RESUMEN

Here we examined whether the Matsuda-DeFronzo insulin sensitivity index (ISI-M) is more efficient than the homeostasis model assessment of insulin resistance (HOMA-IR) for assessing risk of hypertension. Cross-sectional and longitudinal analyses were conducted using normotensive subjects who were selected among 1399 subjects in the Tanno-Sobetsu cohort. In the cross-sectional analysis (n=740), blood pressure (BP) level was correlated with HOMA-IR and with ISI-M, but correlation coefficients indicate a tighter correlation with ISI-M. Multiple linear regression analysis adjusted by age, sex, body mass index (BMI) and serum triglyceride level (TG) showed contribution of ISI-M and fasting plasma glucose, but not of HOMA-IR. In the longitudinal analysis (n=607), 241 subjects (39.7%) developed hypertension during a 10-year follow-up period, and multiple logistic regression indicated that age, TG, systolic BP and ISI-M, but not HOMA-IR, were associated with development of hypertension. In subjects <60 years old, odds ratio of new-onset hypertension was higher in the low ISI-M group (ISI-M, less than the median) than in the high ISI-M group for any tertile of BMI. In conclusion, ISI-M is a better predictor of hypertension than is HOMA-IR. Non-hepatic IR may be a determinant, which is independent of TG, BP level and BMI, of the development of hypertension.

Asunto(s)

Glucemia/metabolismo , Presión Sanguínea , Hipertensión/epidemiología , Resistencia a la Insulina , Insulina/sangre , Modelos Biológicos , Anciano , Biomarcadores/sangre , Índice de Masa Corporal , Estudios Transversales , Ayuno/sangre , Femenino , Prueba de Tolerancia a la Glucosa , Homeostasis , Humanos , Hipertensión/sangre , Hipertensión/fisiopatología , Japón/epidemiología , Modelos Lineales , Modelos Logísticos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Análisis Multivariante , Oportunidad Relativa , Valor Predictivo de las Pruebas , Pronóstico , Medición de Riesgo , Factores de Riesgo , Factores de Tiempo , Triglicéridos/sangre

RESUMEN

OBJECTIVES: Although it is well known that obesity is closely related to insulin resistance, the incidence of the development of insulin resistance in people with obesity is not known. In this study, we investigated the incidence of insulin resistance in citizens of two rural communities in Japan. SUBJECTS AND METHODS: The subjects were 102 men and 126 women over the age of 30 years selected from 1035 citizens who had undergone medical examinations in the towns of Tanno and Sobetsu, Hokkaido, in 1991 and 1998. Those who were on medication for hypertension, diabetes, hyperlipidaemia, coronary heart disease and cerebral vessel disease were excluded. The simple index to determine insulin resistance [i.e. homeostasis model assessment (HOMA-R) > or = 1.73] was used, and subjects who were determined to be positive for insulin resistance according to this index in 1991 were also excluded in order to determine the incidence of insulin resistance in subjects who had no abnormalities other than obesity. The systolic blood pressure (SBP), diastolic blood pressure, body mass index (BMI), triglyceride level, high-density lipoprotein level, blood sugar level, serum insulin value and HOMA-R were measured in all subjects in 1991 and in 1998. Moreover, the subjects were divided into two groups according to BMI, a normal group consisting of subjects with BMI < 25 and an obesity group consisting of subjects with BMI > or = 25. We also compared the incidences of insulin resistance in normal and obesity groups of subjects who were newly determined to be positive for insulin resistance on the basis of data obtained from medical examinations conducted in 1998. RESULTS: The incidence of insulin resistance was significantly higher in the obesity group than in the normal group (25.0 vs. 4.5%). The results of logistic regression analysis showed that obesity was closely related to insulin resistance and that the relative risk of development of insulin resistance adjusted for age, sex, SBP, FPG and HDL was 3.193 (95% CI 1.085-9.401). CONCLUSIONS: The incidence of insulin resistance was significantly higher in the obesity group than in the normal group in this study, suggesting that improvement in obesity is important for prevention of the occurrence of type 2 diabetes or atherosclerotic disease based on insulin resistance.

Asunto(s)

Resistencia a la Insulina , Obesidad/epidemiología , Adulto , Factores de Edad , HDL-Colesterol/sangre , Femenino , Humanos , Incidencia , Insulina/sangre , Japón/epidemiología , Modelos Logísticos , Masculino , Persona de Mediana Edad , Obesidad/sangre , Riesgo , Población Rural

RESUMEN

We examined correlations between the frequency of insulin resistance and the accumulation of coronary risk factors in residents of rural comities in Japanese, using simple criteria for determination of insulin resistance based on evaluation by the euglycaemic-hyperinsulinaemic glucose clamp (GC) method. The subjects were 376 men and 589 women living in two rural communities in Japan. We measured body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), triglyceride (TG), HDL cholesterol (HDL), and homeostasis model assessment (HOMA-R). Correlations between HOMA-R and those parameters were examined. To assess the existence of insulin resistance in these subjects, we used a practical index based on the GC method. The subjects with value of HOMA-R >or= 1.73 have insulin resistance. In addition, the HOMA-R was divided into five quantiles based on the frequency distribution (0.60 or below, from 0.61 to 0.82, from 0.83 to 1.18, from 1.19 to 1.69, and 1.70 or higher), to examine the concentration of risk factors in each group. In total, 74 (19.6%) of the men and 119 (20.3%) of the women had insulin resistance (HOMA-R >or= 1.73). It was found that the higher the HOMA-R, the higher was the number of coronary risk factors, such as hypertension, obesity, hypertriglyceridaemia and hypo HDL cholesterolaemia. The number of coronary risk factors was particular high in subjects with HOMA-R >or= 1.70. HOMA-R in the case of no glucose loading is a useful and practical index for evaluation of insulin resistance and coronary risk factors in the epidemiological study.

Asunto(s)

Enfermedad de la Arteria Coronaria/etiología , Resistencia a la Insulina/fisiología , Adulto , Presión Sanguínea/fisiología , Índice de Masa Corporal , HDL-Colesterol/sangre , Enfermedad de la Arteria Coronaria/sangre , Enfermedad de la Arteria Coronaria/fisiopatología , Femenino , Homeostasis , Humanos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Triglicéridos/sangre

RESUMEN

We describe a complete cytogenetic response to interferon-alpha in a patient with chronic myelogenous leukemia undergoing chronic hemodialysis. Although IFN-alpha therapy has been applied to patients with chronic hepatitis C receiving hemodialysis, the pharmacokinetics of IFN-alpha in patients with poor renal function still remain unclear. In the present patient, the serum IFN-alpha concentration remained high even 48 hours after injection (42.9 IU/ml), and IFN-alpha was almost completely removed by hemodialysis (< 6 UI/ml). The patient was treated with IFN-alpha (3 x 10(6) IU, three times a week), and cytogenetic disappearance (0%) of the Ph-positive clone was confirmed 31 months after the start of therapy. Recombinant human erythropoietin (Epo) was used to treat anemia due to renal failure and IFN-alpha therapy. The anemia was controllable with Epo, and no adverse effect was observed.

Asunto(s)

Interferón-alfa/administración & dosificación , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Anemia/tratamiento farmacológico , Anemia/etiología , Eritropoyetina/uso terapéutico , Hepatitis C/complicaciones , Humanos , Interferón-alfa/efectos adversos , Interferón-alfa/farmacocinética , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/terapia , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Masculino , Persona de Mediana Edad , Cromosoma Filadelfia , Proteínas Recombinantes , Diálisis Renal , Resultado del Tratamiento

RESUMEN

Chromosomal translocation involving the BCL6 gene affects not only immunoglobulin (Ig) genes but also a number of non-Ig genes as partners. The molecular anatomy of the BCL6 gene rearrangements in 39 cases with diffuse large B-cell lymphoma (DLBCL) by long-distance polymerase chain reaction-based assays was determined. The results showed that Ig genes were affected in 21 cases; non-Ig genes, 15 cases; a deletion of more than a 1-kb segment, 2 cases; and a point mutation, 1 case. Comparative studies between the 21 cases with Ig gene partners and the 17 cases with non-Ig gene partners, including 2 cases with the deletion, showed that the overall survival of the latter group of patients was significantly inferior to that of the former (P = .0440), and the estimated 2-year overall survival rates were 58.3% vs 17.6% (P = .005). Non-Ig/BCL6 fusion is a poor prognostic indicator of DLBCL, and DLBCL with BCL6 translocation could be subclassified according to the individual partner locus and/or gene. (Blood. 2000;96:2907-2909)

Asunto(s)

ADN de Neoplasias/genética , Proteínas de Unión al ADN/genética , Genes de Inmunoglobulinas , Linfoma de Células B Grandes Difuso/genética , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética , Translocación Genética , Adulto , Anciano , Cromosomas Humanos Par 1/genética , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 16/genética , Cromosomas Humanos Par 3/genética , Cromosomas Humanos Par 7/genética , Análisis Mutacional de ADN , Femenino , Humanos , Tablas de Vida , Linfoma de Células B Grandes Difuso/mortalidad , Masculino , Persona de Mediana Edad , Mutación Puntual , Reacción en Cadena de la Polimerasa , Pronóstico , Proteínas Proto-Oncogénicas c-bcl-6 , Eliminación de Secuencia , Análisis de Supervivencia , Tasa de Supervivencia

RESUMEN

BCL6 translocations involve not only immunoglobulin (IG) genes but also a number of non-IG loci as partners. Junctional sequences of three IG/BCL6 translocations were readily obtained by long-distance PCR. In cases where partner loci were not determined, we developed a long-distance inverse PCR method, which amplifies unknown fragments flanked by known BCL6 sequences. Using these two long-distance PCR-based approaches, we cloned junctional areas of BCL6 translocations from a total of 58 cases of B-cell tumors. Nucleotide sequencing and database searches revealed that 30 cases involved IGs as partners: IG heavy chain gene in 22, IG kappa light chain gene in 1, and IG lambda light chain gene in 7. In contrast, 23 cases affected non-IG loci, including the H4 histone gene, heat shock protein genes HSP89alpha and HSP90beta, and PIM-1 proto-oncogene. On der(3) chromosomes, complete sets of the promoters of these partner genes replaced that of BCL6 in the same transcriptional orientation. These results suggest that BCL6 gene affected by the translocation is transcriptionally activated by a variety of stimuli, including cell cycle control, changes in the physical environment, and response to cytokines. Break points on BCL6 occurred within the major translocation cluster, and we identified a 120-bp hyper-cluster region a short distance from the 3' end of exon 1. Gel mobility-shift assay suggested the presence of a protein(s) that bound to this particular region.

Asunto(s)

Proteínas de Unión al ADN/genética , Inmunoglobulinas/genética , Linfoma de Células B/genética , Reacción en Cadena de la Polimerasa/métodos , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética , Translocación Genética , Secuencia de Bases , Southern Blotting , Clonación Molecular , Exones , Histonas/genética , Humanos , Modelos Genéticos , Datos de Secuencia Molecular , Mutación , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-bcl-6 , Análisis de Secuencia de ADN , Transcripción Genética , Activación Transcripcional

RESUMEN

PURPOSE: t(8;14)(q24;q32) and/or c-MYC/immunoglobulin heavy-chain (IGH) fusion gene have been observed not only in Burkitt's lymphoma (BL) but also in a proportion of non-BL, diffuse large-cell lymphoma of B-cell type (DLCL). We explored molecular features of DLCL with c-MYC/IGH fusion and the impact of this genetic abnormality on clinical outcome of DLCL. PATIENTS AND METHODS: A total of 203 cases of non-BL DLCL were studied. Genomic DNA extracted from tumor tissues was subjected to long-distance polymerase chain reaction (LD-PCR) using oligonucleotide primers for exon 2 of c-MYC and for the four constant region genes of IGH. RESULTS: Twelve cases (5.9%) showed positive amplification; one had a c-MYC/Cmicro, nine had a c-MYC/Cgamma, and two had a c-MYC/Calpha fusion sequence. Restriction and sequence analysis of the LD-PCR products, ranging from 2.3 to 9.4 kb in size, showed that breakage in the 12 cases occurred within a 1.5-kb region that included exon 1 of c-MYC in combination with breakpoints at the switch regions of IGH (10 of 12). In 10 cases, Myc protein encoded by the fusion genes demonstrated mutations and/or deletions. Six cases had additional molecular lesions in BCL-2 or BCL-6 and/or p53 genes. The age range of the 12 patients was 44 to 86 years, with a median age of 65.5 years. Five patients had stage I/II disease, and seven had stage III/IV disease. Lactate dehydrogenase was elevated in nine of 11 subjects. Seven showed involvement of the gastrointestinal tract. All patients were treated by surgery and/or chemoradiotherapy; six died of the disease within 1 year, resulting in the poorest 1- and 2-year survival rates among DLCL subgroups. CONCLUSION: The c-MYC/IGH fusion gene of DLCL is identical to that of the sporadic type of BL (sBL). DLCL with c-MYC/IGH shares clinical features with sBL but is characterized further by an older age distribution.

Asunto(s)

Genes de Inmunoglobulinas , Genes myc , Cadenas Pesadas de Inmunoglobulina/genética , Linfoma de Células B/genética , Linfoma de Células B Grandes Difuso/genética , Adulto , Anciano , Anciano de 80 o más Años , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 8 , Femenino , Reordenamiento Génico , Humanos , Linfoma de Células B/patología , Linfoma de Células B Grandes Difuso/patología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa/métodos , Translocación Genética

RESUMEN

The complex forming ability of delta-cyclodextrin delta-CD with 7 kinds of macrocyclic compounds (MCCs) with 8-15 carbon atoms in the ring as models of large guest molecules was studied in aqueous solution and compared with the complexation properties of alpha-, beta- and gamma-CD. Both alpha- and beta-CD formed relatively stable complexes with small MCCs, while gamma- and delta-CD were more efficient in binding larger MCCs. The solid MCC/delta-CD complexes were precipitated with the larger MCCs with 11-15 carbon atoms in the ring, while no such precipitates were obtained with smaller MCCs with only 8-10 carbon atoms in the ring. The formation of the solid complexes was confirmed by powder X-ray diffractometry and differential scanning calorimetry. The cell dimensions of cycloundecanone (11 carbon atoms in the ring)/delta-CD complex were determined by X-ray crystallography. The preliminary crystal data were: Monoclinic, P21, a=32.50 (2)A, b=19.02 (3)A, c=16.60 (1) A, beta3=98.37 (5)degrees, V=10148 (16) A3.

Asunto(s)

Ciclodextrinas/química , Cicloparafinas/química , Cetonas/química , Cristalografía por Rayos X

RESUMEN

Chromosomal translocations and/or their molecular equivalents involving the BCL6 gene on 3q27 band have been suggested to be involved in the development of non-Hodgkin's lymphoma of B-cell type (B-NHL). The rearrangement of BCL6 sometimes coexists with other translocations specific to B-NHL. Here, we report a novel B-cell lymphoma cell line, YM, established from a patient with diffuse large cell lymphoma. The YM cells expressed B-cell-associated antigens in addition to mu delta/kappa monoclonal immunoglobulin. Southern blot analysis of DNA from YM cells demonstrated rearrangement of the BCL2 gene within the 5' flanking region (5'-BCL2). Polymerase chain reaction (PCR) using primer pairs for the BCL2 exons 1 and 2, and for the constant region of the immunoglobulin kappa light chain gene (IGkappa) revealed PCR products encompassing the 5'-BCL2/IGkappa fusion, indicating that the YM cells had a t(2;18)(p11;q21) translocation. The BCL6 gene was rearranged at a point within the first intron, and cloning of the rearranged BCL6 revealed unidentified sequences juxtaposed to the 5' side of the gene. The isolated clones were mapped to 16p11.2 by high resolution fluorescence in situ chromosomal hybridization. Thus, the YM cells carried a 3q27 translocation involving 16p11.2 as a partner. Chromosome painting of metaphase spreads confirmed that the YM cells had both t(2;18) and t(3;16). Northern blot analysis using a fragment immediately adjacent to the breakpoint on 16p11.2 revealed transcriptional activity within this locus. The YM cells expressed abundant transcripts with aberrant sizes from BCL2 and BCL6, indicating deregulated overexpression of the two genes resulting from the t(2;18) and t(3;16). The YM cell line will therefore be useful to study whether BCL2 and BCL6 genes collaborate in the pathogenesis of B-NHL.

Asunto(s)

Reordenamiento Génico de Linfocito B , Genes bcl-2 , Linfoma de Células B/genética , Secuencia de Bases , Cromosomas Humanos Par 3/genética , Sondas de ADN , Regulación Neoplásica de la Expresión Génica , Humanos , Cariotipificación , Linfoma de Células B/patología , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/patología , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Translocación Genética , Células Tumorales Cultivadas

RESUMEN

The PAX5 gene encodes the BSAP (B-cell-specific activator protein) which is a key regulator of B-cell development and differentiation. A recurring translocation t(9;14)(p13;q32) in non-Hodgkin's lymphoma moves the PAX5 on 9p13 within close proximity of the immunoglobulin heavy chain gene (IGH). KIS-1 cell line was established from a patient with diffuse large cell lymphoma of B-cell type carrying t(9;14). We analysed PAX5/BSAP expression by Northern and Western blotting in a panel of haematological tumour cell lines with other chromosome abnormalities in comparison with that of KIS-1. PAX5 mRNA and BSAP expression were detected in all B-cell lines tested, and the high level in KIS-1 was confirmed. However, a diffuse large B-cell lymphoma cell line and an acute B-lymphoid/myeloid leukaemia cell line expressed the PAX5/BSAP at levels comparable with KIS-1. PAX5 transcripts were readily detectable in clinical materials with a wide variety of B-cell neoplasms by reverse transcriptase-mediated polymerase chain reaction (PCR). Thus, PAX5/BSAP activation in haematological tumour cells is not necessarily associated with t(9;14). Although binding sites for BSAP have been identified in the promoters of CD19, this study failed to find clear correlation between the level of PAX5/BSAP expression and that of CD19. In contrast to KIS-1 in which the E mu enhancer of IGH was juxtaposed to PAX5, cloning of t(9; 14) from another case by long-distance PCR revealed that the PAX5 promoter was linked to a Cgamma constant region in divergent orientation, suggesting that the mechanism of PAX5 activation through recombination with IGH varies among individual cases. Breakpoints on 9p13 of the two translocations were clustered upstream of PAX5, leaving the PAX5 coding region intact.

Asunto(s)

Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 9/genética , Proteínas de Unión al ADN/genética , Linfoma de Células B/genética , Proteínas Nucleares/genética , Factores de Transcripción , Translocación Genética , Secuencia de Bases , Northern Blotting , Western Blotting , Reordenamiento Génico de Cadena Pesada de Linfocito B , Humanos , Linfoma de Células B/metabolismo , Datos de Secuencia Molecular , Factor de Transcripción PAX5 , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Células Tumorales Cultivadas

RESUMEN

We describe here two cases of diffuse large cell type non-Hodgkin's lymphoma affecting the bilateral breasts. The contralateral tumor in one case appeared 17 months after the first mastectomy, whereas the bilateral tumors occurred concurrently in the other patient who was pregnant and showed widespread dissemination at initial presentation. Lymphoma cells from both cases showed the mature B-cell immunophenotype and had rearrangements of the BCL6 gene. Both patients developed progressive disease despite chemo-radiotherapy and died of leukemic manifestations. There were no apparent pathological features of lymphomas of mucosa-associated lymphoid tissue origin.

Asunto(s)

Neoplasias de la Mama/patología , Linfoma de Células B Grandes Difuso/patología , Complicaciones Neoplásicas del Embarazo/patología , Adulto , Linfocitos B/inmunología , Linfocitos B/patología , Neoplasias de la Mama/genética , Neoplasias de la Mama/inmunología , Terapia Combinada , Cartilla de ADN/química , ADN de Neoplasias/análisis , Proteínas de Unión al ADN/genética , Resultado Fatal , Femenino , Estudios de Seguimiento , Reordenamiento Génico , Humanos , Inmunofenotipificación , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/inmunología , Recurrencia Local de Neoplasia , Reacción en Cadena de la Polimerasa , Embarazo , Complicaciones Neoplásicas del Embarazo/inmunología , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas c-bcl-6 , Factores de Transcripción/genética , Dedos de Zinc/genética

RESUMEN

We developed a novel technique for long-distance polymerase chain reaction (LD-PCR) to detect t(8;14)(q24;q32). LD-PCR can amplify up to 12 kb of DNA encompassing the c-MYC and constant regions of the immunoglobulin heavy chain gene. In this report, we present two patients with B-cell non-Hodgkin's lymphoma. Clinical materials obtained from these patients were examined by LD-PCR. One patient had small noncleaved cell lymphoma (case 1) and the other had diffuse large cell lymphoma (case 2). Both patients showed central nervous system involvement. LD-PCR using appropriate primer pairs and a newly available Taq polymerase for longer product synthesis detected a 9.6 kb (case 1) and a 2.4 kb (case 2) c-MYC/C gamma fusion product indicative of t(8;14) in all materials in which lymphoma cells were shown positive by microscopic examination. LD-PCR provides an advantage in rapid detection of lymphoma cells carrying t(8;14).

Asunto(s)

Cromosomas Humanos Par 14 , Cromosomas Humanos Par 8 , ADN de Neoplasias/genética , Linfoma de Células B/genética , Translocación Genética , Adolescente , Anciano , Humanos , Masculino , Reacción en Cadena de la Polimerasa

RESUMEN

Chromosomal translocations in human leukemias generate fusion transcripts containing messages from two genes involved in the translocation, and these have been the targets for reverse transcriptase-mediated polymerase chain reaction (PCR). In contrast, many of the translocations in B-cell tumors involve immunoglobulin gene (IG) loci, and coding regions of the oncogenes on partner chromosomes are not interrupted by the translocation. Therefore, targets for PCR amplification are single-copy oncogene/IG fusion sequences within the complex genomic DNA. We present here a novel strategy for detection of translocations in B-cell tumors on the basis of long-distance (LD-) PCR that is capable of amplifying up to 30 kb of DNA. LD-PCR is a general method using primer pairs designed for distinctive regions of IG and oncogenes involved in translocations, and amplifying long DNA fragments encompassing the oncogene/IG junction. LD-PCR is capable of detecting virtually all the important translocations in B-cell tumors, including t(8;14)(q24;q32), t(14;18)(q32;q21), t(3;14)(q27;q32) and its variants. We show here that LD-PCR can substitute for time-consuming Southern blot hybridization in the rapid detection of these translocations. Furthermore, as amplified fragments obtained by LD-PCR contained exons and flanking sequences of the oncogenes and IGs, restriction analysis and nucleotide sequencing of the products refined the characteristics of translocations.

Asunto(s)

Mapeo Cromosómico , Cromosomas Humanos , ADN de Neoplasias/análisis , Leucemia de Células B/genética , Linfoma de Células B/genética , Reacción en Cadena de la Polimerasa/métodos , Translocación Genética/genética , Cartilla de ADN , Proteínas de Unión al ADN/genética , Exones , Amplificación de Genes , Humanos , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética

RESUMEN

The t(14;18)(q32;q21) translocation, involving the BCL2 gene and junctional segments (JH) of the immunoglobulin heavy chain gene (IGH), constitutes the most common chromosomal translocation in non-Hodgkin's lymphoma of B-cell type. Although the breakpoints in BCL2 are largely clustered within the major breakpoint region (MBR) and minor cluster region (mcr), it is known that some breakpoints map away from these regions, resulting in negative amplification of the junctional sequence by polymerase chain reaction (PCR) for < 1 kb targets. To circumvent this problem, we applied a novel PCR technology for long DNA targets, long-distance (LD-) PCR, to the detection of t(14;18) in clinical materials. Oligonucleotide primers were designed to be quite distant from the two known cluster regions in BCL2, and those for the corresponding IGH were complementary to the enhancer and constant regions. In all 52 cases identified as carrying BCL2/JH fusion by conventional Southern blot analysis, LD-PCR successfully amplified fragments encompassing the junctions, which were readily identifiable on ethidium bromide-stained gel. The size of the LD-PCR products ranged from 3.9 kb to 10.7 kb in MBR/IGH fusion and 1.9 kb to 16 kb in mcr/IGH fusion. Furthermore, we established an LD-PCR protocol for > 20 kb targets, which covered the intervening region between the MBR and mcr. Restriction analysis of the LD-PCR products revealed that breakpoints in 33 cases fell within the 150 bp-MBR region, and in 3 cases were within the mcr determined previously by others. In contrast, the breakpoints of the remaining 16 cases were distributed over a large region from the MBR through mcr. Nucleotide sequence analysis of a potential cluster region revealed the presence of an Alu repeat sequence. Restriction analysis of LD-PCR products with BstEII demonstrated a predominant usage of the JH6 segment (71%) at the BCL2/JH junctions. LD-PCR using primers for the constant region genes showed that class switch recombination occurred in more than 80% of the IGH genes on the der(14) chromosome. Our study showed that LD-PCR was capable of detecting virtually any t(14;18) that occurred within the approximately 30 kb region downstream of the MBR, and thus is suitable for initial diagnosis of lymphoma tissues. Furthermore, as amplified fragments obtained by the LD-PCR contained distinctive regions of BCL2 and IGH, restriction analysis and nucleotide sequencing of the products refined the characteristics of t(14;18).

Asunto(s)

Cromosomas Humanos Par 14 , Cromosomas Humanos Par 18 , Genes de Inmunoglobulinas , Genes bcl-2 , Cadenas Pesadas de Inmunoglobulina , Translocación Genética , Secuencia de Bases , Desoxirribonucleasas de Localización Especificada Tipo II , Humanos , Cambio de Clase de Inmunoglobulina , Cadenas J de Inmunoglobulina/análisis , Linfoma no Hodgkin/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Mapeo Restrictivo

RESUMEN

3q27 translocations affecting the BCL6 gene can involve not only immunoglobulin genes (IG) but also other as yet uncharacterized chromosomal loci as partners. Here, we describe cloning of the junctional area of a recurring translocation, t(3;6)(q27;p21), in non-Hodgkin's lymphoma of B-cell type and isolation of clones from 6p21; high resolution fluorescence in situ hybridization mapped the clones to sub-band 6p21.3. Nucleotide sequence analysis of a fragment from the junctional area of 6p21 revealed the presence of a novel H4 histone gene that was included in the histone gene cluster on this particular region, and the same fragment detected approximately 380-bp transcripts in hematological tumor cells. Breakpoints on 3q27 of two cases carrying t(3;6) were immediately 3' of the BCL6 exon 1, and the H4 histone gene was substituted for the 5' regulatory elements of BCL6. Because H4 gene expression is tightly coupled to DNA replication, this study suggested an immediate mechanism for deregulated expression of BCL6, leading to the development of non-Hodgkin's lymphoma.

Asunto(s)

Cromosomas Humanos Par 3/genética , Cromosomas Humanos Par 6/genética , Linfoma de Células B/genética , Proteínas Proto-Oncogénicas/genética , Secuencias Reguladoras de Ácidos Nucleicos/genética , Translocación Genética/genética , Secuencia de Bases , Southern Blotting , Mapeo Cromosómico , Cartilla de ADN/genética , Humanos , Cariotipificación , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa

RESUMEN

Variable number of tandem repeat analysis using a ministatellite DNA probe pYNH24 to distinguish variable alleles at a single locus was applied to forensic analysis of DNA extracted from a murder and body abandonment specimen. Two bands detected by pYNH24 correspond to identical DNA from both the upper and lower halves of a separated body, suggesting that these halves were from the same body.

Asunto(s)

Sondas de ADN , Medicina Legal , Homicidio , Secuencias Repetitivas de Ácidos Nucleicos , Femenino , Humanos

Asunto(s)

Cefalosporinas/uso terapéutico , Infecciones Urinarias/tratamiento farmacológico , Adulto , Anciano , Bacterias/efectos de los fármacos , Cefalosporinas/fisiología , Farmacorresistencia Microbiana , Enterobacteriaceae/efectos de los fármacos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos Estructurales , Orina/análisis

Asunto(s)

Ácido Yotalámico , Urografía , Adolescente , Adulto , Niño , Diatrizoato , Femenino , Humanos , Inyecciones Intravenosas , Ácido Yotalámico/efectos adversos , Pelvis Renal/diagnóstico por imagen , Masculino , Métodos , Persona de Mediana Edad , Enfermedades Urológicas/diagnóstico por imagen

Asunto(s)

Neoplasias del Pene/epidemiología , Adulto , Anciano , Humanos , Japón , Masculino , Persona de Mediana Edad