells. To directly address the question whether the 2 protein-kinase-A(PKA) isoforms play different roles in the control of proliferation and cell-cycle distribution, we introduced and over-expressed the different PKA sub-units in Chinese-hamster-ovary (CHO) cells via retroviral-vector-mediated gene transfer. Whereas CHO cells treated with RI alpha anti-sense oligodeoxynucleotides were growth arrested and accumulated in the G0/G1 phases of the cell cycle, infection of CHO cells with a retroviral vector in order to over-express RI alpha determined growth advantages in monolayer conditions and substantially increased their cloning efficiency in soft agar. These events correlated with a sustained percentage of cells in S phase induced by RI alpha over-expression in the infected cells. In contrast, CHO cells infected with retroviral vectors over-expressing either a RII beta sub-unit or a C alpha catalytic sub-unit of PKA exhibited growth arrest within a few days of culture and accumulated in the G2-M phase of the cell cycle. The results of our study demonstrate that the different PKA sub-units play different and specific roles in the control of cell growth and cell-cycle distribution.

Animals↗
PubMed1994

Pilomatrix carcinoma. A case report with immunohistochemical findings, flow cytometric comparison with benign pilomatrixoma and review of the literature.

We report a case of pilomatrix carcinoma in a 42 year old woman. The patient presented a rapidly growing 4-cm subcutaneous mass in the right preauricular area. She was free of recurrence 24 months after surgery. Immunoreactivity was studied for cytokeratins, epithelial membrane antigen, S-100 protein, blood group antigens, Leu-M1, beta-2-microglobulin, vimentin, neuron-specific-enolase and cellular binding for peanut agglutinin. Squamous cells were reactive for cytokeratin AE1/3, epithelial membrane antigen, blood group antigens B and H, and peanut agglutinin. Basaloid cells were focally reactive for cytokeratin AE1/3 and epithelial membrane antigen. Flow cytometry revealed a euploid DNA content and a high proliferative rate of the pilomatrix carcinoma and in benign pilomatrixoma studied for comparison. The pathological diagnosis must be based on histomorphological criteria.

Adult↗
PubMed1994

Flow-cytometric detection of the RI alpha subunit of type I cAMP-dependent protein kinase in human cells.

cAMP-dependent protein kinase (PKA) is composed of two genetically distinct catalytic (C) and regulatory (R) subunits. There are two different classes of PKA, designated as type I and type II, which contain distinct R subunits (RI or RII, respectively) but share a common C subunit. Enhanced expression of type I PKA has been correlated with cell proliferation and neoplastic transformation. Detection of the different PKA subunits is usually performed by photoaffinity labeling with 8-N3-32P-cAMP or by radioimmunolabeling techniques. Both techniques are time consuming and require a high number of cells and the use of radioactive reagents. Using the MCF-10A normal human mammary cell line infected with a recombinant retroviral vector containing the human RI alpha gene (MCF-10A RI alpha), we have developed a flow-cytometric assay to detect the intracellular content of RI alpha protein in human cells. MCF-10A and MCF-10A RI alpha cells were fixed in 1.5% paraformaldehyde at 37 degrees C for 15 min and permeabilized by methanol and acetone (1:1) at -20 degrees C for 5 min before staining with a specific IgG2a MoAb followed by a FITC-conjugate rabbit-anti mouse IgG. This procedure was also successfully utilized to recognize RI alpha protein content in human peripheral blood lymphocytes. Flow-cytometric detection of the RI alpha subunit in human cells is feasible and allows the study of the role of type I PKA in cell growth and neoplastic transformation.

Breast↗
PubMed1993

Chemotherapy with mitomycin C and vinblastine in pretreated metastatic breast cancer.

AIMS: In February 1986 we began a study to test the activity of mitomycin C (12 mg/m2) plus vinblastine (6 mg/m2) on day 1 of a 28-day cycle (MV) as second or third-line chemotherapy for metastatic breast cancer patients. METHODS: As of February 1988 the study was stopped after 26 patients had been enrolled. The median age of the patients was 54 years (range 35-78); all patients were progressive from chemotherapy; 15 (57.7%) patients were treated as second and 11 (42.3%) as third line; 19 (73.1%) patients had received anthracyclines as first (13 patients) or second-line (6 patients) chemotherapy; 18 (69.2%) patients had visceral involvement; 7 (26.9%) had one metastatic site, 11 (42.3%) two sites, 6 (23.1%) three sites and 2 (7.7%) four sites. RESULTS: Overall, 86 cycles were administered, with a median number of 3 cycles per patient. Toxicity was mild; hematologic side effects required discontinuation of treatment in 3 cases. Vomiting occurred in 3 (11.5%) patients, nausea in 5 (19.2%). Moderate neurologic toxicity was recorded in 6 (23%) patients. No complete and 3 partial responses were observed. The objective response rate was 11.5% (exact 95% confidence interval, 2.4-30.1). Responses occurred independently of disease-free interval, dominant metastatic site, response to previous chemotherapy, previous anthracycline and line of treatment; all responses were recorded in patients under 50 years of age. Kaplan-Meier estimated median time to progression and overall survival were 13 and 40 weeks, respectively. CONCLUSION: The MV regimen was well tolerated but showed little activity in pretreated metastatic breast cancer.

Adult↗
PubMed1993

Down-regulation of RI alpha subunit of cAMP-dependent protein kinase induces growth inhibition of human mammary epithelial cells transformed by c-Ha-ras and c-erbB-2 proto-oncogenes.

MCF-10A is a spontaneously immortalized, non-transformed human mammary epithelial cell line. We have recently obtained MCF-10A clones (MCF-10A HE cells) that are transformed following over-expression of both a human point-mutated c-Ha-ras and the c-erbB-2 proto-oncogenes. Two isoforms of the cAMP-dependent protein kinase (cAK) have been described in mammalian cells. Enhanced levels of type-I cAK (cAKI) are generally found in tumor cells. To determine whether inhibition of cAKI expression may interfere with ras and erbB-2 oncogene-induced transformation of human mammary epithelial cells, we have tested the effects of 2 agents that specifically down-regulate cAKI, such as 8-chloro-cAMP and an anti-sense oligodeoxynucleotide targeted against the RI alpha regulatory subunit of cAKI on MCF-10A HE cells. Treatment of MCF-10A HE cells with 8-chloro-cAMP induces a dose-dependent growth inhibition under both monolayer and soft-agar growth conditions, that is correlated with an accumulation of MCF-10A HE cells in G0/G1 phases of the cell cycle and a reduction of the number of cells in S phase. In contrast, 8-chloro-cAMP has no effect on MCF-10A cell growth. Furthermore, 8-chloro-cAMP treatment of MCF-10A HE cells induces a 4- to 6-fold reduction in p185erbB-2 expression and brings p21ras expression to levels comparable to those found in MCF-10A cells. Treatment of MCF-10A HE cells with an RI alpha anti-sense oligodeoxynucleotide determines a comparable inhibition of both anchorage-dependent and anchorage-independent cell growth. Our results suggest that cAKI may act as a mediator of ras and erbB-2 oncogene action in human breast cells and that interference with cAKI action provides a potential tool for inhibiting the growth-promoting effects of these oncogenes.

Antisense Elements (Genetics)↗
PubMed1992

The expression of proliferating cell nuclear antigen in paraffin sections of peripheral, node-negative non-small cell lung cancer.

Cell proliferation of 40 peripheral, node-negative non-small cell lung cancers (NSCLC) treated with surgery alone was investigated by immunohistochemical analysis with the monoclonal antibody (MoAb) PC10, which recognizes a proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded material. Results were correlated with DNA ploidy and S-phase fraction (SPF) analyzed by DNA flow cytometric study. Mitotic count (MC) was analyzed by light microscopic study and histopathologic features. PCNA immunoreactivity was seen in all samples and confined to the nuclei of cancer, but not to the surrounding, tumor-negative cells; its frequency ranged from 0-70% (median, 15%), and tumors expressed either a low (0-25%, n = 25) or intermediate (26-75%, n = 15) proliferative activity. There was no relationship between PCNA immunoreactivity and tumor stage or among size, histologic type, and mitotic count (MC). Tumors with intratumoral blood vessel invasion (BVI) showed a significantly higher (P less than 0.005) PCNA immunoreactivity than BVI-negative tumors. PCNA scores were significantly higher (P less than 0.005) in DNA aneuploid (n = 22) than in DNA diploid (n = 18) tumors and correlated significantly with the SPF of DNA aneuploid tumors (r = 0.825, P less than 0.0001), but not with diploid tumors (r = 0.002, P = 0.9). Intermediate proliferating tumors had a significantly higher (P less than 0.01) MC than their counterparts. In univariate analysis, significant predictors of survival were tumor classification (T1 versus T2), tumor size (less than or equal to 2.6 cm versus more than 2.6 cm), BVI (BVI-negative versus BVI-positive), MC (less than or equal to 8 versus more than 8), and PCNA immunoreactivity (low versus intermediate). DNA ploidy and SPF did not influence survival significantly. Only PCNA immunoreactivity retained its independent level of significance (P = 0.02) by multivariate analysis. It was concluded that PCNA immunostaining is a simple and clinically useful method for estimating cell proliferation in formalin-fixed, paraffin-embedded tissue of resected peripheral, node-negative NSCLC.

Adult↗
PubMed1992

Growth fraction in non-small cell lung cancer estimated by proliferating cell nuclear antigen and comparison with Ki-67 labeling and DNA flow cytometry data.

Results generated by the immunohistochemical staining with PC10, a new monoclonal antibody recognizing PCNA (a nuclear protein associated with cell proliferation) in formalin-fixed and paraffin-embedded tissue were compared with those of Ki-67 labeling and DNA flow cytometry in 47 consecutive non-small cell lung cancer (NSCLC). PCNA reactivity was observed in all samples and confined to the nuclei of cancer cells. Its frequency ranged from 0 to 80% (37.7 +/- 23.6) and larger sized, early-staged and DNA aneuploid tumors expressed a significant higher number of PCNA-reactive cells. The PCNA and Ki-67 labeling rates were closely correlated (r = 0.383, P = 0.009). By flow cytometry, we observed a good correlation among PCNA labeling and S-phase fraction (r = 0.422, P = .0093) and G1 phase (r = 0.303, P = .051) of the cell cycle. Results indicate that PCNA labeling with PC10 is a simple method for assessing the proliferative activity in formalin-fixed, paraffin-embedded tissue of NSCLC and correlates well with Ki-67 labeling and S-phase fraction of the cell cycle.

Aged↗
PubMed1990

[Anesthesia in the diagnostic stages of cerebral arteriovenous malformations in children under 3 years of age].

From 1974 through 1989, 36 patients with arteriovenous malformations were treated in the neuroradiological unit of the Santobono Children's Hospital, in Naples, 7 of these patients were younger than 3 years of age. During such a time-span, dramatic improvements in diagnostic work-up were jelded by introduction of CT-scan and MRI; better angiographic techniques, progress in anaesthesia drugs, equipment and procedures and more rational surgical modalities. Anaesthesia must be tailored in each patient with special consideration of the clinical and neurological conditions of the patients as well of the neuroradiological procedure, specifically, with minimal side effects on cardiovascular and respiratory functions as well on intracranial pressure. The present paper discuss of equipment for cardiovascular assistance and drugs to be employed in the various steps of the diagnostic work-up of such patients.

Anesthesia, General↗
PubMed1990

[Peroperative recovery in surgery of arteriovenous malformations in children under the age of 3 years].

From 1974 through 1989, 36 patients with arteriovenous malformations were studied in the neurosurgical department of the Santobono Children's Hospital in Naples; 7 of these patients were younger than three years of age: 5 underwent surgery. The remaining 2 patients presented, respectively, an aneurysm of the vein of Galen and an arteriovenous malformation of the brain stem. Improvements in surgical and anesthesiological techniques allow to treat lesions considered as not amenable to surgery until recent years, with minimal occurrence of preoperative and postoperative complication and a significant amelioration of prognosis.

Anesthesia, General↗
PubMed1989

Critical density fluctuations in lipid bilayers detected by fluorescence lifetime heterogeneity.

The heterogeneity of the decay of the fluorescence of transparinaric acid in single-component lipid bilayers at temperatures above their gel/liquid crystalline phase transition is shown to be due to the presence of regions of higher local density and higher acyl chain order than the predominant fluid regions. This conclusion is based on selective excitation behavior and the observation of time-resolved fluorescence anisotropies that increase at long times. The fractional amplitude of the long lifetime component of the fluorescence shows a temperature variation that conforms to conventional descriptions of critical behavior. The critical exponent extracted from this variation is 1.1, close to the value of 1.0 that describes ultrasonic data. We therefore conclude that liquid crystalline lipid bilayers exhibit critical behavior with significant density and order fluctuations. This behavior must be taken into account in the interpretation of fluorescence and other spectroscopic measurements of the properties of bilayers.

Kinetics↗
PubMed1989

Analysis of the anisotropy decay of trans-parinaric acid in lipid bilayers.

An analysis is presented of the complex anisotropy behavior of trans-parinaric acid in single component DEPC lipid bilayers. It is shown that a model involving two species with distinct lifetime and motional behavior is required, and is adequate, to explain the observed data. In particular, the observed increase in the anisotropy at long times demonstrates the presence of a species with a long fluorescence lifetime that has a high anisotropy. The time dependence of the anisotropy for these two environments is treated using both a purely mathematical sum of exponentials and a constrained fit based on an approximate solution of the anisotropic diffusion problem. In this latter model the anisotropy is described in terms of the second and fourth rank order parameters, (P2) and (P4), and a single dynamical parameter, D1, the perpendicular diffusion coefficient for this uniaxial probe. The parameters of both models are accurately determined from the fits to the data when two environments coexist and an association is made between lifetime components and distinct rotational sites. The values of the parameters obtained demonstrate the "solid-like" and "fluidlike" nature of these two coexisting environments.

Fatty Acids, Unsaturated↗