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Biomedical subjects

J Sugár

Publications and source records attributed to J Sugár.

At least 19 recordsLinked to original sources

Analysis of T cell receptor AV and BV chain gene expression by infiltrating lymphocytes in Spitz naevi and in halo naevi.

Spitz naevi and halo naevi are benign melanocytic lesions that share many histological features with malignant melanoma. All lesions are characterized by a brisk infiltration of lymphocytes, mainly of the T cell subtype, and halo naevi are known to undergo spontaneous regression. Since the benign nature of Spitz naevi and halo naevi might therefore be caused by specific T cell responses against tumour-associated antigens, it was found of interest to characterize this T cell response in detail. A reverse transcriptase-polymerase chain reaction (RT-PCR)-based method adapted for analysis of paraffin-embedded material combined with Southern blot analysis has been used to analyse the T cell receptor (TCR) AV and BV repertoires of infiltrating lymphocytes in 14 different melanocytic lesions. The results have shown that only a few particular TCRAV and TCRBV regions are expressed in each lesion. To evaluate the T cell response, it is of interest to know the HLA-type of the analysed lesions, since most melanoma-specific effector lymphocytes are CD8+ cytotoxic T cells and therefore HLA class I-restricted. As blood samples were not available from any of these patients, an RT-PCR method using HLA-A2-specific primers was used to analyse for the presence of this allele. The preferentially expressed TCRAV genes were sequenced, and this analysis showed that the high expression of these TCRAV genes was due to a clonal or oligocional expansion of T cells. In summary, the expression of relatively few TCR variable regions indicates a clonal expansion of T cells.

Adolescent

New aspects in the pathology of the preneoplastic lesions of the larynx.

In our 5-year biopsy study (1991 95) 51% of keratoses and 16% of adult-onset papillomas contained certain degrees of dysplasia. Koilocytosis associated with HPV infection was prominent in papillomas and the majority of keratosis but negligible in cancer. Using mouse monoclonal antibody against human p53 we detected by immunohistochemistry mutated p53 overexpression not only in cancer but in some cases of keratoses and papillomas. CD44 is a multifunctional integral cell membrane glycoprotein and is found in many mesenchymal, epithelial and tumor cells. CD44 has a great number of splice variants, isoforms supposedly implicated in development, progression and metastasis formation of tumors. Using monoclonal antibody to CD44 our study suggests that the expression of pan CD44 at successive stages of human laryngeal carcinogenesis gives preliminary information about the selection advantage to the tumor cells.

Adult

Some etio-pathogenetic factors in laryngeal carcinogenesis.

Chemical influences, mainly heavy tobacco smoking, chewing snuff, excessive alcohol consumption, and some occupational hazards, are known to be important etiologic factors in laryngeal carcinogenesis. The synergistic or cooperative interaction of human papilloma virus (HPV) infection with these chemical factors are serious considerations in the development of laryngeal carcinoma. With the development during the last decade of Southern blot hybridization and polymerase chain reaction (PCR), extensive and comprehensive studies have been conducted to determine the presence and biological (etiologic) significance of HPV. Developed cancer, as well as juvenile and adult multiple and single papilloma of the larynx, have been the subject of clinical and molecular-pathological investigation. Our previous study showed that cancer may develop on the basis of leukoplakia and adult-onset papilloma. Extensive kilocytes, an indication of HPV infection, can be seen by histological examination in papillomas and carcinoma. Literary data suggest that in laryngeal squamous cell carcinoma, including varicoses carcinoma, HPV 16, HPV 18, and HPV 33 DNA have been detected. Both in juvenile and adult-onset respiratory papillomatosis, patients could have either HPV type 6 or 11 DNA sequences. Molecular biological and PCR studies indicate that HPV may play an etiologic role in the development of human malignancies of the upper aerodigestive tract and uterine (cervical) origin. However, evidence that unequivocally links HPV infection with laryngeal squamous cell carcinoma is still lacking. In laryngeal cancer, p53 abnormalities are related to smoking-induced mutagenesis rather than HPV. Studies have postulated an interaction between HPV infection and chemical carcinogens and have concluded that HPV possibly are co-adjuvants during the multistage process of neoplastic transformation.

Cocarcinogenesis

Cisplatin containing combination chemotherapy of advanced germ cell line testicular tumours.

One hundred ninety patients with germ cell line testicular tumours were treated according to the modified Einhorn scheme. The response rate was 67.9%. The most favourable results were found in the embryonal histologic type (RR = 76.9%) in the biological markers (beta-HCG and AFP) negative (RR = 97.4%) and in the minimal pulmonary extent group (RR = 94.1%). The authors treated 112 patients with including these VPB-resistant germ cell testicular tumour and those with recurrence after this treatment. The patients' mean age was 28.8 (limits 19 to 44) years. Patients were given Vepeside (100 mg/m in infusion for days 1-5), Adriablastin (40 mg/m in infusion on day 1) and Cisplatin (20 mg/m in infusion) for day 1-5. The treatment resulted in CR with 18 patients (16.1%) and PR with 42 (37.5%) (RR = 53.6%). The best results were obtained with the seminoma patients who were marker-negative and had small-volume metastasis. CR developed in 4 of 7 seminoma patients (57%) and in 7 of 25 marker-negative individuals (28%), and PR developed in 11 patients (44%) (RR = 72%). Out of 12 patients with small volume metastatis four (33%) showed CR and five revealed PR (41.7%), their RR turned out to be 74.6%. The average remission period was 37 (range 4-70) months in CR but merely 6.1 (range 2-38) months in PR. It can be stated that fairly good results can be achieved with second-line VpAP treatment in case of resistance developed to primary VPB therapy or subsequent relapse. The efficacy of combined chemotherapy of Vepesed+Holoxan +/- Adriablastin as third-choice was studied in advanced testicular cancer patients refractory to, or recurrent after, first- and second-line cytostatic therapy. Between September 1981 and January 1988 49 evaluable patients were treated with Vepesid (VP-16213--100 mg/m2 days 1-5), Holoxan (40 ml/kg days 1-5), hydration, urine-alkylation + Uromitexan +/- Adriablastin (40 mg/m day 1). The single dose of Uromitexan was 20% of the daily dose of Holoxan, and the patients received it i.v. just prior to Holoxan administration (h 0), the 4 and 8 h later. Two patients got into CR and 10 to PR. The rate of remission was 24.5%. The most severe side effect was leukopenia. The elevation of BUN and se. creatinine was transient and mild. In those cases where Holoxan was not included in the first- or second-line regimens, when combined with Vepesid and Adriablastin as third-choice therapy one could achieve further improvement. In case of CR the prolongation of life is also noteworthy. The first-, second- and third-line therapy plus salvage RLA and/or pulmonary metastasectomy achieved long-term survival only in one quarter of the patients.

Adult

[The use of multivariate mathematical methods in medical diagnostic systems--a model for the evaluation of cytological smears].

The methods of the multivariate mathematics have been applied in several studies to increase the diagnostic reliability of medical decision support system. In the recent years some new algorithms for decision support (fuzzy logic) and for pattern recognition (neural nets), both specified by nonlinearity, were developed. This paper provides results for the application of this methods in the area of quantitative cytology and the comparison with the traditional classifiers. 21 normal, 15 dysplastic, 23 malignant, Feulgen stained gastric imprint smears were analysed on a Leitz Miamed DNA equipment. The determination of mean DNA content, the 2c deviation index (2cDI), 5c Exceeding rate (RcER), G1,S,G2 phase fraction ratios, cell nucleus area, form factor was performed. The discriminant analysis classified correctly the 95.6% of malignant cases, 86.7% of dysplasias, and 80.7% normal cases. Our diagnostic system using fuzzy logic made the diagnostic borders fine tuneable, and reliable. The back propagation neural net could classify all three diagnostic groups above 95% correctly. The application of nonlinear computational methods made the diagnostic system more reliable. The application of these algorithms are encouraged.

Algorithms

[Quantitative morpho- and densitometric parameters in the evaluation of aspiration cytology smears of the breast].

Quantitative morpho- and densitometric analysis of Feulgen stained breast aspiration smears were performed by absorption cytophotometer and a TV image analyser. The DNA Index, and the ratio of G1-S-G2 phase fraction ratio yielded by the quantitative DNA analysis, showed significant differences between benign and malignant alterations. Based on the 13 morphometric and 5 densitometric parameters the TV-image analyser could classify the examined smears 80% correctly by cluster analysis. Groups of the cytological Grade I.-III. were also differentiable by this method. The application of TV image analyser system in the diagnosis of aspiration cytological smears could be suggested. The complex quantitative morpho- and densitometric analysis could contribute to the correct classification of diploid tumours.

Biopsy, Needle

(-)deprenyl and (-)parafluorodeprenyl-treatment prevents age-related pigment changes in the substantia nigra. A TV-image analysis of neuromelanin.

With the aid of TV-image analysis the number, the area and the density features of melanin granules in neurocytes of the substantia nigra in a group of 3-month-old naive male rats and in a 21-month-old group of male rats treated for 18 months with saline, (-)deprenyl and (-)parafluorodeprenyl, respectively, were determined. According to the Kolmogorov 2-sample test the two drug-treated groups do not differ significantly from each other in the number, total area and area of one pigment granule. In the aged saline-treated group the number of melanin granules decreased significantly and the area of the melanin granules increased in comparison to young controls as well as to the drug-treated groups.

Aging

A procedure for measuring neuromelanin in neurocytes by a TV-image analyser.

With the aid of a Robotron A6471 type TV-image analyser the number, total area, area of one granule and density features (sum, average of gray values and average gray value of one pigment granule) of melanin granules in neurocytes of the substantia nigra in 3-month-old and 3-year-old male rats were determined. The number of cells in sections of identical areas was similar in the young and old rats. A statistically non-significant difference between the two age cohorts in the proportion of neurocytes with and without melanin was found; 773 (48.1%) and 853 (51.8%), in the young rats and 1219 (65.1%) and 652 (34.8%) in the old ones. Within the melanin containing neurocytes, however, statistically conspicuous, age-related differences in the number, area and density features of melanin granules were discovered. The majority of the neurocytes in young rats contained numerous, small sized neuromelanin granules, whereas in the majority of the neurocytes of old rats smaller numbers of large sized, neuromelanin granules were detected.

Aging

Determination of G1-S-G2 phase fraction ratios of tumour subpopulations from DNA histograms.

Authors describe a new mathematical approach based on mixture decomposition and maximum likelihood method for the determination of subpopulations and G1-S-G2 phase fraction ratios in aneuploid, multiploid DNA histograms. A special computer routine was developed for this purpose (DAHSY). The present study relates to DNA histograms obtained by a Leitz Miamed DNA TV image analyser system. Altogether 23 normal, 14 dysplastic and 28 tumours Feulgen stained gastric imprint smears were measured. In addition to determination of subpopulations and G1-S-G2 phase fraction ratios, the authors also performed a morphometric analysis of Feulgen stained nuclei and determined the values of 5c exceeding rate (5cER) and the 2c deviation index (2cDI). More than one subpopulation were found only in tumours. The usual G1-S-G2 phase fraction rates in normal gastric smears were 85.5%, 9.04%, 4.9%, in dysplastic cases 74%, 15.4%, 10.2% and in tumours 53.8%, 22.9%, 18.8%. Significance of the differences were calculated by the T-test. The G1 phase fraction ratio clearly discriminates the malignant lesions from all others. The S+G2 phase fraction ratio, however, distinguishes the normal cases from the dysplastic ones. The 2cDi and 5cEr could support a multivariate diagnostic system. The cell nucleus area was found also to be of diagnostic value.

Algorithms

Influence of luteinizing hormone-releasing hormone agonists on human mammary carcinoma cell lines and their xenografts.

The specific binding of luteinizing hormone-releasing hormone (LH-RH) agonist in estradiol-dependent MCF-7 and estradiol-independent MDA-MB-231 human breast cancer cells has been studied using [3H]Ovurelin [(D-3H-Phe6),des-Gly10-LH-RH- ethylamide]. The results of Scatchard analyses suggest the presence of a single class of receptor sites, both in cell suspensions and membrane fractions. Evaluation of these peptide receptors appears to reflect additional characteristics of biological behaviour of these human breast cancer cells. The synthetic LH-RH agonist Ovurelin [(D-Phe6),des-Gly10-LH-RH-ethylamide] can directly interfere (25-30%) with the proliferation of MDA-MB-231 human breast cancer cells in culture. The inhibitory effect of Ovurelin in vitro was negligible in the MCF-7 cell line. In the in vivo experiments the treated immunosuppressed mice bearing either MCF-7 or MDA-MB-231 xenografts responded to the high-dose LH-RH analogue Zoladex depot and Decapeptyl depot therapy. Since the MDA-MB-231 tumour was found to be ER-negative it seems possible that the regression of this xenograft results from the direct antitumor action of the LH-RH agonist.

Animals

Nucleolar organizer regions in soft tissue tumours.

The diagnostic perspectives of the combination of silver NOR staining method with TV image analysis were tested in 138 soft tissue tumours. In addition to counting the dots per cell, we also measured the summed areas of dots within the nucleus. The so called "malignancy factor" (MF), i.e. the number of dots multiplied by summed areas of dots in one nucleus, was calculated. Tumours were divided into four groups: 1) benign, 2) benign with diagnostic difficulty, 3) intermediate malignant and 4) malignant. The number of dots, the summed area of dots and the MF were separately evaluated by the "t" test. Groups 2 and 3 were not distinguishable at all (p. - 0.788-0.863), while the others showed significant differences in the former values (p less than 3.5 x 10(-6) - 10(-6). When the number of dots was considered in itself, some overlaps occurred between the fourth and the other groups. With the determination of MF, however, no overlap was encountered at all. MF seems to be superior to traditional dot counting when malignant, intermediate and benign soft tissue tumours are to be distinguished.

Humans

Genotypic and phenotypic changes of a mouse lymphoma during long-term cultivation.

NK/Ly mouse lymphoma ascites cells were explanted and established in culture. The cells grew in monolayer and showed fibroblast-like morphology. The original NK/Ly cells contained one large metacentric marker chromosome, while the in vitro growing cells had two metacentrics in the early passages. These so-called bi-armed chromosomes were growing in number, up to seven, along with the number of subcultures. No tumour "take" was observed when the cells were given into mice intraperitoneally. However, a solid tumour developed following injection of cells into the leg muscle. The histological picture of this solid tumour was anaplastic sarcoma. It is concluded that the accumulation of bi-armed chromosomes, which were formed either by anaphase bridges, or by centric fusion of telocentric chromosomes, led to a profound alteration of lymphoma resulting a phenotype of anaplastic sarcoma.

Animals

Lipid-rich cell thyroid adenoma: histopathology with comparative lipid analysis.

A second case of the unique lipid-rich cell thyroid adenoma is described complemented by detailed lipid analysis. New observations were made. The cytoplasm of the tumour cells contained scattered, aggregated sudanophil crystals; under polarized light the frozen, unstained sections exhibited numerous birefringent lipid crystals; electron microscopy provided further evidence that the clear cell appearance was due to intracellular lipid droplets with scanty glycogen particles. Comparative lipid analysis by thin layer chromatography and high-pressure liquid chromatography (HPLC) revealed quantitative and qualitative differences in lipid composition of tumour cells when compared with goitre cells from normal thyroid gland and subcutane fat. Qualitative differences in triglyceride composition (by HPLC) between tumour cells and subcutaneous fat indicated that the fat accumulation in the follicle cells was not a result of simple storage, but an expression of altered intracellular lipid metabolism.

Adenoma

Vinblastine, cisplatin and bleomycin treatment of advanced nonseminomatous testicular tumors.

Between December 1979 and July 1986, 190 patients with nonseminomatous germ-cell testicular tumors were treated according to the modified Einhorn scheme. The response rate was 67.89%. The most favorable results were found in the embryonal histologic type (RR = 76.9%), in the biological marker (AFP, HCG) negative (RR = 97.43%) and in the minimal pulmonary extent group (RR = 94.12%).

Adolescent

Geno- and phenotypic characterization of the de novo arising colon carcinoma in familial polyposis coli.

De novo origin of colon carcinomas has been demonstrated in a case of familial polyposis. Independent origin of the polyps and the synchronous carcinomas was confirmed by their different pheno- and genotypes. Characterization of the phenotype was based on the mucin profile of the polyps and the synchronous carcinoma. The phenotype of each polyp was different and represented either duodenal, colonic or embryonal type of differentiation. The phenotype of carcinoma proved to be adult colonic type. The DNA content of the polyps and carcinoma has been evaluated by cytophotometry. The multiclonal origin of each polyps and synchronous carcinomas has been supported by their different DNA indices. Occurrence of de novo arising carcinoma in familial polyposis calls for a frequent follow-up examination of patients with subtotal colectomy and strongly support the need for total colectomy.

Adenomatous Polyposis Coli

DNA distribution pattern and comparison of quantitative data in breast tumours.

Aspiration cytology smears from 24 breast alterations were subjected to cytophotometry and TV image analysis. The smears were grouped according to histological diagnosis and cytological pattern. Cell cycle parameters and DNA indices were calculated from the decomposition of the DNA histograms. Based on quantitative morphometric features the TV image analyser distinguished the benign cells from the malignant ones. In benign cases its diagnostic accuracy is of 80%. Cytological Grades 1, 2 and 3 are also identifiable by this system. Cytophotometric measurements were also performed with 22 invasive breast cancers with known 5-year survival. The cell cycle parameters were related to the survival data. From the point of view of survival only the ratio of the S plus G2 phase cells of all the parameters examined proved to be significant.

Adenofibroma

[DNA cytometry and morphometry, based on TV image analysis, in the diagnosis of gastric carcinoma].

Cytological smears from gastric biopsies were studied by two types of TAS. Böcking's algorithm and a great number of form factors were used to separate the intact cells from the tumourous ones. The numerically expressed density features and morphometric parameters are able to indicate the possibility of malignant transformation even in dysplasia. These values become much higher in tumours and lead to unequivocal diagnosis. Both types of TAS render valuable help in assessing cases not reliably diagnosed by routine staining procedures.

Biopsy

Extraspinal ependymoma in the sacrococcygeal region. A case report with ultrastructural, immunohistochemical and cytophotometric studies.

We describe on a primary, subcutaneous sacrococcygeal ependymoma presenting in a young female patient. Detailed immunohistochemical and electron microscopic examinations were performed. Out of the 40 similar cases reported in literature this is the first in which the determination of DNA content was also used to predict biological behaviour. The tumour proved to be aneuploid with low proliferative capacity in spite of absent histological signs of malignancy. It is suggested that DNA determination may be helpful in establishing prognosis and that it may contribute to a better understanding of the biological behaviour of this tumour.

Adult