Biomedical subjects

J Bartek

Publications and source records attributed to J Bartek.

At least 163 records · Page 9Linked to original sources

PubMed1992

Studies of clonal cell lines developed from primary breast cancers indicate that the ability to undergo morphogenesis in vitro is lost early in malignancy.

In an attempt to obtain cell lines from the different components found in primary breast cancers, we used a low-calcium medium to culture epithelial cells of mixed phenotype and a recombinant T antigen containing retrovirus to immortalize cells found in these cultures. In each case, the histology of the sample used for culture was examined in detail and the best growth was obtained from samples associated with a substantial in situ or benign component and from lobular rather than ductal carcinomas. Clonal cell lines were developed from each of 4 tumours: 1 infiltrating ductal (tumour number 2), 2 infiltrating lobular (tumours 3 and 5) and 1 mucoid (tumour 6). To try to identify the phenotype and origin of the cell lines, immunohistochemical markers, histological analysis of tissue sections and behavioural markers were used. All the cell lines expressed mainly luminal epithelial cell markers, but the basal epithelial keratin, keratin 14, was also expressed homogeneously or heterogeneously. Growth in agar was seen with some but not all cell lines derived from only 1 tumour (tumour 5) and tumour development in nude mice was observed (with low efficiency) with cell lines from only 1 tumour (tumour 6). The data suggest that the cell lines obtained from the infiltrating ductal carcinoma (tumour 2) developed from cells cultured from the associated benign component, while the cell lines from tumours 3, 5 and 6 may each have developed from a cell in an early stage of malignancy. When tested for their ability to undergo morphogenesis on extracellular matrix components, cell lines from tumour 2 made well-developed ductal-alveolar-like structures, while those from the other tumours did not.

Animals↗

PubMed1992

Distribution of individual components of basement membrane in human colon polyps and adenocarcinomas as revealed by monoclonal antibodies.

Double-label immunofluorescence was used to monitor basement-membrane composition and integrity in 22 human colon polyps, 36 adenocarcinomas and 2 metastases. Cryostat sections were stained with polyclonal anti-laminin anti-serum combined with monoclonal antibodies (MAbs) to all major basement-membrane components (laminin, entactin/nidogen, collagen type IV and large heparan sulfate proteoglycan), as well as to keratin 8. In all adenocarcinomas, including mucinous, basement membranes were altered more at the invasive front than in the parenchyma. The degree of this alteration was inversely correlated with the level of tumor differentiation. An uncoordinated loss of basement membrane components (dissociation of markers), previously described by us in rat colon adenocarcinomas, was also found in human tumors. In the great majority of adenocarcinomas a pronounced stromal reaction was seen. It was manifested by the presence of fibrillar deposits of basement-membrane components, mainly of collagen type IV and/or heparan sulfate proteoglycan. This reaction was never observed in polyps and may be derived from myofibroblasts reported to accumulate in colon cancer stroma. The combined use of antibodies to basement-membrane components and to a specific keratin may constitute an adequate immunohistochemical test for the presence of invasion, and may be useful in the histologic analysis of polyps, especially in dubious cases.

Adenocarcinoma↗

PubMed1992

The preheparin and postheparin lipids of high density lipoprotein subfractions: relation to the serum insulin and postheparin plasma lipase activities in normal human.

The preheparin and postheparin lipoprotein lipids (cholesterol C and triacylglycerols TAG) were related to the serum insulin and to the postheparin plasma activities of lipoprotein lipase (LPL) and hepatic lipase (HL) in normolipemic human. The positive correlation, although not statistically significant, of insulin level of the LPL activity was found, while no correlation to the HL activity was seen. Under preheparin conditions: 1) both insulin level and LPL activity were negatively related to the VLDL-C/TAG ratio indicating an enrichment of VLDL with TAG, 2) moreover, the LPL activity was positively related to the HDL-C and HDL3-C, 3) the HL activity predominated in relation to the HDL, and particularly to the HDL3, as indicated by negative correlations to the both lipids and to the lipid/apoA-I ratios of HDL and HDL3. This lipid depletion of HDL3 was more expressive due to the TAG, while HDL2 appeared to be relatively enriched with TAG, as suggested by correlations of C/TAG ratios with HL activity. The in vivo acceleration of lipoprotein metabolism by heparin resulted in: 1) the reduction of VLDL-TAG and HDL2-TAG, and in the increase of HDL3-TAG, 2) the appearance of positive relation of HDL2-C to the LPL activity and of opposite relation to the HL activity, 3) the lack of HL correlation to the TAG of HDL and HDL3. Even under these conditions no relation of insulin level to any HDL lipid was revealed. The results suggest that in normal human the HL affects more considerably than LPL the lipid metabolism of HDL subfractions and it does not seem to be under insulin control.

Adult↗

PubMed1991

Efficient immortalization of luminal epithelial cells from human mammary gland by introduction of simian virus 40 large tumor antigen with a recombinant retrovirus.

When defined in terms of markers for normal cell lineages, most invasive breast cancer cells correspond to the phenotype of the common luminal epithelial cell found in the terminal ductal lobular units. Luminal epithelial cells cultured from milk, which have limited proliferative potential, have now been immortalized by introducing the gene encoding simian virus 40 large tumor (T) antigen. Infection with a recombinant retrovirus proved to be 50-100 times more efficient than calcium phosphate transfection, and of the 17 cell lines isolated, only 5 passed through a crisis period as characterized by cessation of growth. When characterized by immunohistochemical staining with monoclonal antibodies, 14 lines showed features of luminal epithelial cells and of these, 7 resembled the common luminal epithelial cell type in the profile of keratins expressed. These cells express keratins 7, 8, 18, and 19 homogeneously and do not express keratin 14 or vimentin; a polymorphic epithelial mucin produced in vivo by luminal cells is expressed heterogeneously and the pattern of fibronectin staining is punctate. Although the cell lines have a reduced requirement for added growth factors, they do not grow in agar or produce tumors in the nude mouse. When the v-Ha-ras oncogene was introduced into two of the cell lines by using a recombinant retrovirus, most of the selected clones senesced, but one entered crisis and emerged after 3 months as a tumorigenic cell line.

Animals↗

PubMed1991

Growth and development of the human infant breast.

Seventy-two samples of infant breasts, aged from newborn to 2 years, were collected at necropsy. Whole-mount preparations and histological sections were made. A system of classification was devised to study the extent of the structural development of the ductal system (morphological types I, II, and III) and the functional differentiation of the lining epithelium (functional stages I to V). There was no correlation between the age of the infant and the type of development of the ductal system. In contrast, the epithelial differentiation followed a chronological pattern, starting with secretory changes and apparently going through a period characterized by apocrine metaplasia before post-secretory involution. These epithelial changes were not associated with the morphological type of the ductal system. There were no distinguishing features between the breasts from the two sexes. Immunoperoxidase staining for actin and kappa-casein was carried out to study the myoepithelial cells and secretory cells, respectively. Myoepithelial cells were present at all stages and prominent staining for casein was observed up to 2 months of age. Embryonic-type adipose tissue was seen in 7 cases, in one of which it was associated closely with the developing ductal system. Extramedullary hematopoiesis was observed in the periductal connective tissue until 4 months of age. This paper describes the most extensive anatomical and histological study of the human infant breast to date and lays the foundation for a detailed study of the epithelial and stromal changes that take place during human breast development.

Actins↗

PubMed1991

Human thymic epithelial cells are frequently transformed by retroviral vectors encoding simian virus 40.

The thymic microenvironment contains a mixture of phenotypically distinct epithelial cells of varied functions, some of which are unknown. In an attempt to understand their relevance to T cell differentiation in the thymus, human thymic epithelial cell clones from both fetal (SM3-SM5) and postnatal (SM6) thymus were produced by using a defective recombinant retroviral vector encoding the simian virus 40 large T antigen and the neomycin resistance gene. The presence of keratins 8 and 18, desmosomes, and tonofilaments confirmed the epithelial origin of the cell strains. The cells expressed Thy-1 and HLA-Class I at high levels, showed weak-expression antigens defined by TE3B and A2B5, and low to negligible levels of the MR19-defined molecule. When compared with the phenotype of thymic epithelial cells in situ, the cell strains appear to be derived from neuroendocrine components in the outer cortical region of the human thymus. The use of retroviral vectors to transform human thymic epithelium was considerably more efficient than transfection with a plasmid carrying the origin of replication-defective SV40 large T gene. In the latter case, only two cell strains with subcapsular epithelial phenotypes were derived from fetal thymus. With the retroviral vectors, epithelial cell strains could, for the first time, be generated from human postnatal thymus as well as from fetal thymus.

Cell Division↗

PubMed1990

Selective immortalization of a phenotypically distinct epithelial cell type by microinjection of SV40 DNA into cultured human milk cells.

An immortal cell line, MMSV-1, has been developed which exhibits many features of the common luminal epithelial cell of the human mammary gland. The cell line was developed by microinjection of SV40 DNA into individual cells in selected colonies in cultures of human milk epithelial cells. Immunohistochemical staining shows that the MMSV-1 cells express keratins 7, 8, 18 and 19 homogeneously in organized filaments which lead into well-developed desmosomes. They do not express vimentin or keratins found in stratified epithelia or keratin 14 found in basal cells in the mammary gland. The PEM mucin, recognized by the antibody HMFG-I, is also expressed and appears to be processed normally. Fibronectin is detected but shows the punctate pattern typical of cultured normal milk epithelial cells. MMSV-1 cells show a reduced requirement for added growth factors, including cyclic AMP-elevating agents, but do not grow in agar or form tumours in nude mice. Since the transfected cells could be selected on the basis of an extended in vitro life span, antibiotic resistance markers were not introduced and the cells remain sensitive to hygromycin and neomycin.

Animals↗

PubMed1990

Increased expression of mutant forms of p53 oncogene in primary lung cancer.

Primary lung cancer samples of the major histological types were examined for expression of the tumor suppressor gene p53 by immunohistochemistry. Abnormalities in p53 expression were found in 28 of 40 carcinomas, 14 of 17 squamous tumours showing abnormal p53 expression, whereas no expression of p53 was detectable in 7 carcinoid tumours or in 10 normal lung samples. Direct evidence for homozygous expression of mutant p53 mRNA in representative carcinomas was obtained by means of an asymmetric polymerase chain reaction mRNA sequencing strategy, which allowed sequencing without any cloning step. All the mutations were G to T transversions resulting in mis-sense mutations in aminoacids highly conserved in evolution. Mutation of the p53 gene is the most frequently identified genetic change in human lung cancer; these findings suggest that simple immunohistological methods can provide strong evidence of such mutation.

Adenocarcinoma↗

PubMed1990

Keratin 19 expression in the adult and developing human mammary gland.

In the adult human mammary gland, most of the luminal epithelial cells express keratin 19 (K19+). However, in some small ducts and terminal ductal lobular units where branching would be expected to occur during pregnancy, the pattern of expression of this keratin is heterogeneous. While the keratin 19 negative cells (K19-) appear to have a high proliferative potential in vitro and in vivo, they have a lower secretory activity than the K19+ cells as monitored by expression of secretory component in the resting breast or casein in the pregnant gland. That the K19- cells form a separate proliferative compartment in the luminal cell lineage is suggested by the fact that they are absent in the prepubertal breast and only appear at puberty associated with branching ducts, and newly formed lobules. Our observations are consistent with the hypothesis that the K19- luminal cell is less differentiated than and may be precursor to the K19+ luminal cell, which represents the fully differentiated phenotype able to produce milk in response to a hormonal stimulus.

Aging↗

PubMed1990

Genetic and immunochemical analysis of mutant p53 in human breast cancer cell lines.

The expression of the tumour suppressor gene p53 was analysed in 11 human breast cancer cell lines by immunohistochemistry, immunoprecipitation and cDNA sequencing. We used a panel of anti-p53 monoclonal antibodies for cell staining and found abnormalities in every case. Eight of the cell lines produce a form of p53 which can be immunoprecipitated by the monoclonal antibody PAb240 but not by PAb1620. In the murine system PAb240 only immunoprecipitates mutant p53. We sequenced p53 cDNA directly from four of the PAb240 positive cell lines using asymmetric PCR templates. All four contained missense mutations in p53 RNA, with no detectable expression of the wild type sequence. Different residues were affected in each cell line, but all the mutations changed amino acids conserved from man to Xenopus. These results imply that as in the murine system, the PAb240 antibody reliably detects a wide variety of p53 mutations and that these mutations have a common effect on the structure of p53. Immunohistochemical data suggest that p53 mutation is the commonest genetic alteration so far detected in primary breast cancer.

Amino Acid Sequence↗

PubMed1990

Effect of manganese ions on HDL2 precipitation with dextran sulfate.

The dual precipitation method involving the separation of HDL and HDL3 from normolipemic serum by heparin-MnCl2/dextran sulfate was evaluated with regard to the Mn2+ effect on HDL2 precipitation with dextran sulfate (DS) in the second precipitation step. Increased HDL2-C and HDL2-TAG values brought about by increased Mn2+ concentrations were found at all DS concentrations studied. The comparison of the lipid HDL2 and HDL3 values with those given by polyethylene glycol precipitation method yielded two optimal combinations of DS/Mn2+ concentrations: 1.11 g.l-1/0.076 mol.l-1 and 0.87 g.l-1/0.091 mol.l-1. The obtained results call attention to the importance of final concentration of Mn2+ ions at HDL2 precipitation with DS from heparin-Mn2+ supernate containing HDL.

Adult↗

PubMed1989

Cytokeratins and cytokeratin filaments in subpopulations of cultured human and rodent cells of nonepithelial origin: modes and patterns of formation.

Using immunofluorescence microscopy, we observed that in several established cell culture lines derived from different nonepithelial tissues and species, cells spontaneously emerge, usually at low frequencies, which contain cytoplasmic structures decorated by antibodies specific for cytokeratins 8 and 18. This phenomenon was further examined at both the protein (gel electrophoreses of cytoskeletal proteins, followed by immunoblotting) and the RNA (Northern blots, "nuclear run-on" analysis, in situ hybridization) level. Positive cell lines included simian virus (SV40)-transformed human fibroblasts (HF-SV80, WI-38 VA13), human astrocytic glioma cells (U333 CG/343MG), rat (RVF-SMC) and hamster (BHK-21/13) cells derived from vascular smooth muscle and murine sarcoma MS-180 cells. In two cell lines (HF-SV80 and BHK-21/13), the frequency of the cytokeratin-containing cells and of the cytokeratin fibril arrays per cell was drastically increased upon treatment with 5-azacytidine. The structural appearance of the cytokeratins was variable in the different cell lines but could also differ among cells of the same culture: While small granular or comma-shaped structures or bizarrely shaped filament arrays prevailed in WI-38, RVF and normally grown BHK-21 cells, most of the other lines revealed extended normal-looking, fibrillar arrays. In one line (MS-180), the appearance of cytokeratins was associated with a morphological change, as it was only found in a subpopulation of cells that had lost their typical elongated and spindle-shaped phenotype and assumed a rounded ("coccoid") shape. Our results show that the expression of the genes encoding cytokeratins 8 and 18 is not necessarily restricted to programs of epithelial differentiation and that factors stochastically effective appear in cultured cell lines that allow the synthesis of these cytoskeletal components. Mechanisms possibly involved in this spontaneous and selective advent of cytokeratins 8 and 18 and implications for tumor diagnosis are discussed.

Animals↗

PubMed1989

Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: relation to in vivo phenotypes and influence of medium.

The luminal and basal epithelial cells in the human mammary gland can be distinguished in tissue sections on the basis of the pattern of keratins they express. Moreover, the invasive cells in primary carcinomas show a keratin profile that corresponds to that of the dominant luminal cell (7, 8, 18, 19). When homogeneous populations of luminal epithelial cells from milk or from breast cancer metastases are cultured the profile of keratin expression seen in vivo is maintained. We have therefore used monospecific antibodies reactive with individual keratins to examine the phenotype of cells cultured in three different media from reduction mammoplasty tissue that contains both luminal and basal cells. The phenotype of cells cultured from primary breast cancers in one of these media (MCDB170) has also been examined. In characterizing cell phenotypes, antibodies to a polymorphic epithelial mucin (PEM) expressed in vivo by luminal cells, and to smooth muscle (a) actin, expressed in vivo by basal cells, have also been used. Our results show that proliferation of different cell phenotypes is selected for in different media. In milk mix (MX) developed for growth of luminal cells from milk, only the luminal cell phenotype proliferates (for only 1 or 2 passages). In medium MCDB 170, which was developed for long-term growth of human mammary epithelial cells from reduction mammoplasty organoids, cells from the basal layer proliferate, while in MM medium the basal phenotype dominates, but a few cells with the luminal phenotype are found. Around passage 3, in medium MCDB 170, most cells senesce and a subpopulation of cells proliferates on further passage. These cells retain expression of the basal epithelial keratins but also express some features characteristic of luminal epithelial cells, suggesting that the basal layer may contain a stem cell that can develop along the luminal lineage. In culture, however, they do not express keratin 19, which in vivo is a feature of the fully differentiated luminal cell. The cells cultured from primary breast cancer in medium MCDB 170 have a similar keratin profile to that of the normal cells cultured in this medium. They do not express keratin 19, even though the invasive cells in primary cancers homogeneously express this keratin in vivo. The invasive phenotype, which in its keratin profile corresponds to the differentiated luminal cell and that of the metastatic cancer lines, cannot be cultured from primary breast cancers using MX, which supports proliferation of the corresponding normal cell.

Breast↗

PubMed1989

[Cystic changes in the liver in our 5-year ultrasonographic study].

The authors examined during 1983-1987 by ultrasonography the liver in a group of 13,672 patients with an average age of 54.8 +/- 4.1 years. The group comprised 7830 women (57.27%) and 5842 men (42.73%). Cystic changes were detected in 39 patients (0.28%). During subsequent examinations they revealed that in four patients metastases of tumours were involved. The correctness of the ultrasonographic diagnosis was confirmed in 35 patients (89.7%). Non-inflammatory and non-parasitic cysts were present in 31 patients (0.23%), incl. 20 women (0.26%) and 11 men (0.19%) with a sex ratio of 1.4:1. Concurrent cystic changes on the kidneys were found in 22.6% and on the pancreas in 12.9%. The spleen was not affected by cystic changes in any of the patients. The diagnosis of cysts was supplemented by laboratory, laparoscopic, scintigraphic, computed tomographic peroperative and aimed bioptic examinations, depending on the character of changes. An inflammatory cyst (tuberculous cavity) as well as three cases of echinococcal cysts were treated by operation. In the conclusion the authors discuss contemporary diagnostic and therapeutic methods in cystic liver diseases.

Adolescent↗

PubMed1989

[Initial experience with thin-needle biopsy under ultrasonic control in diffuse liver diseases].

The authors used thin-needle biopsy (Westcott needle) under ultrasonographic control for the morphological determination of ultrasonographic pictures of steatosis in 37 patients. In the case history they reported alcohol consumption of more than 5 years above 60 g of absolute alcohol per day; mostly abuse of beer was involved (32.4%) and beer with liquor (43.3%). A sufficient amount of material for histological examination was taken in all 37 patients, incl. 8 during the second insertion. Histological examination revealed mainly large-droplet steatosis of the mild and medium grade (40.5%) and severe grade (21.6%). Of 37 ultrasonographic pictures of steatosis the latter was confirmed by histological examination in 34 patients (91.2%). The biopsies were made without algic reactions of the patients and no complications were observed. With regard to the non-pretentious, simple and safe character and the high yield of the procedure the authors consider thin-needle biopsy under ultrasonographic control a foremost operation which makes morphological assessment even of diffuse liver diseases possible.

Adult↗

PubMed1989

[Residual urine and phagocytic activity of polymorphonuclear cells in diabetes].

The authors examined by ultrasonography the residual urine in 105 diabetics after passing urine (55 men--53.3% and 49 women--46.7%) aged 20 to 50 years and compared the results with a control group of 103 non-diabetics selected at random (55 men--48.7% and 58 women--51.3%), also aged 20-50 years. Patients with organic diseases affecting the residual urine were not included in the group. In all patients the phagocytic ratio was examined, the urine was analyzed chemically and the sediment was examined, in selected cases also cultivation examinations were 105 diabetics residual urine was found in 34 cases (34.4% and in the control group of 113 non-diabetics in 3 cases (2.7%). The difference is highly significant (p less than 0.001). In the group of diabetic men and women residual urine was found more frequently in type I diabetics with diabetes existing for prolonged periods and the amount of residual urine was larger as compared with type II diabetes. Comparison of the phagocytic ratio in the diabetics and controls of both sexes revealed significant changes at the level of p less than 0.05. In the group of diabetics with residual urine was a significantly higher incidence of pathological findings in urine as compared with diabetics without residual urine which is of pathognomic importance for the development and persistence of urinary infections.

Adult↗

PubMed1988

Rapid selection of cultured cells with increased expression of a membrane marker (secretory component).

This study showed how immunomagnetic monodisperse microspheres (Dynabeads) can be used for rapid selection of stable and homogeneous sublines of cancer cells. The beads were activated with antibody to a specific surface membrane marker. We used a monoclonal antibody (MAb) to human secretory component (SC), the epithelial receptor for polymeric immunoglobulins (pIg). A relatively low number of particles favored rosetting of strongly positive cells. SC-expressing cells were isolated within 30 min from a colonic carcinoma cell line (HT-29.E10). After 13 months of continuous culture the selected cells had a 15 times higher cellular content and a 10 times higher secretory rate of SC than parallel cultures of the original cell line. Immunomagnetic microspheres can be recommended in experimental cancer research, e.g., in order to minimize cellular heterogeneity and to isolate cells with particular surface membrane phenotypes in differentiation studies. The selection method is rapid and simple, and eliminates laborious test systems for conventional subclone screening.

Animals↗

PubMed1987

Ductal adenoma of the breast--a lesion exhibiting a myoepithelial/epithelial phenotype.

A detailed immunocytochemical study has been carried out on a case of ductal adenoma of the breast. The lesion was oestrogen receptor positive and contained focal areas of apocrine metaplasia and marked nuclear atypia. The tumour had a pseudo-infiltrative growth pattern and showed good evidence of both epithelial and myoepithelial differentiation. Islands of tumour cells were surrounded by a clearly defined basement membrane as demonstrated with an anti-type IV collagen antibody. The heterogeneous staining pattern produced by monoclonal antibody BA17, which recognizes keratin 19, was similar to that seen in normal lobules and raises questions concerning the site of origin of this lesion in the breast.

Actins↗