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J Gogusev

Publications and source records attributed to J Gogusev.

At least 37 records · Page 2Linked to original sources
PubMed1996

Potential effect of metabolic acidosis on beta 2-microglobulin generation: in vivo and in vitro studies.

Beta 2-microglobulin (beta 2M) is responsible for dialysis-associated amyloidosis. Level of beta 2M in plasma increase during chronic renal failure; however, retention does not appear to be the sole mechanism responsible. The effect of metabolic acidosis on beta 2M production was examined. Thirty-six patients with stable chronic renal insufficiency, 12 uremic patients before their first dialysis, 8 hemodialysis patients who were assigned to acetate or bicarbonate dialysate and then crossed over to the alternative regimen, and 6 normal subjects given NH4Cl to initiate metabolic acidosis were studied. In vitro studies in the human myeloid cell line U 937 were also performed. beta 2M protein was measured with ELISA, beta 2M mRNA was measured with reverse transcription polymerase chain reaction, and the U 937 cells were studied at two pH levels with FACScan flow cytometry. The cells were exposed in vitro up to 60 min in a buffered incubation medium to either pH 5.10 or pH 7.34. An inverse correlation was found between beta 2M and bicarbonate concentrations in plasma in the stable chronic renal failure patients (r = -0.54; P < 0.05) and in the uremic patients before their first dialysis (r = -0.72; P < 0.05). In hemodialysis patients, blood pH and plasma bicarbonate values were lower (P < 0.05) and beta 2M concentrations in plasma were higher (P < 0.05) with acetate than with bicarbonate dialysate. In normal men, NH4Cl resulted in an increase (P < 0.05) in beta 2M mRNA expression in lymphocytes by an average factor of 1.5 (range, 1.1 to 1.8). In U 937 cells, the cell surface expression of beta 2M and HLA Class I heavy chain assembled with beta 2M decreased at low pH compared with normal pH. Concomitantly, an increase in beta 2M release into the supernatant was observed, possibly as the result of beta 2M dissociation from cell surface HLA Class I complex. The results suggest that metabolic acidosis may enhance cellular beta 2M generation and release.

Acidosis↗
PubMed1995

Expression of bcl-2 oncoprotein in renal cell tumours.

Expression of bcl-2 is associated with inhibition of apoptosis and extension of cell survival. The importance of apoptosis in relation to the development and progression of renal cell neoplasia remains undefined so far. In order to determine the expression of bcl-2 oncoprotein in normal and neoplastic renal cells, 37 renal tumours were investigated by immunolabelling, including 13 clear cell carcinomas, ten tubulopapillary carcinomas, four chromophobic renal cell carcinomas, and ten oncocytomas. Twenty-six samples of adjacent normal renal tissue served as controls. bcl-2 expression was correlated with cell proliferation activity as estimated by Ki67 antigen expression, and p53 protein expression in the tumour samples. The results demonstrate that in the normal kidney, positive bcl-2 immunostaining was present in glomerular parietal epithelial cells, in distal tubular cells, and in sparse proximal tubule cells. Renal cell tumours showed heterogeneous bcl-2 expression according to the tumour cell type. While the majority of carcinomas of clear cell type were usually negative or contained sparsely distributed positive cells, all tubulopapillary carcinomas were consistently positive for bcl-2. In oncocytomas and chromophobic carcinomas, there was a low percentage of bcl-2 immunoreactive tumour cells; some nuclear bcl-2 positivity was detected in one chromophobic tumour. These findings indicate variable bcl-2 oncoprotein expression in different types of renal cell tumours, with the highest level of expression in tubulopapillary carcinomas. No clear relationship was found between nuclear grade, cell proliferation activity, and level of bcl-2 expression. p53 protein was detected in only one tubulopapillary carcinoma.

Biomarkers, Tumor↗
PubMed1994

Effect of erythropoietin on DNA synthesis, proto-oncogene expression and phospholipase C activity in rat vascular smooth muscle cells.

The administration of recombinant human erythropoietin (rHuEpo) to anemic chronic renal failure patients may be associated with an increase in blood pressure, possibly by direct effects on peripheral blood vessels. The experiments of the present study were designed to explore the hypothesis that rHuEpo might exert mitogenic effects on vascular smooth muscle cells (VSMCs), and that pre-existing hypertension might be a predisposing condition. Cultured aortic VSMCs from spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats were studied for DNA synthesis, phospholipase C activity, and cell growth related proto-oncogene expression in the presence of rHuEpo. In cells from both rat strains, rHuEpo dose-dependently increased DNA synthesis and stimulated phospholipase C activity, as indicated by 3H-thymidine incorporation and inositol phosphate formation, respectively. Exposure of VSMCs to rHuEpo for various periods gradually increased the levels of c-myc and JunB mRNAs and transiently induced c-fos mRNA expression as determined by Northern analysis. The hormone-induced DNA synthesis was markedly enhanced in VSMCs from SHR compared to those from WKY. In contrast, rHuEpo-induced phospholipase C activity and proto-oncogene expression did not differ between the two strains. Taken together, these results suggest that rHuEpo may function as a vascular smooth muscle cell growth promoting factor through activation of the phospholipase C cascade and a modulation of proto-oncogene expression. It could thereby contribute to vascular hypertrophy and arterial hypertension.

Animals↗
PubMed1994

Composition of extracellular matrix and distribution of cell adhesion molecules in renal cell tumors.

BACKGROUND: Cell to cell and cell to matrix interactions play a major role in tumor growth and invasion. Therefore, we studied the composition of extracellular matrices and the distribution of cell adhesion molecules in 50 renal cell tumors of various types and various grades of malignancy as compared with nontumoral kidney. EXPERIMENTAL DESIGN: In the present study, we used immunolabeling with specific antibodies directed against the alpha 1, alpha 2, and alpha 3 chains of collagen type IV; laminin; heparan sulfate proteoglycan; fibronectin; collagen I; collagen III; the alpha 1, alpha 2, alpha 3, alpha 5, alpha 6, alpha v, beta 1, and beta 3 subunits of integrins; and ICAM-1, VCAM-1, and ELAM-1 molecules. RESULTS: In clear cell type carcinomas (24 cases) the basal laminae surrounding the tumor islets contained the alpha 1 and alpha 2 chains of collagen type IV and heparan sulfate proteoglycan in all cases, and laminin in 96% of the cases. The alpha 3 chain of collagen IV was present in only one case, whereas fibronectin, collagen I, and collagen III were detected in nearly 50% of the cases. The tumor cells expressed alpha 3, alpha 6, and beta 1 integrin subunits in all cases, alpha 5 in 25%, alpha v beta 3 in 54%, and alpha 2 in none. ICAM-1 was detected in all cases, and VCAM-1 in 58%. The expression of the alpha 6 subunit was weak in 2 tumors of high grade, whereas the alpha v subunit was expressed in 7 of 14 low grade and in 7 of 10 intermediate and high grade tumors. In tubulopapillary carcinomas with chromophilic cells (12 cases), the most prominent findings were the presence of the alpha 3 chain of collagen IV in tumor basal laminae in 66% and the expression of the alpha 2 integrin subunit by the tumor cells in 58% of the cases, both features characterizing distal renal tubules. In chromophobic carcinomas (4 cases), the tumor basement membranes were tenuous and contained no fibronectin or interstitial collagens. The tumor cells expressed the alpha 6, alpha 2, alpha 3, beta 1, and alpha nu beta 3 integrin subunits but neither ICAM-1 nor VCAM-1 molecules. In oncocytomas (10 cases), the tumor basement membranes contained the alpha 1, alpha 2, and alpha 3 chains of collagen IV, laminin, and heparan sulfate proteoglycan. Fibronectin was not detected, whereas interstitial collagens were present in half of the cases. In all tumors, cells expressed the alpha 6 and beta 1 integrin subunits, whereas alpha 2, alpha 3, and alpha nu beta 3 were present in 30%, 60%, and 90% of the cases, respectively. ICAM-1 and VCAM-1 were not detected. The vascular endothelial cells of the stroma expressed the alpha 1, alpha 5, alpha 6, and beta 1 integrin subunits in all 50 of the studied tumors. In addition, ICAM-1 was detected in 84%, VCAM-1 in 50%, and ELAM-1 in 34% irrespective of tumor cell type, growth, or nuclear grade. CONCLUSIONS: These results suggest that each type of renal cell tumor produces particular extracellular matrix components and expresses a characteristic repertoire of cell adhesion molecules, which could provide better understanding of the origin of these tumors. The expression of the alpha nu beta 3 integrin subunit was demonstrated in all types of renal cell tumors and was not found to be related to high grade tumors. Stromal vascular endothelial cells expressed activation molecules VCAM-1 and ELAM-1 in a significant number of cases in both benign and malignant tumors.

Adenocarcinoma, Clear Cell↗
PubMed1994

[Mitogenic effect of erythropoietin on cultured aortic myocytes].

The administration of recombinant erythropoietin (rHuEpo) to anemic chronic renal failure patients may be associated with an increase in blood pressure, possibly by direct effects on peripheral blood vessels. In the present study, experiments were designed to explore the hypothesis that rHuEpo could enhance vascular resistance through mitogenic effect on vascular smooth muscle cells (VSMCs), and that preexisting hypertension might be a predisposing condition. Cultured VSMCs from the thoracic aortae of spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats were studied for DNA synthesis, phospholipase C activity, and cell growth related proto-oncogene expression in the presence of rHuEpo. In cells from both strains, rHuEpo dose-dependently increased DNA synthesis and stimulated phospholipase C activity, as indicated by 3H-thymidine incorporation and 3H-inositol phosphate formation, respectively (EC50 approximately 4 U/ml). Exposure of VSMCs to rHuEpo for various times gradually increased the levels of c-myc and junB and transiently induced c-fos expression, as determined by Northern analysis. rHuEpo-induced DNA synthesis was markedly enhanced in VSMCs from SHR compared to those from WKY. In contrast, rHuEpo-induced phospholipase C activity and proto-oncogene expression did not differ between the two strains. Taken together, these results suggest that rHuEpo may function as a vascular smooth muscle cell growth promoting factor through activation of the phospholipase C cascade and modulation of proto-oncogene expression. It could thereby contribute to vascular hypertrophy and arterial hypertension.

Animals↗
PubMed1993

FLT4 receptor tyrosine kinase gene mapping to chromosome band 5q35 in relation to the t(2;5), t(5;6), and t(3;5) translocations.

FLT4 is a recently cloned receptor tyrosine kinase cDNA, which is characterized by seven immunoglobulin-like loops in its extracellular domain. We have previously mapped the FLT4 gene to chromosome segment 5q33-qter using somatic cell hybrids. Here we have refined the localization to band 5q35 by fluorescence in situ hybridization and show that the gene is translocated to chromosomes 2 and 6 in the t(2;5)(p23;q35) and t(5;6)(q35;p21) translocations, respectively, of Ki-I-positive lymphomas, as well as to chromosome 3 in the t(3;5)(q25.1;q34) translocation, which is occasionally found in myelodysplastic syndromes and acute myeloid leukemia. No evidence was obtained for a rearrangement or deregulation of the translocated FLT4 gene. We further show that abundant FLT4 mRNA expression occurs only in erythroid and megakaryoblastoid cell lines among nine leukemia cell lines studied.

Chromosome Mapping↗
PubMed1993

Interleukin-6 and TNF alpha production in human renal cell carcinoma.

Several clinical and biological features suggest that cytokines implicated in the inflammatory response are produced by renal cell carcinoma (RCC). To define if alterations of tumor necrosis factor alpha (TNF alpha), interleukin 1 alpha (IL-1 alpha), IL-1 beta and IL-6 gene expression are present in this malignancy, samples from 19 tumors as well as samples from seven paired normal renal tissue were examined using Northern blot and immunohistochemical analysis. In addition, the expression of these cytokines was evaluated in seven RCC-derived cell cultures using Northern blot or RT-PCR. TNF alpha and IL-6 proteins were measured in culture supernatants using specific bio- and immunoassays. Consistent levels of IL-6 mRNA were detected in 17 of the 19 tested tumors whereas TNF alpha specific transcripts were present in seven of eight available RNA samples. TNF alpha and IL-6 mRNA were also detected in five of the seven paired normal kidneys. By immunolabeling, IL-6 antigen was not detected in RCC cells in any of the 19 studied samples. In contrast, using anti-TNF alpha antibody a strong labeling of stromal endothelia and macrophage cells was detected in all the 19 cases, and evident TNF alpha staining of the carcinoma cells themselves was observed in eight cases. Spontaneous IL-6 mRNA expression was detected in five RCC cell cultures and TNF alpha mRNA in four. The cultured cells exhibited positive TNF alpha immunolabeling in six of seven cases but were always IL-6 negative. Bioactive IL-6 was detected in all culture supernatants while bioactive TNF alpha was not detected.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Renal Cell↗
PubMed1993

[Expression of IL6 and TNF-alpha in normal and pathological kidney].

The IL6 and TNF alpha specific mRNAs are expressed in normal human kidney; IL6 protein can be detected in normal glomerular mesangium by immunochemistry while TNF alpha is not present. Increased expression of IL6 mRNA is found within the glomeruli in mesangioproliferative glomerulonephritis and IL6 protein is detectable in proliferating mesangial areas. TNF alpha is mainly detected in infiltrating macrophages. During acute rejection episodes de novo expression of TNF alpha appears in renal transplant tubular epithelial cells as well as that of HLA class II antigens, ICAM-1 and VCAM-1 molecules. In renal cell carcinomas, the tumoral cells produce in vivo and in vitro IL6 and TNF alpha at the mRNA and protein levels. Therefore parenchymatous renal cells can produce IL6 and TNF alpha in various pathological conditions. However the mechanisms which regulate the production of these cytokines as well as their role in the genesis or the amplification of tissular damage remain to be elucidated.

Carcinoma, Renal Cell↗
PubMed1993

Modulation of tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6, interleukin-8, and granulocyte/macrophage colony-stimulating factor expression in human monocytes by an endogenous anxiogenic benzodiazepine ligand, triakontatetraneuropeptide: evidence for a role of prostaglandins.

Triakontatetraneuropeptide (TTN) is the major processing product of the endogenous anxiogenic peptide ligand of the benzodiazepine receptor, diazepam binding inhibitor. In the present study, we demonstrated by Northern blot analysis that the mRNA levels for tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, granulocyte/macrophage colony-stimulating factor, IL-6, and IL-8 were significantly increased after 4 hr of incubation of human monocytes with lipopolysaccharide (LPS) and TTN (10(-11) M), compared with cells incubated with LPS alone. Exposure of monocytes for 20 hr to LPS and TTN (10(-11) M) also stimulated TNF-alpha, IL-1 beta and granulocyte/macrophage colony-stimulating factor release by 80%, 110%, and 98%, respectively, relative to the response elicited by LPS alone. Smaller stimulatory effects were observed using the prototypic pharmacological peripheral benzodiazepine Ro5-4864 (10(-11) M) (55%, 72%, and 62%, assessed by means of specific enzyme immunoassays). In contrast, TTN and Ro5-4864 did not modulate LPS-induced IL-6 and IL-8 production. Treatment with the cyclooxygenase inhibitor indomethacin increased IL-1 beta and TNF-alpha secretion but not that of IL-6 or IL-8. The observed stimulatory effects of TTN and indomethacin were not additive. Taken together, these findings suggest a common mechanism of action for TTN and indomethacin, involving PG formation. In this respect, TTN inhibited prostaglandin (PG) E2 production by 30%. The fact that the observed modulatory effects correlated with PG levels suggests the existence of a second-messenger pathway associated with the peripheral-type benzodiazepine receptor. These results indicate that human TTN differentially modulates the LPS-induced expression of proinflammatory cytokines, and they further support the concept that this endogenous psychoactive peptide could be involved in physiological control of the inflammatory response.

Cells, Cultured↗
PubMed1992

Malignant histiocytosis in childhood: a distinctive CD30-positive clinicopathological entity associated with a chromosomal translocation involving 5q35.

The clinicopathological data on 20 cases of malignant histiocytosis (MH) collected over a period of 30 years at the Hôpital des Enfants Malades (Paris) are reported. Childhood MH was characterized by disseminated, frequently tender lymphadenopathy (19/20), skin (8/20), bone (6/20), and soft tissue localizations (7/20). These features were usually accompanied by fever, deterioration of general condition, and hematological abnormalities including anemia, thrombocytopenia, and occasionally fibrinopenia. These manifestations were clinically suggestive of a diagnosis of a severe neoplastic blood disease, although this hypothesis was not entertained for a long time because of the initial absence of abnormal cells in the blood and bone marrow. MH was characterized by the proliferation of large "histiocyte-like," usually mononucleated cells. When suitable material was available, MH cells appeared to react positively with acid-phosphatase, alpha-naphthyl acetate esterase (ANAE), alpha-antichymotrypsin, and antibodies directed against EMA, HLA DR, CD25, CD30, CD68, and CD71. No B- and T-cell antigens (except for one case) have been detected. Due to the frequent abundance of accompanying granulocytes, lymphoid, and plasma cells, and the presence of areas of necrosis, an initial correct diagnosis of MH was often difficult to establish on skin (four cases), bone (two), and soft tissue (three) biopsies. In lymph nodes, the sinusoidal and perifollicular topography of cell proliferation represented a highly reliable morphological feature. A permanent cell line (DEL) was obtained from a pleural effusion showing a t(5;6)(q35;p21) translocation and a monoallelic immunoglobulin (IgjH) rearrangement and consistent levels of expression of c-fms, c-myc, c-myb, c-ki-ras and c-fgr. Since an identical 5q35 breakpoint has been reported in four other MH cell lines with a comparable phenotype and in several isolated published cases, this chromosomal abnormality provides a highly valuable argument for individualizing an authentic malignancy of the mononuclear phagocyte system (MPS) in childhood, among the rather heterogeneous group of the CD30+ anaplastic large cell lymphomas.

Adolescent↗
PubMed1991

Human malignant histiocytosis CD30+ DEL cell line differentiates into macrophage-like cells when treated with a phorbol diester.

The histiocytic or lymphoid origin of human malignant histiocytosis is currently a subject of debate. The aim of this study was to investigate the in vitro effects of 12-O-tetradecanoylphorbol-13-acetate used as a differentiation inducer on the CD30, t(5;6)(q35;p21) DEL cell line, taken to be a reliable representative of the human malignant histiocytosis cell line. Treatment of DEL cells with 33 nM 12-O-tetradecanoylphorbol-13-acetate for 6-24 h resulted in cell surface attachment (up to 80%), decrease in dividing ability, enhancement of nitro blue tetrazolium reducing capacity (from 8 to 42%), occurrence of a limited immunodependent phagocytosis, and transient increase in expression of tumor necrosis factor alpha gene and in production of tumor necrosis factor alpha protein, whereas tumor necrosis factor beta remained undetectable. From these data, we can conclude that the malignant histiocytosis DEL cell line is not of lymphoid origin but stems from a myelomonocyte lineage.

Cell Differentiation↗
PubMed1990

Genotype markers and proto-oncogene analysis in the CD30-positive "malignant histiocytosis" DEL cell line with t(5;6)(q35;p21).

The DEL cell line isolated from a patient who died of malignant histiocytosis exhibits a reciprocal chromosomal translocation t(5;6)(5q35;6p21). The cells were analyzed for Ig(Jh), TCR beta-gene rearrangements and proto-oncogene expression pattern, using a panel of molecularly cloned probes that included c-fms, c-myc, c-myb, c-pim, c-fos, N-myc, c-sis, c-fgr as well as the virally derived probes v-ki-ras and v-src. Consistent levels of expression of c-fms, c-myc, c-myb, c-ki-ras and c-fgr were identified in cells from several in vitro passages as well as from the heterotransplanted tumors in nude mice. Transcripts homologous to the c-fos, c-src and c-sis were not observed. Southern blot study of DNA showed that the banding pattern of the screened proto-oncogenes was not altered. Furthermore, Southern blot analysis demonstrated monoallelic immunoglobulin heavy chain (IgJh) rearrangement but a normal germ-line configuration of the kappa light chain and TCR beta-genes. These results appear to imply that a T- or B-cell origin can be eliminated and that several activated proto-oncogenes, usually expressed in immature MPS cells (c-fms) and myeloblastic cells (c-fgr), may be implicated in the proliferative activity of the DEL cell line, the stem of which may be a primitive, ancestral myelomonocytic cell.

Antigens, CD↗
PubMed1990

DEL cell line: a "malignant histiocytosis" CD30+ t(5;6)(q35;p21) cell line.

A new cell line DEL, established in vitro, was isolated from a pleural effusion of a boy who died of malignant histiocytosis. Its principal characteristics are: strong positivity with monoclonal antibodies (MAbs) to CD25, CD30, CD45R, KiM7, EMA, HLA Cl I and II; constant presence of acid phosphatase, ANAE, alpha-anti-trypsin, alpha-anti-chymotrypsin and NBT reductase activity; rearrangement of the immunoglobulin heavy-chain gene (JH) and a germ-line configuration of the T-chain gene; and finally a translocation between chromosomes 5-6 with a breakpoint in 5q35. The DEL cell line is appropriate for studying the role of the 5q localized c-fms oncogene and of the genes of the mononuclear phagocyte growth factor (CSFI) and of their receptors in the dynamics and etiology of malignant hemopathies associated with a 5q35 breakpoint.

Antigens, Neoplasm↗
PubMed1990

Expression of the human nephron differentiation molecules in renal cell carcinomas.

The authors tested frozen sections from 28 renal cell carcinomas (RCC)--21 clear, 1 eosinophilic, 4 basophilic, and 2 spindle-shaped cell type--with monoclonal antibodies (MAb) reacting against cytokeratin, vimentin, CD24, CALLA/CD10, villin, CD26, and HLA class I and class II molecules. These molecules are markers of specific segments of the mature kidney, and their loss or acquisition reflects the different steps of human nephrogenesis. KI67 MAb was used to evaluate cell-proliferating activity. All RCC cases expressed cytokeratin. Coexpression of vimentin was observed in 21 of 28 cases. Whether of clear or chromophilic type, all tumoral cells strongly expressed CD24 molecule, present on primitive blastema cells. All clear-type RCCs expressed CALLA/CD10 and 60% were also villin positive; some were faintly positive for CD26. CALLA, villin, and CD26 were not detected in basophilic cell type. HLA class I molecules were variably expressed in almost all cases, but HLA class II were never detected on tumoral cells. Except for the spindle-shaped population, cell-proliferating activity was low. These results favor the hypothesis that RCCs derive from cells that have 'recovered' the different options of metanephric differentiation. Clear cells show evidence of maturation toward proximal type, while basophilic cells do not. It would be of interest to evaluate the usefulness of serum measurements of villin and/or CALLA as markers in clear cell-type RCC.

Aged↗
PubMed1989

Major histocompatibility complex class II (Ia-like) antigens on chicken embryo fibroblasts: effect of transformation by Rous sarcoma virus.

In this study, three monomorphic monoclonal antibodies to chicken MHC class II molecules (B-L) were tested for reactivity in normal and RSV-transformed embryo fibroblasts. The immunocytochemical staining, the cell-bound ELISA assay, and the immunoprecipitation analysis showed that all three antibodies reacted with the B-L (Ia-like) molecules on normal cells of different genotypes. Conversely, the expression of these antigens was not detected in fibroblasts cultured from feather follicles of adult birds. The level of expression of B-L molecules as well as the class II specific RNA increased consistently after transformation of the cells by the SR-RSV and infection with the avian leukosis virus RAV-1. Analysis of genomic DNA by the Southern blot technique, performed after digestion with several restriction endonucleases, showed that the restriction pattern of B-L genes was not altered in cells transformed by Rous sarcoma virus.

Animals↗