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Many studies have been devoted to adapting the design of gold nanoparticles to efficiently exploit their promising capability to enhance the effects of radiotherapy. In particular, the addition of magnetic resonance imaging modality constitutes an attractive strategy for enhancing the selectivity of radiotherapy since it allows the determination of the most suited delay between the injection of nanoparticles and irradiation. This requires the functionalization of the gold core by an organic shell composed of thiolated gadolinium chelates. The risk of nephrogenic systemic fibrosis induced by the release of gadolinium ions should encourage the use of macrocyclic chelators which form highly stable and inert complexes with gadolinium ions. In this context, three types of gold nanoparticles (Au@DTDOTA, Au@TADOTA and Au@TADOTAGA) combining MRI, nuclear imaging and radiosensitization have been developed with different macrocyclic ligands anchored onto the gold cores. Despite similarities in size and organic shell composition, the distribution of gadolinium chelate-coated gold nanoparticles (Au@TADOTA-Gd and Au@TADOTAGA-Gd) in the tumor zone is clearly different. As a result, the intravenous injection of Au@TADOTAGA-Gd prior to the irradiation of 9L gliosarcoma bearing rats leads to the highest increase in lifespan whereas the radiophysical effects of Au@TADOTAGA-Gd and Au@TADOTA-Gd are very similar.
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BACKGROUND: Although K14E6 transgenic mice develop spontaneous tumors of the skin epithelium, no spontaneous reproductive tract malignancies arise, unless the transgenic mice were treated chronically with 17beta-estradiol. These findings suggest that E6 performs critical functions in normal adult cervix and skin, highlighting the need to define E6-controlled transcriptional programs in these tissues. METHODS: We evaluated the expression profile of 14,000 genes in skin or cervix from young K14E6 transgenic mice compared with nontransgenic. To identify differentially expressed genes a linear model was implemented using R and the LIMMA package. Two criteria were used to select the set of relevant genes. First a set of genes with a Log-odds > or = 3 were selected. Then, a hierarchical search of genes was based on Log Fold Changes. RESULTS: Microarray analysis identified a total of 676 and 1154 genes that were significantly up and down-regulated, respectively, in skin from K14E6 transgenic mice. On the other hand, in the cervix from K14E6 transgenic mice we found that only 97 and 252 genes were significantly up and down-regulated, respectively. One of the most affected processes in the skin from K14E6 transgenic mice was the cell cycle. We also found that skin from transgenic mice showed down-regulation of pro-apoptotic genes and genes related to the immune response. In the cervix of K14E6 transgenic mice, we could not find affected any gene related to the cell cycle and apoptosis pathways but did observe alterations in the expression of immune response genes. Pathways such as angiogenesis, cell junction and epidermis development, also were altered in their gene expression profiles in both tissues. CONCLUSION: Expression of the HPV16 E6 oncoprotein in our model alters expression of genes that fell into several functional groups providing insights into pathways by which E6 deregulate cell cycle progression, apoptosis, the host resistance to infection and immune function, providing new opportunities for early diagnostic markers and therapeutic drug targets.
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Cuello del Útero/fisiología , Cuello del Útero/virología , Perfilación de la Expresión Génica , Proteínas Oncogénicas Virales/genética , Proteínas Represoras/genética , Piel/virología , Animales , Femenino , Papillomavirus Humano 16 , Hibridación in Situ , Ratones , Ratones Transgénicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The complete human genome sequence will facilitate the identification of all genes that contribute to disease. We propose that the functional classification of disease genes and their products will reveal general principles of human disease. We have determined functional categories for nearly 1,000 documented disease genes, and found striking correlations between the function of the gene product and features of disease, such as age of onset and mode of inheritance. As knowledge of disease genes grows, including those contributing to complex traits, more sophisticated analyses will be possible; their results will yield a deeper understanding of disease and an enhanced integration of medicine with biology.
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Enfermedades Genéticas Congénitas , Proyecto Genoma Humano , Enfermedad , Genes , HumanosRESUMEN
Systematic detection of inborn errors of metabolism (IEM) has usually encountered difficulties in developing countries. We present our experience in a high-risk population in Mexico between 1973 and 1998 with particular reference to the last 10 years, during which time infrastructure and support were considerably improved. Only disorders of intermediary metabolism were sought. The total number of patients studied is not available, but in the last 10 years, patients numbered 5,186. Routine metabolic screening was performed on all patients, with additional tests according to the clinical picture and screening results. The referral criteria have increasingly diversified, one-third being neurological conditions. Of the referrals, 33.8% were from pediatricians (31.1% of whom were at critical medicine departments) and the remainder from specialists. The number of diagnosed patients has increased to 1 per 43.9 patients studied. Amino acid defects have been the most prevalent, the proportion of organic acid and carbohydrate disorders having increased in the last 10 years, associated with improved diagnostic facilities. The most frequently diagnosed diseases were PKU, type 1a glycogen storage, and maple syrup urine disease (MSUD), their frequency apparently varying among different regions of Mexico. Other results of our program include training of specialists and technicians, development of the Latin American Metabolic Information Network, a procedure to locally prepare a special food product low in phenylalanine for the treatment of PKU patients, and extension of approaches for these disorders to the investigation metabolic derangements of infant malnutrition. This work demonstrates that inherited metabolic diseases constitute a significant load in pediatric pathology and that their study can and should be pursued in developing nations.
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Errores Innatos del Metabolismo/diagnóstico , Técnicos Medios en Salud/educación , Niño , Preescolar , Países en Desarrollo , Educación Médica , Femenino , Pruebas Genéticas , Genética Médica/educación , Genética Médica/métodos , Genética Médica/organización & administración , Accesibilidad a los Servicios de Salud , Humanos , Lactante , Recién Nacido , Discapacidad Intelectual/epidemiología , Discapacidad Intelectual/etiología , Discapacidad Intelectual/genética , Discapacidad Intelectual/metabolismo , Hepatopatías/epidemiología , Hepatopatías/etiología , Hepatopatías/genética , Hepatopatías/metabolismo , Masculino , Errores Innatos del Metabolismo/epidemiología , Errores Innatos del Metabolismo/genética , México/epidemiología , Hipotonía Muscular/epidemiología , Hipotonía Muscular/etiología , Hipotonía Muscular/genética , Hipotonía Muscular/metabolismo , Tamizaje Neonatal , Fenilcetonurias/diagnóstico , Fenilcetonurias/epidemiología , Derivación y Consulta/estadística & datos numéricos , Estudios Retrospectivos , Recursos HumanosRESUMEN
Mammalian cells typically contain hundreds of peroxisomes but can increase peroxisome abundance further in response to extracellular stimuli. We report here the identification and characterization of two novel human peroxisomal membrane proteins, PEX11alpha and PEX11beta. Overexpression of the human PEX11beta gene alone was sufficient to induce peroxisome proliferation, demonstrating that proliferation can occur in the absence of extracellular stimuli and may be mediated by a single gene. Time course studies indicated that PEX11beta induces peroxisome proliferation through a multistep process involving peroxisome elongation and segregation of PEX11beta from other peroxisomal membrane proteins, followed by peroxisome division. Overexpression of PEX11alpha also induced peroxisome proliferation but at a much lower frequency than PEX11beta in our experimental system. The patterns of PEX11alpha and PEX11beta expression were examined in the rat, the animal in which peroxisome proliferation has been examined most extensively. Levels of PEX11beta mRNA were similar in all tissues examined and were unaffected by peroxisome-proliferating agents. Conversely, PEX11alpha mRNA levels varied widely among different tissues, were highest in tissues that are sensitive to peroxisome-proliferating agents, and were induced more than 10-fold in response to the peroxisome proliferators clofibrate and di(2-ethylhexyl) phthalate. Taken together, these data implicate PEX11beta in the constitutive control of peroxisome abundance and suggest that PEX11alpha may regulate peroxisome abundance in response to extracellular stimuli.
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Proteínas Fúngicas/metabolismo , Proteínas de la Membrana/metabolismo , Microcuerpos/fisiología , Secuencia de Aminoácidos , Animales , Línea Celular , Cartilla de ADN , ADN Complementario , Técnica del Anticuerpo Fluorescente Indirecta , Proteínas Fúngicas/genética , Expresión Génica , Humanos , Proteínas de la Membrana/genética , Datos de Secuencia Molecular , Peroxinas , Ratas , Homología de Secuencia de AminoácidoRESUMEN
We surveyed Delta1-pyrroline 5-carboxylate dehydrogenase genes from four patients with hyperprolinemia type II using RT-PCR amplification, genomic PCR amplification and direct sequencing. We found four mutant alleles, two with frameshift mutations [A7fs(-1) and G521fs(+1)] and two with missense mutations (S352L and P16L). To test the functional consequences of three of these, we expressed them in a P5CDh-deficient strain of Saccharomyces cerevisiae . In contrast to wild-type human P5CDh, yeast expressing S352L and G521fs(+1) failed to grow on proline and had no detectable P5CDh activity. The P16L allele, however, produced fully functional P5CDh and subsequent analysis suggests that it is polymorphic in the relevant (Spanish) population. Interestingly, the G521fs(+1) allele segregates in the large Irish Traveller pedigree used to define the HPII phenotype. To our knowledge, this is the first description of the molecular basis for this inborn error.
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Errores Innatos del Metabolismo de los Aminoácidos/enzimología , Errores Innatos del Metabolismo de los Aminoácidos/genética , Mutación , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/genética , Prolina/metabolismo , 1-Pirrolina-5-Carboxilato Deshidrogenasa , Alelos , Errores Innatos del Metabolismo de los Aminoácidos/clasificación , Secuencia de Aminoácidos , Secuencia de Bases , Línea Celular , Cartilla de ADN/genética , ADN Complementario/genética , Femenino , Mutación del Sistema de Lectura , Expresión Génica , Humanos , Masculino , Linaje , Mutación Puntual , Reacción en Cadena de la Polimerasa , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismoRESUMEN
The 70-kDa peroxisomal membrane protein (PMP70) is a member of a family of half-ATP-binding cassette (ABC) transporter proteins located in the human peroxisomal membrane. Other members include the PMP70-related peroxisomal membrane protein, the adrenoleukodystrophy protein (ALDP), and the adrenoleukodystrophy-related protein. The functions of ABC transporters in the peroxisomal membrane are poorly understood. Evidence from yeast and human mutants suggests that they are involved in the peroxisomal import of fatty acids and/or fatty acyl-CoAs into the organelle. We report the cloning and characterization of the human PMP70 structural gene (gene symbol: PXMP1) localized on human chromosome 1p21-p22. PXMP1 is approximately 65 kb in length, contains 23 exons, and is quite different in structure from the gene (ALD) that encodes the related protein, ALDP. We also analyzed the 5' flanking region of the human PXMP1 gene and the corresponding region of murine Pxmp-1. Both promoters have features of housekeeping genes, including a high GC content and multiple consensus Sp1 binding sequences. In more than 3 kb of Pxmp-1 5' flanking sequence we did not identify a consensus peroxisomal proliferator responsive element.
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Transportadoras de Casetes de Unión a ATP , Proteínas de la Membrana/genética , Microcuerpos/genética , Animales , Secuencia de Bases , Clonación Molecular , Exones , Humanos , Intrones , Ratones , Microcuerpos/química , Datos de Secuencia Molecular , Regiones Promotoras Genéticas/genética , Alineación de Secuencia , Homología de Secuencia de Ácido NucleicoRESUMEN
Three half ATP-binding cassette transporters (ALDP, ALDR, PMP70) are known to be present in the human peroxisome membrane. Mutations in the gene encoding ALDP cause X-linked adrenoleukodystrophy; the role of ALDR and PMP70 in human disease is unclear. We report the cloning and characterization of a fourth human gene encoding a peroxisomal half ABC transporter. The gene, designated P70R, maps to chromosome 14q24, encodes a 73 kDa transporter most similar to PMP70, and is expressed in all human tissues examined. Because half ABC transporters heterodimerize to form functional transporters, the identification of a fourth member of this family in the peroxisome membrane has implications for our understanding of mammalian peroxisomes and the genetic disorders of peroxisomal function.
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Transportadoras de Casetes de Unión a ATP/genética , Cromosomas Humanos Par 14 , Proteínas de la Membrana , Microcuerpos/metabolismo , Transportadoras de Casetes de Unión a ATP/metabolismo , Secuencia de Aminoácidos , Animales , Línea Celular , Membrana Celular/metabolismo , Clonación Molecular , ADN Complementario , Expresión Génica , Humanos , Immunoblotting , Datos de Secuencia Molecular , Fracciones SubcelularesRESUMEN
Plasma free fatty acid profiles from patients suffering from various mitochondrial beta-oxidation deficiencies were analyzed by gas chromatography-mass spectrometry. cis-4-Decenoic acid (10:1n-6) in medium-chain acyl-CoA dehydrogenase deficiency and cis-5-tetradecenoic acid (14:1n-9) in very-long-chain and 3-hydroxy-long chain acyl-CoA dehydrogenase deficiencies are characteristic of these diseases. In addition, patients with 3-hydroxy-long chain acyl-CoA dehydrogenase deficiency showed a specific increase of 3-hydroxy-long chain fatty acids. The study of plasma free fatty acids is an easy and useful methodology for the diagnostic approach of some mitochondrial beta-oxidation deficiencies, allowing us to establish a quick differentiation between medium- and long-chain defects.
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Acil-CoA Deshidrogenasa de Cadena Larga/deficiencia , Ácidos Grasos no Esterificados/sangre , Ácidos Grasos/metabolismo , Mitocondrias/metabolismo , Acil-CoA Deshidrogenasa , Adolescente , Adulto , Carnitina/sangre , Carnitina/orina , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Espectrometría de Masas , Mitocondrias/enzimología , Oxidación-ReducciónAsunto(s)
ADN Complementario/genética , Microcuerpos/química , Animales , Secuencia de Bases , Clofibrato/farmacología , Clonación Molecular , Dietilhexil Ftalato/farmacología , Regulación Enzimológica de la Expresión Génica , Biblioteca de Genes , Genes , Hipolipemiantes/farmacología , Datos de Secuencia Molecular , Ratas , Regulación hacia ArribaRESUMEN
Hypoalbuminemia has been recently informed by us as a risk factor in aminoglycoside nephrotoxicity. Since amikacin has a low serum binding capacity to albumin, the present study was designed to determine if the higher risk of amikacin nephrotoxicity in patients with hypoalbuminemia was due to low serum albumin per se or to malnutrition. One-hundred and thirteen ward patients who received endovenous amikacin for greater than 36 hours were studied prospectively. All were evaluated for the following factors: age, sex, diagnosis, serum creatinine, serum albumin, and nutritional status. They were followed with serum creatinine twice a week until cessation of therapy. Amikacin pharmacokinetics was studied in 11 subjects: 6 patients had a serum albumin less than 3.0 g/dL and 5 greater than 3.0 g/dL, but there were no differences in age, sex, weight, diagnosis, arterial pressure and nutritional status. The overall incidence of toxicity was 11%. In patients with serum albumin less than 3.0 g/dL it was 17.3% and in those greater than 3.0 g/dL it was 2.2%, p less than 0.05. There was no difference in the nutritional status between toxicity and non-toxicity groups. In the pharmacokinetic study, the peak levels obtained one hour after amikacin administration were higher in patients with serum albumin less than 3.0 g/dL than in those with normal serum albumin (12.7 +/- 1.6 vs 9.0 +/- 1.2, p less than 0.002). In conclusion hypoalbuminemia is a risk factor in aminoglycoside nephrotoxicity regardless of the nutritional status.
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Amicacina/efectos adversos , Enfermedades Renales/inducido químicamente , Estado Nutricional , Albúmina Sérica/análisis , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factores de RiesgoRESUMEN
We studied 113 patients treated with intravenous amikacin to determine the value of determining serial trough and peak amikacin levels in plasma for predicting nephrotoxicity. Thirteen patients (11.5%) developed renal toxicity, with significant increases from 48 to 96 h in both peak and trough amikacin levels (6.7 +/- 4.7 [standard deviation] days before the serum creatinine rose). The nontoxicity group had no change or even showed decrements in amikacin levels in plasma. A higher nephrotoxicity risk was seen in patients with increments greater than 1 microgram/ml between 48 and 96 h, with odds ratios of 16.4 for trough, 8 for peak, and 7.2 for both levels. We suggest that an increment of at least 1 microgram/ml in amikacin levels in plasma from 48 to 96 h may predict the appearance of renal toxicity.