RESUMEN
BACKGROUND: Repeated exposure to allergens induces chronic allergic lesions in the skin and a shift in the cutaneous cytokine milieu to T helper (Th)2. AIM: To assess the relationships between Th17 and Th2 response during allergic contact dermatitis (ACD) in mice. METHODS: ACD was induced in C57BL/6 mice by single or repeated epicutaneous challenge of 2,4,6-trinitro-1-chlorobenzene. Relationships between Th17 and Th2 response were analyzed by immunohistochemical observations and activity of cytokines on days 8 (first challenge), 18 (11th challenge), 28 (21st challenge) and 38 (31st challenge). RESULTS: On day 8, tissue levels of interleukin (IL)-17 and IL-22 were high, whereas tissue levels of IL-4 and serum IgE concentration were low. Following acute contact dermatitis, mice developed chronic eczematous lesions on day 18, and gradually improved on days 28 and 38. Tissue IL-4 and serum IgE levels corresponded to the development and improvement of chronic eczematous lesions. Numbers of Th17 cells and tissue levels of IL-17 and IL-22 rapidly decreased as IL-4 and IgE levels increased on day 18. As levels of IL-4 and IgE decreased, the number of Th17 cells and tissue levels of IL-17 and IL-22 increased again on days 28 and 38. On day 18, tissue levels of Th17 response-inducing cytokines (IL-6, IL-23 and transforming growth factor-ß) were high, and IL-23-expressing cells appeared in abundance, when Th2 response was extremely high. IL-17 injection decreased tissue IL-4 and serum IgE levels. CONCLUSIONS: Th17 correlates closely with Th2 in murine chronic ACD induced by repeated epicutaneous challenge.
Asunto(s)
Citocinas/metabolismo , Dermatitis Alérgica por Contacto/inmunología , Células Th17/metabolismo , Células Th2/metabolismo , Enfermedad Aguda , Alérgenos/inmunología , Alérgenos/toxicidad , Animales , Dermatitis Alérgica por Contacto/patología , Modelos Animales de Enfermedad , Inmunoglobulina E/metabolismo , Ratones , Ratones Endogámicos C57BL , Cloruro de Picrilo/inmunología , Cloruro de Picrilo/toxicidadRESUMEN
Basal-supported oral therapy (BOT) is often used to treat poorly controlled type 2 diabetes. However, patients sometimes experience nocturnal and early morning hypoglycemia. Thus, maintaining targeted glycemic control by BOT is limited in some patients. We assessed the efficacy and safety of replacing basal insulin by sitagliptin therapy in Japanese type 2 diabetes patients on BOT. Forty-nine subjects were sequentially recruited for the 52-week, prospective, single arm study. Patients on BOT therapy were switched from basal insulin to sitagliptin. The primary endpoint was change in HbA1c in 52 weeks. The secondary endpoints were dropout rate, changes in body weight, frequency of hypoglycemia, and relationship between change in HbA1c and insulin secretion capacity evaluated by glucagon loading test. The average dose of basal insulin was 15.0±8.4 units. Sixteen subjects (31.3%) were dropped because replacement by sitagliptin was less effective for glycemic control. In these subjects, diabetes duration was longer, FPG and HbA1c at baseline were higher, and insulin secretion capacity was lower. Change in HbA1c in 52 weeks was - 4 mmol/mol (95% CI - 5 to - 4 mmol/mol) (p<0.05). Change in body weight was - 0.71 kg (95% CI - 1.42 to - 0.004 kg) (p<0.05). Frequency of hypoglycemia was decreased from 1.21±1.05 to 0.06±0.24 times/month. HbA1c level was improved if C-peptide index (CPI) was over 1.19. In conclusion, basal insulin in BOT can be replaced by sitagliptin with a decrease in HbA1c level and frequency of hypoglycemia in cases where insulin secretion capacity was sufficiently preserved.
Asunto(s)
Pueblo Asiatico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Insulina/efectos adversos , Insulina/uso terapéutico , Pirazinas/efectos adversos , Pirazinas/uso terapéutico , Triazoles/efectos adversos , Triazoles/uso terapéutico , Anciano , Índice de Masa Corporal , Peso Corporal/efectos de los fármacos , Péptido C/sangre , Demografía , Diabetes Mellitus Tipo 2/complicaciones , Relación Dosis-Respuesta a Droga , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Hipoglucemia/complicaciones , Hipoglucemia/tratamiento farmacológico , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/efectos adversos , Hipoglucemiantes/farmacología , Insulina/administración & dosificación , Insulina/farmacología , Japón , Masculino , Pirazinas/administración & dosificación , Pirazinas/farmacología , Curva ROC , Fosfato de Sitagliptina , Resultado del Tratamiento , Triazoles/administración & dosificación , Triazoles/farmacologíaRESUMEN
BACKGROUND: Enzastaurin, an oral serine-threonine kinase inhibitor, was initially developed as an ATP-competitive selective inhibitor against protein kinase Cß. However, the mechanism by which enzastaurin contributes to tumourigenesis remains unclear. METHODS: We analysed the anti-tumour effects of enzastaurin in 22 lung cancer cell lines to ascertain the potential for enzastaurin-based treatment of lung cancer. To identify molecules or signalling pathways associated with this sensitivity, we conducted a gene, receptor tyrosine kinases phosphorylation and microRNA expression profiling study on the same set of cell lines. RESULTS: We identified eight genes by pathway analysis of molecules having gene-drug sensitivity correlation, and used them to build a support vector machine algorithm model by which sensitive cell lines were distinguished from resistant cell lines. Pathway analysis revealed that the JAK/STAT signalling pathway was one of the main ones involved in sensitivity to enzastaurin. Overexpression of JAK1 was observed in the sensitive cells by western blotting. Simultaneous administration of enzastaurin and JAK inhibitor inhibited enzastaurin-induced cell growth-inhibitory effect. Furthermore, lentiviral-mediated JAK1-overexpressing cells were more sensitive to enzastaurin than control cells. CONCLUSION: Our results suggested that the JAK1 pathway may be used as a single predictive biomarker for enzastaurin treatment. The anti-tumour effect of enzastaurin should be evaluated in lung cancer with overexpressed JAK pathway molecules.
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Antineoplásicos/farmacología , Indoles/farmacología , Janus Quinasa 1/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Resistencia a Antineoplásicos , Perfilación de la Expresión Génica , Humanos , Indoles/uso terapéutico , Janus Quinasa 1/antagonistas & inhibidores , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , MicroARNs/metabolismo , Inhibidores de Proteínas Quinasas/farmacologíaRESUMEN
AIMS: To assess the efficacy and safety of combination therapy with sitagliptin and low dosage sulphonylureas on glycaemic control and insulin secretion capacity in Japanese type 2 diabetes. METHODS: Eighty-two subjects were sequentially recruited for the 52-week, prospective, single arm study. Sitagliptin was added on to sulphonylureas (glimepride or gliclazide) with or without metformin. The primary endpoint was a change in A1C. The secondary endpoints were changes in BMI, insulin secretion capacity, blood pressure and urinary albumin excretion, unresponsive rate, and hypoglycaemia. Insulin secretion capacity was evaluated by glucagon loading test. RESULTS: Change in A1C was -0.80% (95% CI -0.90 to -0.68) (p < 0.001). Change in BMI, systemic and diastolic blood pressure, and urinary albumin excretion were -0.38 kg/m(2) (95% CI -0.72 to -0.04) (p < 0.05), -6.7/-3.6 mmHg (95% CI -10.0 to -3.4/-4.8 to -2.4) (p < 0.001), and -43.2 mg/gCr (95% CI -65.7 to -20.8) (p < 0.001) respectively. Mild hypoglycaemia was observed in three cases. The unresponsive rate was 6.1%. Glucagon loading test showed that 0-min and 6-min CPR at baseline and 52-week were not significantly changed: 0-min CPR, 1.58 ± 0.58-1.71 ± 0.73 ng/ml; 6-min CPR, 3.48 ± 1.47-3.58 ± 1.21 ng/ml. Insulin secretion capacity, CPI and SUIT index at baseline did not predict the efficacy of the combination therapy. The final dosages of glimepiride and gliclazide were 1.44 ± 0.90 mg and 34.5 ± 15.3 mg respectively. The dosage of sitagliptin was increased from 50 mg to 69.0 ± 24.5 mg in 52-week. CONCLUSIONS: The combination therapy with sitagliptin and low dosage sulphonylureas was safe and effective for glycaemic control. Glucagon loading test indicated that 1 year administration of sitagliptin and sulphonylureas preserved insulin secretion capacity.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hipoglucemiantes/administración & dosificación , Pirazinas/administración & dosificación , Compuestos de Sulfonilurea/administración & dosificación , Triazoles/administración & dosificación , Anciano , Albuminuria/etiología , Glucemia/metabolismo , Presión Sanguínea/fisiología , Índice de Masa Corporal , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatología , Relación Dosis-Respuesta a Droga , Quimioterapia Combinada , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Hipoglucemia/inducido químicamente , Hipoglucemiantes/efectos adversos , Insulina/metabolismo , Secreción de Insulina , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Pirazinas/efectos adversos , Fosfato de Sitagliptina , Compuestos de Sulfonilurea/efectos adversos , Resultado del Tratamiento , Triazoles/efectos adversosRESUMEN
BACKGROUND: The present study observed effects of the histamine H(4) receptor on chronic allergic contact dermatitis induced by repeated challenge in mice. METHODS: Acute contact dermatitis was induced by single epicutaneous challenge of 2,4,6-trinitro-1-chlorobenzene (TNCB) to the ear. Chronic allergic contact dermatitis was developed by repeated epicutaneous challenge using TNCB on the dorsal back skin. H(4) receptor antagonist JNJ7777120 was administered to wild-type mice, while H(4) receptor agonist 4-methylhistamine was administered to histidine decarboxylase (HDC) (-/-) mice that synthesized no histamine. RESULTS: HDC (-/-) mice did not differ phenotypically from HDC (+/+) mice, and H(4) receptor antagonist/agonist did not have clinical effects in terms of acute contact dermatitis reactions. H(4) receptor antagonist ameliorated skin eczematous lesions induced by repeated TNCB challenge in HDC (+/+) mice. On the contrary, H(4) receptor agonist exacerbated skin lesions exclusively in HDC (-/-) mice. Application of H(4) receptor agonist induced migration of mast cells and eosinophils in skin lesions, and H(4) receptor antagonist suppressed these changes. H(4) receptor was immunohistochemically detected on mast cells in eczematous lesions. Levels of interleukin (IL)-4, -5, and -6 in lesions were decreased, whereas levels of interferon-gamma and IL-12 were increased by H(4) receptor antagonistic activity. Serum Immunoglobulin E levels rapidly increased with repeated challenge, but decreased with H(4) receptor antagonist. CONCLUSION: Because chronic allergic contact dermatitis is developed by H(4) receptor stimulation, H(4) receptor antagonists might represent new candidate drugs for treating chronic allergic contact dermatitis.
Asunto(s)
Dermatitis Alérgica por Contacto/tratamiento farmacológico , Antagonistas de los Receptores Histamínicos/farmacología , Receptores Acoplados a Proteínas G/inmunología , Receptores Histamínicos/inmunología , Animales , Quimiotaxis de Leucocito/inmunología , Enfermedad Crónica , Citocinas/biosíntesis , Dermatitis Alérgica por Contacto/inmunología , Dermatitis Alérgica por Contacto/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Agonistas de los Receptores Histamínicos/farmacología , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Cloruro de Picrilo/inmunología , Cloruro de Picrilo/toxicidad , Receptores Acoplados a Proteínas G/metabolismo , Receptores Histamínicos/metabolismo , Receptores Histamínicos H4RESUMEN
In the neurological mutant mouse reeler, the histological organization of the neocortex develops abnormally and essentially results in an inversion of the relative positions of the cortical layers. The reeler mutation, therefore, provides an insight into the molecular mechanisms underlying the formation of the cortical layers. We have generated a monoclonal antibody (CR-50) that probes a distinct allelic antigen present in wild-type but not in reeler mutant mice. CR-50 reacted specifically with Cajal-Retzius neurons, one of the first cortical neurons to differentiate in the neocortex, but whose functional role is not known. When dissociated cerebral cortical cells were incubated with CR-50 in reaggregation culture, the genotype-dependent histogenetic assembly of wild-type cortical cells resembled that of reeler mutants. These findings revealed that the selective expression of a distinct molecule on Cajal-Retzius neurons is critical for the normal lamination of cortical neurons in the mammalian neocortex.
Asunto(s)
Moléculas de Adhesión Celular Neuronal/análisis , Corteza Cerebral/citología , Proteínas de la Matriz Extracelular/análisis , Mutación , Proteínas del Tejido Nervioso/análisis , Neuronas/citología , Animales , Anticuerpos Monoclonales , Adhesión Celular , Comunicación Celular , Espacio Extracelular/metabolismo , Hibridomas/inmunología , Inmunohistoquímica , Ratones , Ratones Mutantes Neurológicos , Neuronas/química , Proteína Reelina , Serina Endopeptidasas , Distribución TisularRESUMEN
Idiopathic pulmonary fibrosis (IPF) seems to be closely associated with lung carcinogenesis. To identify the genetic characteristics of precancerous IPF lesions in the peripheral lung, we performed PCR-based microsatellite analysis with DNA extracted from microdissected tissues; fluorescent in situ hybridization (FISH) analysis of the fragile histidine triad (FHIT) gene and immunohistochemical analysis of Fhit protein expression in samples of metaplasias and bronchiolar epithelia obtained from patients with IPF. We used four microsatellite markers of the FHIT gene within or flanking the FHIT gene on chromosome 3p for loss of heterozygosity (LOH) analysis. LOH of the FHIT locus was frequently found among the lesions of metaplasias and bronchiolar epithelia in the patients with IPF [62 (52%) of 119 informative lesions]. Fifty-four (73%) of the 74 lesions of metaplasias and bronchiolar epithelia obtained from the IPF patients with lung carcinoma and 8 (17%) of the 46 samples obtained from the IPF patients without lung carcinoma showed LOH at the FHIT gene (P < 0.0001). We confirmed allelic loss in the metaplasias and bronchiolar epithelia of IPF by FISH analysis of the FHIT gene. Additionally, the level of Fhit protein expression in the metaplastic cells of IPF was frequently reduced. Our findings suggest that allelic loss of the FHIT gene may be involved in carcinogenesis in the peripheral lung of patients with IPF.
Asunto(s)
Ácido Anhídrido Hidrolasas , Pérdida de Heterocigocidad , Neoplasias Pulmonares/genética , Proteínas de Neoplasias/genética , Fibrosis Pulmonar/genética , Cromosomas Humanos Par 3/genética , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Lesiones Precancerosas/genéticaRESUMEN
The velocity of the aggregation of human erythrocytes was examined in the range of 5-43 degrees C with a rheoscope combined with a video camera, an image analyzer and a computer. (1) With increasing temperature, the velocity of erythrocyte aggregation induced by fibrinogen, immunoglobulin G and artificial macromolecules (dextran of 70 kDa and poly(glutamic acid) of 50 kDa) increased. However, the relationship between the velocity of erythrocyte aggregation and the temperature was different among these macromolecules. (2) In 70% autologous plasma, the velocity of erythrocyte aggregation was minimum at 15-18 degrees C, and increased at both higher and lower temperatures. (3) The shape of erythrocyte aggregates in 12 mumol/l fibrinogen (containing 770 mumol/l albumin) and in 70% autologous plasma was dependent on temperature: three-dimensional below 15-18 degrees C and one-dimensional (mainly rouleaux) above 15-18 degrees C. However, the shape of aggregates in 27 mumol/l immunoglobulin G (containing 770 mumol/l albumin) was three-dimensional in all temperature ranges. (4) The temperature dependency of erythrocyte aggregation was discussed in terms of the changes of medium viscosity, of erythrocyte properties and of bridging macromolecules.
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Eritrocitos/citología , Adulto , Agregación Celular , Eritrocitos/ultraestructura , Fibrinógeno , Humanos , Inmunoglobulina G , Cinética , Microscopía Electrónica de Rastreo , Valores de Referencia , TemperaturaRESUMEN
The contribution of membrane glycoproteins to the velocity of fibrinogen-induced erythrocyte aggregation was examined using a rheoscope combined with a video camera, an image analyzer and a computer. The structure of glycoproteins was modified with proteolytic enzymes, trypsin or alpha-chymotrypsin. (1) Mild enzymatic treatment of erythrocytes decreased the velocity of erythrocyte aggregation, but more intense treatment increased the velocity remarkably. (2) The erythrocyte aggregation was affected not only by the density of surface negative charge of erythrocytes, but also by the structural changes of glycoproteins. (3) Erythrocyte deformability and the morphological characteristics were not altered by these enzymatic treatments. The physiological significance of glycoproteins of erythrocyte surface for the survival of erythrocytes and for the suspension stability of blood was discussed.
Asunto(s)
Agregación Eritrocitaria/efectos de los fármacos , Fibrinógeno/farmacología , Glicoproteínas de Membrana/metabolismo , Adulto , Carbohidratos/análisis , Quimotripsina , Deformación Eritrocítica , Humanos , Hidrólisis , Indicadores y Reactivos , Masculino , Ácidos Siálicos/análisis , TripsinaRESUMEN
The effect of fibrinogen and fibrinogen-derived products on the velocity of rouleau formation of human erythrocytes was quantitatively examined with a rheoscope combined with a video-camera, an image analyzer and a computer. (i) The velocity of rouleau formation by naturally occurring low-molecular-weight fibrinogen of 305 kDa and by desialylated fibrinogen was the same as that by native fibrinogen of 340 kDa. (ii) Concerning fibrinogen degradation products by plasmin, the velocity of rouleau formation decreased upon going from fibrinogen greater than fragment X greater than fragment Y (the ratio of molar concentration of fibrinogen, fragment X and fragment Y for giving a certain velocity of rouleau formation was approx. 1:2:5). The effect of fragments X and Y on the fibrinogen-induced rouleau formation was additive. (iii) Fragments D and E could not induce rouleau formation and did not affect the fibrinogen-, fragment X- and fragment Y-induced rouleau formation. (iv) Fibrinopeptides A and B and artificial tetrapeptides (Gly-Pro-Arg-Pro and Gly-His-Arg-Pro) did not affect the fibrinogen-induced rouleau formation. (v) The possible erythrocyte-binding site in fibrinogen molecule for leading to rouleaux was proposed to be in A alpha-chain (probably, around residues No. 207-303) near the terminal domain of the trinodular structure of fibrinogen.
Asunto(s)
Agregación Eritrocitaria/efectos de los fármacos , Productos de Degradación de Fibrina-Fibrinógeno/farmacología , Fibrinógeno/farmacología , Sitios de Unión , Fibrinógeno/metabolismo , Fibrinolisina/metabolismo , Fibrinopéptido A/farmacología , Fibrinopéptido B/farmacología , Humanos , Masculino , Peso Molecular , Ácido N-Acetilneuramínico , Ácidos SiálicosRESUMEN
The p16INK4 gene, which is a tumor suppressor gene, is frequently altered in lung cancers. Hypermethylation of the promoter region of the p16INK4 gene seems to be the major mechanism through which p16INK4 become inactivated. Hypermethylation of the p16INK4 gene was reported to occur at an early stage in lung cancer. To determine whether the change in p16INK4 methylation status occurs at the late stage in the progression of primary lung cancers, we analyzed the primary and metastatic tumor tissues and normal lung samples from 29 cases of advanced lung cancer with distant metastasis. In each tissue sample, we analyzed the p16INK4 and p15INK4b genes for mutations and the methylation status of both genes using PCR-single strand conformation polymorphism, direct sequencing, and methylation-specific PCR analysis. We also analyzed a subset of the samples for p16INK4 protein expression. Genetic mutations in the coding region of the p16INK4 and p15INK4b genes were not found in any of the examined specimens. The promoter region of the p16INK4 gene was hypermethylated in the tumor samples of the primary or metastatic site of 37.0% (10 of 27) of the subjects. The promoter region of the p16INK4 gene was hypermethylated at both the primary and metastatic sites in two of the 10 cases and at only the metastatic site in 8 cases. By immunohistochemical analysis, we confirmed the presence of p16INK4 protein at the primary site of all cases in which the promoter region of the p16INK4 gene was hypermethylated at only the metastatic site. Interestingly, all 8 cases with a hypermethylated p16INK4 promoter region, at only the metastatic site, did not have p53 mutation. The results of this study indicate that tumor cells in which the p16INK4 gene has been inactivated by hypermethylation of the promoter region could have an advantage in progression and metastasis in non-small cell lung cancers, especially in the tumors with normal p53, and that the frequency of p16INK4 gene inactivation by hypermethylation could vary in clinical course.
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Proteínas de Ciclo Celular , Metilación de ADN , Genes p16 , Genes p53 , Neoplasias Pulmonares/genética , Proteínas Supresoras de Tumor , Adenocarcinoma/genética , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Células Escamosas/genética , Inhibidor p15 de las Quinasas Dependientes de la Ciclina , Femenino , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Mutación , Metástasis de la Neoplasia , Factores de Transcripción/genéticaRESUMEN
Non-small cell lung cancer is associated with approximately 85% mortality due to its high metastatic potential. Therapeutic efforts have failed to produce a significant improvement in prognosis. In this situation, a better understanding of the key factors of metastasis may be useful for designing new molecular targets of therapy. In order to identify these factors, we compared the expression profiles of two subpopulations of an adenocarcinoma cell line with a high metastatic potential, PC9/f9 and PC9/f14, with the parent cell line, PC9, using a cDNA array. The expression of 15 genes was found to be significantly enhanced or reduced in the highly metastatic subpopulations. The expression of matrix metalloproteinase-2 (MMP-2), plasminogen activator inhibitor-1 (PAI-1) and interleukin-1 (IL-1 alpha) were upregulated in the highly metastatic subpopulations, while the expression of carcinoembryonic antigen (CEA), caspase-5, Fas ligand, Prk/FNK, cyclin E, cyclin B1, Ki-67, proliferating cell nuclear antigen (PCNA), Smad4, macrophage proinflammatory human chemokine-3 alpha (MIP-3 alpha)/LARC, Met and CD44 were downregulated. Data from the literature suggest that the altered expression of MMP-2, PAI-1, IL-1 alpha, CEA, caspase-5, Fas ligand, Prk/FNK and Smad4 promotes the highly metastatic phenotype. The differential expression of these genes was confirmed by Northern blot analysis, standard reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR. This analysis in subpopulations of a lung cancer cell line indicated that the highly metastatic potential of lung cancer may be induced not by an alteration in the expression of a single gene, but by the accumulation of alterations in the expression of several genes involved in extracellular matrix (ECM) adhesion disruption, ECM degradation, escape from apoptosis, and resistance to transforming growth factor-beta(1) (TGF-beta(1)). Strategies for inhibiting metastasis of pulmonary adenocarcinoma should be designed accordingly.
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Adenocarcinoma/genética , Neoplasias Pulmonares/genética , Oncogenes/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/secundario , Animales , Northern Blotting , Regulación hacia Abajo , Matriz Extracelular/metabolismo , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Regulación hacia ArribaRESUMEN
The mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R) is involved in activating the transforming growth factor beta(1) (TGF-beta(1)), an inhibitor of the cell proliferation, and limiting the insulin-like growth factor 2 mediated-growth stimulation. The M6P/IGF2R gene has been reported to be mutated and deleted in various cancers, and is a candidate tumor suppressor gene. We studied the genomic structure of the M6P/IGF2R gene and designed the intron primers to detect mutations in the M6P/IGF2R gene of genomic DNA samples. The M6P/IGF2R gene consists of 48 exons. The previously reported 23 mutations of the M6P/IGF2R gene in human cancers, liver, breast, and gastrointestinal tumors, are located in five exons, exon 27, 28, 31, 40, 48. Using the intron primers designed in this study, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, and direct sequencing, we performed an initial analysis of the complete coding sequences of the M6P/IGF2R gene in 21 human cell lines resistant to growth inhibition by TGF-beta(1). An adenine-to-guanine transition, resulting in an asparagine-to-serine amino acid substitution, was found in one lung adenocarcinoma cell line at exon 40 where the mutation has been previously reported in human cancers. This is the first report of a mutation of the M6P/IGF2R gene in lung tumor. These results indicated that the mutation in M6P/IGF2R may be involved in human lung cancinogenesis.
Asunto(s)
Mutación , Receptor IGF Tipo 2/genética , Factor de Crecimiento Transformador beta/farmacología , Línea Celular , Análisis Mutacional de ADN , Cartilla de ADN , ADN de Neoplasias/genética , Resistencia a Antineoplásicos/genética , Exones , Genoma Humano , Humanos , Intrones , Reacción en Cadena de la Polimerasa , Factor de Crecimiento Transformador beta1 , Células Tumorales CultivadasRESUMEN
Some of the many human cancers that exhibit chromosomal instability also carry mutations in mitotic checkpoint genes and/or reveal reduced expression of some of those genes, such as hMAD2. To facilitate investigation of alterations of hMAD2, we determined its genomic structure and intronic primers designed to amplify the entire coding region. Since general impairment of the mitotic checkpoint is frequently reported in lung cancers, and reduced expression of hMAD2 has been reported in breast cancers as well, we searched for mutations throughout the coding sequence of this gene in the genomic DNA of 30 primary lung tumors, 30 lung-cancer cell lines and 48 primary breast cancers. Our approach, which involved polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis and direct sequencing, revealed nucleotide variants in only two of the 108 specimens. One was a cytosine-to-adenine substitution 3 bp upstream of exon 4 that occurred in one lung cancer cell line and one primary breast tumor, a change that did not alter transcriptional sequence. The other was an adenine-to-guanine substitution within exon 4, of the same lung cell line; this change already had been reported as a polymorphism. The results suggested that the hMAD2 gene is not commonly mutated in either lung nor breast cancers. Further studies should focus on other mechanisms that might account for reduced expression of the hMAD2 gene, and/or pursue analyses of other mitotic checkpoint genes for mutations in human cancer. Nevertheless, the genomic structure, the intronic primer sequences, and polymorphisms of the hMAD2 gene presented here will facilitate future studies to determine the full spectrum and frequency of the genetic events that can affect expression of the hMAD2 gene in human tumors.
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Neoplasias de la Mama/genética , Proteínas de Unión al Calcio/genética , Proteínas Portadoras , ADN de Neoplasias/análisis , Proteínas Fúngicas/genética , Neoplasias Pulmonares/genética , Proteínas de Unión al Calcio/biosíntesis , Proteínas de Ciclo Celular , Análisis Mutacional de ADN , Femenino , Proteínas Fúngicas/biosíntesis , Humanos , Masculino , Proteínas Nucleares , Polimorfismo Genético , Polimorfismo Conformacional Retorcido-Simple , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
The PTEN/MMAC1 gene located at 10q23, has been proposed to be a tumor suppressor gene. To determine the involvement of alteration of the PTEN/MMAC1 gene in carcinogenesis and the progression of primary lung cancers, we analyzed tumor samples of primary and distant metastatic sites and normal lung tissue samples of 30 patients with advanced lung cancer with distant metastasis. The tissues were analyzed for allelic deletion and mutational inactivation of PTEN/MMAC1 by loss of heterozygosity (LOH) analysis, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), and direct sequence analysis. LOH of the PTEN/MMAC1 locus was common in each histologic type of primary lung cancer. In this study, the overall allelic deletion rate was 33.3% (7/21). Allelic loss at the primary site and that at the metastatic site of each patient, were identical; in most cases, it seemed that the allelic loss had occurred before metastasis. Sequence analysis of the PTEN/MMAC1 gene revealed a G to C substitution located 8 bp upstream of the coding region of exon 1 and which seems to be a polymorphism, in 4 of the 30 cases. Somatic mutations of the PTEN/MMAC1 gene were not identified in any of the tumors at the primary and metastatic sites. These data indicate that point mutations in the PTEN/MMAC1 gene are probably not an important factor in tumorigenesis and the progression of a major subset of lung cancers. Due to frequent allelic loss at the PTEN/MMAC1 locus occurring at a stage earlier than the metastatic process, alternative mechanisms in which the remaining allele is inactivated such as methylation or homozygous deletion of a small region of the gene that can not be detected by the usual analysis, or alteration of other important tumor suppressor genes lying close to the PTEN/MMAC1 gene on 10q23, may be involved in the tumorigenesis of lung cancers of all histologic subtypes.
Asunto(s)
Eliminación de Gen , Genes Supresores de Tumor/genética , Neoplasias Pulmonares/genética , Metástasis de la Neoplasia/genética , Monoéster Fosfórico Hidrolasas/genética , Mutación Puntual , Proteínas Supresoras de Tumor , Alelos , Humanos , Pérdida de Heterocigocidad , Neoplasias Pulmonares/patología , Fosfohidrolasa PTEN , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
A population of neurons in the area of human thalamic nucleus ventralis caudalis (Vc) respond to noxious heat stimuli. In the cutaneous core of Vc 6% (6/108) of recorded neurons had a significantly greater response to noxious heat stimuli than to innocuous control stimuli. Half of these neurons (n = 3) also responded to innocuous cold stimuli. Within the region posterior and inferior to the cutaneous core of Vc 5% (4/77) of neurons responded exclusively to noxious heat stimuli. Cells responding to noxious heat were recorded at a greater proportion (66%) of sites where painful sensations were evoked by microstimulation than at sites where nonpainful sensations were evoked (1.5%). The results suggest that neurons in the region of human Vc mediate the sensory aspect of pain.
Asunto(s)
Calor , Neuronas/fisiología , Dolor/fisiopatología , Núcleos Talámicos/fisiología , Estimulación Eléctrica , Electrofisiología , Humanos , Trastornos del Movimiento/fisiopatología , Estimulación Física , Umbral Sensorial , Núcleos Talámicos/citologíaRESUMEN
A 64-year-old man suffering from crescendo brainstem symptoms due to acute total occlusion of the vertebrobasilar artery was successfully treated by cerebral artery stent placement. The total occlusion of a long segment of the vertebrobasilar artery was completely recanalized by implanting two flexible, balloon-expandable coronary stents. The patient's clinical outcome 30 days later was favorable. No complications occurred during or after the procedure. This therapeutic option may prove to be a useful means to revascularize an acute total occlusion of the vertebrobasilar artery.
Asunto(s)
Arteriopatías Oclusivas/terapia , Trastornos Cerebrovasculares/etiología , Stents , Insuficiencia Vertebrobasilar/complicaciones , Enfermedad Aguda , Humanos , Masculino , Persona de Mediana Edad , Cuidados Posoperatorios/métodos , Resultado del TratamientoRESUMEN
The genetic mechanisms involved in lung cancer development and progression are beginning to be understood. Many studies have documented frequent loss of heterozygosity (LOH) at specific chromosomal regions in cancer cells; this implies that tumor suppressor genes (TSG) are usually present in those regions. Recently, it has been reported that LOH or chromosomal deletions at chromosome 8p21-23 represent early events frequently occurring in lung cancer. In addition, the size of these chromosome 8 deletions, as well as their frequency, was also reported to increase during lung cancer progression. To determine the spectrum and frequency of alterations of chromosome 8p21-23 in human lung cancer and whether these increase with progression of the tumors, we performed LOH analysis of chromosome 8p and 3p in the genomic DNA from cells from primary and metastatic sites of lung cancer, as well as from normal lung. We studied 35 subjects with primary lung cancer including 30 tumors with distant metastasis. Detection of allelic deletion utilized a PCR-based approach of microsatellite polymorphism analysis, which was performed using the microsatellite markers D8S1130, D8S1106, D8S511, D8S1827, D8S549, D8S261, LPL, D8S258, D8S136, NEFL, D3S1295, D3S1313, D3S1234, D3S1300, D3S1351, D3S1339, and D3S1340. The overall allelic deletion rates were 10 of 28 (35.7%) at 8p and 13 of 33 (39.4%) at 3p. The allelic deletions in the primary cancer and its metastatic sites were in each case identical in both frequency and size of the deleted regions. In our analysis, 8p21-23 deletions were not always associated with 3p deletions in primary lung cancer. These results therefore suggest that allelic deletion at chromosome 8p21-23 is an early and frequent event in the carcinogenesis and development of lung cancer, independent of chromosome 3p deletion. However, a continuing increase in the frequency of LOH at 8p21-23 and in the size of the deleted region rarely occurs during the process of metastasis.
Asunto(s)
Deleción Cromosómica , Cromosomas Humanos Par 8/genética , Pérdida de Heterocigocidad/genética , Neoplasias Pulmonares/genética , Adenocarcinoma/genética , Adenocarcinoma/patología , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Cromosomas Humanos Par 3/genética , ADN de Neoplasias/genética , Frecuencia de los Genes , Humanos , Neoplasias Pulmonares/patología , Repeticiones de Microsatélite , Metástasis de la Neoplasia/genéticaRESUMEN
The effect of pH, temperature and osmotic pressure on velocity of erythrocyte aggregation was quantitatively examined with a rheoscope combined with a video-camera, an image analyzer and a computer, (a) in an artificial medium containing fibrinogen and albumin and (b) in diluted autologous plasma. (1) With increasing pH of the medium, the velocity of erythrocyte aggregation increased. (2) The velocity of erythrocyte aggregation in the artificial medium increased as the temperature rose. However, in 70% autologous plasma the velocity was minimum at 15-18 degrees C, increasing both above and below this temperature (above 30 degrees C, the velocity saturated). (3) The velocity of erythrocyte aggregation decreased in hypotonic medium, while it increased in hypertonic medium (at osmotic pressures higher than 400 mOsm, the velocity decreased). The mechanism of erythrocyte aggregation is discussed with special reference to the morphological changes produced by pH, temperature and osmotic pressure, and the implications of the phenomena for oxygen transport to tissues in (patho)physiological situations are considered.
Asunto(s)
Velocidad del Flujo Sanguíneo , Eritrocitos/citología , Adulto , Agregación Celular , Humanos , Concentración de Iones de Hidrógeno , Técnicas In Vitro , Cinética , Masculino , Presión Osmótica , Valores de Referencia , TemperaturaRESUMEN
A 53-year-old woman was admitted because of Raynaud's phenomenon, polyarthralgia and polymyalgia. Biopsy specimens of the liver and thyroid gland revealed characteristic findings of primary biliary cirrhosis (PBC) (stage I by Scheuer's classification) and chronic thyroiditis. Her clinical features were also complicated by scleroderma (type I by Barnett's classification) and Sjögren's syndrome (Sjs) with keratoconjunctivitis sicca. Thyroid hormone replacement therapy led to improvement in thyroid function, normalization of the biliary tract enzymes and alleviation of subjective symptoms.