RESUMEN
Rice storage protein glutelin genes are coordinately regulated during seed development. A previous 5' deletion analysis using transgenic tobacco revealed that the minimum 5' region necessary for endosperm specificity was within -245 bp of the transcription start site, and included the AACA and GCN4 motifs that are highly conserved in the 5'-flanking regions of all glutelin genes. In this paper, the sequence elements essential for endosperm-specific expression are characterized in stable transgenic tobacco plants by both loss-of-function and gain-of-function experiments using this minimum promoter. Base substitution analysis shows that the proximal AACA motif between -73 and -61, and the GCN4 motif between -165 and -158 act as critical elements. An ACGT motif between -81 and -75, and Skn-I-like elements between -173 and -169 also play important roles in controlling the seed-specific expression. When the distal region between -245 and -145 containing the AACA and the GCN4 motifs or the proximal region between -113 and -46 containing the ACGT and AACA motifs is fused to a truncated promoter (-90 to +9) of the CaMV 35S gene fused to the beta-glucuronidase (GUS) reporter gene, high levels of seed-specific expression are observed in these fusions, thereby indicating that either pair of motifs is sufficient to confer seed expression in these fusions. However, when substituted for by the CaMV 35S core promoter (-46 to +1), seed expression is abolished, suggesting that the sequence between -90 and -46 of the CaMV 35S promoter containing G-box-like motif (as-1 element) is required for such specific expression in addition to AACA and GCN4 motifs. Therefore, we conclude that at least three cis-regulatory elements, the AACA motif, GCN4 motif and ACGT motif, are necessary to mediate endosperm expression of the GluB-1 glutelin gene.
Asunto(s)
Genes de Plantas , Glútenes/genética , Oryza/genética , Proteínas de Plantas , Regiones Promotoras Genéticas , Semillas/genética , Secuencia de Bases , Sitios de Unión , Expresión Génica , Genes Reporteros , Glucuronidasa/genética , Glútenes/biosíntesis , Histocitoquímica , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oryza/metabolismo , Plantas Modificadas Genéticamente , Plantas Tóxicas , Plásmidos , Semillas/metabolismo , Eliminación de Secuencia , Distribución Tisular , Nicotiana/genética , Transformación GenéticaRESUMEN
The clone pAS"C" is a sequence related to M13 phage minisatellite sequences isolated from bovine DNA. It contains TGG repeats, and related sequences are abundant in the Citrus genome. Fluorescent in situ hybridization (FISH) signals detected using this clone as a probe showed that the TGG repeat-related sequences are found on all the Citrus chromosomes, but are not found in Citrus-specific CMA+/DAPI- heterochromatic regions located at the extreme ends of each chromosome. These data suggest that the TGG-repeated sequences are evolutionarily conserved and that the CMA+/DAPI- heterochromatic regions were added to the chromosome ends at a recent stage in Citrus evolution.
RESUMEN
To investigate the mechanism of sugar accumulation in fruit vacuoles, a full length cDNA (CitVATP-A) encoding the vacuolar H+-ATPase 69-kDa catalytic subunit was isolated from a cDNA library constructed from citrus fruit (Citrus unshiu Marc.). A 2304-bp insert of CitVATP-A was coded for a 623 amino acid polypeptide with a predicted molecular mass of 68.68 kDa. The deduced amino acid sequence for CitVATP-A showed a 96.5% homology with the carrot homologue. Genomic Southern blot analysis suggested that CitVATP-A is a low-copy number gene. Northern blot analysis of leaves and fruits during the developing stages showed that the level of expression is high in young leaves and is low in mature leaves, and that it increased in both the edible parts and the peel, during fruit growth and maturity.
Asunto(s)
Citrus/genética , ADN Complementario/química , ATPasas de Translocación de Protón/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Southern Blotting , Citrus/enzimología , Clonación Molecular , Expresión Génica , Biblioteca de Genes , Datos de Secuencia Molecular , ATPasas de Translocación de Protón/química , Alineación de SecuenciaRESUMEN
Interspecific somatic fusion was performed between 'Seminole' tangelo (Citrus reticulata Blanco xC. paradisi Macf.) protoplasts isolated from embryogenic callus and rough lemon (C. jambhiri Lush.) mesophyll protoplasts. Eight plants out of ten randomly selected regenerants had 18 chromosomes and the same nuclear rDNA fragment patterns as that of the mesophyll parent. The remaining two plants showed rDNA fragment patterns from both parents and had 36 chromosomes. For the analysis of mitochondrial DNA (mtDNA),rrn26 derived from pea was used to probeBamHI digests of the regenerants. All plants showed mtDNA band patterns identical to that of the callus parent, suggesting that eight plants were cybrids and the remaining two plants were somatic hybrids. In addition to the callus parent band patterns, additional fragments from the mesophyll parent and/or a novel band fragment were revealed in some of the putative cybrids by peaatpA probe after digestion withDraI andPstI. These results suggest the occurrence of mtDNA recombination/rearrangement inCitrus cybrids produced by somatic fusion in this interspecific combination.
RESUMEN
A successful, efficient system for multiple soybean shoot induction of soybean [Glycine max (L.) Merr.] is reported. Multiple shoots were induced from cotyledonary nodes and hypocotyl segments cultured on media supplemented with 2 mg/l thidiazuron (TDZ) or 1.15 mg/l benzyladenine (BA). It was found that TDZ induced adventitious shoots more efficiently than BA and that hypocotyl segments promoted more adventitious shoots than cotyledonary nodes. The optimal TDZ concentrations for shoot organogenesis from hypocotyl segments were between 1 and 2 mg/l. Basal media also influenced the efficiency of shoot organogenesis. The frequency of adventitious shoot formation tended to increase when the salt concentration in the basal media supplemented with 2 mg/l TDZ was reduced. Two media (1/2B5 and 1/2L2) stimulated shoot organogenesis efficiently from hypocotyl segments. This method can thus be advantageously applied in the production of transgenic soybean plants.
RESUMEN
CMA/DAPI double staining and FISH methods using rDNA and telomere specific repeated sequences as probes were applied to characterize the large amount of heterochromatin in the telomeric regions of 'Trovita' orange (Citrus sinensis Osbeck) chromosomes. As a result, all the heterochromatic regions were CMA+/DAPI-, and thus were GC rich. Three CMA+/DAPI- heterochromatic regions were positive to the 26S rDNA probe. Telomere arrays consisting of Tm(A)Gn were detected on the extreme ends of each chromosome and most of the CMA+/DAPI- heterochromatic regions were ordered close inside the telomere-specific repeated sequences. The data obtained in this study will be useful for karyotyping, systematics, and molecular analysis of orange chromosomes.
Asunto(s)
Frutas/genética , Heterocromatina/ultraestructura , Sondas de ADN , ADN de Plantas , ADN Ribosómico/genética , Hibridación Fluorescente in Situ , ARN RibosómicoRESUMEN
Three partial cDNA clones (pSPS1, pSPS2 and pSPS3) encoding sucrose phosphate synthases (SPS) were isolated by Reverse Transcription (RT)-PCR using first-strand cDNA prepared from the leaf or fruit of citrus (Citrus unshiu Marc.). The nucleotide and deduced amino acid sequences of the three clones showed significant similarities to SPS previously isolated in other plants. A full-length, cDNA clone, CitSPS1, was isolated from a fruit (juice sacs and pulp segment) cDNA library using one (pSPS1) of the three partial clones as a probe. The 3539-bp CitSPS1 clone coded for a 1057-amino acid polypeptide with a predicted molecular mass of 117.8 kDa. The amino acid sequence deduced from the CitSPS1 clone showed homology with SPS from maize (55.8% identity) and spinach (74.0% identity). Genomic Southern blot analysis suggested that CitSPS1 clone represents a lowcopy-number gene. RNA blot analysis of leaf, flower and fruit showed that CitSPS1 and pSPS2 were expressed in all organs. However, the levels of expression of CitSPS1 in young leaves, flowers and immature fruits were low, but high in mature leaves, and fruit. pSPS2 transcripts were barely detectable in young leaves and immature fruits, low in mature leaves, and high in flowers and mature fruits. pSPS3 transcripts were only detected in young and in mature leaves.
Asunto(s)
Citrus/enzimología , Glucosiltransferasas/genética , Isoenzimas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Sondas de ADN , ADN Complementario , ADN de Plantas , Expresión Génica , Glucosiltransferasas/aislamiento & purificación , Isoenzimas/aislamiento & purificación , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Mensajero , ARN de Planta , Homología de Secuencia de AminoácidoRESUMEN
Nucleotide-sequence analysis showed that the gene for cytochrome oxidase subunit I (coxI) from tomato mitochondrial DNA has an ACG codon at a conserved position corresponding to an ATG initiation codon in other higher-plant coxI genes. cDNA-sequence analysis of the coxI transcripts showed that 15 positions in the genomic DNA were converted from C to U in the transcripts by RNA editing. One of the editing events is observed at the indicated ACG codon, producing an ATG initiation codon. The nucleotide sequences of 37 cDNA clones showed that the initiation codon was created in 32 out of the 37 clones, while nucleotide positions 254 and 11 were edited in 37 and 34 of the 37 clones examined, respectively, suggesting that creation of the initiation codon is a post-transcriptional event. The BamHI site at nucleotide position 757-762 within the coxI genomic DNA was altered in all 97 cDNA clones examined, demonstrating that RNA editing at this site in the transcripts is very common. RNA editing takes place to a lesser extent at the initiation codon, compared with editing at internal position 254. This indicates that editing is either a random process or that it involves a mechanism favoring less RNA editing in the initiation codon than in internal sites.
Asunto(s)
Codón Iniciador , Complejo IV de Transporte de Electrones/genética , Edición de ARN , Solanum lycopersicum/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Codón de Terminación , ADN Complementario , ADN Mitocondrial , Complejo IV de Transporte de Electrones/química , Solanum lycopersicum/enzimología , Mitocondrias/enzimología , Datos de Secuencia Molecular , ARN Mensajero/genética , Homología de Secuencia de AminoácidoRESUMEN
Renal and electrolyte disturbances in 91 patients with chronic myelogenous leukemia (CML) were analyzed over a period of these twenty years. At diagnosis, renal and electrolytes were studied in 72 patients including 65 in chronic cases, 5 in accelerated phase and 2 in blastic crisis. There were 8 cases of hypocalcemia among 62 patients and 5 cases of hyperphosphatemia among 48 patients. The cases of hyperphosphatemia and renal dysfunction had short median survival. There were no significant differences of renal and electrolyte disturbances between before and after chemotherapy. Various electrolyte disturbances, that is, hyponatremia, hypo-, hyperkalemia, hypocalcemia, hypo-, hyperphosphatemia, were found in the blastic crisis of CML. In the last admission, renal dysfunction and various electrolyte disturbances were present in almost half of the cases. Pathological studies were performed in 18 autopsy cases. Acute tubular insufficiency or necrosis, hypercalcemic nephropathy, and renal infiltration of leukemic cells were recognized in patients who had renal dysfunction.
Asunto(s)
Enfermedades Renales/etiología , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Desequilibrio Hidroelectrolítico/etiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
One hundred patients with diffuse non-Hodgkin's lymphoma were treated with combination chemotherapy including adriamycin. Seventy-one patients were under age 70 and 29 were 70 years or older. The clinical characteristics of the two groups were similar, except that the patients showed elevated serum BUN were in the elderly group. The complete remission rate in the elderly patients (66%) was slightly lower than that in the younger patients (75%) (p = NS). The average length of survival in the elderly patients was significantly shorter than in the younger patients (p less than 0.05) (projected 5-year survival: 28 +/- 11% vs 52 +/- 7%). The duration of remission for all patients in the elderly patients did not differ from that in the younger patients (projected 2-year survival: 40 +/- 10% vs 45 +/- 6%). Death during the induction chemotherapy from causes other than lymphoma occurred in 14% of patients over 70-year-old and in one percent of younger patients. At relapse, the response rate was significantly lower in the elderly patients than in the younger patients (CR+PR: 28% vs 78%, p = 0.03). To prevent toxic death in remission induction therapy, drug dose elderly patients should be attenuated according to their general conditions and performance status.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfoma no Hodgkin/tratamiento farmacológico , Factores de Edad , Anciano , Anciano de 80 o más Años , Ciclofosfamida/administración & dosificación , Doxorrubicina/administración & dosificación , Femenino , Humanos , Linfoma no Hodgkin/mortalidad , Masculino , Persona de Mediana Edad , Prednisolona/administración & dosificación , Inducción de Remisión , Estudios Retrospectivos , Tasa de Supervivencia , Vincristina/administración & dosificaciónRESUMEN
The nucleotide sequences of the tRNASer (trnS), pseudo-tRNA, NADH dehydrogenase subunit 3 (nad3), and ribosomal protein S12 (rps12) genes from rice mitochondrial DNA (mtDNA) were determined. Both trnS and nad3 were confirmed to be single copy genes by Southern blot analysis. The nad3 and rps12 genes were arranged in tandem, and the two were co-transcribed. The order of the above four genes in rice mtDNA differed from the linear order observed for the wheat and maize genes. In rice mitochondria, the trnS and pseudo-tRNA genes were found upstream of the cytochrome c oxidase subunit I gene, instead of the nad3 and rps12 genes as observed in maize and wheat. Additionally, while the rice nad3 and rps12 genes remain paired, they too are in a different sequence environment from the wheat and maize genes. The apparent split of the two pairs of genes indicates the occurrence of a mitochondrial intramolecular recombinational event. Another peculiarity is that the sequence upstream of the translational initiation codon of the rice nad3 gene is different from that of the wheat and maize versions. The ATG initiation codon of wheat and maize nad3 is replaced by TTG in the rice nad3. A subsequent deduction of the amino acid sequence, accompanied by a primer extension analysis, indicates that the predicted rice NAD3 protein has an additional 37 amino acid residues at its N-terminus compared to the wheat and maize NAD3 proteins. cDNA sequence analysis showed no introns or the occurrence of RNA editing at the newly replaced TTG codon.
Asunto(s)
ADN Mitocondrial/genética , Oryza/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Clonación Molecular , ADN/genética , Datos de Secuencia Molecular , NADH Deshidrogenasa/genética , Sistemas de Lectura Abierta , ARN/genética , ARN Mitocondrial , Mapeo Restrictivo , Alineación de Secuencia , Transcripción GenéticaRESUMEN
The clinical significance of immunophenotyping of the non-Hodgkin's lymphomas (NHL) is still controversial. Therefore, we evaluated the prognostic significance of T- and B-cell phenotype in 102 patients having diffuse non-Hodgkin's lymphoma who had been treated in a majority with Adriamycin-containing regimens. The significant differences in pretreatment clinical variables between the two patient groups were the higher frequency of markedly elevated LDH (greater than or equal to X 2 normal) and the lower frequency of decreased cholesterol level in B-NHL (p less than 0.05). Patients with B-NHL had a higher complete remission (CR) rate (77% vs 59%: p = NS), a significantly better overall survival (62% vs 34% at 3-years: p less than 0.05) and longer duration of remission for all patients (50% vs 27% at 3-years: p less than 0.05). Among forty-five patients with stages III and IV disease of nodal origin (excluding those with diffuse small cleaved histology), the B-cell group was associated with a better prognosis; a higher CR rate (68% vs 41%: p = NS) and a longer duration of remission for all patients (44% vs 12% at 2-years: p less than 0.05). Furthermore, for patients with T-cell phenotype, the primary site was the only prognostic factor; the patients whose disease originated from Waldeyer's ring, nasal cavity, and paranasal sinuses, achieved a better CR rate (92% vs 40%: p less than 0.05) and a longer duration of remission for all patients (48% vs 15% at 2-years: p less than 0.05). We conclude that advanced T-cell lymphoma of nodal origin is a subgroup of patients with very poor prognosis if treated with less intensive chemotherapeutic regimens.
Asunto(s)
Linfoma no Hodgkin/patología , Linfoma de Células T/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biomarcadores de Tumor/análisis , Doxorrubicina/administración & dosificación , Humanos , Inmunofenotipificación , Linfoma no Hodgkin/tratamiento farmacológico , Linfoma no Hodgkin/mortalidad , Linfoma de Células T/tratamiento farmacológico , Linfoma de Células T/mortalidad , Masculino , Persona de Mediana Edad , Pronóstico , Tasa de SupervivenciaRESUMEN
Ten patients with non-Hodgkin's lymphoma originated in the nasal cavity (four patients) and in the paranasal sinuses (six patients) were treated mainly with irradiation and combination chemotherapy including adriamycin. According to the TNM AJC staging system, four patients were in stage T1-T2, and six patients were in stage T3-T4. Nine patients, other than one with stage IV (Ann Arbor) disease, achieved complete remission. Death due to lymphoma occurred in four patients, 4 to 39 months following diagnosis. Three of these patients developed systemic extranodal dissemination, and died in a short time after relapse. Death due to second malignancies occurred in two patients. One died of acute myelogenous leukemia, and the other died of colon cancer, 26 and 53 months after diagnosis, respectively. Four patients were alive and disease-free, from 23 to 68 months following diagnosis (median 40 months). Out of four patients who died of disease, three were in stage T3-T4, and one was in stage T1. Two patients with stage T1 originated in the nasal cavity were both alive and disease-free. Except for lymphomas with stage T1 originated in the nasal cavity, more intensive chemotherapy should be instituted in an attempt to achieve better disease-free survival.
Asunto(s)
Linfoma no Hodgkin/patología , Neoplasias Nasales/patología , Neoplasias de los Senos Paranasales/patología , Adulto , Anciano , Terapia Combinada , Femenino , Humanos , Linfoma no Hodgkin/terapia , Masculino , Persona de Mediana Edad , Neoplasias Nasales/terapia , Neoplasias de los Senos Paranasales/terapia , Pronóstico , Inducción de RemisiónRESUMEN
A 42-year-old woman was admitted to our hospital because of easy fatigability in Jan. 1976. Laboratory examination revealed severe macrocytic anemia and slight lymphocytosis. She was diagnosed as having pure red cell aplasia (PRCA). She went into hematological remission 6 weeks following 40 mg/day treatment with prednisolone, but the anemia relapsed frequently when the dosage was lessened. She was then treated with 50 mg/day of cyclophosphamide, 50 mg/day of azathioprine, splenectomy, and methylprednisolone pulse therapy, but the recovery from anemia was temporary after each treatment. Since 1984, peripheral lymphocyte counts were 1-30,000/microliters, and reticulocyte counts were 0. She died of sepsis of Listeria in Sep. 1986. Peripheral lymphocytes had large azurophilic granules and an immunophenotype of OKT3+8+11+Ia1+Leu7+.
Asunto(s)
Trastornos Linfoproliferativos/complicaciones , Aplasia Pura de Células Rojas/complicaciones , Adulto , Azatioprina/uso terapéutico , Terapia Combinada , Ciclofosfamida/uso terapéutico , Femenino , Humanos , Metilprednisolona/uso terapéutico , Prednisolona/uso terapéutico , Aplasia Pura de Células Rojas/terapia , Inducción de Remisión , Esplenectomía , Linfocitos TRESUMEN
A 42-year-old male treated with carbamazepine developed a skin eruption in the trunk, followed by acute renal failure. Laboratory data revealed leukocytosis (15,700/mm3) with eosinophilia (37%). A renal biopsy disclosed granulomatous necrotizing angiitis that differed from classic periateritis nodosa and hypersensitivity angiitis. All drugs were discontinued immediately, and there was a gradual improvement of the renal function. A carefully performed provocation test using carbamazepine was positive. We made a diagnosis of carbamazepine-induced granulomatous necrotizing angiitis. Only 6 cases of drug-induced granulomatous angiitis have been reported prior to the present case. Drug-induced granulomatous angiitis is often fatal. The present report suggests the importance of making a diagnosis quickly, and providing adequate treatment in order to prevent a fatal course.
Asunto(s)
Lesión Renal Aguda/inducido químicamente , Carbamazepina/efectos adversos , Síndrome de Churg-Strauss/inducido químicamente , Hipersensibilidad a las Drogas/etiología , Lesión Renal Aguda/patología , Adulto , Síndrome de Churg-Strauss/patología , Humanos , Riñón/patología , MasculinoRESUMEN
One patient with gastric carcinoma and secondary myelofibrosis due to disseminated bone marrow metastasis had markedly elevated lysozyme (LZM) levels in serum and urine with the intense presence of LZM within tumor tissues. It is considered to be a case of gastric carcinoma with LZM secreting functional capacities. To date, there were many reported cases to verify the LZM positive cells by LZM staining in the tissue of gastric carcinoma and to demonstrate the elevated serum levels of LZM in malignant tumor bearing patients, whereas no papers to disclose the elevated levels of LZM in serum and urine originated from the productions and secretions of gastric carcinoma cells. So, this report might be the first reported case of LZM secreting tumor verified by LZM staining of carcinoma cells except for haematological malignancies such as acute monocytic leukemia or myelomonocytic leukemia.
Asunto(s)
Médula Ósea , Neoplasias Óseas/secundario , Muramidasa/metabolismo , Mielofibrosis Primaria/etiología , Neoplasias Gástricas/metabolismo , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Médula Ósea/patología , Neoplasias Óseas/patología , Humanos , Masculino , Persona de Mediana Edad , Muramidasa/sangre , Muramidasa/orina , Neoplasias Gástricas/patologíaRESUMEN
A 54-year-old male had monoclonal IgM-kappa light chains in the serum and free monoclonal kappa light chains in the urine. Renal biopsy revealed nodular glomerulosclerosis associated with the accumulation of kappa light chains. Isolated microscopic hematuria was present for over 1 year. He also showed a defect in urine concentration for which the light chains deposited along the basement membrane in the inner medullary tubules were judged to be responsible. When the glomerular filtration rate fell to 30 ml/min, the syndrome of mild renal failure, decreased potassium excretion, and hyporeninemic hypoaldosteronism developed, suggesting that kappa light chain nephropathy is a cause of this syndrome. Chemotherapy appeared to have some beneficial effects in maintaining renal function.