Proceedings of the Korean Society of Plant Pathology Conference
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1995.06b
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pp.27-53
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1995
In plant pathology there is an increasing necessity for improved cytological techniques as basis for the localization of cellular substances within the dynamic fine structure of the host-(plant)-pathogen-interaction. Low temperature (LT) preparation techniques (shock freezing, freeze substitution, LT embedding) are now successfully applied in plant pathology. They are regarded as important tools to stabilize the dynamic plant-pathogen-interaction as it exists under physiological conditions. - The main advantage of LT techniques versus conventional chemical fixation is seen in the maintenance of the hydration shell of molecules and macromolecular structures. This results in an improved fine structural preservation and in a superior retention of the antigenicity of proteins. - A well defined ultrastructure of small, fungal organisms and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. From analyses of the compatible interaction between Erysiphe graminis f. sp. hordei on barley (Hordeum vulgare), various steps in the infection sequence can be identified. Infection sites of powdery mildew on primary leaves of barley are analysed with regard to the fine structural preservation of the haustoria. The presentation s focussed on the ultrastructure of the extrahaustorial matrix and the extrahaustorial membrane. - The integration of improved cellular preservation with a molecular analysis of the infected host cell is achieved by the application of secondary probing techniques, i.e. immunocytochemistry. Recent data on the characterization of freeze substituted powdery mildew and urst infected plant tissue by immunogold methodology are described with special emphasis on the localization of THRGP-like (threonine-hydrxyproline-rich glycoprotein) epitopes. Infection sites of powdery mildew on barley, stem rust as well as leaf rust (Puccinia recondita) on primary leaves of wheat were probed with a polyclonal antiserum to maize THRGP. Cross-reactivity with the anti-THRGP antiserum was observed over the extrahaustorial matrix of the both compatible and incompatible plant-pathogen interactions. The highly localized accumulation of THRGP-like epitopes at the extrahaustorial host-pathogen interface suggests the involvement of structural, interfacial proteins during the infection of monocotyledonous plants by obligate, biotrophic fungi.
Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.
Kim Young-Sook;Kim Mi-Young;Kang Tae-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yang Moon-Sik
Journal of Plant Biotechnology
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v.7
no.2
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pp.97-103
/
2005
This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.
Proceedings of the Korean Nutrition Society Conference
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2002.05a
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pp.29-41
/
2002
While the sequencing of several genomes was underway, several advanced techniques in genetics, molecular biology and protein chemistry emerged. Within the nutritional sciences, while the focus on nutrition education, epidemiology and public health aspects remains essential; it is crucial to incorporate the new advances in gene and protein discovery in nutritional studies. Nutrition is a discipline that has always integrated social, biochemical and physiological sciences from the studies at the molecule level to studies at the population level. For this reason, nutritionists are in a prime position to readily incorporate the current genomics approaches in nutrition research, All the available analytical techniques can and should be used in modern nutritional sciences. These include genetics, genomics, proteomics and metabolomics which also require integration and use of bioinformatics and computational methods for data analysis and management. These applications will be briefly reviewed with a primary focus on what the genomics and genetics approaches offer to nutritionists. We will use one of our research focus areas to illustrate uses of some of these applications in obesity-hypertension research. Our central hypothesis is that adipose tissue is an endocrine organ that plays a major role in obesity and related hypertension. We are primarily studying the renin angiotensin system (RAS). We provide evidence from our own studies and others for the paracrine as well as endocrine role of adipocyte-derived angiotensin II in adipocyte gene expression, adiposity and blood pressure regulation. Both cell culture studies as well as knockout and transgenic mice models are used to test our hypothesis. Genomics and proteomics technologies are currently developed to complement our physiological and molecular studies on the RAS and for a fine analysis of this system and its function in health and disease.
Proceedings of the Korean Nutrition Society Conference
/
2002.06a
/
pp.598-603
/
2002
While the sequencing of several genomes was underway, several advanced techniques in genetics, molecular biology and protein chemistry emerged. Within the notritional sciences, while the focus on nutrition education, epidemiology and public health aspects remains essential; it is crucial to incorporate the new advances in gene and protein discovery in nutritional studies. Nutrition is a discipline that has always integrated social, biochemical and physiological sciences from the studies at the molecule level to studies at the population level. for this reason, nutritionists are in a prime position to readily incorporate the current genomics approaches in nutrition research. All the available analytical techniques can and should be used in modem nutritional sciences. These include genetics, genomics, proteomics and metabolomics which also require integration and use of bioinformatics and computational methods for data analysis and management. These applications will be briefly reviewed with a primary focus on what the genomics and genetics approaches offer to nutritionists. We will use one of our research focus areas to illustrate uses of some of these applications in obesity-hypertension research. Our central hypothesis is that adipose tissue is an endocrine organ that plays a major role in obesity and related hypertension. We are primarily studying the renin angiotensin system (RAS). We provide evidence from our own studies and others for the paracrine as well as endocrine role of adipocyte-derived angiotensin II in adipocyte gene expression, adiposity and blood pressure regulation. Both cell culture studies as well as knockout and transgenic mice models are used to test our hypothesis. Genomics and proteomics technologies are currently developed to complement our physiological and molecular studies on the RAS and for a fine analysis of this system and its function in health and disease.
The herbicide bialaphos is a potent inhibitor of glutamine synthetase in higher plants. A bialaphos resistance (bar) gene encoding for an acetyltransferase was isolated from genomic DNA of Pseudomonas syringae pv tabaci. The bar gene was ligated to the binary vector pBI121. Pistils of tobacco plane were heated with the bar gene containing plasmid DNA at various times after pollination. When the treatment was applied at 30 and 40 h after pollination, a number of transgenic plants were obtained. Premary transformation (T$_{0}$ generation) and their progenies (T$_1$T$_2$) were resistant to both bialaphos and kanamycin at a dosage lathal to untransformed control plants. Stable integration of bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from T$_1$progenies. These results show that the bialaphos resistant plane could be obtained by treatment to pistils with the exgenous bar gene through the fertilization cycle of tobacco.o.
Acupuncture works by stimulating peripheral sensory nerves and their endings causing an increase in cutaneous blood flow and microcirculation, as well as release of neurotransmitters, neuropeptides, and hormones. SP6 Patch is a nanoscale nontransdermal device that mildly stimulates Stomach 36 (zusanli) and other acupuncture points. As stimulation of these points has been indicated to have an effect on hypothalamic function, it is of great research interest to investigate the effect of SP6 Patch on the physiology of organs that are affected by hypothalamic regulation. Bioelectrical tissue impedance data indicative of cellular physiologic organ function, using an Electro Interstitial Scan (EIS) system, were acquired from hypothalamus, pancreas, liver, intestines, kidneys, thyroid and adrenal glands in 10 (1 male, 9 females) volunteers while wearing the SP6 Patch daily for 1 week. EIS testing was performed at baseline with no patch, 30 min after wearing the patch, and after wearing the patch 12 h/day for 1 week. Subjects were instructed to keep well hydrated during the study period. All subjects served as their own control. The hypothesis was: The SP6 Patch worn 12 h/day on the Stomach 36 acupuncture point for 1 week, may significantly improve cellular physiologic functional status of different organs measured by EIS. All tested organs achieved significant improvement in their functional physiologic status after wearing the SP6 Pach 12 h/day for 1 week compared to baseline with an overall average statistical power > 89%. Based upon these results the hypothesis was accepted as true.
This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing $N^6$-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.
Adaptive lengthening or shortening of the nervous system is essential in order for a part of the body to move without restriction or resistance. Up until now, most treatment concepts have focused on normalizing muscle tone and preventing contractures in muscles and joint, but, as Butler and Gifford have brought to the attention of therapists, when a part of the body moves, nerves are required to move as well, so that the integration of nervous system mobilization in treatment is equally important. As the main function of the nervous system is impulse conduction, it is obvious therefore that adaptive lengthening, both centrally and peripherally, is essential in order to accommodate the enormous variety and extent of body movements and postures used in everyday life impeding news conduction. In present day manual therapy, most physical therapist mobilize the nervous system inadvertently. Probably what may make it more difficult is that, for most physical therapist it is a relatively new tissue to contemplate, at least in biomechanical terms, and basic knowledge of structure is generally less than if joints and muscles. The purpose of this study was to introduce the nervous system mobilization to a physical therapist who is primary a clinician and who has concerned in the areas of biomechanics and pathology looking far answers to the clinical Problems.
Digital subtraction radiography may be one of the most precise and noninvasive methods for assessing subtle density changes in peri-implant bone, providing additional diagnostic information on implant tissue integration in overall maintenance. The aims of this study were to evaluate density changes after first, second surgery of dental implant and to measure the amount of marginal bone loss 9 months after second surgery using digital subtraction radiography. Bone change around 30 screw-shaped implants in 16 patients were assessed on radiographs. 17 Branemark implants of 3.75mm in diameter(Nobel Biocare, Goteborg, Sweden), 2 Branemark implants of 5.0mm in diameter, 11 $Replace^{TM}$ implants of 4.3mm in diameter(Nobel Biocare, Goteborg, Sweden) were used. To standardize the projection geometry of serial radiographs of implants, customized bite block was fabricated using XCP film holder(Rinn Corporation, Elgin, IL.) with polyether impression material of Impregum(ESPE, Germany) and direct digital image was obtained. Qualitative and quantitative changes on radiographs were measured with Emago software(The Oral Diagnostic System, Amsterdam, Netherlands). The results were as follows: 1. The peri-implant bone density of 69.2% implants did not change and the peri-implant bone density of 30.8% implants decreased after 3 months following first surgery. 2. The crestal bone density of 53.9% implants decreased first 3 months after second surgery. The crestal bone density of 58.8% implants increased 9 months after second surgery. No density change was observed around the midportion of the implants after second surgery, 3. The amount of marginal bone loss between different kinds of implants showed no statistically significant differences (p>0.05). 4. More than 90% of total marginal bone loss recorded in a 9-month period occurred during the first 3 months.
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