• The Plant Pathology Journal
• /
• 제26권1호
• /
• pp.25-31
• /
• 2010

Papaya ringspot virus (PRSV) causes the most widespread and devastating disease in papaya. Isolates of PRSV originating from different geographical regions in south India were collected and maintained on natural host papaya. The entire coat protein (CP) gene of Papaya ringspot virus-P biotype (PRSV-P) was amplified by RTPCR. The amplicon was inserted into pGEM-T vector, sequenced and sub cloned into a bacterial expression vector pRSET-A using a directional cloning strategy. The PRSV coat protein was over-expressed as a fusion protein in Escherichia coli. SDS-PAGE gel revealed that CP expressed as a ~40 kDa protein. The recombinant coat protein (rCP) fused with 6x His-tag was purified from E.coli using Ni-NTA resin. The antigenicity of the fusion protein was determined by western blot analysis using antibodies raised against purified PRSV. The purified rCP was used as an antigen to produce high titer PRSV specific polyclonal antiserum. The resulting antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) assay and compared its sensitivity levels with ELISA based assays for detection of PRSV isolates. IC-RT-PCR was shown to be the most sensitive test followed by dot-blot immunobinding assay (DBIA) and plate trapped ELISA.

• Journal of Microbiology and Biotechnology
• /
• 제21권10호
• /
• pp.1026-1035
• /
• 2011

Vibrio parahaemolyticus, which owes its origin to the marine environment, is considered as one of the most common causes of infectious diarrhea worldwide. The present study investigated the pathogenicity of V. parahaemolyticus against the model organism, Caenorhabditis elegans. Infection in the host was localized with GFP-tagged V. parahaemolyticus using confocal laser scanning microscopy. The times required for causing infection, bacterial load in intestine, chemotactic response, and alteration in pharyngeal pumping were analyzed in the host system. In addition, the regulation of innate immune-related genes, lys-7, clec- 60, and clec-87, was analyzed using real-time PCR. The role of immune-responsible pmk-1 was studied using mutant strains. The pathogenicity of environmental strain CM2 isolated from the Gulf of Mannar, India was compared with that of a reference strain obtained from ATCC. The pathogen infected animals appeared to ward off infection by up-regulating candidate antimicrobial genes for a few hours after the exposure, before succumbing to the pathogen. For the first time, the pathogenicity of V. parahaemolyticus at both the physiological and molecular levels has been studied in detail using the model organism C. elegans.

• Proceedings of the SCSK Conference
• /
• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
• /
• pp.590-600
• /
• 2003

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

• Journal of Environmental Science International
• /
• 제23권6호
• /
• pp.1165-1173
• /
• 2014

For the field application of dielectric barrier discharge plasma reactor in nutrient solution culture, a filtration-DBD (dielectric barrier discharge) plasma reactor was investigated for the Ralstonia solanacearum which causes bacterial wilt in aquiculture. The filtration-DBD plasma reactor system of this study was consisted of filter, plasma reactor, reservoir. The DBD plasma reactor consisted of a quartz dielectric tube, discharge electrode (inner) and ground electrode (outer). The experimental results showed that the inactivation of R. solanacearum with filter media type in filter reactor ranked in the following order: anthracite > fiber ball > sand > ceramic ball > quartz ceramic. In filtration + plasma process, disinfection effect with the voltage was found to small. In disinfection time of 120 minutes, residual R. solanacearum concentration was 1.17 log (15 CFU/mL). When the continuous disinfection time was 120 minute, disinfection effect was thought to keep the four days. In sporadic operation mode of 30 minutes disinfection - 24 hours break, residual R. solanacearum concentration after five days was 0.3 log (2 CFU/mL). It is considered that most of R. solanacearum has been inactivated substantially.

• Journal of Plant Biotechnology
• /
• 제45권1호
• /
• pp.1-8
• /
• 2018

Meiosis is a specialized cell division, essential in most reproducing organisms to halve the number of chromosomes, thereby enabling the restoration of ploidy levels during fertilization. A key step in meiosis is homologous recombination, which promotes homologous pairing and generates crossovers (COs) to connect homologous chromosomes until their separation at anaphase I. These CO sites, seen cytologically as chiasmata, represent a reciprocal exchange of genetic information between two homologous non-sister chromatids. RAD51, the eukaryotic homolog of the bacterial RecA recombinase, plays a central role in homologous recombination (HR) in yeast and animals. Loss of RAD51 function causes lethality in the flowering plant, Arabidopsis thaliana, suggesting that RAD51 has a meiotic stage-specific function that is different from homologous pairing activity.

• Journal of Microbiology and Biotechnology
• /
• 제12권5호
• /
• pp.849-853
• /
• 2002

Metal ions have an adverse effect on anaerobic digestion. In an acetate degradation test of upflow of anaerobic sludge blanket granules with $Cu^{2+}$, not all of the acetate that disappeared was stoichiometrically converted to methane. In the presence of 400 mg/g-VSS (volatile suspended solids) $Cu^{2+}$, only 26% of the acetate consumed was converted to methane. To study acetate conversion by other anaerobic microorganisms, sulfate and nitrate reductions were investigated in the presence of $Cu^{2+}$ Sulfate and nitrate reductions exhibited more resistance to $Cu^{2+}$than methanogenesis, and the granules reduced 2.2 mM and 5.4 mM of nitrate and sulfate, respectively, in the presence of 400 mg/g-VSS copper ion. However, the acetate degraded by sulfate and nitrate reductions was only 24% of the missing acetate that could have been stoichiometrically converted to $CO_2$. Accordingly, 76% of the acetate consumed appeared to have been converted to other unknown compounds.

• Journal of Chest Surgery
• /
• 제24권1호
• /
• pp.1-7
• /
• 1991

This report describes our 10-year experience with intracardiac repair in 42 patients older than 16 year with tetralogy of Fallot. The mean age was 22.0$\pm$5.18 years[range 16~41]. The preoperative clinical manifestations were cyanosis & clubbing[93%], frequent URI [55%], anoxic spell[40%], pulmonary tuberculosis[21%], tuberculous empyema[7%], chronic renal failure[7%], congestive heart failure[7%] and subacute bacterial endocarditis [2%], etc. The previous shunt procedure for palliation had been performed in 7 patients. The type of VSD were typical perimembranous type[67%], total canal defect[28%] and combined type[5%]. The right ventricular outflow tract obstruction were combined type[69%], infundibular type[21%] and valvular type[10%]. Transannular patch was used in 50% of patients. Associated cardiovascular anomalies were ASD[33%], PFO[31%], Rt. aortic arch[10%], Lt. SVC[10%], single Rt. pulmonary artery, single Lt. coronary artery, ASI, proximal stenosis of Rt. pulmonary artery and anomalous systemic venous return, etc. Hospital mortality was 7.1%[3 cases]in overall. The causes of hospital deaths were revealed low cardiac output & acute renal failure[2 cases], postoperative bleeding[1 case]. There were 2 late deaths 3 & 68 months after surgery. Residual intracardiac shunt was detected in 2 patients. one patient was successfully reoperated and another patient had Qp /Qs ratio less than 1.5.

• Journal of The Korean Society of Inherited Metabolic disease
• /
• 제13권1호
• /
• pp.43-47
• /
• 2013

Introduction: ALT/AST enzymes are present inside the cells. AST is found in cardiac and skeletal muscle and red blood cells but the ALT is checked mainly in the liver. In general, the rise of these two indicators shows liver damage. The usual measurements of these enzymes are used in liver function tests, but the levels of AST and ALT do not always reflect liver function. Method and Cases: 17 cases of liver dysfunction transiently were evaluated clinically, biochemically, and imaging study of sonogram in pediatric in-patients for 3 years. Result: Most common causes of transient liver dysfunction were infection, especially viral gastroenteritis, and bacterial infection interfering oral food intake. More often occurred in the children who have infant hyperbilirubinemia, positive history of mitochondrial dysfunction or hypoglycemia. Fasting study in one case of hypoglycemia patient showed reversible liver dysfunction during fasting over 20 hours fasting. Discussion: A significant increase in AST and ALT with normal bilirubin can be observed in clinically healthy people during blunt trauma, viral infection, severe pain, metabolic syndrome, fasting or accidental health screening.

• Mycobiology
• /
• 제44권2호
• /
• pp.105-111
• /
• 2016

Paclitaxel (taxol) has long been used as a potent anticancer agent for the treatment of many cancers. Ever since the fungal species Taxomyces andreanae was first shown to produce taxol in 1993, many endophytic fungal species have been recognized as taxol accumulators. In this study, we analyzed the taxol-producing capacity of different Colletotrichum spp. to determine the distribution of a taxol biosynthetic gene within this genus. Distribution of the taxadiene synthase (TS) gene, which cyclizes geranylgeranyl diphosphate to produce taxadiene, was analyzed in 12 Colletotrichum spp., of which 8 were found to contain the unique skeletal core structure of paclitaxel. However, distribution of the gene was not limited to closely related species. The production of taxol by Colletotrichum dematium, which causes pepper anthracnose, depended on the method in which the fungus was stored, with the highest production being in samples stored under mineral oil. Based on its distribution among Colletotrichum spp., the TS gene was either integrated into or deleted from the bacterial genome in a species-specific manner. In addition to their taxol-producing capacity, the simple genome structure and easy gene manipulation of these endophytic fungal species make them valuable resources for identifying genes in the taxol biosynthetic pathway.

• IMMUNE NETWORK
• /
• 제15권1호
• /
• pp.9-15
• /
• 2015

Streptococcus pneumoniae (the pneumococcus) causes a major upper respiratory tract infection often leading to severe illness and death in the elderly. Thus, it is important to induce safe and effective mucosal immunity against this pathogen in order to prevent pnuemocaccal infection. However, this is a very difficult task to elicit protective mucosal IgA antibody responses in older individuals. A combind nasal adjuvant consisting of a plasmid encoding the Flt3 ligand cDNA (pFL) and CpG oligonucleotide (CpG ODN) successfully enhanced S. pneumoniae-specific mucosal immunity in aged mice. In particular, a pneumococcal surface protein A-based nasal vaccine given with pFL and CpG ODN induced complete protection from S. pneumoniae infection. These results show that nasal delivery of a combined DNA adjuvant offers an attractive potential for protection against the pneumococcus in the elderly.