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Biomedical subjects

A Calugi

Publications and source records attributed to A Calugi.

At least 19 recordsLinked to original sources

Effects of 50 Hz magnetic fields on mouse spermatogenesis monitored by flow cytometric analysis.

Flow cytometry (FCM) was performed to monitor the cellular effects of extremely-low-frequency magnetic field on mouse spermatogenesis. Groups of five male hybrid F1 mice aged 8-10 weeks were exposed to 50 Hz magnetic field. The strength of the magnetic field was 1.7 mT. Exposure times of 2 and 4 h were chosen. FCM measurements were performed 7, 14, 21, 28, 35, and 42 days after treatment. For each experimental point, a sham-treated group was used as a control. The possible effects were studied by analyzing the DNA content distribution of the different cell types involved in spermatogenesis and using the elongated spermatids as the reference population. The relative frequencies of the various testicular cell types were calculated using specific software. In groups exposed for 2 h, no effects were observed. In groups exposed for 4 h, a statistically significant (P < 0.001) decrease in elongated spermatids was observed at 28 days after treatment. This change suggests a possible cytotoxic and/or cytostatic effect on differentiating spermatogonia. However, further studies are being carried out to investigate the effects of longer exposure times.

Animals

Cellular effects of image diagnostic ultrasound on murine spermatogenesis monitored by flow cytometry.

A study has been carried out to evaluate the possible cellular effects induced by image diagnostic ultrasound on murine spermatogenetic cells. Exposure to ultrasound was carried out using a commercial diagnostic instrument that operates in B-mode. Male hybrid F1 mice, aged 8-10 weeks, were exposed to ultrasound for 30 min and observed from 7 to 35 days after treatment. Flow cytometric analysis has been used to monitor the relative frequency of the different types of spermatogenetic cells. This analytical approach showed changes in cell frequency in the compartment containing elongated spermatids which was used as an endpoint. A statistically significant decrease in the frequency of this cell type was observed 21, 28 and 35 days after exposure. These changes suggest that there may be a cytotoxic and/or cytostatic effect on spermatocytes and spermatogonia. These results showed that image diagnostic ultrasound induces effects on murine spermatogenesis at cellular level and that the flow cytometric approach makes it possible to identify quantitative cellular changes with reference to specific cell type.

Animals

Flow cytometric and cytogenetic analyses in human spontaneous abortions.

Cytogenetic and flow cytometric analyses were performed on 38 human spontaneous abortions in an attempt to obtain information on karyotype abnormalities and to compare the two approaches of analysis. In 19 cases, it was not possible to perform cytogenetic analysis because too long a time had passed between surgical sampling and cell culture, and in vitro culture failed. Of the 19 cases analyzed, 10/19 showed a normal karyotype and 5/19 showed a single trisomy (2/5 trisomies involved chromosome 16, 1/5 trisomy involved chromosome 18, 1/5 trisomy involved chromosome 20, and 1/5 was Klinefelter syndrome). Of the remaining 4/19 cases, 2/19 showed a polyploid condition (1 tetraploidy and 1 triploidy), 1/19 a double trisomy (chromosomes 13 and 21), and 1/19 a pentasomy of the sex chromosomes (49,XXXXY). Flow cytometric analysis was performed on all abortive samples. The samples were subdivided, when possible, into two portions conventionally named "amniotic" and "chorionic", using the amniotic membrane as an anatomical reference. Maternal blood lymphocytes were used as a diploid standard for each sample. In the 19 cases not analyzed by the cytogenetic approach, flow cytometric analysis showed 9 diploid and 10 aneuploid DNA distributions. In the remaining 19 cases, analyzed with both approaches, the comparison of DNA estimations using cytogenetic and flow cytometric analyses showed good agreement. In the cases with karyotype abnormalities, flow cytometric measurement provided evidence of an alteration of DNA content with respect to the diploid standard. Flow cytometric analysis showed a diploid distribution, whereas cytogenetic analysis revealed chromosomal abnormalities in only 4/19 cases. These discordant results could be related to mosaic conditions or maternal cell contamination. Moreover, cytogenetic and flow cytometric analyses were performed on 2 amniotic cell cultures, and concordant results were obtained. The results obtained suggest that a combination of these techniques is beneficial in attempts to obtain information about DNA content alterations, even when cultures fail, and in screening studies of human abortions.

Abortion, Spontaneous

Cellular effects of diagnostic Doppler ultrasound on murine spermatogenic cells monitored by flow cytometry.

A study has been carried out to investigate the possible effects caused by Doppler diagnostic ultrasound on murine spermatogenesis. The frequency of the different types of cells has been analyzed using flow cytometry. Exposure to ultrasound was carried out using a commercial device used in diagnostic conditions. Male hybrid mice were exposed to ultrasound for 30 min and observed from 7 to 35 days after treatment. Flow cytometrical analysis showed changes in the relative frequency of the elongated spermatids and this was used as an end-point. A statistically significant decrease in the frequency of this cell type was observed after 7 and 35 days with both pulsed and continuous ultrasound. With the former, a decrease was also seen in this compartment after 14 and 21 days. Our results have shown that diagnostic ultrasound used in the Doppler technique induces effects on specific cell types of murine spermatogenesis.

Animals

Flow cytometric nuclear DNA content of fresh and paraffin-embedded tissues of breast carcinomas and fibroadenomas.

Flow cytometric analysis of DNA ploidy level in fresh tissue samples of human breast tumors has been carried out extensively. Recently, investigations regarding the prognostic value of nuclear DNA content have been facilitated by using nuclei isolated from paraffin-embedded tissues. The aim of our study was to monitor the possible differences between the analysis in fresh and fixed paraffin-embedded tissues in the same tumor and to investigate the possible prognostic implications obtained with this new approach. Nuclei suspensions were obtained, according to the method proposed by Hedley et al. with minor modifications, from 45 carcinomas and 5 fibroadenomas. Flow cytometric analysis revealed diploidy in 57% of carcinomas, while the remaining 43% showed cytometric aneuploidy. Corresponding results were observed between fresh and paraffin-embedded tissue in 26/35 cases. Moreover, a fairly good correlation between the DNA indices of fresh and paraffin-embedded carcinoma samples was observed. Furthermore, the frequency of recurrence was higher in the aneuploid group. Finally, 4 fibroadenomas were diploid and one was aneuploid. Our results confirm that this approach permits retrospective studies to evaluate the potential prognostic significance of nuclear DNA content monitored by flow cytometry.

Adenofibroma

Flow cytometric DNA analysis of the human cervix affected by human papillomavirus and/or intraepithelial neoplasia.

The relative DNA contents of 164 cellular samples from 59 patients affected by the viral cytopathic effects (VCE) of human papillomavirus (HPV) infection and/or by cervical intraepithelial neoplasia (CIN) and 12 cellular samples from 12 normal donors were analyzed by flow cytometry (FCM) with the aim of correlating the cytometric measurements with the morphologic and etiologic parameters. The unselected group of 59 patients was found to be characterized by statistically significant differences in average ages in the VCE and CIN (31.4 years) and CIN only (44.8 years) subgroups. Of the pathologic samples, 32 (54%) exhibited at least one cell subpopulation with an abnormal DNA content; in all but 2 of those cases, a diploid cell subpopulation was also present. The results indicate a relationship between the FCM ploidy and the morphologic classification, as shown by the increase in the occurrence of subpopulations with abnormal DNA contents from VCE only (38%) to VCE + CIN 1 (57%), to VCE + CIN 2/3 (70%). These results suggest that cytometric parameters, in association with the determination of the HPV types and in parallel to the colpocytohistopathologic criteria, can contribute to a more accurate characterization of cervical lesions in diagnostic and prognostic terms.

Adolescent

Flow cytometrically determined DNA content of breast carcinoma and benign lesions: correlations with histopathological parameters.

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p less than 0.01) were also observed for the percentage of S cells between normal tissues (6.2 +/- 3.2 SD) and benign lesions (11.1 +/- 6.6 SD), normal tissues (6.2 +/- 3.2 SD) and aneuploid tumors (19.7 +/- 10.3 SD), benign lesions (11.1 +/- 6.6 SD) and aneuploid tumors (19.7 +/- 10.3 SD), and diploid (7.9 +/- 4.0 SD) and aneuploid tumors (19.7 +/- 10.3 SD).

Adult