RESUMEN
BACKGROUND: Although asthma is a common cause of morbidity in adults, relatively few objectively measured population studies of asthma prevalence in adult populations have been conducted. OBJECTIVE: To evaluate the prevalence of asthma, based on both a questionnaire and methacholine bronchial provocation test, and to determine the risk factors of asthma prevalence in an adult population. METHODS: A total of 2,467 adults, who were randomly selected from metropolitan urban, non-metropolitan urban and rural areas, responded to the modified ISAAC questionnaire, and underwent methacholine bronchial provocation tests and skin prick tests to locally common aeroallergens. RESULTS: The prevalence of current asthma based on the questionnaire and the methacholine challenge was 2.0% in adults younger than 40, 3.8% in 40- to 54-year-olds, 7.7% in 55- to 64-year-olds and 12.7% in those aged 65 or higher. For subjects of 55-64 years, active smoking was found to be significantly related with the prevalence of current asthma and bronchial hyper-responsiveness, although smoking was positively associated with percentage predictive value of forced expiratory volume of 1 s (FEV1). CONCLUSION: The prevalence of current asthma is common among the elderly, and active smoking may play an important role in the development of asthma and bronchial hyper-responsiveness among the elderly.
Asunto(s)
Asma/epidemiología , Asma/etiología , Fumar/efectos adversos , Adulto , Distribución por Edad , Anciano , Asma/fisiopatología , Pruebas de Provocación Bronquial , Broncoconstrictores , Femenino , Humanos , Hipersensibilidad Inmediata/epidemiología , Corea (Geográfico)/epidemiología , Modelos Logísticos , Pulmón/fisiopatología , Masculino , Cloruro de Metacolina , Persona de Mediana Edad , Prevalencia , Salud Rural , Pruebas Cutáneas , Fumar/epidemiología , Salud UrbanaAsunto(s)
Compuestos de Calcio/administración & dosificación , Difosfonatos/administración & dosificación , Fracturas Espontáneas/prevención & control , Terapia de Reemplazo de Hormonas/métodos , Osteoporosis Posmenopáusica/tratamiento farmacológico , Osteoporosis Posmenopáusica/prevención & control , Anciano , Anciano de 80 o más Años , Densidad Ósea/fisiología , Intervalos de Confianza , Densitometría , Femenino , Humanos , Persona de Mediana Edad , Prevención Primaria/métodos , Pronóstico , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del TratamientoRESUMEN
BACKGROUND: Insulin-dependent diabetes mellitus (IDDM) is caused by the autoimmune destruction of pancreatic beta-cells. Susceptibility to IDDM appears to depend on more than one genetic locus. Evidence of a genetic linkage for IDDM2 was found in male meioses from French and North American populations. It is linked to maternal imprinting (i.e. monoalleleic expression of the insulin gene) that is considered the most likely cause of these gender-related differences. IGF2 is expressed only in the paternal allele and, therefore, is considered a candidate gene for IDDM2 transmission because of its important autocrine/paracrine effects on the thymus, lymphocytes and pancreas. Nevertheless, it remains controversial whether the parental origin of IDDM2 influences IDDM susceptibility. METHODS: Using PCR and semi-quantitative RT-PCR, we analyzed the INS/Pstl + 1127 and IGF2/Apal polymorphisms and RNA expression level between Pstl (+/-) and Pstl (+/+) to determine genotype and allele-specific expression of the INS and IGF2 genes. RESULTS: INS/Pstl (+/+) and IGF2/Apal (+/-) were observed in 36 (97.3%) of 37 IDDM patients and in 29 (72.5%) of 40 IDDM patients, respectively. The presence of both IGF2 alleles in RNA was observed in 21 (91.6%) of 24 IDDM patients. Our results show a 3-fold increase in RNA expression from Pstl (+/-) allele over Pstl (+/+) allele. CONCLUSION: Our conclusion does not entirely exclude IGF2 as the gene involved in IDDM2, even though the parental effect of IDDM2 transmission is not related to IGF2 maternal imprinting. The INS genotype appeared mostly in the Pstl (+/+) homozygote and, therefore, we could not explain the INS imprinting pattern in Korean type 1 diabetic patients. Genetic differences between populations may account for the discrepancy between Korean type I diabetic patients and American or French type I diabetic patients.
Asunto(s)
Diabetes Mellitus Tipo 1/genética , Impresión Genómica , Factor II del Crecimiento Similar a la Insulina/genética , Insulina/genética , Adolescente , Niño , Femenino , Humanos , Corea (Geográfico) , Masculino , Factores SexualesAsunto(s)
Acetolactato Sintasa/metabolismo , Disulfuros/metabolismo , Nicotiana/enzimología , Plantas Tóxicas , Acetolactato Sintasa/química , Acetolactato Sintasa/genética , Disulfuros/análisis , Ácido Ditionitrobenzoico/metabolismo , Cinética , Mutación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Compuestos de Sulfhidrilo/análisis , Compuestos de Sulfhidrilo/metabolismo , Reactivos de Sulfhidrilo/metabolismo , Tiocianatos/metabolismoRESUMEN
Acetolactate synthase (ALS) is the common enzyme in the biosynthesis of valine, leucine, and isoleucine. The role of four cysteinyl residues in tobacco ALS was determined using site-directed mutagenesis and cysteine-specific cleavage. The C411A mutation abolished the enzymatic activity, as well as the binding affinity for the cofactor FAD. The activation constant of C411S for FAD is approximately 50-fold higher than that of wALS. The C607S mutation did not significantly affect the kinetic parameters. The IC(50) values of C411S and C607S for ALS-inhibiting herbicides are not much different from those of wALS. Two mutants, C163S and C309S, are labile and readily degraded to peptide fragments. The treatment of wALS with 2-nitro-5-thiocyanobenzoic acid, specific for cleavage of the N-terminal side of cysteine, yielded three peptides of 37.0, 22. 0, and 7.0 kDa. This fragmentation pattern is consistent with that deduced from the amino acid sequence of tobacco ALS, assuming the disulfide bond between Cys163 and Cys309. These results suggest that Cys411 is involved in the binding of FAD and that the intrachain disulfide bond between Cys163 and Cys309 plays a key role in maintaining the correct conformation of tobacco ALS.
Asunto(s)
Acetolactato Sintasa/química , Cisteína/química , Nicotiana/enzimología , Plantas Tóxicas , Acetolactato Sintasa/genética , Disulfuros/química , Electroforesis en Gel de Poliacrilamida , Inhibidores Enzimáticos/farmacología , Flavina-Adenina Dinucleótido/metabolismo , Herbicidas/farmacología , Cinética , Mutagénesis Sitio-Dirigida , Fragmentos de Péptidos/química , Unión Proteica/genética , Conformación Proteica , Espectrofotometría , Tiocianatos/químicaRESUMEN
PURPOSE: Angiostatin is a potent angiogenesis inhibitor that has been identified as a cryptic fragment of plasminogen molecule containing the first four kringle domain. Angiogenin, a 14-kDa monomeric protein, a potent blood vessel inducer, is expressed in tumors and present in mammalian plasma. The purpose of this study was to determine whether recombinant kringle 1-3 (rKI-3) of human plasminogen could interfere with angiogenesis induced by angiogenin and to evaluate the role of angiogenin in corneal angiogenesis in rabbit. METHODS: A Hydron polymer pellet containing 2.0 microg of angiogenin was implanted intrastromally into the superior cornea of each of 44 rabbit eyes. All eyes received an intrastromal pellet and were randomized into either one group treated with 12.5 microg of rKI-3 (n = 25) or the other group treated with phosphate-buffered saline (PBS; n = 19). Both pellets were positioned in parallel at the site 1.2 mm from the superior limbus. Two masked observers kept the angiogenesis score daily, based on the number and the length of new vessels. The corneas with induced angiogenesis also were examined histologically. RESULTS: On the third day of the angiogenin pellets implantation, the eye treated with rKI-3 had less angiogenesis (mean score, 4.2 +/- 6.6) than eye treated with PBS (mean score, 16.1 +/- 17.1; p < 0.05, Mann-Whitney U test). The cornea treated with PBS also showed much more leukocyte adhesion than the cornea treated with rKI-3. CONCLUSION: Recombinant kringle 1-3 appears to inhibit angiogenin-induced angiogenesis in a rabbit corneal pocket assay. Recombinant kringle 1-3 may have therapeutic potential as an antiangiogenic agent.
Asunto(s)
Inductores de la Angiogénesis/toxicidad , Antineoplásicos/administración & dosificación , Córnea/patología , Neovascularización de la Córnea/tratamiento farmacológico , Fragmentos de Péptidos/administración & dosificación , Plasminógeno/administración & dosificación , Ribonucleasa Pancreática/toxicidad , Animales , Adhesión Celular/efectos de los fármacos , Córnea/efectos de los fármacos , Neovascularización de la Córnea/inducido químicamente , Neovascularización de la Córnea/patología , Modelos Animales de Enfermedad , Implantes de Medicamentos , Leucocitos/efectos de los fármacos , Leucocitos/fisiología , Masculino , Conejos , Distribución Aleatoria , Proteínas RecombinantesRESUMEN
Kringle 1-3 of human plasminogen is a potent inhibitor of endothelial cell proliferation. To understand a possible role for the unique cystine bridge between kringle 2 and kringle 3, we disrupted the interkringle disulfide bond by mutating Cys(169) and Cys(297) to serine residues. The yield of the mutant during the refolding process was decreased significantly. Anti-endothelial cell proliferative activity of the mutant was similar to that of the wild type. There was no significant difference in in vivo antiangiogenic activity between the wild type and the mutant in chorioallantoic membrane assay. However, in the mutant, the weak lysine binding capability of kringle 2 was not detected and its mobility in nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis is different from that of the wild type. These results support the notion that the overall antiangiogenic function of angiostatin is mediated by individual kringles, and suggest that the lysine binding capability of kringle 2 is likely not important for the antiangiogenic activity of kringle 1-3.
Asunto(s)
Disulfuros/metabolismo , Kringles/fisiología , Lisina/metabolismo , Neovascularización Fisiológica , Plasminógeno/química , Plasminógeno/metabolismo , Sustitución de Aminoácidos/genética , Inhibidores de la Angiogénesis/química , Inhibidores de la Angiogénesis/genética , Inhibidores de la Angiogénesis/metabolismo , Inhibidores de la Angiogénesis/farmacología , Animales , Bovinos , División Celular/efectos de los fármacos , Embrión de Pollo , Corion/citología , Corion/efectos de los fármacos , Corion/fisiología , Cisteína/genética , Cisteína/metabolismo , Electroforesis en Gel de Poliacrilamida , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiología , Humanos , Kringles/genética , Ligandos , Mutación/genética , Neovascularización Fisiológica/efectos de los fármacos , Plasminógeno/genética , Plasminógeno/farmacología , Unión Proteica , Pliegue de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , TermodinámicaRESUMEN
Human malignant gliomas are highly vascularized and aggressive tumors. Angiogenesis inhibitors have been shown to induce regression of a variety of primary and metastatic tumors in vivo. However, their usefulness in treating brain tumors is not well understood. Angiostatin, a multiple kringle (1-4 of 5)-containing fragment of plasminogen, is one of the highly effective natural cryptic angiogenesis inhibitors. In our study, the therapeutic efficacy of non-glycosylated and small molecular size recombinant kringles 1-3 (rPK1-3) was examined in the treatment of brain tumors generated by stereotactic intracerebral implantation of U-87 human glioma cells in nude mice. Mice bearing tumors 7 days post-implant were treated daily with rPK1-3 (100 mg/kg) s.c. for 21 days. Treated animals showed suppressed brain tumor growth by greater than 71.2% along with a 3-fold increase of apoptotic index and suppressed vascularization by 78.9%, without any observable signs of toxicity. Analysis of bFGF and VEGF expression in the tumors of treated animals using immuno-histochemical methods showed near complete absence of growth factors. Our results indicate that the non-glycosylated, small molecular size rPK1-3 is an efficient tumoristatic agent for the treatment of intracranial human glioma xenografts in mice and might provide new strategies for the treatment of brain tumors.
Asunto(s)
Glioma/patología , Kringles/genética , Neoplasias Experimentales/prevención & control , Plasminógeno/uso terapéutico , Animales , Apoptosis , Neoplasias Encefálicas/patología , División Celular/efectos de los fármacos , Factores de Crecimiento Endotelial/biosíntesis , Factores de Crecimiento de Fibroblastos/biosíntesis , Humanos , Linfocinas/biosíntesis , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neovascularización Patológica , Fragmentos de Péptidos/uso terapéutico , Plasminógeno/genética , Proteínas Recombinantes/uso terapéutico , Células Tumorales Cultivadas , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine. ALS is the target of three classes of herbicides, the sulfonylureas, the imidazolinones, and the triazolopyrimidines. Five mutants (W266F, W439F, W490F, W503F, and W573F) of the ALS gene from Nicotiana tabacum were constructed and expressed in Escherichia coli, and the enzymes were purified. The W490F mutation abolished the binding affinity for cofactor FAD and inactivated the enzyme. The replacement of Trp573 by Phe yielded a mutant ALS resistant to the three classes of herbicides. The other three mutations, W266F, W439F, and W503F, did not significantly affect the enzymatic properties and the sensitivity to the herbicides. These results indicate that the Trp490 residue is essential for the binding of FAD and that Trp573 is located at the herbicide binding site. The data also suggest that the three classes of herbicides bind ALS competitively.
Asunto(s)
Acetolactato Sintasa/química , Nicotiana/enzimología , Plantas Tóxicas , Triptófano/química , Acetolactato Sintasa/genética , Escherichia coli/enzimología , Cinética , Mutagénesis Sitio-Dirigida , Mutación , Proteínas Recombinantes de Fusión/genética , Espectrometría de Fluorescencia , EspectrofotometríaRESUMEN
Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of L-leucine, L-isoleucine, and L-valine. Triazolopyrimidine sulfonamide (TP) is a mixed-type inhibitor of ALS with respect to both pyruvate and thiamine pyrophosphate. In this study, we synthesized new substituted quinoline-linked TP analogues and several TP analogues which contained either unsubstituted aminoquinolines or amino isoquinolines. In addition, we examined the interactions of both the wild-type and the sulfonylurea-resistant recombinant tobacco ALS enzymes in a highly pure and active form with the quinoline-linked TP analogues, respectively. The wild-type tobacco ALS was extremely sensitive to inhibition by the quinoline-linked TP analogues. In contrast, the mutant tobacco ALS was insensitive to both the quinoline-linked triazolopyrimidine and the sulfonylurea herbicides. The results indicate that the ability of the quinoline-linked TP analogues to inhibit ALS is highly sensitive to substitution at the ortho position (C-7) and to the position of the ring nitrogen around the sulfonamide functionality (C-8).
Asunto(s)
Acetolactato Sintasa/química , Nicotiana/enzimología , Plantas Tóxicas , Quinolinas/química , Proteínas Recombinantes/química , Triazoles/síntesis química , Acetolactato Sintasa/genética , Herbicidas/química , Mutación , Triazoles/químicaRESUMEN
Angiogenin is a potent angiogenic factor secreted by cultured tumor cells and is found in various normal organs and tissues. The ovary is one of the adult organs in which angiogenesis normally occurs during the female reproductive cycle. In this study, we examined whether angiogenin protein is localized and if angiogenin mRNA is expressed in the normal bovine ovary by immunohistochemistry using polyclonal rabbit anti-bovine angiogenin IgG and by in situ hybridization using bovine angiogenin probe, respectively. The localization and mRNA expression of angiogenin in the ovarian follicle and in the corpus luteum were different in their developmental stages. The intensities of immunoreactivities and angiogenin transcripts in the follicle increased from the primordial to the tertiary (or Graafian) follicle. The early corpus luteum contained strong immunoreactivities and mRNA expression of angiogenin but these intensities weakened during regression. The results suggest that angiogenin is involved in morphological changes and angiogenesis in the ovary.
Asunto(s)
Neovascularización Fisiológica , Ovario/irrigación sanguínea , Ovario/crecimiento & desarrollo , Proteínas/metabolismo , Ribonucleasa Pancreática , Animales , Bovinos , Cuerpo Lúteo/citología , Cuerpo Lúteo/crecimiento & desarrollo , Cuerpo Lúteo/metabolismo , Endotelio Vascular/citología , Endotelio Vascular/metabolismo , Factor VIII/análisis , Femenino , Regulación del Desarrollo de la Expresión Génica , Células de la Granulosa/metabolismo , Sueros Inmunes , Inmunohistoquímica , Hibridación in Situ , Células Lúteas/metabolismo , Músculo Liso Vascular/citología , Músculo Liso Vascular/metabolismo , Oocitos , Folículo Ovárico/citología , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Ovario/citología , Ovario/metabolismo , Proteínas/genética , ARN Mensajero/metabolismoRESUMEN
Endostatin, a carboxyl-terminal fragment of collagen XVIII is known as an anti-angiogenic agent, that specifically inhibits the proliferation of endothelial cell and the growth of several primary tumor. We report here the purification and characterization of the recombinant murine endostatin (rmEndostatin) which was expressed in a prokaryotic expression system. This rmEndostatin has similar physiochemical properties of yeast-produced recombinant endostatin, and it also specifically inhibits the proliferation and migration of bovine capillary endothelial cells stimulated by basic fibroblast growth factor. The biological activity of rmEndostatin was also shown by its anti-angiogenic ability on the chorioallantoic membrane of chick embryo in vivo. In this article, we demonstrate the refolding and purification of rmEndostatin, expressed using E. coli system, to a biologically active and soluble form. In addition, these results confirm the activity of endostatin as a potent anti-angiogenic agent.
Asunto(s)
Inhibidores de la Angiogénesis/genética , Inhibidores de la Angiogénesis/farmacología , Colágeno/genética , Colágeno/farmacología , Escherichia coli/genética , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/farmacología , Inhibidores de la Angiogénesis/aislamiento & purificación , Animales , Western Blotting , Bovinos , Movimiento Celular/efectos de los fármacos , Embrión de Pollo , Corion/efectos de los fármacos , Corion/patología , Dicroismo Circular , Colágeno/aislamiento & purificación , Colágeno Tipo XVIII , Electroforesis en Gel de Poliacrilamida , Endostatinas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Factor 2 de Crecimiento de Fibroblastos/farmacología , Ratones , Neovascularización Fisiológica/efectos de los fármacos , Fragmentos de Péptidos/aislamiento & purificación , Pliegue de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/farmacología , Solubilidad , Levaduras/genéticaRESUMEN
Angiostatin is a potent angiogenesis inhibitor that is composed of the first four kringles of plasminogen fragment. Angiostatin with one less kringle molecule (kringle 1 to 3) was recently demonstrated to be an effective angiogenic inhibitor. To determine whether recombinant plasminogen kringle 1-3 (rPK1-3) can inhibit the corneal neovascularization induced by potent angiogenic factors; angiogenin, bFGF, or VEGF, hydron polymer discs each containing 2.0 microg of angiogenin, 500 ng of bFGF, or 500 ng of VEGF respectively were implanted into the corneal stroma of 138 rabbit eyes, and then discs each containing 10 microg, 12.5 microg, 20 microg or 30 microg of rPK1-3 were implanted randomly. Discs containing phosphate buffered saline were also implanted as a control. The angiogenesis score on number and length of newly formed vessels on the each of the rabbit's cornea were recorded daily by two observers (blinded). The treated corneas were also examined histologically. Recombinant PK1-3 treated corneas showed less neovascularization induced by all angiogenic factors (p < 0.05). and the extent of inhibition of neovascularization was proportional to the concentration of rPK1-3 (p < 0.05). Histologic examination showed leukocyte infiltration into the corneal stroma on the PBS treated eyes whereas rPK1-3 treated eyes showed only traces of leukocytes. These results of the effective rPK1-3 inhibition of corneal neovascularization induced by angiogenin, bFGF, or VEGF suggest that this angiostatin related fragment, rPK1-3, may be useful in the treatment of various neovascular diseases.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Córnea/irrigación sanguínea , Neovascularización Patológica/tratamiento farmacológico , Plasminógeno/genética , Plasminógeno/farmacología , Inhibidores de la Angiogénesis/genética , Animales , Embrión de Pollo , Corion/irrigación sanguínea , Corion/efectos de los fármacos , Córnea/efectos de los fármacos , Córnea/patología , Factores de Crecimiento Endotelial/farmacología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Kringles/genética , Linfocinas/farmacología , Microscopía/métodos , Conejos , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacología , Ribonucleasa Pancreática/farmacología , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
Several human imprinted genes have been identified and are implicated in genetic diseases and tumorigenesis. We studied alterations of two imprinted genes, the paternally imprinted H19 and maternally imprinted IGF2, in 15 ovarian tumors with various cell types. To know allele-specific expression of the two genes, we analyzed restriction fragment length polymorphisms (RFLPs) at the 3'-untranslated region (UTR) in their cDNA, compared with those in the respective genomic DNA. As a result, biallelic H19 and IGF2 expression was observed in 8 (62%) of 13 informative (heterozygous) ovarian cancers and in 6 of 11 informative cases, respectively. H19 loss of imprinting (LOI) was most frequently observed in malignant serous cystadenocarcinoma (in four of six cases), whereas IGF2 LOI was not common in malignant epithelial cancers because three of six such LOI events occurred in benign mucinous cystadenomas and non-cancerous endometriotic cyst. Our data suggest that the alteration of H19 and IGF2 imprinting plays differential roles in tumorigenesis and progression of ovarian tumors, depending on the tissue type as well as the developmental stage. Our data may argue against tumor suppressor activity of H19 in ovarian cancers.
Asunto(s)
Impresión Genómica , Factor II del Crecimiento Similar a la Insulina/genética , Proteínas Musculares/genética , Neoplasias Ováricas/genética , ARN no Traducido , Alelos , ADN de Neoplasias/análisis , ADN de Neoplasias/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Genes Supresores de Tumor/genética , Humanos , Neoplasias Ováricas/patología , Polimorfismo de Longitud del Fragmento de Restricción , ARN Largo no CodificanteRESUMEN
Despite the fact that Hodgkin's and Reed-Sternberg (H-RS) cells are morphological hallmarks of Hodgkin's disease (HD), the nature of H-RS cells still remains to be resolved. Here we report that downregulation of CD99 (Mic2) leads to the generation of cells with an H-RS phenotype. IM9 and BJAB B-cell lines that were transfected with an antisense CD99 expression construct showed the morphological and immunological characteristics of H-RS cells such as multinuclearity, expression of CD15, decreased expression of major histocompatibility complex (MHC) class I and CD45RB, and deregulated secretion of cytokines. The reduced expression of CD99 was also confirmed in H-RS cells of patient's lymph nodes and three HD-derived cell lines, L428, KM-H2, and HDLM-2. Moreover, features characteristic of H-RS cells were completely abolished by forced expression of CD99 and by a constitutively active form of Rac, which functions downstream of CD99. We suggest that CD99 molecules play a crucial role in regulating functions and morphology of cells through a Rac-Rho signaling pathway and that the loss of CD99 expression is a significant molecular event to generate H-RS cells.
Asunto(s)
Antígenos CD/genética , Antígenos CD/inmunología , Moléculas de Adhesión Celular/genética , Moléculas de Adhesión Celular/inmunología , Enfermedad de Hodgkin/patología , Células de Reed-Sternberg/inmunología , Células de Reed-Sternberg/patología , Antígeno 12E7 , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Antígenos de Histocompatibilidad Clase I/genética , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Antígenos Comunes de Leucocito/genética , Antígenos Comunes de Leucocito/inmunología , Oligonucleótidos Antisentido/genética , TransfecciónRESUMEN
OBJECTIVE: To provide an overview of key pharmacotherapeutic issues in epilepsy for the woman of childbearing potential. DATA SOURCES: A MEDLINE search (1966-1997) was done to identify pertinent literature. Chapters in epilepsy textbooks, pregnancy registries, and their respective bibliographies were also evaluated. STUDY SELECTION AND DATA EXTRACTION: All identifiable sources written in English were evaluated. DATA SYNTHESIS: Epilepsy is a common neurologic disorder. It is estimated that nearly 1 million American women of childbearing age have epilepsy. There are many women's health issues in epilepsy. These include menstrual cycle influences on seizure activity, contraceptive-antiepileptic drug interactions, pharmacokinetic changes during pregnancy, teratogenicity of antiepileptic drugs, breast-feeding, and quality of life. These issues challenge both the woman with epilepsy and the many healthcare providers involved in her care. This article reviews these issues and makes recommendations. It addresses both the first-generation antiepileptic drugs (phenobarbital, phenytoin, carbamazepine, valproic acid) and the newer or second-generation agents (felbamate, gabapentin, lamotrigine, topiramate, tiagabine). CONCLUSIONS/RECOMMENDATIONS: Drug interactions between enzyme-inducing antiepileptic drugs and contraceptives are well documented. Higher doses of oral contraceptives or a second contraceptive method are suggested if epileptic women use an enzyme-inducing antiepileptic drug. Planned pregnancy is highly recommended and counseling before conception is crucial. Prepregnancy counseling should include, but is not limited to, folic acid supplementation, optimal control of seizure activity, monotherapy with the lowest effective antiepileptic drug dose, and medication adherence. Patient information should be provided about the risk of teratogenicity and the importance of prenatal care. Antiepileptic drug dosage adjustments may be necessary and should be based on clinical symptoms, not solely on serum drug concentrations. While the future holds promise for many aforementioned women's issues in epilepsy, many questions remain to be answered.
Asunto(s)
Anticonvulsivantes/farmacología , Anticonvulsivantes/uso terapéutico , Epilepsia/tratamiento farmacológico , Adolescente , Adulto , Anticonvulsivantes/efectos adversos , Anticonceptivos/efectos adversos , Interacciones Farmacológicas , Femenino , Humanos , Embarazo , Complicaciones del Embarazo/tratamiento farmacológico , Complicaciones del Embarazo/prevención & controlRESUMEN
Transactivating factor PuF which interacts with a nuclease hypersensitive element locates upstream from the c-myc gene. PuF was recently identified as being encoded by nm23-H2/ NDP kinase gene [Postel, E. H., Berberich, S. J., Flint, S. J., and Ferrone, C. A. (1993) Science 261, 428-429]. Here we have studied the correlation of expression between c-myc and nm23 genes in vitro. By a comparative study of the expression of 2 genes in cell lines, no direct correlation of kinetics was found. A plasmid containing the human c-myc fragment (c-myc CAT) was cloned upstream from the bacterial chloramphenicol acetyltransferase (CAT) gene. When the murine melanoma cell line was cotransfected with a nm23-M2/ NDP kinase expression vector and c-myc CAT, CAT activity was elevated. In addition, cell cycle phases in the murine cell lines transfected with nm23/NDP kinase were estimated; an alteration of the cell cycle, prolonged S-phase was found in the cell lines transfected with nm23-M2/NDP kinase, whereas human nm23-H2/NDP kinase did not play a role in transactivating the c-myc gene or in S-phase prolongation in murine cell lines. From these results we conclude that the murine nm23-M2 gene transactivates the c-myc gene and controls the cell cycle, S-phase, indirectly via a cellular cofactor in the murine cell line, which does not work with the human nm23-H2 gene.
Asunto(s)
Genes myc , Proteínas de Unión al GTP Monoméricas , Proteínas de Neoplasias/fisiología , Nucleósido-Difosfato Quinasa/fisiología , Factores de Transcripción/fisiología , Animales , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Genes myc/efectos de los fármacos , Melanoma , Ratones , Nucleósido Difosfato Quinasas NM23 , Proteínas de Neoplasias/biosíntesis , Nucleósido-Difosfato Quinasa/biosíntesis , Factores de Transcripción/biosíntesis , Transfección , Células Tumorales CultivadasRESUMEN
Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of L-leucine, L-isoleucine, and L-valine. The wild-type ALS gene from Nicotiana tabacum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS2 was used to transform Escherichia coli strain XL1-Blue, and the wild-type tobacco ALS (wALS) was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). The fusion product GST-wALS was purified in a single step on a glutathione-Sepharose column. The purified GST-wALS was sensitive to a sulfonylurea herbicide, and was lost its sensitivity to end products, L-valine, L-leucine and L-isoleucine. These results suggest that the purified recombinant tobacco ALS was functionally active, and that the sulfonylureas may not bind to the feedback regulatory site on the plant ALS.