Biomedical subjects

R Kiss

Publications and source records attributed to R Kiss.

At least 289 records · Page 16Linked to original sources

PubMedDate not supplied

[Initial results of a study on morphonuclear parameters at the level of benign and primary neoplastic hepatic tissue].

This study characterises morphonuclear parameters from normal, cirrhotic, tumoral benign and malignant liver specimens obtained by computerized morphonuclear images analyses. The material is obtained by cytological aspirations or by imprint smears from liver biopsy. The monoparametric analysis differentiates a benign and a malignant cellular populations. In the malignant populations, two types are individualised: the well differentiated and the pleiomorphic hepatocarcinoma. The treatment of the data by canonical projections onto the factorial plan and by discriminant analysis allowed the selection of homogeneous cell populations belonging to each of the pathological groups described above.

Carcinoma, Hepatocellular↗

PubMedDate not supplied

Influence of suramin alone or in combination with DHT and PDGF on the cell proliferation of benign and malignant human prostatic tissues in organ cultures.

We studied the suramin-induced influence on the cell proliferation of 16 benign and 6 malignant lesions of the human prostate maintained in vitro as organ cultures. The cell proliferation was assessed by nuclear labeling with tritiated thymidine autoradiography. We also studied the dihydrotestosterone (DHT)-and platelet-derived growth factor (PDGF)-induced modulation of suramin influence on such prostate organ culture cell proliferation. Our results indicate that more than half of the benign prostatic tissues showed cell proliferation which was modulated by DHT and/or PDGF, while none of the six carcinomas responded to such hormonal stimulation. Suramin alone inhibited the cell proliferation of only 19% of the prostate organ culture under study, while in combination with DHT and/or PDGF this inhibition level reached 48%. However, we occasionally observed that S alone or in combination with DHT and/or PDGF was also able to stimulate prostate cell proliferation. We think that organ cultures of human prostatic tissues might represent a helpful pre-clinical tool to study the anti-tumoral influence of suramin, which is a new antineoplastic generative compound.

Adult↗

PubMedDate not supplied

The combination of the tetrazolium derivative reduction (MTT) and digital cell image analysis to monitor in vitro the cytotoxicity of anti-neoplastic drugs.

We assessed the in vitro drug-induced cytotoxicity by means of the rapid low-cost but weakly sensitive Thiazolyl blue (MTT) test, and the less rapid, higher cost, but highly sensitive cell image analysis (CIA) test. We studied the influence of three drugs, i.e. a vinca-alkaloid (Navelbine), an alkylating investigational agent (PE1001), and an intercalating drug, i.e. adriamycin, on the proliferation (MTT test) and cell kinetics (CIA test) of the mouse MXT and the MCF-7 mammary cancer cell lines. Adriamycin and PE1001 decreased MXT and MCF-7 cell proliferation in a dose-dependent manner as assessed by both the MTT and CIA tests. Navelbine was highly cytotoxic in a dose-dependent manner. We demonstrated that Navelbine arrests cells in the M phase without altering the G2 phase. In sharp contrast, PE1001 arrest cells in the G2 phase without altering the M phase. An adriamycin-induced effect was apparent on S phase. Thus, the MTT test allows the screening of a great number of drugs analyzed at the cell proliferation level and, in the case of MTT positive drug-induced response, the CIA test enabled the drug-induced effect at cell cycle kinetic level to be investigated.

Adenocarcinoma↗

PubMedDate not supplied

Comparison of biochemical and immunoenzymatic macromethods and a new immunocytochemical micromethod for assaying estrogen receptors in human breast carcinomas.

Estrogen receptors (ERs) were assayed in 23 breast carcinomas by: (1) the conventional biochemical assay with dextran-coated charcoal (DCC); (2) the immunoenzymatic assay using a monoclonal antibody (MAb), ER-EIA (Abbott); and (3) an original cytochemical method using another MAb, ER-ICA (Abbott). The first two techniques were performed on biopsy samples, whereas the last was carried out on fine needle aspiration (FNA) samples. The ER contents in aspirates were evaluated by: (1) scaled proportions of colored neoplastic cells; (2) scaled coloration intensity; (3) total grading (= proportion plus intensity); (4) product grading (= proportion times intensity); and (5) a new index (NI) described in this paper. The ER-EIA assay correlated best, with a high statistical significance, with the NI (P less than .001); NI was also the only index that significantly correlated (P less than .05) with the DCC results. The results show that the ER-ICA assay offers the great advantages of being applicable to FNA specimens and of producing rapidly available results. This new technique enriches the panel of MAbs for the diagnosis of adenocarcinomas and offers a new tool for the therapeutic follow-up of breast cancer patients. Our preliminary results suggest that the anti-ER MAbs might be helpful for measuring the hormone dependence of small lesions not assayable by DCC, even under endocrine therapy, thus avoiding false-negative assays.

Antibodies, Monoclonal↗

PubMedDate not supplied

In vitro influence of estradiol or progesterone on the thymidine labeling indices of human benign breast tumors.

Using an in vitro tritiated thymidine (3H-dThd) nuclear labeling followed by autoradiography, the effects of 17-beta-estradiol (E2) or progesterone (Pg) on cell proliferation were studied in 22 human benign breast tumors, i.e. 7 fibroadenomas 8 fibrocystic dysplasias 4 gynecomastias and 3 phyllodas. Small tumor fragments were incubated in a chemically-defined medium without serum and were hormonally stimulated in vitro. The procedure used allowed discrimination between weak labeling, suggestive of DNA repair mechanism, and strong labelling, suggesting a true DNA synthesis (S phase). Taking this into account, our results have shown that both estradiol and progesterone can induce in vitro cell replication of both ER+PgR+ and ER-PgR- human breast fibroadenoma, without affecting those of fibrocystic dysplasia, gynecomastia and phylloda. Progesterone, but not estradiol, might significantly decrease DNA repair mechanism in fibrocystic dysplasia, according to the Pg-induced decrease of nuclear incorporation of small amounts of 3H-dThd. We have thus characterized a dynamic test of hormone dependence which permits in vitro study of the hormonal sensitivity of human benign breast tumors, as well as that of any other human neoplasm. This test could be of great value to help clinicians to diagnose and treat hormone-dependent neoplasms properly. Its clinical relevance is now under study.

Autoradiography↗

PubMedDate not supplied

MXT mammary tumor treated by low dose chemotherapy. A 31P-NMR and 1H-MRI study.

The effects induced by a low dose cyclophosphamide treatment (CPA) along with an attempt of estradiol-17-beta (E2) cell recruitment, on the bioenergetics and the phosphomonoesters/phosphodiesters (PME/PDE) ratio in the transplantable MXT mouse mammary tumor have been studied by 31P-NMR spectroscopy in vivo. Our results show that neither ovariectomy (OO) nor the additional low dose E2 + CPA treatment significantly decreased tumor growth, whilst E2 + CPA treatment induced significant modifications compared to values recorded in tumors grafted on intact or ovariectomized animals, of all peak height ratios but PME/PDE, which further remained at a constant value during tumor growth.

Animals↗

PubMedDate not supplied

Effect of estradiol -17- beta (E2) on cell proliferation in the uterus and the MXT mammary tumor borne by intact, ovariectomized and/or hypophysectomized mice.

The aim of the present work was to study the effects of estradiol (E2) on cell proliferation in well-differentiated mammary MXT tumors borne either by intact (INT), or by castrated (OVX), or by hypophysectomized (HX), or by castrated and hypophysectomized mice) OV-X-HX). Ovariectomy and hypophysectomy were performed, respectively, 6 days and 5 days before the sacrifice of animals bearing tumors during a 28 day period. Cell proliferation was measured by using an in vivo nuclear labeling with tritiated thymidine (3H-TdR) followed by autoradiography. Uteri were used as controls for the methodology. Our data demonstrate that E2 exerts a significant and transient stimulatory effect on cell proliferation in the uterine luminal and glandular epithelia of OVX or OVX-HX adult mice. This mitogenic enhancement is significantly higher in OVX-HX mice than in the other experimental groups. In MXT tumors, a significant increase of cell proliferation is induced by E2 in INT or OVX mice; however, no enhancement occurs after HX performed either in intact or in castrated mice. In the latter situation (HX), a dramatic increase of basal cell proliferation rates is consistently observed in tumors, as compared to those levels observed in INT or in OVX animals. The paradoxical increase in basal proliferation rates of MXT mammary tumors in HX animals was not suppressed by administration of ovine prolactin. This suggests that other hypothalamo-hypophysial factors might be endowed with direct inhibitory influence on growth of the MXT tumors.

Animals↗

PubMedDate not supplied

Hormonal regulation of uterine epithelial cell proliferation. I. Effects of estradiol or progesterone administered separately.

Using an in vivo tritiated thymidine (3H-TdR) labeling followed by autoradiography, the effects at different times before sacrifice of single or paired injections of 17-beta-estradiol (E2) or progesterone (Pg) at various concentrations were investigated in adult B6D2FI ovariectomized mouse luminal and glandular uterine epithelium. With regard to the luminal epithelium, E2 (0.25, 2.50 or 25.00 micrograms/animal) exerts a mitogenic influence which is dose-related. Pg (125, 600 or 5000 micrograms/animal) exerts a significant proliferative effect only at a high dose. With regard to glandular epithelium, E2 and Pg have an almost identical mitogenic influence which is dose-related. A two-step Pg administration (2 x 125 micrograms) inhibits cell proliferation in glandular epithelium as compared to the promoting action exerted by a single injection. No refractory response was observed after a two-step E2 administration in either the luminal or the glandular epithelium. These data demonstrate that cell proliferation in the uterine luminal epithelium of an adult B6D2F1 mouse is very sensitive to E2 but not to Pg, whereas these two steroids have a similar mitogenic influence on the cell proliferation of the glandular epithelium. Moreover, a second Pg administration seems to have an inhibiting effect on cell proliferation as compared to that exerted by the first injection performed 12 or 24 hours earlier. In contrast, the repetition of E2 treatment would seem to exert an additive effect on uterine epithelial cell proliferation. In conclusion, epithelial components of the endometrium are complex steroid targets. The two major cell types (luminal, glandular) differ in their abilities to respond to steroid stimulation by entering into DNA synthesis.

Animals↗

PubMedDate not supplied

Hormonal regulation of uterine epithelial cell proliferation. II. Effects of estradiol and progesterone administered in combination.

The effects of estradiol (E2) and progesterone (Pg) administered in combination on the cell proliferation of the ovariectomized B6D2F1 mouse uterine epithelium were assessed by thymidine autoradiography. In the luminal and glandular cell populations Pg inhibits the mitogenic effect of E2 when administered simultaneously or 24 hours after E2; when Pg is given 12 hours after E2 it does not block but slows down the proliferative effect of E2, at least at the concentrations studied here. Given prior to E2, Pg rather exerts an additive (synergistic) effect on uterine epithelia. It is concluded that Pg can exert different actions on the E2- mitogenic effect induced at both luminal and glandular levels. Further investigations are needed to understand better the precise biochemical mechanisms involved in the modulating actions of these steroids on the mouse endometrium epithelial cells.

Animals↗

PubMedDate not supplied

Binding of fluorescein isothiocyanate conjugated lectins to MXT mouse mammary neoplasm and their relation to steroid receptor status.

As previous studies have suggested a hormone dependence of binding sites for peanut agglutinin in mammary neoplasm, this feature has been thought to be correlated to steroid receptor status. The present investigation was undertaken on a well-established ovarian-dependent cancer model in order to check this hypothesis. Sections of primitive tumor transplants as well as of tumors induced in vivo by injection of cell clones were analyzed with the use of three fluorescent lectins. The lectin binding sites were evaluated semi-quantitatively and compared with estrogen and progesterone receptor levels. Using non-parametric statistical tests, the results revealed a strong correlation between the expression of peanut agglutinin (PNA) binding sites and steroid receptor status, but only in primitive tumor transplants. No such correlation was observed in tumors induced in vivo, by injection of cell clones. No correlation between the steroid receptor status and the two other lectins (Concanavalin A and Dolichos biflorus) was observed. These data suggest that PNA can be used as a valuable histochemical tool in steroid hormone dependence study.

Animals↗

PubMedDate not supplied

A new assay to evaluate cell growth and drug sensitivity in culture using a cell image processor.

The use of a cell image processor for the in vitro assessment of drug effects on cell growth, cell kinetics and chromatin organization is described. We have studied the influence of two well documented cytotoxic drugs, i.e. BCNU and vincristine (VIN), on the above mentioned parameters of the P-388 mouse leukemia, MCF-7 human mammary and HBL human melanoma cell lines. The cells were cultured for 1 to 4 days on glass coverslips put in Petri dishes containing or not (control) 10, 1 or 0.1 microgram/ml medium of the drug, after which they were fixed for histology, Feulgen-stained and analyzed through a cell image processor, i.e. the System for Analytical Microscopic Biomedical Applications (SAMBA 200). Our results showed that both BCNU and VIN exerted a well-known antineoplastic effect that was assessed at three different but highly complementary levels on the same sample of cells in a very rapid and simple procedure.

Animals↗

PubMedDate not supplied

Phenotypic change of the transplantable MXT mammary adenocarcinoma into mixed bone producing sarcoma-like tumors.

The B6D2F1 mouse mammary adenocarcinoma was adapted to grow in vitro as monolayer. After in vitro passaging of tumor cells, phenotypic changes occurred that were expressed in vivo. Following intraperitoneal inoculation of tumor cells, bone-forming tumors developed. These tumors consisted of undifferentiated adenocarcinoma mixed with large amount of cartilagenous and osseous tissue. The etiology of these phenotypic changes was not yet determined. However, hypothesis of the possible origin of the cartilage and bone forming tissue was formulated. The biologic characterization of the intraperitoneally bone-forming tumor was achieved and the experimental conditions to preserve and induce the reproducible sarcoma-like bone forming tumors were defined. Our data support the usefulness of this new original model for fundamental research as well as for screening of anticancer drugs.

Adenocarcinoma↗

PubMedDate not supplied

Effect of estradiol on proliferation and on cell loss in the MXT mouse mammary tumor.

The MXT tumor is an experimental mammary tumor, maintained by transplantation in B6D2F1 mice, which contains significant amounts of estrogen and progesterone receptors. The aim of the present work was to study the effect of estradiol (E2) administration on the cell kinetic parameters (thymidine labeling index: TLI, S phase duration: TS, cell cycle duration: Tc, growth fraction: GF, potential doubling time: T, cell loss factor: luminal diameter) of an MXT anaplastic carcinoma strain. These parameters were analyzed using single-, double- or repetitive-tritiated thymidine (3H-TdR) labeling techniques. Uteri were used as controls for the methodology. Our data clearly demonstrate that E2 exerted a marked stimulatory effect on the tumor cell proliferation. This effect is the net result of a shortening of the mean cell cycle duration (Tc) and of a 3-fold increase of the number of cells produced per time unit in 89% of the total cancer cell population. Beside this stimulatory effect, E2 produces a marked increase of the tumoral cell loss. This loss seems to affect a cell fraction which has reached the quiescent state (Q) for a short time.

Animals↗

PubMedDate not supplied

Influence of inoculation sites and tumor cell culture techniques on the phenotype of a mixed cartilage- and bone-producing mouse mammary tumor.

We have previously reported the characterization of an intraperitoneally (IP) transplantable bone-forming MXT tumor. However, the question was unresolved as whether the bone-forming cells originated from either the host animal or from the neoplasm itself. The present work attempts to answer this question by studying the influences of inoculation sites (subcutaneously, SC; intraperitoneally, IP; in the brain, IB; intracranially, ICR) on both the cartilage- and bone-forming tumor phenotypes. Furthermore the influence of cell culture procedures (two- and three- dimensional cultures) on these phenotypes was investigated. SC administered MXT cancer cells never produce bone-forming tumors, suggesting the existence in the dermis of substance(s) inhibitory to the formation of cartilage or bone. On the contrary, our data clearly demonstrate that bone-forming tumors can be obtained by either IP route, in a way which mimics endochondral ossification, or in the brain (IB), a region usually devoid of connective tissue. This observation substantiates the hypothesis according to which the tumor itself is able to produce osseous tissue. Another main finding is the increasing occurrence of skeletal tissues produced by cells proceeding from three-dimensional culture. Finally, ICR and IB tumors exerted a bone-lytic action against the host skull suggesting that tumor cells either produce osteolytic substances (prostaglandins, enzymes) and/or that they contain various cell types exhibiting different properties toward osteogenesis. This model offers new perspectives for studying the mechanisms of both normal and pathologic osteogenesis.

Animals↗

PubMedDate not supplied

Digital cell image analysis of Feulgen-stained nuclei from human papillary, medullary, colloid, lobular and comedocarcinomas of the breast.

The morphonuclear characteristics (nuclear size and chromatin pattern), the proliferation index and the ploidy level were characterized in a series of 46 breast tumors including medullary (5 cases), papillary (6 cases), lobular (27 cases), colloid (4 cases) and comedo- (4 cases) carcinomas. The quantitative assessments were carried out by means of digital cell image analyses of Feulgen-stained nuclei from imprint smears. The results show that monovariate analyses (one-way variance analyses) were much less potent than multivariate analyses (principal components analyses followed by the canonical transformation of the data and discriminant analyses) in assessing the morphonuclear characteristics of these breast tumors. The multivariate analyses indicated that there might be a level of malignancy which increases according to the sequence papillary and medullary and colloid carcinomas-->comedocarcinomas-->lobular carcinomas. This assertion is corroborated by the ploidy-level-related results which revealed a higher proportion of highly aneuploid cases in the group of lobular carcinomas than in the group which included medullary, papillary and colloid carcinomas. However, since highly aneuploid cases were also encountered in this latter low malignancy level group, we expressed the hypothesis firstly that aneuploidy reflects two distinct biological properties, i.e. the aggressiveness of a tumor and its age, and secondly that a highly aneuploid but low malignancy tumor should correspond to old degenerating tumors.

Adenocarcinoma, Mucinous↗

PubMedDate not supplied

Introduction of the cytological grading, the nuclear area, the DNA index and the DNA histogram type in the setting up of a score for ductal breast carcinoma.

The present study describes the setting up of a new score which makes it possible objectively to grade ductal breast carcinomas, i.e. not-otherwise-specified (NOS) cancers, on cellular material from fine-needle aspirations (FNAs). For this purpose FNAs from 252 patients--with NOS breast cancers--were smeared onto histological slides, fixed in an ethanol-formolacetic acid mixture, Feulgen-stained and analysed by means of a cell image processor. Four parameters were taken into account in setting up the score, namely the cytological prognostic grade (CPM) of malignancies similar to the Scarff-Bloom-Richardson (SBR) grading, the nuclear area (NA), the DNA index (DI) and the DNA histogram type (DHT). Each of these four parameters was considered as a "sub-score" which may take three values, i.e. 1, 2 and 3. The final result may thus range from 4 to 12. Subscores of 4 and 5 correspond to a cytological score of I, subscores of 6, 7 and 8 to a cytological score of II, sub-scores of 9 and 10 to a cytological score of III, and sub-scores of 11 and 12 to a cytological score of IV. In the present study, the results show 17% of CPM grade 1.52% of CPM grade II and 31% of CPM grade III cancers. All the cases exhibiting a cytological score of IV (5%) fully fit in with the CPM grade III cancers. In the same way, none of the cases exhibiting a score of I fit in with CPM grade III cancers. The cancers with a CPM grade II fit in with the scores of II and III. It thus seems possible to convert a three-value malignancy grading system (CPM and/or SBR grading) into a four-value one (cytological score). The main advantage in this latter type of system is that it becomes possible to split up the over-large group of CPM grade II cancers. As things stand, we are unable to give any prognostic value for the score proposed here because our study is prospective only. A study of this type has been necessary so as to provide against problems connected with ways of preserving specimens that might be used in a retrospective study. The bank of clinical and biological data now in existence must be allowed to mature for a number of years before the prognostic worth of the cytological score can be established, always assuming that such a value exists.

Biopsy, Needle↗

PubMedDate not supplied

[Diagnostic and/or prognostic value of the nuclear level determination of desoxyribonucleic acid (DNA) in nervous system tumors].

The diagnostic and prognostic value of the DNA ploidy level (nuclear DNA content) was studied in a series of 847 tumours of the nervous system. This series included 93 nerve sheath tumours, 224 meningiomas, 389 neuro-epithelial tissue tumours, 46 primitive neuroectodermal tumours (retinoblastomas, medulloblastomas, neuroblastomas, etc.) and 95 brain metastases. The DNA ploidy level determination was carried out by means of the computer-assisted microscope analysis (digital cell image analysis) of Feulgen-stained nuclei. The results show that the DNA ploidy level does not contribute significant diagnostic information when tumours are analyzed individually. Indeed, some tumours which are definitely benign like certain schwannomas and meningiomas can exhibit very high levels of aneuploidy, while some highly malignant tumours like certain glioblastomas and neuroblastomas can be diploid. In contrast to its weak diagnostic value, the DNA ploidy level appears to be a powerful prognostic factor with respect to the supratentorial astrocytic tumours of the adult. Indeed, patients with hypertriploid astrocytic tumours exhibit a survival period which is significantly longer when compared to that of patients with non-hypertriploid astrocytic tumours. These hypertriploid astrocytic tumours could be involved in a process of biological degeneration when reference is made to their proliferation activity which is significantly weaker than that of non-hypertriploid tumours.

Aneuploidy↗

PubMedDate not supplied

The combination of a decision tree technique with the computer-assisted microscope analysis of Feulgen-stained nuclei to assess aggressiveness in lipomatous and smooth muscle tumors.

The present study describes a computer-assisted methodology whose purpose is to reduce the degree of subjectivity in the diagnosis of soft tissue tumors. This methodology associates three complementary techniques, namely digital cell image analysis, the discretisation of numerical data and a Decision Tree technique (DT). The first technique relies on the use of the digital cell image analysis of Feulgen-stained nuclei, a technique which makes possible a quantitative and thus objective description of nuclei with the help of 24 numerical parameters (15 morphonuclear and 9 DNA content- (ploidy level and proliferation activity) related). The second technique transforms each numerical parameter into an ordinal one with a small number of values (2 to 4) so that only the relevant physical significance of the parameters is retained. The Decision Tree technique generates classification rules on the basis of the discretised parameters quoted above. This methodology was applied to 53 human soft tissue tumors which included 26 lipomatous tumors (13 malignant liposarcomas and 13 benign lipomas) and 27 smooth muscle tumors (11 malignant leiomyosarcomas and 16 benign leiomyomas). The results show that a distinction between benign (lipoma) and malignant (liposarcoma) lipomatous tumors can easily be made by means of simple logical rules depending on only four discretised cytological parameters (two ploidy- and two morphonuclear-related). In contrast, no stable or predictive characterisation can be obtained with respect to the difference between leiomyosarcomas and the leiomyomas. Hence, while lipomas and liposarcomas appeared to be two completely distinct biological entities, leiomyomas and leiomyosarcomas seem to involve a continuous biological process.

Cell Nucleus↗