RESUMEN
OBJECTIVE: Disruption of the third zinc finger domain of specificity protein 6 (SP6) presents an enamel-specific defect in a rat model of amelogenesis imperfecta (AMI rats). To understand the molecular basis of amelogenesis imperfecta caused by the Sp6 mutation, we established and characterized AMI-derived rat dental epithelial (ARE) cells. MATERIALS AND METHODS: ARE cell clones were isolated from the mandibular incisors of AMI rats, and amelogenesis-related gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Localization of wild-type SP6 (SP6WT) and mutant-type SP6 (SP6AMI) was analyzed by immunocytochemistry. SP6 transcriptional activity was monitored by rho-associated protein kinase 1 (Rock1) promoter activity with its specific binding to the promoter region in dental (G5 and ARE) and non-dental (COS-7) epithelial cells. RESULTS: Isolated ARE cells were varied in morphology and gene expression. Both SP6WT and SP6AMI were mainly detected in nuclei. The promoter analysis revealed that SP6WT and SP6AMI enhanced Rock1 promoter activity in G5 cells but that enhancement by SP6AMI was weaker, whereas no enhancement was observed in the ARE and COS-7 cells, even though SP6WT and SP6AMI bound to the promoter in all instances. CONCLUSION: ARE cell clones can provide a useful in vitro model to study the mechanism of SP6-mediated amelogenesis imperfecta.
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Amelogénesis Imperfecta/patología , Células Epiteliales/patología , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Amelogénesis Imperfecta/genética , Amelogénesis Imperfecta/metabolismo , Animales , Células Cultivadas , Expresión Génica , Incisivo/patología , Regiones Promotoras Genéticas , Ratas , Quinasas Asociadas a rho/genéticaRESUMEN
OBJECTIVE: This study examined the association between cardiac function and pulmonary function in hypertensive patients. METHODS: Hypertensive patients without overt cardiovascular disease were enrolled (n=43; mean±SD age 71±9 years). Pulmonary function was measured by the percentage of predicted forced vital capacity (%FVC) and the ratio of 1 s forced expiratory volume (FEV1) to FVC (FEV1/FVC ratio). Left ventricular ejection fraction (LVEF) and the ratio of peak early diastolic transmitral flow (E) to peak early diastolic mitral annular velocity (e') (E/e' ratio) were assessed using echocardiography. RESULTS: Multiple linear regression analysis revealed that E/e' was independently associated with %FVC and that LVEF was independently associated with FEV1/FVC ratio. Both LVEF and FEV1/FVC ratio were significantly lower in hypertensive former or current smokers than in hypertensive never smokers. CONCLUSIONS: Subclinical cardiac dysfunction was independently associated with reduced pulmonary function in hypertensive patients. Hypertensive patients with decreased pulmonary function may need preventive care to prevent the progression of heart failure.
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Pruebas de Función Cardíaca , Corazón/fisiopatología , Hipertensión/fisiopatología , Pulmón/fisiopatología , Anciano , Anciano de 80 o más Años , Demografía , Femenino , Humanos , Hipertensión/diagnóstico por imagen , Modelos Lineales , Masculino , Persona de Mediana Edad , Pruebas de Función Respiratoria , Fumar , UltrasonografíaRESUMEN
We present a novel concept for x-ray waveguiding based on electromagnetism in photonic crystals, using a waveguide consisting of a pair of claddings sandwiching a core with a periodic structure. By confining the x rays undergoing multiple interference in the core by total reflection, a characteristic waveguide mode whose field distribution matches the periodicity of the core is formed. The distinctively low propagation loss enables the single-mode propagation of x rays. This concept opens broad application possibilities in x-ray physics from coherent imaging to x-ray quantum optics.
RESUMEN
Left ventricular (LV) hypertrophy (LVH) may be eccentric or concentric (2 × LV posterior wall thickness relative to LV end-diastolic dimension ≤ 0.42 or > 0.42, respectively). The LV diastolic function between age-matched hypertensive patients with eccentric and concentric LVH was compared in the present study. Echocardiography was used to measure LV mass index (LV mass/body surface area; LVMI) as an index of LVH. LV diastolic function was assessed by measurements of peak early transmitral flow velocity (E)/peak late transmitral flow velocity (A) (the E/A ratio), peak early diastolic mitral annular velocity (e') and the E/e' ratio. Although LVMI, E/A and e' did not differ between the two groups, E/e' was significantly higher (worse) in patients with concentric LVH (13.4 ± 5.4) than in those with eccentric LVH (11.1 ± 3.6). Among hypertensive patients with LVH, those with concentric LVH may, therefore, have more severe LV diastolic dysfunction than those with eccentric LVH even if their LVMIs, which reflect the degree of LVH, are similar.
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Diástole , Hipertensión/fisiopatología , Sístole , Disfunción Ventricular Izquierda/fisiopatología , Anciano , Ecocardiografía , Femenino , Humanos , Hipertensión/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Disfunción Ventricular Izquierda/diagnóstico por imagenRESUMEN
Consensus is lacking about the clinical importance of aortic root dilatation in assessment of the risk of cardiovascular disease. In this study, correlations between aortic root diameter and echocardiographic features of left ventricular (LV) diastolic function were investigated in 333 patients with at least one cardiovascular risk factor (hypertension, diabetes or dyslipidaemia) and preserved LV systolic function. Aortic root diameter was measured by M-mode echocardiography, and LV diastolic function was evaluated by measuring the peak velocity of early (E) and late (A) diastolic transmitral blood flow and peak early diastolic mitral annular velocity (E') by Doppler echocardiography. Linear regression analysis showed that, in men, age was not related to aortic root diameter but hypertension and LV hypertrophy were, whereas the converse was true in women. The parameters E, E/A ratio and E', were related to aortic root diameter in both sexes. Stepwise multiple regression analysis confirmed that E in women and E' in men were independently associated with aortic root diameter. It is concluded that aortic root dilatation might be a useful marker of subclinical LV diastolic dysfunction. Patients with preserved systolic function showing aortic root dilatation should, therefore, be given preventative therapy against LV diastolic heart failure.
Asunto(s)
Aorta/fisiopatología , Complicaciones de la Diabetes , Dilatación Patológica/complicaciones , Dislipidemias/complicaciones , Hipertensión/complicaciones , Disfunción Ventricular Izquierda/etiología , Anciano , Aorta/diagnóstico por imagen , Biomarcadores , Diabetes Mellitus/diagnóstico por imagen , Diabetes Mellitus/fisiopatología , Diástole , Dilatación Patológica/diagnóstico por imagen , Dislipidemias/diagnóstico por imagen , Dislipidemias/fisiopatología , Ecocardiografía Doppler , Femenino , Humanos , Hipertensión/diagnóstico por imagen , Hipertensión/fisiopatología , Masculino , Factores de Riesgo , Sístole , Disfunción Ventricular Izquierda/diagnóstico por imagen , Disfunción Ventricular Izquierda/fisiopatologíaRESUMEN
Apoptosis regulates inflammatory cell survival in allergic inflammation, and decreased apoptosis contributes to the chronicity of inflammation. To investigate the mechanisms of onset and remission of mite-sensitive childhood asthma, we evaluated peripheral blood mononuclear cell apoptosis in patients with asthma and in remission. There was a similar percentage of hypodiploid cells in unstimulated mononuclear cell cultures from patients with active asthma (29.5+/-5.0%) and normal individuals (25.9+/-4.9%). In contrast, the percentage increased in patients in remission (44.5+/-3.2%). In Dermatophagoides farinae (Df) antigen-stimulated mononuclear cell, the stimulation index was lower in patients with active asthma (0.95+/-0.06%) than in normal individuals (1.31+/-0.16%). In contrast to active patients, the proportion of hypodiploid cells stimulated with Df in patients with remission was equivalent to that of normal controls. After phytohemaglutinin (PHA) stimulation, the percentage of hypodiploid cells in patients with active asthma (35.1+/-3.2%) was also lower than in normal individuals (48.5+/-4.3%) or patients in remission (49.5+/-5.7%). Apoptosis occurred predominantly in CD8+, but not CD4+, cells in patients in remission. Interleukin IL-2 inhibited apoptosis in Df-activated cells in normal individuals, whereas IL-2 did not inhibit apoptosis in cells from patients in remission as well as with active asthma. The expression of Fas receptors on resting mononuclear cells was similar in the three groups. However, Fas receptor expression in Df-stimulated mononuclear cells was greater in patients with active asthma than in healthy individuals. In patients with remission that was equivalent to healthy controls. The PHA increased Fas expression to a similar degree in the three groups. With regard to Fas ligand, the expression was lower in unstimulated cultured mononuclear cells from patients than in normal individuals. In patients in remission that was comparable to normal individuals. The Df stimulation upregulated the Fas ligand in patients with active asthma, and downregulated it in patients in remission. In conclusion, apoptosis in Df-stimulated mononuclear cells is impaired in patients with active asthma, while spontaneous apoptosis of CD8+ cells in vivo is augmented in patients in remission, and may be involved in the onset and remission of mite-sensitive asthma.
Asunto(s)
Apoptosis , Asma/inmunología , Leucocitos Mononucleares/inmunología , Adolescente , Animales , Niño , Dermatophagoides farinae/inmunología , Femenino , Humanos , Masculino , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Receptor fas/metabolismoAsunto(s)
Linfoma de Células B/complicaciones , Pericarditis Constrictiva/etiología , Humanos , Leucemia Linfocítica Crónica de Células B/complicaciones , Leucemia Linfocítica Crónica de Células B/diagnóstico por imagen , Linfoma de Células B/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Pericarditis Constrictiva/diagnóstico por imagen , Tomografía Computarizada por Rayos X/métodosRESUMEN
BACKGROUND: Whether normal peripheral blood mononuclear cells (PBMCs) transferred to severe combined immunodeficient (SCID) mice produce specific IgE remains unclear. METHODS: Mice received injections of Dermatophagoides farinae antigen (Df)-stimulated PBMCs from healthy persons (IgE RAST score of 0). RESULTS: High titers of Df-specific IgE were detected. The Df-specific IgE activity produced was comparable to or higher than that produced by cells from patients with asthma although the time to maximal production was longer. IgE derived from PMBCs of healthy persons or patients with asthma induced histamine release from cultured human basophils that had been stimulated with Df antigen or an anti-IgE antibody. Treatment of Df-stimulated PBMCs with a high dose, but not a low dose, of interleukin-4 stimulated production of Df-specific IgE by PMBCs from healthy persons or patients with asthma. In contrast, intravenous injection of IFN-gamma into reconstituted SCID mice decreased Df-specific IgE production by PBMCs from patients with asthma. In PMBCs from healthy persons, IgE class-switching may occur later and block the effects of treatment with IFN-gamma. CONCLUSIONS: PBMCs from healthy persons and persons with asthma have clones reactive to allergen and produce functional IgE specific for relevant antigens in mite-sensitive bronchial asthma.
Asunto(s)
Alérgenos/inmunología , Asma/inmunología , Glicoproteínas/inmunología , Inmunoglobulina E/análisis , Leucocitos Mononucleares/inmunología , Animales , Antígenos Dermatofagoides , Basófilos/inmunología , Niño , Preescolar , Femenino , Liberación de Histamina , Humanos , Interferón gamma/farmacología , Interleucina-4/farmacología , Leucocitos Mononucleares/efectos de los fármacos , Masculino , Ratones , Ratones SCID , Factores de TiempoRESUMEN
Kawasaki disease (KD) is a childhood-onset vascular disease. We assessed the concentrations of macrophage-colony stimulating factor (M-CSF) and those of lipids in sera from patients with KD. The M-CSF concentration in patients with acute-phase KD was 2,914+/-159 U/ml, significantly higher than that in control subjects with Infectious diseases (1,241+/-96 U/ml). The elevated levels of this cytokine in the acute phase fell to 1,319+/-138 U/ml in the convalescent phase. Total and high-density lipoprotein cholesterol concentrations in acute phase KD (113.8+/-8.4 and 21.5+/-2.3 mg/dl, respectively) were lower than in the infectious disease controls (195.8+/-7.0 and 62.5+/-1.8 mg/dl). The elevation of M-CSF correlated with the decrease of total and high-density lipoprotein cholesterol. Overproduction of macrophage-colony stimulating factor activates macrophages and monocytes and may disturb the lipid metabolism. Both effects could contribute to vasculitis in KD.
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HDL-Colesterol/sangre , Colesterol/sangre , Factor Estimulante de Colonias de Macrófagos/sangre , Síndrome Mucocutáneo Linfonodular/sangre , Protección a la Infancia , Preescolar , HDL-Colesterol/efectos de los fármacos , Femenino , Humanos , Inmunoglobulinas Intravenosas/uso terapéutico , Lactante , Bienestar del Lactante , Japón/epidemiología , Recuento de Leucocitos , Factor Estimulante de Colonias de Macrófagos/efectos de los fármacos , Masculino , Monocitos/citología , Monocitos/metabolismo , Síndrome Mucocutáneo Linfonodular/tratamiento farmacológico , Estadística como AsuntoRESUMEN
This study investigated the effect of pioglitazone, an insulin sensitizer, on metabolic abnormalities and oxidative stress as a cause of myocardial collagen accumulation in prediabetic rat hearts. Twenty male diabetic rats and 9 male nondiabetic age-matched rats were used. The diabetic rats were divided into two groups: diabetic treated and untreated. Pioglitazone was mixed in rat chow fed to the diabetic treated group (0.01%). Treatment duration was 5 weeks. At baseline (15 weeks) and 20 weeks of age, blood glucose, lipid, insulin, and plasma malondialdehyde-thiobarbituric acid (MDA) levels were measured and Doppler echocardiography was tracked. At 20 weeks of age, left ventricular collagen content was studied. Blood glucose, plasma insulin, and triglyceride levels in the diabetic treated group were significantly lower than those in the untreated diabetic group. Deceleration time (ms) of early diastolic inflow in the treated diabetic group decreased significantly compared with the untreated diabetic group (65 +/- 8 vs. 77 +/- 8, p < 0.01). Ratio of left ventricular weight to body weight (mg/g) and ratio of left ventricular collagen content to dry weight (mg/100 mg) were decreased in the treated diabetic group (1.5 +/- 0.1, 1.3 +/- 0.3) compared with the untreated diabetic group (1.7 +/- 0.2, p < 0.01; 1.7 +/- 0.3, p < 0.05). Plasma MDA concentration (nmol/ml) significantly decreased (2.9 +/- 0.3 at baseline to 2.3 +/- 0.3 at 20 weeks, p = 0.001) in the treated diabetic group, and was lower than that in the untreated diabetic group (3.2 +/- 0.7 at 20 weeks, p < 0.05). Pioglitazone improved glucose and lipid metabolism and reduced oxidative stress in the left ventricle, which decreased left ventricular collagen accumulation and improved left ventricular diastolic function of prediabetic rat hearts.
Asunto(s)
Colágeno/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Hipoglucemiantes/farmacología , Estado Prediabético/metabolismo , Tiazoles/farmacología , Tiazolidinedionas , Disfunción Ventricular Izquierda/tratamiento farmacológico , Animales , Glucemia/metabolismo , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/fisiopatología , Diástole/efectos de los fármacos , Ecocardiografía Doppler , Hemodinámica/efectos de los fármacos , Hipoglucemiantes/uso terapéutico , Insulina/sangre , Cinética , Lípidos/sangre , Masculino , Malondialdehído/sangre , Estrés Oxidativo/efectos de los fármacos , Pioglitazona , Estado Prediabético/tratamiento farmacológico , Estado Prediabético/fisiopatología , Ratas , Ratas Endogámicas OLETF , Ratas Long-Evans , Tiazoles/uso terapéutico , Sustancias Reactivas al Ácido Tiobarbitúrico/análisisRESUMEN
The previously isolated cDNA encoding human adenylate kinase (AK) isozyme 3 was recently renamed AK4. Consequently, human AK3 cDNA remains to be identified and we have little information about the functional relationship between human AK3 and AK4. In pursuit of the physiological roles of both the AK3 and AK4 proteins, we first isolated an authentic human AK3 cDNA and compared their expression. Nucleotide sequencing revealed that the cDNA encoded a 227-amino-acid protein, with a deduced molecular mass of 25.6 kDa, that shares greater homology with the AK3 cDNAs isolated from bovine and rat than that from human. We named the isolated cDNA AK3. Northern-blot analysis revealed that AK3 mRNA was present in all tissues examined, and was highly expressed in heart, skeletal muscle and liver, moderately expressed in pancreas and kidney, and weakly expressed in placenta, brain and lung. On the other hand, we found that human AK4 mRNA was highly expressed in kidney, moderately expressed in heart and liver and weakly expressed in brain. Western-blot analysis demonstrated expression profiles of AK3 and AK4 that were similar to their mRNA expression patterns in each tissue. Over expression of AK3, but not AK4, in both Escherichia coli CV2, a temperature-sensitive AK mutant, and a human embryonic kidney-derived cell line, HEK-293, not only produced significant GTP:AMP phosphotransferase (AK3) activity, but also complemented the CV2 cells at 42 degrees C. Subcellular and submitochondrial fractionation analysis demonstrated that both AK3 and AK4 are localized in the mitochondrial matrix.
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Adenilato Quinasa/química , Mitocondrias/enzimología , Adenilato Quinasa/metabolismo , Animales , Northern Blotting , Western Blotting , Bovinos , Línea Celular , Clonación Molecular , ADN Complementario/química , ADN Complementario/metabolismo , Escherichia coli/enzimología , Humanos , Iones , Riñón/enzimología , Hígado/enzimología , Ratones , Mitocondrias/metabolismo , Datos de Secuencia Molecular , Músculo Esquelético/enzimología , Miocardio/enzimología , Plásmidos/metabolismo , Isoformas de Proteínas , ARN Mensajero/metabolismo , Ratas , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Análisis de Secuencia de ADN , Fracciones Subcelulares , Distribución Tisular , TransfecciónRESUMEN
The objectives of the present study were (1) to determine whether oxidized low-density lipoprotein (LDL) and lysophosphatidylcholine (lyso-PC), a major phospholipid component of oxidized LDL, stimulate the production of endothelin-1 (ET)-1 in cultured human coronary artery smooth muscle cells (SMCs), and (2) to examine the possible effect of an antiatherogenic agent, eicosapentaenoic acid (EPA), on oxidized-LDL- and lyso-PC-stimulated ET-1 production in these cells. Oxidized LDL (10-50 microg/ml) and lyso-PC (10(-7) to 10(-5) mol/l) stimulated ET-1 production in a concentration-dependent manner. By contrast, the effects of native LDL and phosphatidylcholine were modest or absent. Lyso-PC (10(-7) to 10(-5) mol/l) and oxidized LDL (10-50 microg/ml) significantly induced particulate protein kinase C (PKC) activation. Lyso-PC- and oxidized-LDL-stimulated ET-1 production was significantly inhibited by PKC inhibitor, PKC (19-36). EPA (80-160 micromol/l) clearly suppressed ET-1 production stimulated by oxidized LDL and lyso-PC in a concentration-dependent manner. Furthermore, EPA (160 micromol/l) significantly inhibited lyso-PC (10(-5) mol/l)- and oxidized LDL (50 microg/ml)-induced particulate PKC activation. Results suggest that oxidized LDL and lyso-PC stimulate ET-1 production by a mechanism involving activation of PKC, and that EPA suppresses ET-1 production stimulated by lyso-PC as well as oxidized LDL probably through the modulation of PKC in human coronary artery SMCs. EPA may exert an antiatherosclerotic effect, in part, through these mechanisms.
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Vasos Coronarios/efectos de los fármacos , Ácido Eicosapentaenoico/farmacología , Endotelina-1/biosíntesis , Lipoproteínas LDL/farmacología , Lisofosfatidilcolinas/farmacología , Músculo Liso Vascular/efectos de los fármacos , Angiotensina II/farmacología , Células Cultivadas , Vasos Coronarios/metabolismo , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Humanos , Músculo Liso Vascular/metabolismo , Fragmentos de Péptidos/farmacología , Fosfatidilcolinas/farmacología , Factor de Crecimiento Derivado de Plaquetas/farmacología , Proteína Quinasa C/antagonistas & inhibidores , Proteína Quinasa C/metabolismo , Proteína Quinasa C/farmacología , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Clinical evidence suggests that roxithromycin (RXM) may be an effective additional therapy for bronchial asthma. However, how it interferes with allergic responses is unclear. To investigate the mechanisms of action of RXM, lymphocyte transformation and interferon (IFN)-gamma, interleukin (IL)-4 and IL-5 synthesis associated with Dermatophagoides farinae (Df), mite antigen in patients with bronchial asthma were evaluated in vitro in the presence of RXM. T cell proliferation in Df antigen-stimulated patients' lymphocytes was suppressed by 50-100 microg/ml of RXM. Production of IL-4 and IL-5 was similarly decreased by 1-10 microg/ml RXM, whereas, IFN-gamma production, which was reduced by Df-stimulation alone, was increased by 50 microg/ml RXM. Our results suggest that skewed cytokine profiles of patients with mite antigen-induced bronchial asthma may be corrected with RXM, which may mimic those of patients in remission, who are tolerant of Df antigen.
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Antibacterianos/farmacología , Asma/inmunología , Citocinas/biosíntesis , Activación de Linfocitos/efectos de los fármacos , Ácaros/inmunología , Roxitromicina/farmacología , Linfocitos T/efectos de los fármacos , Adolescente , Adulto , Animales , Niño , Femenino , Humanos , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Interleucina-5/biosíntesis , Masculino , Linfocitos T/inmunologíaAsunto(s)
Adenosina Trifosfato/metabolismo , Insuficiencia de la Válvula Aórtica/metabolismo , Proteínas de Transporte de Membrana , Proteínas Mitocondriales , Miocardio/metabolismo , Proteínas/metabolismo , Desacopladores/metabolismo , Animales , Factor Natriurético Atrial/genética , Factor Natriurético Atrial/metabolismo , Northern Blotting , Gasto Cardíaco Bajo/fisiopatología , Modelos Animales de Enfermedad , Humanos , Canales Iónicos , Masculino , Mitocondrias Cardíacas/metabolismo , Músculo Esquelético/fisiología , Fosfocreatina/metabolismo , Proteínas/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Proteína Desacopladora 2RESUMEN
Abdominal aortic aneurysms (AAAs) are characterized by structural alterations of the aortic wall resulting from degradation of collagen and elastin. Matrix metalloproteinases (MMPs), particularly MMP-2 and MMP-9, show strong elastinolytic activity. We examined the levels of mRNA for MMP-2, MMP-9, membrane type (MT)-MMP-1, tissue inhibitor of metalloproteinase-1 (TIMP-1), and TIMP-2 in AAAs (n = 8), atherosclerotic occlusive diseases (AOD) (n = 8), and normal subjects (n = 8) using the reverse transcription-polymerase chain reaction (RT-PCR). We also analyzed the gelatinolytic activity of these metalloproteinases using gelatin zymography. The levels of MMP-2 and MMP-9 mRNA were increased in the AAA group compared with those in the AOD group and normal subjects. The levels for TIMP-1 and TIMP-2 mRNA in the AAA group were also higher than those in the AOD and normal groups. Only in the case of MT-MMP-1 was the difference between AAA and AOD not statistically significant. By gelatin zymography with the same samples used for RT-PCR, gelatinolytic activity of MMP-9 was elevated in all AAA tissues. The 62-kDa form of MMP-2 was elevated in both the AAA and AOD groups and did not differ significantly between them. Linear regression analysis demonstrated a significant positive correlation between mRNA levels of MMPs and those of TIMPs. These observations suggest that aneurysm formation in patients with atherosclerosis is related to the degree of MMP-9 expression.
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Aneurisma de la Aorta Abdominal/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Anciano , Arteriopatías Oclusivas/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Inhibidor Tisular de Metaloproteinasa-1/metabolismoRESUMEN
To investigate the relationship between the telomerase activity levels and clinicopathological features of tumors, we quantified the telomerase activities of 23 renal cell carcinomas (RCCs) and four non-cancerous tissues, using a modified telomeric repeat amplification protocol assay, and assessed the hTERT mRNA levels of these samples by reverse transcription-polymerase chain reaction analysis. Elevated levels of telomerase activity had correlation with tumor stages as well as the degree of nuclear grades. Our findings suggested that telomerase activity is a useful indicator for tumor aggressiveness in RCCs. However, hTERT mRNA levels in RCCs had no correlation with nuclear grades and tumor stages. The telomerase activities and the hTERT mRNA levels in cancer cells were not always in parallel. These results suggested that telomerase activity is regulated in a posttranscriptional manner as well as a post-translational manner in tumor cells.
Asunto(s)
Carcinoma de Células Renales/enzimología , Neoplasias Renales/enzimología , ARN , Telomerasa/genética , Telomerasa/metabolismo , Adulto , Anciano , Empalme Alternativo , Carcinoma de Células Renales/patología , Proteínas de Unión al ADN , Femenino , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Renales/patología , Masculino , Persona de Mediana Edad , Neoplasias de la Próstata , Procesamiento Postranscripcional del ARN , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Neoplasias de la Vejiga UrinariaRESUMEN
Troglitazone, an oral antidiabetic agent, has hypoglycemic effects in insulin-resistant animal models and humans. This study was conducted to evaluate its effect on left ventricular diastolic dynamics of a spontaneous diabetic (DM) rat model. Twenty DM rats and 20 age-matched nonDM rats were used, and 10 of each group were treated with troglitazone as a 0.2% food admixture for 10 weeks. At 5 and 15 weeks of age, Doppler echocardiography and M-mode echocardiography were performed. Troglitazone treatment significantly improved the left ventricular diastolic dynamics of DM rats: deceleration time (msec) of early diastolic inflow decreased significantly (treated 52 +/- 3 vs untreated 64 +/- 5, p = 0.0002), and peak velocity of early transmitral inflow (cm/sec) increased significantly (treated 96 +/- 7 vs untreated 86 +/- 8, p = 0.0216). The data suggest that troglitazone improves left ventricular diastolic dynamics of a DM rat model at prediabetic stage.
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Cromanos/farmacología , Diástole/fisiología , Hipoglucemiantes/farmacología , Estado Prediabético/tratamiento farmacológico , Tiazoles/farmacología , Tiazolidinedionas , Función Ventricular Izquierda/efectos de los fármacos , Animales , Presión Sanguínea/efectos de los fármacos , Cromanos/uso terapéutico , Diástole/efectos de los fármacos , Modelos Animales de Enfermedad , Ecocardiografía Doppler , Frecuencia Cardíaca/efectos de los fármacos , Hipoglucemiantes/uso terapéutico , Masculino , Distribución Aleatoria , Ratas , Ratas Endogámicas OLETF , Tiazoles/uso terapéutico , TroglitazonaRESUMEN
IL-18 is a novel cytokine that induces interferon (IFN)-gamma secretion and plays an important role in antitumor immunity. In the present study, we constructed plasmid vectors encoding the murine mature IL-18 cDNA linked with the Igkappa leader sequence and the pro-IL-18 cDNA to estimate the efficacy of the mature IL- 18 vector and to evaluate IL-18--producing tumor cells as a tumor vaccine. Colon 26 cells were transfected with the abovementioned vectors or with vector alone (mock). Reverse transcription-polymerase chain reaction analysis showed increased expression of murine IL-18 cDNA in both mature IL-18 and pro-IL-18 transfectants in comparison to that in mock transfected cells. The ability of the culture supernatants of mature IL-18 transfectants to induce IFN-gamma secretion was extremely high (40-140 pg/10(6) cells) in comparison to that of pro-IL-18 transfectants (4-18 pg/10(6) cells). When injected into syngeneic BALB/c mice, the growth of mature IL-18 transfectants, but not pro-IL-18 transfectants, was significantly less than that in mock transfected cells ( P< .01, by ANOVA and analysis of covariance). In addition, injection of colon 26 or Meth-A cells into mice immunized with a mature IL-18 transfectant revealed acquired immunity. Depletion of natural killer cells did not affect the growth of transfectants. However, the growth inhibitory effects were partially abrogated following treatment with anti-CD4+ and anti-CD8+ antibodies. These data suggest that the rejection of mature IL-18/colon 26 cells was mediated through T-cell activation. Gene therapy using mature IL-18 transfectants containing a plasmid vector and the Igkappa leader sequence may be a useful tumor vaccine.
Asunto(s)
Neoplasias del Colon/terapia , Fibrosarcoma/terapia , Terapia Genética/métodos , Vectores Genéticos , Inmunoglobulinas/genética , Interleucina-18/genética , Adenoviridae/genética , Animales , Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Antígeno B7-2 , Antígenos CD4/metabolismo , Antígenos CD8/metabolismo , Cartilla de ADN/química , Fibrosarcoma/inducido químicamente , Expresión Génica , Genes MHC Clase I/fisiología , Genes MHC Clase II/fisiología , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Técnicas para Inmunoenzimas , Inmunoglobulina G/inmunología , Inmunoglobulinas/metabolismo , Interferón gamma/metabolismo , Interleucina-18/metabolismo , Células Asesinas Naturales/metabolismo , Glicoproteínas de Membrana/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transfección/métodos , Células Tumorales CultivadasRESUMEN
There is considerable evidence to indicate that humoral factors play an important role in the development of left ventricular hypertrophy. Cardiotrophin-1 (CT-1) is a cytokine that has been shown to induce cardiac hypertrophy in a dose-dependent manner. The aim of the present study was to investigate the acute effect of CT-1 on hemodynamic parameters in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) and to study the relationship between the plasma concentration of CT-1 and its hemodynamic effect. Ten-week-old SHR and age-matched WKY were used. Blood pressure (BP), heart rate (HR) and plasma concentration of CT-1 were measured both before and for 60 min after intravenous bolus injection of human CT-1 (10 microg/kg). CT-1 injection significantly decreased BP and significantly increased HR in SHR and WKY. There were significant differences in BP and HR between the two groups at all time points after injection. The lowest BP, highest HR and maximal plasma concentrations of CT-1 were observed in both groups within 10 min after injection. However, after converting the values into the percentage change from their respective baselines, there were no significant differences between the two groups in BP or HR at any time point. There was also no significant difference between the two groups at any time point in the plasma concentration of CT-1. This study indicates that CT-1 decreases BP and increases HR in both SHR and WKY. The most obvious change occurred within 10 min after injection. However, there was no significant difference in the hypotensive effect of CT-1 on 10-week-old SHR and WKY.