Grapes (Vitis spp. L.) are the third most produced fruit in the world. Crown gall disease caused by Agrobacterium vitis forms galls in the stems of the grapevines and reduces the vitality of the fruit trees, resulting in reduced yields. This pathogen has occurred in vineyards worldwide and caused serious economic losses. It is a soil-borne disease, so Agrobacterium vitis can survive for several years in vineyards and is difficult to control. Additionally, since there is no effective chemical control method, the most effective control method is the breeding of resistant varieties. To make the resistant variety, marker-assisted selection (MAS) enables fast breeding with low cost. In this study, we applied a genome-wide association study (GWAS), by combining phenotyping and genotyping-by-sequencing (GBS), for the development of a single nucleotide polymorphism (SNP) marker set related to crown gall disease using 350 grapevine varieties. As a result of the GBS based genotyping analysis, about 58,635 SNPs were obtained. In addition, the phenotypic analysis showed 35.2% resistance, 73% moderate susceptibility and 16.4% highly susceptibility. Moreover, after confirmation, two genes (VvARF4 and VvATL6-like) were shown to be related to crown gall disease based on the results of GWAS analysis, using the phenotypic data, and GBS. High-resolution melting analysis (HRMA) was performed using the Luna® Universal Probe with real-time PCR to distinguish the melting peaks of the resistant and susceptible varieties. Our data show that these SNP markers are expected to be helpful in evaluating resistance against grapevine crown gall disease and in breeding.
K-RAS is frequently mutated in human lung adenocarcinomas (ADCs), and the p53 pathway plays a central role in cellular defense against oncogenic K-RAS mutation. However, in mouse lung cancer models, oncogenic K-Ras mutation alone can induce ADCs without p53 mutation, and loss of p53 does not have a significant impact on early K-Ras-induced lung tumorigenesis. These results raise the question of how K-Ras-activated cells evade oncogene surveillance mechanisms and develop into lung ADCs. RUNX3 plays a key role at the restriction (R)-point, which governs multiple tumor suppressor pathways including the p14ARF-p53 pathway. In this study, we found that K-Ras activation in a very limited number of cells, alone or in combination with p53 inactivation, failed to induce any pathologic lesions for up to 1 year. By contrast, when Runx3 was inactivated and K-Ras was activated by the same targeting method, lung ADCs and other tumors were rapidly induced. In a urethane-induced mouse lung tumor model that recapitulates the features of K-RAS-driven human lung tumors, Runx3 was inactivated in both adenomas (ADs) and ADCs, whereas K-Ras was activated only in ADCs. Together, these results demonstrate that the R-point-associated oncogene surveillance mechanism is abrogated by Runx3 inactivation in AD cells and these cells cannot defend against K-Ras activation, resulting in the transition from AD to ADC. Therefore, K-Ras-activated lung epithelial cells do not evade oncogene surveillance mechanisms; instead, they are selected if they occur in AD cells in which Runx3 has been inactivated.
$\beta$-Xylosidase B was produced by Escherichia coli HB101/pKMG12 carrying the xylB gene of Bacillus stearothermophilus No.236 on its recombinant plasmid. The $\beta$-xylosidase B produced was purified by ammonium sulfate fractionation, DEAE-Sepharose CL-6B, Sephacryl S-200 and Superdex 200 HR gel filtration. The purified enzyme showed the highest activity at pH 6.5 and 5$0^{\circ}C$. But, the enzyme was observed to be very sensitive to the pH and temperature of the reaction mixture. The enzyme was activated about 35% of its original activity in the presence of 1 mM of $Mn^{2+}$ but it was completely inhibited by $Ag^{+}$, $Cu^{2+}$and $Hg^{2+}$ions. In contrast with the $\beta$-xylosidase A, the B enzyme was found to have $\alpha$-arabinofuranosidase activity though the activity was fairly low compared with the $\alpha$-arabinofuranosidase produced from the arfI gene of the same Bacillus stearothermophilus. Therefore, $\beta$-xylosidase B is considered to be more suitable than $\beta$-xylosidase A at least for the biodegradation of arabinoxylan. The $K_{m}$ and V$_{max}$ values of the $\beta$-xylosidase B for o-nitrophenyl-$\alpha$-D-xylopyranoside were 6.43 mM and 1.45 $\mu$mole/min, respectively. Molecular mass of the enzyme was determind to be about 54 kDa by SDS-PAGE and 160 kDa by Superdex 200HR gel filtration, indicating that the functional $\beta$-xylosidase B was composed of three identical subunits.s.
From May 1, 1993 to May 31 1995, the authers studied retrospectively 211 patients who underwent cardiovascular operation with cardiopulmonary bypass(CPB). Because we were interested in new development of ARF(prevalence, mortality rate, and main risk factors), we performed a multivariate statistical analysis about data of patients with preoperative serum creatinine values of less than 1.5 mg/dL. Normal renal function before operation(serum creatinine level less than 1.5 mg/dL) was registered in 198(74%) patients. Of these, 27(14%) patients showed postoperative renal complication, including 20(10%) patients classified as renal dysfunction(serum creatinine level between 1.5 and 2.5 mg/dL) and 7(4%) patients as acute renal failure(serum creatinine level higher than 2.5 mg/dL). The mortality rate was 5.8% in normal patients, 5% in patients with renal dysfunction, and 43% when acute renal failure developed(p=0.036). Indeed, the renal impairment proved to be an independent predictor of mortality(odd ratio 2.52∼11.25), along with cardiovascular(odd ratio 4.20) and respiratory(odd ratio 2.18) complications. Multivariate analysis identified the following variables as independent risk factors for postoperative renal impairment : advanced age(odd ratio 1), need for emergency operation(odd ratio 3.78), low-output syndrome(odd ratio 3.66), respiratory complication(odd ratio 1.30), need for deep hypothermic circulatory arrest(odd ratio 1.4). The 13 patients(7%) with preoperative renal failure showed a significantly higher morbidity and mortality rate than those without renal complications before operation. We concluded that the likelihood of severe renal complications is resonably low in the patients undergoing cardiac operation without preexisting renal dysfunction, but associated mortality remains high. A prominant role of hemodynamic factor in the development of postoperative acute renal failure must be recognized during preoperative, intraoperative, and postoperative periods.
The use of the lung assist device (LAD) would be well suited for acute respiratory failure (ARF) patients, combining the simplicity of mechanical ventilation with the ability of extracoporeal membrane oxygenators (ECMO) to provide temporary relief for the natural lungs. This study's specific attention was focused on the effect of membrane vibration in the LAD. Quantitative experimental measurements were performed to evaluate the performance of the device, and to identify membrane vibration dependence on blood hemolysis. We tried to decide upon excited frequency band of limit hemolysis when blood hemolysis came to through a membrane vibration action. The excited frequency of the module type 5, consisted of 675 hollow fiber membranes, showed the maximum gas transfer rate. We concluded that the maximum oxygen transfer rate seemed to be caused by the occurrence of maximum amplitude and the transfer of vibration to hollow fiber membranes. It was excited up to $25{\pm}5$ Hz at each blood flow rate of module type 5. We found that this frequency became the 2nd mode resonance riequency of the flexible in blood flow. Blood hemolysis was low at the excited frequency of $25{\pm}5$ Hz. Therefore, we decided that limit hemolysis frequency of this LAD was $25{\pm}5$ Hz.
p53 undergoes various post-translational modifications, including phosphorylation, ubiquitination, sumoylation, acetylation, methylation, and poly(ADP-ribosyl)ation. Modification of p53 widely affects to various functions of p53. Acetylation and phosphorylation of p53 have been studied for regulating its transcriptional activity which is observed in various stress condition. Otherwise, ubiquitination of p53 by Mdm2 has been well-studied as a canonical ubiquitin-mediated proteasomal degradation pathway. Moreover several investigators have recently reported that ubiquitination of p53 modulates not only its proteasome-dependent degradation by poly-ubiquitination but also its localization and transcriptional activity by mono-ubiquitination which usually does not serve the proteasome dependent degradation. Here we review recent studies on the cellular functions of p53 regulated by post-translational modifications, particularly focusing on mechanisms of ubiquitination.
The aim of this study was to evaluate the early and midterm results of off-pump total arterial revascularization using the skeletonized right gastroepiploic artery (RGEA) as a third arterial conduit. Material and Method: We prospectively analyzed 103 patients who underwent off-pump total arterial revascularization using bilateral internal thoracic arteries (ITAS) and RGEA. The RGEA was used as in situ graft in 88 patients, composite graft in 10 patients, and free graft in 5 patients. Postoperative coronary angiographies were performed before discharge in 100 patients, and at postoperative one year in 88 patients. Result: The RGEA showed a significantly higher free flow (130$\pm$95 ml/min) than that of right ITA(113$\pm$57 ml/min) or left ITA (107$\pm$55 ml/min), which was measured before anastomosis (p < 0.05). The total number of distal anastomoses was 3.8$\pm$0.7. The number of distal anastomoses per bilateral ITAs was 2.8$\pm$0.7 and the number of distal anastomosis per RGEA was 1.0. There were two morialities including one operative mortality. The late mortality was not related to cardiac events. Early postoperative morbidities were atrial fibrillation in 15 patients, bleeding reoperation in 4 patients, mediastinitis in 1 patients, perioperative myocardial infarction in 2 patient, and transient ARF in 3 patients. Postoperative coronary angiographies showed the early patency rate of 98.6% (272/276) for ITAs and 97.0% (97/100) for RGEA, respectively (p=ns), and the one-year patency rate of 95.9% (234/244) for ITAs and 88.6% (78/88) for RGEA, respectively (p=0.07). Flow competition between the RGEA and NCA (native coronary artery) was seen in 5 of the 100 patients (5.0%) immediate postoperatively and 7 of the 88 patients (8.0%) 1 year after surgery. Since July, 2000, we measured transit time flow intraoperatively and could reduce flow competition significantly Conclusion: The skeletonized RGEA demonstrated excellent early and midterm patency rates and could be used as a third arterial graft following the bilateral ITAs.
Lee Jee-Hyun;Jeon Ga-Won;Park Sung-Eun;Jin Dong-Kyu;Paik Kyung-Hoon
Childhood Kidney Diseases
/
v.9
no.1
/
pp.38-45
/
2005
Purpose : The purpose of this study was to analyze the therapeutic effect of plasmapheresis in various pediatric diseases. Methods : Therapeutic plasmapheresis was performed by COBE Spectra centrifugation. Nine cases were included in this study. The number an[;. method of plasmapheresis, together with the progress and prognosis of each case were retrospectively reviewed. Results : The patients' ages ranged from 26 mont]Is to 16 years of age, and the mean age was 9.9 years. There were S males and 4 females. The underlying diseases requiring plasmapheresis included 2 cases of hemolytic uremic svndrome(HUS), 1 case of lupus nephritis, 2 cases of rapidly Progressive glomerulonephritis(RPGN), 1 case of focal segmental glomorulosclerosis(FSGS), 1 case of systemic vasculitis after pulmonary hemorrhage, 1 case of acute renal failure associated with pulmonary hemoIThage, and 1 case of acute rejection after renal transplantation. The average number of plasmapheresis performed was 6.2 times with a range of 3 to 13 times. The patients with HUS, lupus nephritis, ANCA positive systemic vasculitis induced by pulmonary hemorrhage and ARF-associated pulmonary hemorrhage showed a good response to therapeutic plasmapheresis, but the patients with RPGN, refractory FSGS, and acute rejection after renal transplantation were not responsive to treatment. The most common side effect was hypocalcemia which was rarely symptomatic. Vital signs were not compromised. Conclusion : Although it is presumptuous to generalize the therapeutic effects of plasma pheresis in different diseases due to the small number of study subjects, this study shows that plasmapheresis may be an effective therapeutic modality in various pediatrics diseases and should be considered as a therapeutic option.
Between June 1994 to August 1996, 13 patients underwent emergency coronary artery bypass operations. There were 3 males and 10 females and ages ranged from 56 to 80 years with the mean of 65.5 years. The indications for emergency operations were cardiogenic shock in 12 cases and intractable polymorphic VT(ve'ntricular tachycardia) in 1 case. The causes of cardiogenic shock were acute evolving infarction in 6 cases, PTCA failure in 4 cases, acute myocardial infarction in 1 case, and post-AMI VSR(ventricular septal rupture) in 1 case. Pive out of 13 patients could go to operating room within 2 hours. However, the operations were delayed from 3 to 10 hours in 8 patients due to non-medical causes. In 12 patients, 37 distal anastomoses were constructed with only 3 LITA's(left internal thoracic arteries) and 34 saphenous veins. In a patient with post-AMI VSR, VSR repair was added. In a patient with intractable VT and critical sten sis limited to left main coronary artery, left main coronary angioplasty was performed. Pive patients died after operation with the operative mortality of 38.5%. Three patients died in the operating room due to LV pump failure, one patient died due to intractable ventricular tachycardia on postoperative second day, and one patient died on postoperative 7th day due to multi-organ failure with complications of mediastinal bleeding, low cardiac output syndrome, ARF, and lower extremity ischemia due to IABP. In 8 survived patients, 3 major complications (mediastinitis, PMI, UGI bleeding) developed but eventually recovered. We think that the aggressive approach to critically ill patients will salvage some of such patients and the most important factor for patient salvage is early surgical intervention before irreversible damage occurs.
From January 1993 to April 1995, 27 neonates (under age of 30 days underwent open heart surgery in the Department of Thoracic and Cardiovascular Surgery, Dong-A Medical Center. Mean age and weight were 12.1 days(2days∼306ays) and 3.29 kg(2.6kg∼4.1 kg) respectively. Cardiac anomalies were simple complete transposition of great arteries(TGA) in 11 neonates, TGA with coarctation of aorta(COA) in 1 , total anomalous pulmonary venous connection(TAPVC) in 5, double inlet right ventricle with TAPVC in 1, interrupted aortic arch(IAA) with ventricular septal defect(VSD) in 3, pulmonary atresia(PA) with intact ventricular septum(IVS) in 3, pulmonary stenosis with IVS in 1, Taussig-Bing anomaly with IAA in 1, and hypoplastic left heart syndrome(HLHS) in 1 . Postoperative complications were myocardial and/or pulmonary edema which caused open sternum in 13 patients(54.2%), acute renal failure( RF) in 10(37.0%), Intractable low cardiac output syndrome (LCOS) including weaning failure from cardiopulmonary bypass in 7(25.9%), bronchopulmonary dysplasia in 1, wound infection in 1, and paroxysmal supraventricular tachycardia in 1. Nine of 13 patients with postoperative open sternum were recovered with delayed sternal closure, and seven of 10 patients survived postoperative ARF with peritoneal dialysis. There were 8 operative deaths(29.6%): 3 in the patients with simple complete TGA, 1 In TCA with COA, 1 in PA with IVS, 1 in Taussig-Bing anomaly with IAA, 1 in DIRV with TAPVC, and 1 in HLHS. One late death occurred after arterial switch operation in simple TGA. The mosts common cause of death was low cardiac output syndrome. Our initial experience of open heart surgery in neonates showed high operative mortality and morbidity, especially in complex anomalies.
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