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BACKGROUND: Patients with incarcerated obturator hernia are usually elderly, frail, and physically inactive women with serious comorbidities. Although a laparotomy is standard surgical intervention for emergency incarcerated or strangulated obturator hernia, it is invasive particularly for these high-risk patients. The aim of this study is to show the feasibility of minimum open inguinal approach to reduce surgical risk for preoperatively diagnosed incarcerated obturator hernia. METHODS: Between April 2008 and July 2012, 3 consecutive incarcerated obturator hernia patients at Kamitsuga General Hospital who were diagnosed preoperatively by computed tomography underwent the following procedure. First a 4 cm inguinal hernia incision and preperitoneal dissection through the opening of the deep inguinal ring are made. The obturator hernia can be easily found 2 cm dorsally from the Cooper's ligament extraperitoneally. A small incision is made at medial sharp edge of the hernia defect. The hernia sac and its content can then be reduced. If the incarcerated bowel is viable, a prosthetic mesh is placed as a patch. If the bowel is necrotic, the damaged bowel loop is withdrawn through the wound and easily reconstructed extra-abdominally. RESULTS: All operations were successfully completed with this procedure. All patients recovered without incident. CONCLUSIONS: Minimal incision transinguinal repair for diagnosed incarcerated obturator hernia is feasible and provides an improved option to more invasive procedures.
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Hernia Obturadora/cirugía , Herniorrafia/métodos , Anciano , Anciano de 80 o más Años , Femenino , Hernia Obturadora/complicaciones , Humanos , Obstrucción Intestinal/etiología , Obstrucción Intestinal/patología , Obstrucción Intestinal/cirugía , Intestinos/irrigación sanguínea , Procedimientos Quirúrgicos Mínimamente Invasivos , Mallas QuirúrgicasRESUMEN
The effects of soil moisture and pH, and pathogen resting spore density, on the effectiveness of the biological control of clubroot by the fungal endophyte Heteroconium chaetospira was evaluated in greenhouse and field experiments. Conditions favoring disease development included low pH (5.5) and high soil moisture content (80%), with significant reductions in the disease being observed at a higher pH (6.3 and 7.2) and lower soil moisture content (40 and 60%). In greenhouse tests, H. chaetospira effectively controlled clubroot (reducing the disease by 90 to 100%) at pathogen resting spore densities of 104 and 105 spores/g of soil at all soil pHs tested (5.5, 6.3, and 7.2). However, when the resting spore density was 106 spores/g of soil, plants were severely diseased, regardless of treatment, and H. chaetospira had no effect on disease. At a soil moisture content of 40%, disease occurrence was low, regardless of pathogen spore density, but disease was significantly lower in H. chaetospira-treated plants at pathogen spore density of 105 spores/g of soil. At 60% soil moisture content, H. chaetospira significantly could affect at pathogen spore densities of 104 and 105 but not 104/g of soil. At 80% soil moisture content, there was no effect of H. chaetospira at pathogen density. In situ, the soil moisture contents were constantly adjusted to relatively low to moderate (pF 2.2 to 2.4 and pF 2.0 to 2.2) and high (pF 1.6 to 1.8). Other environmental conditions, such as resting spore density and soil pH, were maintained at constant levels. Control plants (not treated with H. chaetospira) showed uniformly high disease levels and proportions of diseased plants across all three moisture treatments (disease index = 72 to 80, proportion of diseased plants 85 to 97%). In the field, H. chaetospira-treated plants at low soil moisture (pF 2.2 to 2.4, plot 1) had 68% disease reduction compared with untreated controls and 49% reduction at moderate moisture pF (pF 2.0 to 2.2, plot 2). There was no effect on disease by H. chaetospira at high soil moisture (pF 1.6 to 1.8, plot 3). Based on our results, H. chaetospira is an effective biocontrol agent against clubroot in Chinese cabbage at a low to moderate soil moisture range and a pathogen resting spore density of 105 (or lower resting spores per gram of soil in situ.
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OBJECTIVES: We have tried to approach the aqueduct less invasively with the endoscope in combination with a small suboccipital craniectomy, especially for lesions of the aqueduct close to the fourth ventricle. METHODS: The patient is placed in the prone position and a small suboccipital craniectomy is performed. After elevating the bilateral tonsils with retractors, the sheath of the endoscope is inserted from a small skin incision made on the posterior midline of the neck, far from the craniectomy site. The skin incision for endoscopic insertion is planned on the linear extension connecting the aqueduct and the foramen of Magendi on the craniocervical MRI. A rigid endoscope is inserted through the fourth ventricle to the aqueduct for exploration and surgical manipulation. RESULTS: Two cases with hydrocephalus due to aqueductal stenosis, with gait disturbance were operated. After exploration of the aqueduct via the fourth ventricle, endoscopic aqueductal plasty was performed. The postoperative courses were uneventful. The patients' symptoms disappeared. CONCLUSIONS: This approach can be applied for less invasive endoscopic exploration and surgery around the aqueduct close to the fourth ventricle with a rigid endoscope, without overflexion of the neck, or a large craniectomy, or overretraction of the tonsils, or incision of the inferior vermis.
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Acueducto del Mesencéfalo/cirugía , Craneotomía/métodos , Cuarto Ventrículo/cirugía , Hidrocefalia/cirugía , Neuroendoscopía/métodos , Hueso Occipital/cirugía , Humanos , Masculino , Microcirugia/métodos , Persona de Mediana EdadRESUMEN
ABSTRACT Three hundred forty-nine fungal endophytes were obtained from a total of 1,214 root segments of eggplant, melon, barley, and Chinese cabbage grown as bait plants in a mixed soil made up of samples from different forest soils in Alberta and British Columbia, Canada. Three of the 349 isolates, when inoculated in axenically reared Chinese cabbage seedlings grown in petri dishes, almost completely suppressed the effects of a postinoculated and virulent strain of Verticillium longisporum. Two isolates effective against the pathogen were Phialocephala fortinii, which had been obtained from the roots of eggplant and Chinese cabbage. The third isolate was a dark septate endophytic (DSE) fungus obtained from barley roots. Hyphae of P. fortinii grew along the surface of the root and formed microsclerotia on or in the epidermal layer. Hyphae of the DSE fungus heavily colonized root cells of the cortex. Seedlings grown for 1 week in the presence of the endophytes were then challenged with the Verticillium pathogen. In DSE-treated roots, some of cell walls in the epidermal and cortical layers showed cell wall appositions and thickenings, which appeared to limit the ingress of the pathogen into adjacent cells. Such marked host reactions were not observed in the root cells colonized by P. fortinii. Chinese cabbage preinoculated with the above endophytes and, for comparison, a previously reported disease-suppressive fungal endophyte, Heteroconium chaetospira, were transplanted into the field and disease symptoms were assessed. The DSE could most effectively inhibit the development of Verticillium yellows, with reductions in the percentages of external and internal disease symptoms of 84 and 88%, respectively. The protective values against the disease are extremely high compared with those of other isolates. Most of the DSE-treated plants in the plots achieved marketable quality.
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The complete nucleotide sequence of the linear DNA plasmid (pRS224-1) from the plant-pathogenic fungus Rhizoctonia solani isolate H-16 was determined; and its unique RNA transcripts were characterized. The pRS224-1 DNA consists of 4,986 nucleotides. A computer-based study of the folding of pRS224-1 at both termini predicted hairpin-loop structures. The hairpin loops consisted of the left and right termini of 236 and 264 nucleotides, respectively, and share no sequence homology. Unique poly(A) RNAs, 4.7 kb and 7.4 kb in length and hybridizing with the pRS224 DNA, were found in mycelial cells of R. solani H-16. Transcript product-mapping allowed the prediction of the locations of different expression signals. The 7.4-kb transcript is generated from the left terminal region of the complementary strand, via the full-length sense-strand, to the right terminal region of the complementary strand. The 4.7-kb transcript is generated from the center region of the sense strand to the right terminal region of the complementary strand. One open reading frame (ORF) found in pRS224-1 is 887 amino acids long and has a potential coding capacity of 102 kDa. The ORF contains the highly conserved domains characteristic of reverse transcriptase sequences, including the highly conserved YXDD sequence.
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ADN de Hongos/genética , Plásmidos/genética , Rhizoctonia/genética , Secuencia de Aminoácidos , Secuencia de Bases , ADN de Hongos/química , ADN de Hongos/ultraestructura , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Hibridación de Ácido Nucleico , Plásmidos/ultraestructura , Poli A/genética , ARN de Hongos/metabolismo , ARN Mensajero/metabolismo , Mapeo Restrictivo , Rhizoctonia/ultraestructura , Análisis de Secuencia de ADN , Transcripción GenéticaRESUMEN
Heme oxygenase-1 (HO-1) is an inducible heat shock protein that regulates heme metabolism to form bilirubin, ferritin and carbon monoxide. Based on recent evidence that HO-1 is involved in the resolution of inflammation by modulating apoptotic cell death or cytokine expression, the present study examined whether overexpression of exogenous HO-1 gene transfer provides a therapeutic effect on a murine model of acute lung injury caused by the type A influenza virus. We demonstrate herein that the transfer of HO-1 cDNA resulted in (1) suppression of both pathological changes and intrapulmonary hemorrhage; (2) enhanced survival of animals; and (3) a decrease of inflammatory cells in the lung. TUNEL analysis revealed that HO-1 gene transfer reduced the number of respiratory epithelial cells with DNA damage, and caspase assay suggested that HO-1 suppressed lung injury via a caspase-8-mediated pathway. These findings suggest the feasibility of HO-1 gene transfer to treat lung injury induced by a pathogen commonly seen in the clinical setting. Since oxidative stress and lung injury are involved in many lung disorders, such as pneumonia induced by a variety of microorganisms and pulmonary fibrosis, HO-1 may be useful for wider clinical applications in gene therapy targeting lung disorders including acute pneumonia and pulmonary fibrosis.
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Terapia Genética/métodos , Hemo Oxigenasa (Desciclizante)/genética , Virus de la Influenza A , Pulmón/metabolismo , Infecciones por Orthomyxoviridae/terapia , Adenoviridae/genética , Animales , Caspasas/metabolismo , Fragmentación del ADN , Expresión Génica , Vectores Genéticos/administración & dosificación , Hemo-Oxigenasa 1 , Etiquetado Corte-Fin in Situ , Proteínas de la Membrana , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/metabolismo , Infecciones por Orthomyxoviridae/patología , Estrés OxidativoRESUMEN
By using a direct, intratracheal inoculation of an adenovirus encoding heme oxygenase 1 (Ad.HO-1), model gene therapy for acute lung injury induced by inhaled pathogen was performed. Data demonstrated that Ad.HO-1 administration is as effective as the pharmacologic upregulation of the endogenous HO-1 gene expression by hemin to attenuate neutrophilic inflammations of the lung after aerosolized lipopolysaccharide (LPS) exposure. Interestingly, immunohistochemical analysis revealed that the HO-1 gene was transferred not only to the airway epithelium, but to the alveolar macrophages (AMs). Moreover, overexpression of exogenous HO-1 in the macrophages provided a high level of endogenous interleukin 10 (IL-10) production from the macrophages, and additional experiments using IL-10 knockout mice demonstrated that the increase in IL-10 in the macrophages was critical for the resolution of neutrophilic migration in the lung after LPS exposure. These results suggest that AMs not only are barriers for efficient gene transfer to the respiratory epithelium, but also represent logical targets for Ad-mediated, direct, in vivo gene therapy strategies for inflammatory disorders in humans.
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Adenoviridae/genética , ADN Complementario/metabolismo , Técnicas de Transferencia de Gen , Hemo Oxigenasa (Desciclizante)/genética , Hemo Oxigenasa (Desciclizante)/metabolismo , Interleucina-10/biosíntesis , Interleucina-10/genética , Lipopolisacáridos/metabolismo , Lesión Pulmonar , Macrófagos Alveolares/metabolismo , Aerosoles , Animales , Lavado Broncoalveolar , Línea Celular , Relación Dosis-Respuesta a Droga , Células Epiteliales/metabolismo , Hemo-Oxigenasa 1 , Peróxido de Hidrógeno/farmacología , Inmunohistoquímica , Interleucina-8/biosíntesis , Pulmón/metabolismo , Proteínas de la Membrana , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Peroxidasa/biosíntesis , Factores de Tiempo , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
A cDNA clone encoding a putative EIN3-like protein (DC-EIL1) was obtained from total RNA isolated from senescing carnation (Dianthus caryophyllus L.) petals using RT-PCR and RACE techniques. The cDNA (2382 bp) contained an open reading frame of 1986 bp corresponding to 662 amino acids. The amino acid sequence of the N-terminal half of the protein, ranging from 80-300 amino acid residues, had 84% identity with that of the corresponding regions of Arabidopsis EIN3 and tobacco TEIL, although the overall identity was 49% and 52%, respectively. Northern blot analysis revealed that the amount of mRNA corresponding to DC-EIL1 decreased in flower tissues, especially in petals, during natural senescence and senescence induced by exogenously applied ethylene or ABA.
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Proteínas de Arabidopsis , Senescencia Celular/fisiología , Magnoliopsida/genética , Proteínas Nucleares/genética , Factores de Transcripción , Secuencia de Aminoácidos , Northern Blotting , Senescencia Celular/genética , Clonación Molecular , ADN de Plantas/genética , Proteínas de Unión al ADN , Magnoliopsida/metabolismo , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Proteínas de Plantas , Tallos de la Planta/genética , Tallos de la Planta/metabolismo , ARN Mensajero/metabolismo , ARN de Planta/metabolismo , Alineación de SecuenciaRESUMEN
The cDNA encoding a novel member (NT-ERS1) of ethylene receptor family of tobacco (Nicotiana tabacum L.) was obtained by a combination of RT-PCR and 5'-/3'-RACE cloning. The cDNA was 2,092 nucleotides long and had an open reading frame of 1,911 bp encoding 637 amino acids. The deduced polypeptide lacked a response regulator domain, indicating that the ethylene receptor belongs to an ERS-group. The amino acid sequence was similar to respective members of the tobacco ethylene receptor family: 67.8% to NT-ETR1, 39.1% to NTHK1 and 31.1% to NTHK2. Comparison of amino acid sequence suggested that NT-ERS1 is the counterpart of Nr in the ethylene receptor family of tomato, which belongs to Solanaceae as does tobacco. Northern blot analysis showed that mRNA of NT-ERS1 was present in leaf, shoot and root tissues, and accumulated in leaves treated with exogenous ethylene. A mutated NT-ERS1 cDNA transgene, obtained by introducing one nucleotide substitution into NT-ETR1 cDNA, conferred ethylene insensitivity in tobacco plants, indicating that the translation product of the cDNA actually functioned in the plants.
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Etilenos/metabolismo , Nicotiana/genética , Proteínas de Plantas/genética , Plantas Tóxicas , Receptores de Superficie Celular/genética , Northern Blotting , Clonación Molecular , ADN Complementario , Mutagénesis , Fenotipo , Plantas Modificadas Genéticamente , ARN Mensajero , ARN de Planta , Semillas , Transformación Genética , TransgenesRESUMEN
Carnation petals exhibit autocatalytic ethylene production and wilting during senescence. The autocatalytic ethylene production is caused by the expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase genes, whereas the wilting of petals is related to the expression of the cysteine proteinase (CPase) gene. So far, it has been believed that the ethylene production and wilting are regulated in concert in senescing carnation petals, since the two events occurred closely in parallel with time. In the present study, we investigated the expression of these genes in petals of a transgenic carnation harboring a sense ACC oxidase transgene and in petals of carnation flowers treated with 1,1-dimethyl-4-(phenylsulfonyl)semicarbazide (DPSS). In petals of the transgenic carnation flowers, treatment with exogenous ethylene caused accumulation of the transcript for CPase and in-rolling (wilting), whereas it caused no or little accumulation of the transcripts for ACC oxidase and ACC synthase and negligible ethylene production. In petals of the flowers treated with DPSS, the transcripts for ACC synthase and ACC oxidase were accumulated, but no significant change in the level of the transcript for CPase was observed. These results suggest that the expression of ACC synthase and ACC oxidase genes, which leads to ethylene production, is differentially regulated from the expression of CPase, which leads to wilting, in carnation petals.
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An 86-year-old male presented with progressive myelopathy due to retro-odontoid massive deposits of calcium pyrophosphate dihydrate (CPPD) crystals. Magnetic resonance imaging revealed a non-enhanced isointense extradural mass on the T1-weighted image and heterogeneously intense mass on the T2-weighted image. Computed tomography showed typical punctate and linear calcifications within the mass. The mass was resected via a lateral approach resulting in marked improvement of the symptoms. Histological examination revealed birefringent rhomboid crystals consistent with CPPD. CPPD deposition should be considered in the differential diagnosis of retro-odontoid extradural mass because surgical therapy is beneficial even for elderly patients.
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Calcinosis/patología , Pirofosfato de Calcio/metabolismo , Apófisis Odontoides/patología , Compresión de la Médula Espinal/patología , Anciano , Anciano de 80 o más Años , Cristalización , Humanos , Imagen por Resonancia Magnética , Masculino , Tomografía Computarizada por Rayos XRESUMEN
We developed a serum-free coculture model of benign prostatic hyperplasia (BPH) to clarify whether stromal cells stimulate growth of epithelial cells from BPH tissues. Epithelial and stromal cells from freshly isolated BPH tissue were cultured separately in defined serum-free WAJC 404/RPMI 1640 medium supplemented with insulin, transferrin, selenium, hydrocortisone, bovine serum albumin, epidermal growth factor, basic fibroblast growth factor and keratinocyte growth factor. (3)H-Tdr incorporation into epithelial cells and stromal cells was used as a measure of proliferation. When epithelial cells were cocultured with stromal cells, (3)H-Tdr incorporation into epithelial cells was increased in comparison to that in epithelial cells cultured alone. Dihydrotestosterone significantly increased this effect. It is likely that the in vitro coculture model reported here will be useful for isolating and understanding stromal cell-derived paracrine growth factor(s).
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Factores de Crecimiento de Fibroblastos , Sustancias de Crecimiento , Queratinocitos , Hiperplasia Prostática/patología , División Celular , Células Cultivadas , Medios de Cultivo Condicionados , Células Epiteliales/patología , Factor 10 de Crecimiento de Fibroblastos , Factor 7 de Crecimiento de Fibroblastos , Humanos , Masculino , Células del Estroma/patologíaRESUMEN
For gender determination of preimplantation embryos or circulating fetal cells in maternal blood, we developed a multiplex polymerase chain reaction assay from a single cell. This assay which co-amplifies X (DXZ1)- and Y (DYZ1)-specific repeat sequences, yields a 308-bp band in females and two bands of 154 and 308 bp in males. In a randomized, blinded assay of 100 isolated single amniocytes, 99 (99%) were amplified successfully. All 50 of the XY cells were correctly diagnosed as male (100%), whereas 49 of the 50 XX cells were diagnosed as female (98%). This accurate and efficient assay may be applicable in these clinical settings.
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Feto/citología , Reacción en Cadena de la Polimerasa/métodos , Procesos de Determinación del Sexo , Líquido Amniótico/citología , Femenino , Humanos , Cariotipificación , Masculino , Embarazo , Diagnóstico Preimplantación , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
A 68-year-old woman underwent surgical treatment for renal cell carcinoma associated with tumor thrombus extending into the right atrium. Although the tumor thrombus reached the level of the right atrium, there were no other apparent metastases. Combination therapy with interferon alfa plus tegafur/uracil (UFT) was attempted with the expectation of reducing the tumor thrombus, but there was no change. Successful management was achieved with right radical nephrectomy, right auriculotomy, and partial cavectomy using cardiopulmonary bypass under high-grade hypothermia. After removal of the tumor and thrombus, blood loss was 13,900 ml during the patient's recovery. She had mild heart failure for about two weeks after the operation, but recovered. She was discharged on the 40th day after the operation. Proper preparation for blood transfusion is the key point of this operation.
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Carcinoma de Células Renales/cirugía , Circulación Extracorporea , Atrios Cardíacos/cirugía , Hipotermia Inducida/métodos , Neoplasias Renales/cirugía , Células Neoplásicas Circulantes , Anciano , Transfusión Sanguínea , Femenino , Humanos , Cuidados Intraoperatorios , Nefrectomía , Resultado del Tratamiento , Vena Cava Inferior/cirugíaRESUMEN
PURPOSE: The effect of paternal age on the nondisjunction of sex chromosomes is controversial. Also, the prevalence of chromosomal anomalies in infertile patients is controversial, it has been reported that the sex chromosomal aneuploidy rate following treatment with intracytoplasmic sperm injection (ICSI) is higher than in naturally conceived pregnancies. We investigated the influence of paternal age and oligozoospermia on the nondisjunction of spermatozoa. METHODS: We determined the rate of aneuploidy for gonosomes and autosomes, using two-color fluorescence in situ hybridization (FISH) of the X and Y chromosomes and chromosomes 12 and 18 in 10 donors under 25 years of age who had a normal sperm count (> or = 20 x 10(6)/ml), 10 donors over the age of 39 years with idiopathic infertility and normozoospermia (> or = 20 x 10(6)/ml), and 5 oligozoospermic donors (< 20 x 10(6)/ml). RESULTS: There was no obvious relationship between increasing age and autosomal disomy (disomy 12 and disomy 18). Neither autosomal disomy nor diploidy was increased in any group. The frequency of X-, Y-, XX-, and YY-bearing sperm did not differ significantly among groups, but the frequency of XY-bearing sperm was significantly higher in the older infertile group than in the control donors. CONCLUSIONS: The incidence of nondisjunction of paternal sex chromosome in meiosis I was higher in older men with idiopathic infertility. The present results suggest that the risk of producing XXY fetuses is higher among men > 39 years of age with idiopathic infertility.
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Aberraciones Cromosómicas , Infertilidad Masculina/genética , Recuento de Espermatozoides , Espermatozoides/fisiología , Adulto , Factores de Edad , Aneuploidia , Cromosomas Humanos Par 12 , Cromosomas Humanos Par 18 , Humanos , Hibridación Fluorescente in Situ , Masculino , Meiosis , Oligospermia/genética , Oligospermia/patología , Cromosoma X , Cromosoma YRESUMEN
Replication-deficient adenovirus vector (Ad) is one of the most efficient gene transfer vehicles for human gene therapy. However, Ad is antigenic, known to evoke prominent inflammatory responses in vivo, and there are concerns that using Ad in patients with immune-mediated disorders (allergy and autoimmune diseases) may affect the status of the diseases. To evaluate this concept in a manner close to clinical scenarios, a mouse model of airway eosinophilic inflammation was developed by administering intraperitoneal injections and inhalations of chicken ovalbumin (OA), with Ad administered intranasally 5 days after the OA sensitization. The administration of Ad resulted in a significant suppression of eosinophil counts in peripheral blood as well as in the bronchoalveolar lavage fluid (BALF), and a decrease in OA-specific IgE. The decrease in the number of eosinophils in BALF was associated with a marked upregulation of interferon gamma (IFN-gamma) expression. In contrast, the Ad-specific, delayed-type hypersensitivity response and efficacy of reporter gene expression mediated by Ad were only marginally affected in animals sensitized with OA. Together, these data support the idea that Ad administration in patients with Th2-mediated immune disorders does not exacerbate the parameters of ongoing inflammations or gene transfer efficiency, and with its ability to induce prominent type 1 immune response to the antigen in vivo, Ad could potentially be used as an efficient adjuvant to control immune disorders where Th2 cell-mediated mechanisms are involved.
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Adenoviridae/genética , Técnicas de Transferencia de Gen , Hipersensibilidad/inmunología , Adenoviridae/inmunología , Animales , Líquido del Lavado Bronquioalveolar/citología , Pollos , Eosinófilos/patología , Genes Reporteros , Vectores Genéticos/efectos adversos , Humanos , Hipersensibilidad/metabolismo , Hipersensibilidad/patología , Hipersensibilidad Tardía/inmunología , Inmunoglobulina E/sangre , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Interferón gamma/metabolismo , Recuento de Leucocitos , Ratones , Ratones Endogámicos BALB C , Ovalbúmina , Células Th2/inmunologíaRESUMEN
Although the role of the gynoecium in natural senescence of the carnation flower has long been suggested, it has remained a matter of dispute because petal senescence in the cut carnation flower was not delayed by the removal of gynoecium. In this study, the gynoecium was snapped off by hand, in contrast to previous investigations where removal was achieved by forceps or scissors. The removal of the gynoecium by hand prevented the onset of ethylene production and prolonged the vase life of the flower, demonstrating a decisive role of the gynoecium in controlling natural senescence of the carnation flower. Abscisic acid (ABA) and indole-3-acetic acid (IAA), which induced ethylene production and accelerated petal senescence in carnation flowers, did not stimulate ethylene production in the flowers with gynoecia removed (-Gyn flowers). Application of 1-aminocyclopropane-1-carboxylate (ACC), the ethylene precursor, induced substantial ethylene production and petal wilting in the flowers with gynoecia left intact, but was less effective at stimulating ethylene production in the -Gyn flowers and negligible petal in-rolling was observed. Exogenous ethylene induced autocatalytic production of the gas and petal wilting in the -Gyn flowers. These results indicated that ethylene generated in the gynoecium triggers the onset of ethylene production in the petals of carnation during natural senescence.
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Magnoliopsida/fisiología , Envejecimiento/fisiología , Etilenos/metabolismo , Estructuras de las Plantas/fisiologíaRESUMEN
PURPOSE: In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary polymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes. METHODS: Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZ1 and a 154-bp DYZ1 repeat sequence on the X and Y chromosomes, respectively. RESULTS: All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate in diagnosing the gender of single amniocytes. No DNA contamination was observed in any samples. CONCLUSIONS: The multiplex PCR assay was rapid and accurate in diagnosing gender in single cells and may be clinically applicable in PGD.