RESUMEN
The paper presents a case history of multiple myeloma in a female patient. Its diagnosis was established only 10 years after the onset of the disease, which results in severe invalidity (she was treated with calcium preparations in accordance with the diagnosis of generalized osteoporosis. Persistent therapy with calcium preparations was ineffective; the number of pathological fractures increased. Further bone study and current instrumental imaging methods could make a precise diagnosis and use etiotropic treatment, which yielded a positive clinical effect. The data on high-technology studies, body magnetic resonance imaging used in the diagnosis of the diseases accompanied by bone destructive changes are given. The problems in the diagnosis of multiple myeloma are outlined.
Asunto(s)
Imagen por Resonancia Magnética , Mieloma Múltiple/diagnóstico , Tomografía Computarizada Espiral , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Diagnóstico Diferencial , Femenino , Humanos , Mieloma Múltiple/diagnóstico por imagen , Mieloma Múltiple/tratamiento farmacológico , Mieloma Múltiple/patología , Invasividad Neoplásica , Resultado del TratamientoRESUMEN
New comparative genome hybridization technology on NotI-microarrays is presented (Karolinska Institute International Patent WO02/086163). The method is based on comparative genome hybridization of NotI-probes from tumor and normal genomic DNA with the principle of new DNA NotI-microarrays. Using this method 181 NotI linking loci from human chromosome 3 were analyzed in 200 malignant tumor samples from different organs: kidney, lung, breast, ovary, cervical, prostate. Most frequently (more than in 30%) aberrations--deletions, methylation,--were identified in NotI-sites located in MINT24, BHLHB2, RPL15, RARbeta1, ITGA9, RBSP3, VHL, ZIC4 genes, that suggests they probably are involved in cancer development. Methylation of these genomic loci was confirmed by methylation-specific PCR and bisulfite sequencing. The results demonstrate perspective of using this method to solve some oncogenomic problems.
Asunto(s)
Cromosomas Humanos Par 3/metabolismo , Epigénesis Genética , Perfilación de la Expresión Génica/métodos , Regulación Neoplásica de la Expresión Génica , Proteínas de Neoplasias/biosíntesis , Neoplasias Glandulares y Epiteliales/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Cromosomas Humanos Par 3/genética , Femenino , Humanos , Masculino , Proteínas de Neoplasias/genética , Neoplasias Glandulares y Epiteliales/genética , Especificidad de Órganos , Sitios de Carácter Cuantitativo/genéticaRESUMEN
Not I linking clones contain sequences flanking Not I recognition sites and were previously shown to be tightly associated with CpG islands and genes. To directly assess the value of Not I clones in genome research, high density grids with 50 000 Not I linking clones originating from six representative Not I linking libraries were constructed. Altogether, these libraries contained nearly 100 times the total number of Not I sites in the human genome. A total of 3437 sequences flanking Not I sites were generated. Analysis of 3265 unique sequences demonstrated that 51% of the clones displayed significant protein similarity to SWISSPROT and TREMBL database proteins based on MSPcrunch filtering with stringent parameters. Of the 3265 sequences, 1868 (57.2%) were new sequences, not present in the EMBL and EST databases (similarity < or =90%). Among these new sequences, 795 (24.3%) showed similarity to known proteins and 712 (21.8%) displayed an identity of >75% at the nucleotide level to sequences from EMBL or EST databases. The remaining 361 (11.1%) sequences were completely new, i.e. <75% identical. The work also showed tight, specific association of Not I sites with the first exon and suggest that the so-called 3' ESTs can actually be generated from 5'-ends of genes that contain Not I sites in their first exon.
Asunto(s)
Desoxirribonucleasas de Localización Especificada Tipo II , Genoma Humano , Secuencia de Bases , Clonación Molecular , Islas de CpG , Cartilla de ADN/genética , Bases de Datos Factuales , Etiquetas de Secuencia Expresada , Biblioteca de Genes , Humanos , Datos de Secuencia Molecular , ARN Mensajero/genética , Mapeo RestrictivoRESUMEN
A plasmid expression vector (pCEQ3), using temperature-regulated transcription from the p'R promoter of bacteriophage lambda, has been constructed. The vector is derived from pBR327 in which the EcoRI-ClaI fragment of plasmid DNA is replaced with a 2.2-kb DNA module cI857-pR-Q-p'R-qut-t'R, consisting of two regions of the lambda genome. The first region contains the repressor gene cI857 and promoter pR; the second one contains gene Q and the late promoter p'R. When the repressor protein, product of the cI857 gene, becomes temperature-inactivated, it allows the promoter pR to initiate the transcription of the Q gene. The product of the Q gene, in turn, acts as a positive regulator of transcription from promoter p'R. The promoter activity of pR is fully repressed at a low temperature (30 degrees C) and transcription from p'R is terminated in the absence of Q gene product, but the shift of temperature up to 37 degrees C is sufficient to make the transcription from the p'R promoter highly active. Foreign genes can be inserted into the single EcoRI site downstream from the p'R promoter. The resultant constructions express extremely high levels of the cloned gene product in Escherichia coli.
Asunto(s)
Bacteriófago lambda/genética , Regulación de la Expresión Génica , Vectores Genéticos , Plásmidos , Clonación Molecular , Escherichia coli/genética , Operón Lac , Regiones Promotoras Genéticas , Temperatura , Transcripción Genética , beta-Lactamasas/biosíntesisRESUMEN
Plasmid expression vector using the temperature-regulated promoter P'R of bacteriophage lambda is described. The vector carries a combination of two regions of lambda cI857indgenome, that contain: 1) gene cI and promoter PR, and 2) gene Q and promoter P'R. Transcription or gene Q is initiated at promoter PR, which is controlled by cI857 repressor. The Q gene product acts as a positive regulator of RNA synthesis from P'R. At 37 degrees C, sufficient amounts of protein Q are synthesised to initiate the expression of the cloned gene from P'R. Inactivation of Q gene (by elimination of a single NcoI site) results in the loss of P'R expression activity in the vector. E. coli beta-galactosidase gene (lacZ) and human leukocyte interferon alpha 2 gene (ifn alpha 2) were cloned into a single EcoRI cleavage site under the control of P'R. These constructs express high levels of beta-galactosidase and interferon alpha 2 in E. coli at 37 degrees C.