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1.
Cell Biol Toxicol ; 24(1): 55-62, 2008 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-17610031

RESUMEN

The heavy metal cadmium, an environmental pollutant, has been widely demonstrated to be toxic, in particular for liver. In murines, cadmium induces apoptosis of hepatocytes and hepatomas. In human cells, apoptosis induced by cadmium has been exclusively demonstrated in tumoral cell lines. Nothing was known in normal liver, in vitro or in vivo. In the present study, we examined the effects of cadmium in nonmalignant human hepatocytes. For that purpose, we investigated whether cadmium was able to induce apoptosis of normal human hepatocytes (NHH) in primary culture and of a SV40-immortalized human hepatocyte (IHH) cell line. Treatment of IHH and NHH with cadmium induced the presence of a sub-G(1) population at 10 and 100 micromol/L, respectively. DAPI staining of both cell types treated with cadmium 100 micromol/L revealed the induction of nuclear apoptotic bodies, supporting the hypothesis of apoptosis. In IHH and NHH, cadmium 100 micromol/L induced PARP cleavage into a 85 kDa fragment. In order to investigate the involvement of mitochondria in cadmium-induced apoptosis, we measured the mitochondrial membrane potential (Delta(Psim)). We observed that in IHH and NHH, cadmium 100 micromol/L induced a decrease of Delta(Psim). As expected, cadmium under the same conditions enhanced caspase-9 and caspase-3 activities. In addition, cadmium from 1 to 100 micromol/L induced the expression of p53 and phosphorylation of its Ser15 in IHH and NHH. In conclusion, we showed in this study that human hepatocytes were sensitive to cadmium and apoptosis induced at concentrations suggested in the literature to inhibit p53 DNA-binding and DNA repair.


Asunto(s)
Apoptosis/efectos de los fármacos , Cadmio/toxicidad , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Línea Celular Transformada , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Activación Enzimática/efectos de los fármacos , Fase G1/efectos de los fármacos , Hepatocitos/enzimología , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/enzimología , Fosfoserina/metabolismo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Proteína p53 Supresora de Tumor/metabolismo
2.
Diabetes Metab ; 31(1): 47-54, 2005 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-15803113

RESUMEN

OBJECTIVES: In type 2 diabetic patients with no cardiac history or symptoms, 1) to evaluate whether the soluble forms of Fas (sFas) and Fas-ligand (sFasL), involved in apoptosis, may be markers of silent coronary disease or related to hypertension or microangiopathic complications; 2) to examine the effect of short-term glycemic control on sFas and sFasL. METHODS: (1) sFas and sFasL were measured with the ELISA method in 44 asymptomatic diabetic patients, 33 with hypertension, and with a normal myocardial scintigraphy (n=14), with silent myocardial ischemia (SMI) and without (n=15) or with (n=15) significant coronary stenoses; and in 14 controls; (2) sFas and sFasL were measured in 15 poorly controlled diabetic patients before and after 7 days of CSII treatment. RESULTS: (1) sFas and sFasL differed in the four groups of patients (p=0.003 each). sFas was significantly higher in the patients with SMI without (p=0.035) and with coronary stenoses (p=0.002) than in the control group. sFasL was lower in the three groups of diabetic patients (p<0.05 each) than in control group. In the diabetic population, sFas correlated positively with hypertension (p=0.021), and sFasL negatively with hypertension (p=0.027) and HOMA index in the non-insulin treated patients (p=0.049); (2) sFas did not differ before or after CSII, and there was a marginal decrease in sFasL. CONCLUSION: Fas-mediated apoptosis is involved in type 2 diabetes and might be associated with hypertension and/or its vascular consequences. sFasL might be affected by insulin resistance. sFas and sFasL are not effective markers of SMI.


Asunto(s)
Complicaciones de la Diabetes/inmunología , Diabetes Mellitus Tipo 2/inmunología , Hipertensión/inmunología , Resistencia a la Insulina/inmunología , Glicoproteínas de Membrana/fisiología , Receptor fas/fisiología , Adulto , Glucemia/metabolismo , Presión Sanguínea , Índice de Masa Corporal , Enfermedad Coronaria/sangre , Enfermedad Coronaria/inmunología , Complicaciones de la Diabetes/sangre , Proteína Ligando Fas , Femenino , Humanos , Lípidos/sangre , Masculino , Glicoproteínas de Membrana/sangre , Pulso Arterial , Receptor fas/sangre
3.
J Bone Miner Res ; 5(5): 425-35, 1990 May.
Artículo en Inglés | MEDLINE | ID: mdl-1695060

RESUMEN

We report the presence of reverse transcriptase activity in the supernatant of long-term culture of mononuclear blood cells (monocytes and lymphocytes) isolated from a 27-year-old patient suffering from benign osteopetrosis. The enzyme was purified to homogeneity according to the technique of Chandra and Steel, by chromatography, first on DEAE-cellulose (DE 52) and then on phosphocellulose (P11). After purification, the enzyme was characterized biochemically for its template specificity and ionic requirements. The purified enzyme was able to transcribe poly(rA).(dT)12-18 and poly(rC).(dG)12-18 very efficiently and had a marked preference for Mg2+ ions over Mn2+ ions. The pattern of ionic dependency for this enzyme is similar to that of reverse transcriptases purified from human lymphotropic viruses. The patient was tested and found sero-negative for HIV-1, HIV-2, and HTLV-I and seropositive (immunoglobulin G) for cytomegalovirus. Epstein-Barr virus nuclear antigens (EBNA) were detected in the patient's B lymphocytes. Since reverse transcriptase is the hallmark of retroviruses, we suggest that a retrovirus may be involved in the etiology of osteopetrosis.


Asunto(s)
Leucocitos Mononucleares/microbiología , Osteopetrosis/microbiología , Retroviridae/aislamiento & purificación , Adulto , Cromatografía , Femenino , Humanos , Leucocitos Mononucleares/enzimología , Osteopetrosis/enzimología , Osteopetrosis/etiología , Polinucleótidos , ADN Polimerasa Dirigida por ARN/sangre , Retroviridae/enzimología , Retroviridae/patogenicidad , Especificidad por Sustrato
4.
Invest Ophthalmol Vis Sci ; 40(3): 619-30, 1999 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-10067965

RESUMEN

PURPOSE: The aim of this study was to investigate the action of benzalkonium chloride (BAC), used as a preservative in most ophthalmic topical solutions, on epithelial conjunctival cells in vitro. METHODS: A continuous human conjunctival cell line (Wong-Kilbourne derivative of Chang conjunctiva) was exposed to BAC solutions at various concentrations (0.1%-0.0001%) during a period of 10 minutes. Cells were examined before treatment and 3, 24, 48, and 72 hours later, after reexposure to normal cell culture conditions. Cell number and viability were assessed with crystal violet and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide colorimetric assays. The expression of the apoptotic marker Apo 2.7, nuclear antigen p53, membrane proteins Fas and Fas ligand, and DNA content was studied by flow cytometry. Morphologic aspects of cell nuclei were analyzed on slides with a nucleic acid-specific dye, 4',6'-diamidino-2-phenylindole dihydrochloride. Cytoskeleton was labeled with a monoclonal anti-pancytokeratin antibody. In addition, apoptosis was measured by DNA electrophoresis assays in agarose gel. RESULTS: Cell exposure to 0.1% and 0.05% BAC induced cell lysis immediately after treatment. All cells (100%) treated with 0.01% BAC died in a delayed manner within 24 hours, with most of the characteristics of apoptosis (chromatin condensation and DNA fragmentation, reduction in cell volume, expression of the apoptotic marker Apo 2.7, and apoptotic changes in DNA content). Aliquots of 0.005%, 0.001%, 0.0005%, and 0.0001% BAC induced growth arrest and apoptotic cell death in a dose-dependent manner between 24 and 72 hours after treatment. The expressions of Fas and p53 did not vary after BAC treatment. Fas ligand was always negative. CONCLUSIONS: These results suggest that BAC induces cell growth arrest and death at a concentration as low as 0.0001%. The mode of BAC-induced cell death is dose-dependent. Cells die by necrosis after BAC treatment at high concentrations and by apoptosis if low concentrations of BAC are applied. This new aspect of in vitro toxicity of BAC could in part explain some ocular surface disorders observed in patients undergoing long-term topical treatments with preservative-containing drugs.


Asunto(s)
Compuestos de Benzalconio/farmacología , Conjuntiva/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Conservadores Farmacéuticos/farmacología , Apoptosis/efectos de los fármacos , Recuento de Células , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Conjuntiva/citología , Conjuntiva/metabolismo , ADN/análisis , Relación Dosis-Respuesta a Droga , Células Epiteliales/citología , Células Epiteliales/metabolismo , Proteína Ligando Fas , Citometría de Flujo , Humanos , Glicoproteínas de Membrana/metabolismo , Necrosis , Proteína p53 Supresora de Tumor/metabolismo , Receptor fas/metabolismo
5.
Hum Pathol ; 32(11): 1190-6, 2001 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-11727257

RESUMEN

Alterations of CD44 proteins, a family of cell adhesion molecule, have been linked with tumorigenesis, carcinogenesis, and prognosis in various neoplasms. Our aims were to evaluate and compare CD44 isoforms expression patterns in normal myometrium, uterine leiomyomas, and leiomyosarcomas and to correlate CD44 expression with clinicopathologic parameters. Fresh (n = 15) and formalin-fixed, paraffin-embedded (n = 76) tissues samples of myometrium, leiomyomas, and leiomyosarcomas were used for immunoblotting and immunohistochemistry, respectively. Semiquantitative evaluation was made after immunostaining. Monoclonal antibodies were used. By immunoblotting in myometrium and leiomyomas samples, we observed a band at 85 kd, corresponding to the apparent molecular weight of CD44s, and bands at 140 kd with the monoclonal antibodies against CD44v3 and CD44v6. In leiomyosarcomas, CD44s and CD44v6 were detected, but not CD44v3. By immunohistochemistry, decreased CD44s expression was found in leiomyomas and leiomyosarcomas (73.9% +/- 16.6% and 82.1% +/- 20.7%, respectively) compared with myometrium (97.3% +/- 6.2%; P < .0001). No CD44v6 staining was detected in myometrium, leiomyomas, and leiomyosarcomas. No CD44v3 expression was detected in leiomyosarcomas, whereas myometrium and leiomyomas expressed CD44v3. For the diagnosis of leiomyosarcoma, the absence of CD44v3 staining had a sensitivity, specificity, and positive and negative predictive values of 100%. In patients with recurrence of leiomyosarcomas, CD44s expression was decreased (P = .03). We conclude that CD44s immunostaining in leiomyosarcomas may have prognostic significance. The loss of CD44v3 expression could be used as a putative diagnostic tool for uterine leiomyosarcomas.


Asunto(s)
Glicoproteínas/metabolismo , Receptores de Hialuranos/metabolismo , Leiomioma/metabolismo , Leiomiosarcoma/metabolismo , Neoplasias Uterinas/metabolismo , Adulto , Anciano , Western Blotting , Femenino , Glicoproteínas/inmunología , Humanos , Receptores de Hialuranos/inmunología , Técnicas para Inmunoenzimas , Antígeno Ki-67/inmunología , Antígeno Ki-67/metabolismo , Leiomioma/diagnóstico , Leiomioma/patología , Leiomiosarcoma/diagnóstico , Leiomiosarcoma/patología , Masculino , Persona de Mediana Edad , Miometrio/metabolismo , Pronóstico , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/patología
6.
Diabetes Metab ; 27(3): 315-21, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11431596

RESUMEN

OBJECTIVE: The aim of this study was to investigate circulating soluble Fas (sFas) and Fas ligand (sFasL), two transmembrane glycoproteins involved in apoptosis, in the serum of diabetic patients. MATERIAL AND METHODS: We assessed sFas and sFasL serum levels in normal controls (n=15), and in both 42 diabetic patients without complications, or with predominant retinopathy or neuropathy, using sFas and sFasL specific ELISA method. RESULTS: sFasL serum levels were less than 0.1 ng/ml in normal controls and in each group of diabetic patients. In diabetic patients with a predominant neuropathy, sFas serum levels were significantly increased not only when compared with normal controls (13.5 +/- 3.6 ng/ml vs 7.1 +/- 1.1 ng/ml, p<0.001), but also when compared with patients without complications (vs 9.1 +/- 1.8 ng/ml, p<0.001) or with a predominant retinopathy (vs 8.7 +/- 1.9 ng/ml, p<0.001). CONCLUSIONS: These preliminary data suggest that a dysregulation of the Fas system in peripheral neuronal cells may be involved in the increase of sFas observed in diabetic patients with neuropathy.


Asunto(s)
Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 2/sangre , Neuropatías Diabéticas/sangre , Retinopatía Diabética/sangre , Receptor fas/sangre , Adulto , Anciano , Albuminuria , Biomarcadores/sangre , Glucemia/metabolismo , Creatinina/sangre , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 2/inmunología , Neuropatías Diabéticas/inmunología , Retinopatía Diabética/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valores de Referencia
7.
Clin Chim Acta ; 133(2): 215-22, 1983 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-6627683

RESUMEN

Affinity chromatography purification was combined with a radioimmunoassay for 'Tamm-Horsfall-like' glycoprotein. This enabled serum concentrations to be established and to demonstrate its presence in cerebrospinal fluid for the first time. This assay method used in different circumstances suggests a multifocal synthesis. Nevertheless, urinary Tamm-Horsfall glycoprotein so far must be distinguished from the serum or cerebrospinal fluid Tamm-Horsfall-like glycoprotein.


Asunto(s)
Mucoproteínas/análisis , Radioinmunoensayo , Adulto , Cromatografía de Afinidad , Humanos , Mucoproteínas/sangre , Mucoproteínas/líquido cefalorraquídeo , Enfermedades del Sistema Nervioso/metabolismo , Valores de Referencia , Uromodulina
8.
Biomed Pharmacother ; 48(2): 103-11, 1994.
Artículo en Inglés | MEDLINE | ID: mdl-7919103

RESUMEN

We report here the spontaneous in vitro transformation of blood monocytes into fibroblasts in a patient who developed pulmonary fibrosis following ciclosporin-mediated immunosuppression, necessitated by heart transplantation. The blood monocytes with this capacity expressed HLA-DR specificity. Monocytes/macrophages were identified by immunofluorescence using monoclonal antibodies against a specific monocyte/macrophage antigen, while the neo-fibroblasts were identified by electron microscopy and immunofluorescence using monoclonal antibodies against a cytoplasmic enzyme specifically involved in the synthesis of collagen. The secretion of collagen was demonstrated using antibodies against collagen. Both the monocytes/macrophages and the neo-fibroblasts express macrophage and fibroblast markers and are able to synthesize collagen. The all-trans retinoic acid derivative (all-trans RA) inhibits this in vitro transformation of HLA-DR monocytes/macrophages into neo-fibroblasts. Therefore, the possible therapeutic role of all-trans RA in controlling the development of fibrosis remains open to investigation. Until now, no efficient therapy is known for fibrotic diseases which are often lethal when affecting the lungs.


Asunto(s)
Antígenos HLA-DR/inmunología , Monocitos/patología , Fibrosis Pulmonar/patología , Tretinoina/farmacología , Anciano , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/patología , Células Cultivadas , Ciclosporina/efectos adversos , Fibroblastos/patología , Humanos , Huésped Inmunocomprometido , Técnicas In Vitro , Masculino , Monocitos/inmunología , Fibrosis Pulmonar/inmunología
9.
Biomed Pharmacother ; 46(2-3): 91-108, 1992.
Artículo en Inglés | MEDLINE | ID: mdl-1384752

RESUMEN

Previous studies led us to demonstrate in pathological situations that the fibroblast, not the macrophage, was the terminal maturation step of the HLA-DR monocyte and that the entire process came under T-lymphocyte control. Fibrosis which developed under immunosuppressive treatment (cyclosporin) after organ transplantation is an illustration of these in vitro observations. The present in vitro study was undertaken in order to investigate whether or not this transformation process takes place under physiological conditions and if so, the nature of the T-lymphocyte control. We report that normal HLA-DR monocytes/macrophages are able to secrete type 1 collagen and to differentiate into neo-fibroblasts. However, contrarily to what happened in pathology, only a few neo-fibroblasts developed transiently. The addition of conditioned medium (CM) from activated T-lymphocytes greatly enhanced the transformation process. Counteracting this CM effect, cell-to-cell contact between neo-fibroblasts and T-cells resulted in the loss of fibroblastic shape. The 'end-result' macrophage engulfed numerous lymphocytes giving rise to a multinucleated cell. This giant cell no longer adhered to the slide and died. The question is raised as to whether the process observed in vitro is involved in vivo in tissue repair. We also report that HLA-DR monocytes and the neo-fibroblasts which derive from them are able to secrete, in addition to type 1 collagen, a variety of proteins such as uromodulin, amyloid-beta peptide, alpha-fetoprotein and carcinoembryonic antigen. In cystic fibrosis we previously reported a high level of uromodulin production by HLA-DR monocytes differentiating towards the fibroblastic phenotype. Pathologies characterized by excessive production of either alpha-feto-protein, carcinoembryonic antigen, beta-amyloid protein (Alzheimer's disease) should be investigated, taking into account the involvement of HLA-DR monocytes and their possible uncontrolled differentiation into neo-fibroblasts.


Asunto(s)
Antígenos HLA-DR/análisis , Monocitos/inmunología , Linfocitos T/inmunología , Péptidos beta-Amiloides/metabolismo , Antígeno Carcinoembrionario/inmunología , Diferenciación Celular , Fibroblastos/citología , Humanos , Monocitos/citología , Monocitos/metabolismo , Mucoproteínas/metabolismo , Uromodulina , alfa-Fetoproteínas/metabolismo
10.
Biomed Pharmacother ; 51(2): 79-93, 1997.
Artículo en Inglés | MEDLINE | ID: mdl-9161472

RESUMEN

Nodules and multilayered areas composed of fibroblasts and chondrocyte-like cells embedded in an abundant extracellular matrix appeared spontaneously in in vitro culture of mononucleated blood cells taken from a patient with chondrosarcoma. Using specific antibodies it was demonstrated that the neo-fibroblasts which developed in the culture resulted from a direct transdifferentiation of monocytes expressing HLA-DR specificity. The experiment was carried out twice, once before surgery and then two years later. In both cases the spontaneous transdifferentiation of HLA-DR monocytes into neo-fibroblasts was observed. Previously it was shown that normal monocytes were also able to give rise in vitro to neo-fibroblasts. However, the latter are normally rapidly destroyed by cell-cell contact with T-cells. Normal T-cells adhere to normal neo-fibroblasts by which they are finally engulfed. As a result, the neo-fibroblasts lose their fibroblastic shape, no longer adhere to their support and die. Therefore the abnormal proliferation and persistence of neo-fibroblasts in pathological situations such as the present case may result either from an intrinsic defect in monocytes, T-cells or both. The question is whether or not this transdifferentiation process observed in vitro accounts for the development of chondrosarcoma in vivo. The present results suggest that in vivo chondrosarcoma may start in a necrotic zone (resulting for instance from trauma) and attract HLA-DR monocytes, where they accumulate and transdifferentiate into neo-fibroblasts and chondrocyte-like cells. The uncontrolled transdifferentiation of these HLA-DR monocytes resulting from a dysregulation of the immune system is probably linked to the malignant process which may have a retroviral origin. The question is raised regarding the embryologic origin of this special sub-population of blood monocytes in which pluripotential capabilities are retained; its origin may differ from that of the other circulating monocytes.


Asunto(s)
Cartílago/patología , Condrosarcoma/sangre , Fibroblastos/patología , Antígenos HLA-DR/inmunología , Monocitos/patología , Diferenciación Celular , Condrosarcoma/metabolismo , Femenino , Fibroblastos/inmunología , Fibroblastos/metabolismo , Técnica del Anticuerpo Fluorescente Indirecta , Histocitoquímica , Humanos , Técnicas In Vitro , Microscopía Electrónica , Persona de Mediana Edad , Monocitos/inmunología , Monocitos/metabolismo
11.
Biomed Pharmacother ; 45(9): 387-401, 1991.
Artículo en Inglés | MEDLINE | ID: mdl-1786344

RESUMEN

We report here the spontaneous in vitro transformation of blood monocytes into fibroblasts, in a patient suffering from cystic fibrosis (CF). The blood monocytes with this capacity express HLA-DR specificity. Monocytes were identified by non-specific esterase activity and by immunofluorescence using monoclonal antibodies against monocytes/macrophages antigens. Neo-fibroblasts were identified by electron microscopy and immunofluorescence using monoclonal antibodies against a cytoplasmic enzyme specifically involved in the synthesis of collagen. The secretion of collagen was evidenced using antibodies against type I collagen. Both monocytes/macrophages and neo-fibroblasts express the monocytic and the fibroblastic markers and synthesize type I collagen. This transformation observed in vitro might mimick the process of fibrosis development which takes place in vivo, particularly in pancreatic acini, lungs and intestine of patients with CF. Interestingly, the whole process in vitro is inhibited when T lymphocytes are properly stimulated by IL2. In addition, both monocytes and neo-fibroblasts secrete high quantities of uromodulin-like glycoprotein. The significance of this finding is discussed in relation to the thick mucus secretion which characterizes the disease. In addition, from a fundamental point of view, it confirmed in a large series of patients that this observation may have significant implications, since CF mutation impairs the gene coding for cAMP-regulated Cl- channel and that it has been proposed that uromodulin might be implicated in Cl- transport. Therefore the question of the relationships between uromodulin and the cAMP-regulated Cl- channel arises.


Asunto(s)
Fibrosis Quística/sangre , Antígenos HLA-DR/análisis , Monocitos/patología , Mucoproteínas/sangre , Adolescente , Línea Celular Transformada , Fibroblastos/inmunología , Fibroblastos/patología , Técnica del Anticuerpo Fluorescente , Humanos , Técnicas In Vitro , Masculino , Microscopía Electrónica , Monocitos/inmunología , Fenotipo , Uromodulina
12.
Biomed Pharmacother ; 45(7): 289-99, 1991.
Artículo en Inglés | MEDLINE | ID: mdl-1760520

RESUMEN

We describe here two pathological situations, osteomyelosclerosis and Engelmann's disease, in which HLA-DR blood monocytes modulate to the fibroblastic class, in long-term culture. Monocytes/macrophages were identified by immunofluorescence, using monoclonal antibodies against surface markers (Leu M3, CD 68, and HLA-DR) and the neo-fibroblasts by electron microscopy and immunofluorescence using monoclonal antibodies against a cytoplasmic enzyme specifically involved in the synthesis of collagen (5B5). Macrophages makers were found on the neo-fibroblasts, whereas HLA-DR macrophages expressed the cytoplasmic marker 5B5. Since osteoblasts are classically derived from fibroblasts, the significance of the in vitro differentiation of monocytes/macrophages into fibroblasts to the in vivo mechanism leading to excessive osteoblastic proliferation in both osteomyelosclerosis and Engelmann's disease, is discussed. The possible involvement of this pathway leading from monocytes to fibroblasts and osteoblasts in the normal process of bone modeling and remodeling in questioned.


Asunto(s)
Síndrome de Camurati-Engelmann/patología , Monocitos/patología , Mielofibrosis Primaria/patología , Células Cultivadas , Femenino , Fibroblastos/patología , Fibroblastos/ultraestructura , Humanos , Macrófagos/patología , Masculino , Microscopía Electrónica , Microscopía Fluorescente , Persona de Mediana Edad , Bazo/patología
14.
C R Seances Soc Biol Fil ; 179(1): 27-40, 1985.
Artículo en Francés | MEDLINE | ID: mdl-2411366

RESUMEN

A comparative analysis of urine and serum THG is carried out using physico-chemical and radioimmunological methods. It demonstrates a real similarity in the antigenicity, the glycan structure and the polymerization-depolymerization. The presence of THG in various biological fluids and tissues is an extra argument in favor of such an identity. A multifocal synthesis of THG and not an exclusively renal one is admitted. Such a conception seems to be more consistent with the membrane function of THG towards some ions in various tissues.


Asunto(s)
Mucoproteínas/inmunología , Cromatografía de Afinidad , Cromatografía en Gel , Epítopos , Humanos , Lectinas/farmacología , Cloruro de Mercurio/farmacología , Mucoproteínas/sangre , Mucoproteínas/orina , Radioinmunoensayo , Temperatura , Uromodulina
15.
Clin Orthop Relat Res ; (313): 239-52, 1995 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-7543835

RESUMEN

Reverse transcriptase activity is reported in the mononuclear blood cells isolated from a patient in whom paraarticular ossification developed after surgery for an aneurysmal bone cyst. The enzyme was purified to apparent homogeneity by chromatography before being characterized biochemically for its template specificity and ionic requirement. The enzyme was able to transcribe poly(rA).(dT)12-18 very efficiently in the presence of Mn++ ions. Viral particles were observed in the HuT-78 cell line, cocultured with the mononuclear cells of the patient. No viral particles were observed in HuT-78 cells before the coculture. The patient was found seronegative for HIV-1, HIV-2, and HTLV-1. These results suggest that a new retrovirus infecting mononuclear blood cells may be involved in the development of ectopic ossification. This hypothesis is strengthened by the previous finding of a retrovirus in the mononuclear blood cells of a patient with benign osteopetrosis, and by the fact that HTLV-1 infected T-lymphocytes acquire the ability to secrete factors responsible for the lytic bone lesions observed in the patients. A family of human bone diseases that reflect T-cell dysfunction(s) and are caused by lymphotropic viruses may exist.


Asunto(s)
Leucocitos Mononucleares/virología , Osificación Heterotópica/virología , Complicaciones Posoperatorias/virología , ADN Polimerasa Dirigida por ARN/sangre , Infecciones por Retroviridae/diagnóstico , Infecciones Tumorales por Virus/diagnóstico , Quistes Óseos Aneurismáticos/cirugía , Articulación del Codo , Cabeza Femoral/cirugía , Articulación de la Cadera , Humanos , ADN Polimerasa Dirigida por ARN/aislamiento & purificación , Infecciones por Retroviridae/virología , Infecciones Tumorales por Virus/virología , Virión/aislamiento & purificación
16.
Cell Mol Biol (Noisy-le-grand) ; 46(1): 13-28, 2000 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-10726968

RESUMEN

Fas-induced apoptosis is one form of programmed cell death responsible for hepatocyte demise. However, the role of this cell surface receptor in the death of tumoral hepatic cells is still being debated. It has been shown that some hepatoma cell lines may escape apoptosis because of abnormal Fas localization correlated with non-functionality of the Fas protein or dysfunctionality in the Fas pathway cascade. The aim of this study was to investigate the behaviour of four hepatoma cell lines, HepG2, Hep3B, SKHep1 and Chang-Liver and two extrahepatic cell lines, MCF7, a mammary tumoral cell line and OVCAR-3, an ovarian tumoral cell line, when they were treated with an agonistic anti-Fas antibody alone, with interferon gamma (IFNgamma), an up-regulator of Fas protein expression, alone or with a combination of both agents. We first performed immunofluorescence and flow cytometry to confirm that Fas was present on the cell surface of each cell line in the normal state. Apoptosis was then investigated after induction with the various treatments, by DAPI staining, agarose gel DNA electrophoresis and PARP cleavage. Caspase 8 and 3 expression, as well as two anti-apoptotic proteins Bcl-2 and HSP70, and one proapoptotic protein Bax were also investigated by immunoblot allowing identification of several apoptotic pathways based on the behaviour of the different studied proteins. HepG2 and OVCAR-3 cells were sensitive to the anti-Fas antibody alone. Hep3B was resistant to Fas-induced apoptosis but sensitive to IFNgamma-induced apoptosis. MCF7 was resistant to anti-Fas antibody and IFNgamma Chang-Liver and SKHep1 were sensitive to IFNgamma and anti-Fas antibody but at different degrees. Chang-Liver used the Fas and IFNgamma pathways, while SKHep1 involved mostly the Fas pathway. These results show that each tumor cell line is characterized by different apoptotic behaviour in relation to Fas and/or IFNgamma-induced apoptosis. In addition, despite the high level of Bcl-2 and HSP70 proteins in the tumoral cells investigated here, they were not fully protected against apoptosis, except for MCF7. This emphasizes the necessity to analyse the different proteins responsible for apoptosis to adapt anti-tumoral therapeutics.


Asunto(s)
Apoptosis , Proteínas Proto-Oncogénicas c-bcl-2 , Receptor fas/metabolismo , Anticuerpos Monoclonales/farmacología , Caspasas/metabolismo , Cromatina/metabolismo , Fragmentación del ADN/efectos de los fármacos , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Regulación Neoplásica de la Expresión Génica , Humanos , Interferón gamma/farmacología , Hígado/metabolismo , Poli Adenosina Difosfato Ribosa/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Transducción de Señal , Células Tumorales Cultivadas , Proteína X Asociada a bcl-2
17.
Eur Neurol ; 23(2): 65-72, 1984.
Artículo en Inglés | MEDLINE | ID: mdl-6723716

RESUMEN

A protein antigenically similar to Tamm-Horsfall glycoprotein ( THLG ) was determined in CSF by affinity chromatography followed by a radioimmunological assay. THLG seems not to be a constituent of normal CSF or an autoantigen. It probably originates from nervous tissue and not from plasma proteins. Its mean is higher in the CSF of multiple sclerosis patients than in the CSF of degenerative disorders patients. THLG seems to be a chemical component of neuronal membrane and its physiological functions remain unknown.


Asunto(s)
Enfermedades Desmielinizantes/líquido cefalorraquídeo , Mucoproteínas/líquido cefalorraquídeo , Barrera Hematoencefálica , Enfermedades del Sistema Nervioso Central/patología , Enfermedades Desmielinizantes/inmunología , Femenino , Humanos , Leucocitos/patología , Masculino , Mucoproteínas/sangre , Esclerosis Múltiple/líquido cefalorraquídeo , Esclerosis Múltiple/inmunología , Necrosis/metabolismo , Uromodulina
18.
Cell Biol Toxicol ; 16(3): 185-200, 2000.
Artículo en Inglés | MEDLINE | ID: mdl-11032362

RESUMEN

The in vitro occurrence of apoptosis in hepatic cells has not been well characterized because it depends on apoptosis inducing-agents and culture conditions. Furthermore, for a given hepatic cell and the same agent, discrepant results have been reported depending on the technique used to evaluate the proportion of apoptotic cells. In this study, we compared the effects of several apoptosis-inducing agents - transforming growth factor beta1 (TGF-beta1), retinoic acid (RA), okadaic acid (OA), and cycloheximide (CY) - on two types of hepatic cells, the human hepatoma cell line Hep3B and normal rat hepatocytes, maintained either plated for 24 to 48 h or in suspension for 20 h. Chromatin condensation and/or nucleus fragmentation were investigated morphologically by DAPI staining. DNA fragmentation was investigated biochemically by agarose gel electrophoresis and poly(ADP-ribose) polymerase (PARP) cleavage was studied by western blot. Apoptotic cells were quantified either by counting cells on UV microscopy after DAPI staining or by flow cytometry. Nuclear changes, the ladder pattern on DNA electrophoresis and PARP cleavage were observed in plated cells, hepatoma cells and normal rat hepatocytes, with all inducers but especially with OA. Semiquantification confirmed that OA was a strong inducer in plated cells under the present conditions, since about 14% and 30% of Hep3B cells (with DAPI staining and flow cytometry, respectively) were apoptotic after 48 h treatment, while, with the other inducers, apoptosis was weaker and discrepancies were also observed between the two counting methods (TGF-beta1; 4% and 12%; RA, 7% and 12%; CY, 4% and 16%, with DAPI staining and flow cytometry, respectively). OA induced a moderate apoptosis in cultured hepatocytes (13% with DAPI staining), while TGF-beta1, RA and CY were found to be weakly apoptotic (respectively 4% for the first two and 6% for the last ) after 48 h. In contrast, in suspension cells, apoptosis was observed neither in Hep3B cells nor in normal hepatocytes, whatever the apoptotic inducer and whatever the techniques used to detect apoptosis. In conclusion, our results show that induction of apoptosis in hepatic cells depends not only on the apoptosis-inducing agent but also on the culture conditions.


Asunto(s)
Apoptosis/fisiología , Carcinoma Hepatocelular , Hepatocitos/citología , Neoplasias Hepáticas , Animales , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Carcinógenos/farmacología , Técnicas de Cultivo de Célula/métodos , Cicloheximida/farmacología , Colorantes Fluorescentes , Hepatocitos/metabolismo , Humanos , Indoles , Ácido Ocadaico/farmacología , Poli(ADP-Ribosa) Polimerasas/metabolismo , Inhibidores de la Síntesis de la Proteína/farmacología , Ratas , Factor de Crecimiento Transformador beta/farmacología , Factor de Crecimiento Transformador beta1 , Tretinoina/farmacología , Células Tumorales Cultivadas
19.
Cell Mol Biol (Noisy-le-grand) ; 46(1): 187-97, 2000 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-10726984

RESUMEN

We have analyzed the type of cell death occurring in human normal ejaculated spermatozoa. Sperm cells were prepared either by centrifugation alone (group 1) or by density gradient centrifugation (group 2) and were cultured for 24 hrs. Cells were examined after 4 and 24 hrs. By comparison unprepared spermatozoa were used as a control group. Necrosis was investigated by intra-cellular vital stain penetration and electron microscopy. Apoptosis was researched by DAPI staining, annexin V-binding, electron microscopy, DNA fragmentation and PARP cleavage. In group 1, after 4 hrs., there was a mixture of spermatozoa dead either by necrosis or apoptosis while after 24 hrs., necrosis was prominent. Similar findings were observed in the control group. In contrast, in group 2 apoptosis was the major form of cell death of spermatozoa after 24 hrs. of culture. These findings suggest that apoptosis can be an important factor when spermatozoa are used for assisted reproductive technologies.


Asunto(s)
Apoptosis , Espermatozoides/química , Adulto , Anexinas/metabolismo , Supervivencia Celular , Centrifugación por Gradiente de Densidad , Fragmentación del ADN , Proteínas HSP70 de Choque Térmico/análisis , Humanos , Indoles , Masculino , Microscopía Electrónica , Microscopía Fluorescente , Persona de Mediana Edad , Necrosis , Propidio , Unión Proteica , Proteínas/análisis , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Espermatozoides/citología
20.
Hum Reprod ; 16(7): 1376-81, 2001 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-11425816

RESUMEN

BACKGROUND: The aim of our study was to measure concentrations of vascular endothelial growth factor (VEGF), platelet endothelial cell adhesion molecule-1 (PECAM-1) CD31 and vascular cell adhesion molecules (VCAM-1) in the follicular fluid of women treated with assisted reproduction technology to determine whether these proteins might be outcome markers. METHODS: Follicular fluid was collected from 75 patients < or =40 years undergoing oocyte retrieval procedures at our tertiary hospital during 1997 and 1998: 50 with tubal disease, 12 with endometriosis, and 13 whose partners had been diagnosed with severe oligoasthenoteratozoospermia. This retrospective analysis considered age and information about treatment and outcome for all these women who had undergone fewer than three assisted reproduction attempts. RESULTS: Nineteen women became pregnant (defined by human chorionic gonadotrophin concentrations and embryonic cardiac activity 1 month after follicular aspiration); 56 did not. Women did not differ significantly in their follicular fluid concentrations of VEGF, sCD31 and VCAM-1 according to cause of infertility, or assisted reproduction outcome, or age. Follicular fluid concentrations of VEGF were significantly correlated with the number of gonadotrophin ampoules administered (P < 0.012), and follicular fluid concentrations of sVCAM-1 with the fertilization rate (P < 0.01). Follicular fluid concentrations of VEGF and sVCAM-1 were also correlated (P < 0.007). CONCLUSIONS: Our results do not suggest that VEGF, CD31, or sVCAM-1 in follicular fluid predict assisted reproduction outcome, especially among patients < or =40 years old. The correlation of a high fertilization rate and sVCAM-1 in follicular fluid suggests that sVCAM-1 might be a marker of fertilization.


Asunto(s)
Factores de Crecimiento Endotelial/análisis , Fertilización In Vitro , Líquido Folicular/química , Linfocinas/análisis , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/análisis , Molécula 1 de Adhesión Celular Vascular/análisis , Aborto Espontáneo , Adulto , Factores de Edad , Gonadotropina Coriónica/administración & dosificación , Estradiol/sangre , Femenino , Humanos , Infertilidad/etiología , Masculino , Embarazo , Resultado del Tratamiento , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial Vascular
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