Purpose : To observe the histopathological changes in the pulp tissues of mandibular molars in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes-irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in control and irradiation groups were injected with citrate buffer only. After 5 days, the head and neck region of the rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10 Gy. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the mandibular molars were sectioned and observed using a histopathological method. Results : In the diabetes group, capillary dilatation was observed. However, there was no obvious morphologic alteration of the odontoblasts. In the irradiation group, generalized necrosis of the dental pulp tissues was observed. Vacuolation of the odontoblasts and dilatation of the capillaries were noted in the early experimental phases. In the diabetes-irradiation group, generalized degeneration of the dental pulp tissues was observed. Vacuolation of the dental pulp cells and the odontoblasts was noted in the late experimental phases. Conclusion : This experiment suggests that dilatation of the capillaries in the dental pulp tissues is induced by diabetic state, and generalized degeneration of the dental pulp tissues is induced by irradiation of the diabetic group.
A rapid, user-friendly and simple immune-chromatographic dipstick kit named 'rapid immune-gold strip' (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein-A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.
The $\beta$-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5$\alpha$ with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3AI and ligated into pUC19 for the transformation of Escherichia coli DH5$\alpha$. Positive clones of $\beta$-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3AI insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5$\alpha$ (pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0$\sim$5.0 and 55$^{\circ}C$, respectively. The cloned enzyme was stable at 55$^{\circ}C$ or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).
A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.
Kim, Eun-Ah;Lee, Kyung-Wook;Boo, Won-Back;Lee, Hyung-Hoan;Kwak, Hae-Soo
Korean Journal of Food Science and Technology
/
v.23
no.4
/
pp.452-458
/
1991
Psychrotrophs producing protease were isolated during ripening periods of Cheddar cheese and one of them containing the highest protease activity was identified as Pseudomonas fluorescens 65. The extracelluar proteolytic enzyme was partially purified from P. fluorescens 65 through the Sephadex G-100 gel filtration. The protease was eluted between 190 ml and 230 ml of elution volume of sodium phosphate buffer. The purified protease showed a single band in SDS-PAGE and its molecular weight was 47,000. The composition of amino acid for the protease was determined and the most abundant amino acids were glutamic acid (14.96%) and serine (13.86%). The optimum temperature and pH for the activity was $45{\sim}50^{\circ}C$ and 6.0, respectively.
Purpose: To observe the histologic changes and clusterin expression in the acinar cells of the submandibular gland in streptozotocin-induced diabetic rat following irradiation. Materials and Methods: Mature Sprague-Dawley rats were divided into three groups: control, diabetic, and diabetic-irradiated groups. Diabetes mellitus was induced in the Sprague-Dawley rats by injecting streptozotocin, while the control rats were injected with citrate buffer only. After 5 days, rats in diabetic-irradiated group were irradiated with single absorbed dose of 10 Gy to the head and neck region. The rats were killed at 1, 3, 7, 14,21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histologic and immunohistochemical methods. Results : Morphologic change of acinar cells was remarkable in the diabetic group, but was not observed in the diabetic-irradiated group. Necrotic tissues were observed in the diabetic-irradiated group. Coloring of toluidine blue stain was most increased at 14 days in the diabetic group, however there were no significant change throughout the period of the experiment in the diabetic-irradiated group. Expression of clusterin was most significant at 14 days in the diabetic group, but gradually decreased with time after 7 days in the diabetic-irradiated group. Degeneration of clusterin was observed in the diabetic-irradiated group. Conclusion : This experiment suggests that the acinar cells of submandibular gland in rats are physiologically apoptosed by the induction of diabetes, but that the apoptosis is inhibited and the acinar cells necrotized after irradiation.
Purpose To observe the histopathological changes in the periodontal tissues of mandibular molars in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes- irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only After 5days, the head and neck region of the rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10Gy. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the mandibular molars were sectioned and observed using a histopathological method. Results In the diabetes group, osteoclastic activity was observed in the alveolar bone and the root throughout the period of experiment. Also, osteoblastic and fibroblastic activities were markedly decreased. In the irradiation group, the osteoclasts were observed in the alveolar bone and the dilated capillaries were increased in the early experimental phases. However, vigorous osteoblastic activity was noted in the late experimental phases. In the diabetes-irradiation group, osteoblastic activity in the alveolar bone and the root was observed in the early experimental phases. However, there were no resorption and osteoblastic activity in the alveolar bone and the root in the late experimental phases, and obvious atrophic change of fibrous tissues was noted. Conclusion : This experiment suggests that osteoblastic activity was caused by irradiation in the late experimental phases, but atrophic change of the periodontal ligament tissues was induced after irradiation in diabetic state.
Purpose : To observe the histopathological changes and caspase-3 expression in the submandibular gland in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes-irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only. After 5days, rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10 Gy to the head and neck region. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histopathological and immunohistochemical methods. Results : In the irradiation group, the condensed nucleus, karyolysis, and degeneration of the acinar cells and atrophy of the duct cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 28 days after irradiation. In the diabetes group, the condensed nucleus, karyolysis, atrophy, and degeneration of the acinar cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 21 days after diabetic state induction. In the diabetes-irradiation group, the ductal epithelial cells were predominant in their glandular tissues at 28 days after irradiation. In all of the experimental groups, the most prominent change of the acinar cells and ductal cells were observed at 14 days after diabetic state induction and irradiation. Conclusion The expression of caspase-3 in the acinar cells and ductal cells of the submandibular gland was weak after irradiation, but that in the acinar cells, ductal cells, and fibrous cells of the submandibular gland was prominent after diabetic state induction.
Thioltransferase, also known as glutaredoxin, was previously purified and characterized from Chinese cabbage (Brassica campestris ssp. napus var. pekinensis). However, in the process of gel filtration on Sephadex G-75, there were two activity peaks. In this study, a second thioltransferase (TTase CC-2) in the minor peak of the Sephadex G-75 elution profile was further purified using affinity chromatography on an S-hexylglutathione-agarose column by eluting with buffer solution containing 2.5 mM S-hexylglutathione. It showed a single band on SDS-PAGE indicating that TTase CC-2 is electrophoretically homogeneous. The molecular weight of TTase CC-2 was estimated to be about 22,000 daltons, and its isoelectric point was determined to be 6.73. Its size appears to be atypical and much larger than that of the first thioltransferase (TTase CC-1) from Chinese cabbage, and it can utilize 2-hydroxyethyl disulfide, S-sulfocysteine, and insulin as substrates. S-sulfocysteine was found to be a superior substrate for TTase CC-2. TTase CC-2 also displayed the reducing activity for non-disulfides such as dehydroascorbic acid. Its optimum pH was 8.5, which was consistent with that of TTase CC-1. TTase CC-2 activity was greatly activated by L-cysteine and reduced glutathione, and was found to be less heat-stable compared with TTase CC-1. Molecular and physiological differences between TTase CC-1 and TTase CC-2 remain to be elucidated. Chinese cabbage is the first plant which is known to contain two kinds of thioltransferases.
The bond strength of three types of $TiO_2$ coatings onto fluorine-doped $SnO_2$ (FTO) glass was investigated with the aid of a tape test according to ASTM D 3359-95. Transmittance was then measured using an UV-vis spectrophotometer in the wavelength range of 300 nm to 800 nm to evaluate the extent of adhesion of $TiO_2$ nanorods/nanoparticles on FTO glass. A sharp interface between the coating layer and the substrate was observed for single $TiO_2$ coating ($TiO_2$ nanorods/FTO glass), which may be detrimental to the bonding strength. In multicoating sample ($TiO_2$ nanorod/$TiO_2$ nanoparticle/$TiO_2$ nanoparticle/FTO glass), the tape test was not performed due to severe peeling-off prior to the test. On the other hand, the dual coating sample ($TiO_2$ nanorod/$TiO_2$ nanoparticle/FTO glass) showed minimum variation of transmittance (4%) after the test, suggesting that the topcoat adheres well with the FTO substrate due to the presence of the $TiO_2$ nanoparticle buffer layer. The use of a $TiO_2$ nanorod electrode layer with good adhesion may be attributed to the excellent dye sensitized solar cell performance.
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