Purpose : To observe direct effect of irradiation on cariogenic Streptooccus mutans. Materials and Methods : S. mutans GS5 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Viability and changes in antibiotic sensitivity, morphology, transcription of virulence factors, and protein profile of bacterium after irradiation were examined by pour plate, disc diffusion method, transmission electron microscopy, RT-PCR, and SDS-PAGE, respectively. Results : After irradiation with 10 and 20Gy, viability of S. mutans was reduced. Further increase in irradiation dose, however, did not affect the viability of the remaining cells of S. mutans. Irradiated 5. mutans was found to have become sensitive to antibiotics. In particular, the bacterium irradiated with 40Gy increased its susceptibility to cefotaxime, penicillin, and tetracycline. Under the transmission electron microscope, number of morphologically abnormal cells was increased as the irradiation dose was increased. S. mutans irradiated with 10 Gy revealed a change in the cell wall and cell membrane. As irradiation dose was increased, a higher number of cells showed thickened cell wall and cell membrane and Iysis, and appearance of ghost cells was noticeable. In RT-PCR, no difference was detected in expression of gtfB and spap between cells with and without irradiation of 40Gy. In SDS-PAGE, proteins with higher molecular masses were gradually diminished as irradiation dose was increased. Conclusion : These results suggest that irradiation affects the cell Integrity of S. mutans, as observed by SDS-PAGE, and as manifested by the change in cell morphology, antibiotic sensitivity, and eventually viability of the bacterium.
Purpose: The aim of this study was to observe a direct effect of irradiation on the periodontopathic Porphyromonas gingivalis (P. gingivalis). Materials and Methods: P. gingivalis 2561 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Changes in viability and antibiotic sensitivity, morphology, transcription, and protein profile of the bacterium after irradiation were examined by pour plating method, disc diffusion method, transmission electron microscopy, RT-PCR, and immunoblot, respectively. Results: Viability of irradiated P. gingivalis drastically reduced as irradiation dose was increased. Irradiated P. gingivalis was found to have become more sensitive to antibiotics as radiation dose was increased. With observation under the transmission electron microscope, the number of morphologically abnormal cells was increased with increasing of irradiation dose. In RT-PCR, decrease in the expression of fimA and sod was observed in irradiated P. gingivalis. In immunoblot, change of profile in irradiated P. gingivalis was found in a number of proteins including 43-kDa fimbrillin. Conclusion: These results suggest that irradiation may affect the cell integrity of P. gingivalis, which is manifested by the change in cell morphology and antibiotic sensitivity, affecting viability of the bacterium.
Crude sap, which was extracted from six Amaranthaceae plants, inhibited local lesion formation on Nicotiana glutinosa by tobacco mosaic virus(TMV) infection. Especially the remark. able inhibitory effect to TMV infection was shown on leaves of N. glutinosa precoated with the sap from Amaranthus mangostanus. The inhibitory activity of the sap from A. mangostanus was stable to storage in vitro for I day and to dilution 1/4 of the sap with distilled water. However, its activity was lost when the sap was heated at $70^{\circ}C\;to\;100^{\circ}C$ for 10 minutes. When the leaves of N. glutinosa precoated with the sap were sprayed with water, the inhibitory effect to TMV infection was maintained for 2 days. The A. mangostanus sap readjusted pH 3, pH 5, or pH 9 with 1 N HCl or 1 N NaOH did not decline the inhibitory action but the sap absorbed with $5\%\;to\;15\%$ charcoal completely lost their action. The protein components purified from A. mangostanus sap revealed three major bands by $5\%\;to\;15\%$ polyacrylamide gel electrophoresis and the top component of which showed the inhibitory action to TMV infection.
The classical concept for iron uptake into mammalian cells has been the endocytosis of transferrin( $T_{f}$ )-bound F $e^{3+}$ via the $T_{f}$ - $T_{f}$ receptor cycle. In this case, we could not explain the uptake of F $e^{2+}$ ion and the export of iron from endosome. Studies on iron transport revealed that other transport system exists in epithelial cells of the intestine. One of non- $T_{f}$ -receptor-mediated transport systems is Nramp2/DMT1/DCT1 which transports M $n^{++}$, $Mg^{++}$, Z $n^{++}$, $Co^{++}$, N $i^{++}$ or C $u^{++}$ ion as well as F $e^{+2}$ ion. DMT1 was cloned from intestines of iron-deficient rats and shown to be a hydrogen ion-coupled iron transporter and a protein regulated by absorbed dietary iron. DMT1 is founded in other cells such as cortical and hippocampal glial cells as well as endothelial cells in duodenum. Two F $e^{3+}$ ion bound to transferrin( $T_{f}$ ) are taken up via the $T_{f}$ - $T_{f}$ receptor cycle in the intestinal epithelial cell. F $e^{3+}$ in endosome was converted to F $e^{2+}$ ion, and then exported to cytosol via DMT1. F $e^{2+}$ ion is taken up into cytosol via DMT1. Several other transporters such as FET, FRE, CCC2, AFT1, SMF, FTR, ZER, ZIP, ZnT and CTR have been reported recently and dysfunction of the transporters are related with diseases containing Wilson's disease, Menkes disease and hemochromatosis. Evidences from several studies strongly suggest that DMT1 is the major transporter of iron in the intestine and functions critically in transport of other metal ions.
Choi, Moon Sil;Han, Bok Kyung;Choi, Hyuk Joon;Park, Young-Seo
Food Engineering Progress
/
v.15
no.3
/
pp.276-281
/
2011
The optimum condition for the extraction and purification processes of phytic acid from defatted rice bran was examined. The phytic acid was efficiently extracted when the defatted rice bran was treated with 10 volumes of 0.5% HCl for 1 hr. For the neutralization of acid-treated extract, 0.5% NaOH was the most acceptable. To purify phytic acid, Diaion HP20 resin was used to remove impurities from the extract. The flow-through was then loaded onto ion exchange columns packed with various resins and among them, Amberlite IRA-416 resin showed highest recovery yield. When the phytic acid was absorbed onto Amberlite IRA-416 resin and then eluted with 0.5% NaOH, 89% of applied phytic acid was eluted. Most proteins were removed from the purified phytic acid and total protein content of the phytic acid was 0.14%(w/w).
Purpose: Adsorption properties of lysozyme and albumin according to physiochemical properties of commercial contact lens classified with the FDA categories and a contact lens fabricated in the laboratory were investigated. Methods: The contact lens were prepared using HEMA(2-hydroxyethyl methacrylate) and TRIM(3-(trimethoxysilyl) propyl methacrylate) in a cast mold. Artificial tears containing lysozyme and albumin were prepared. We measured the amounts of protein adsorbed on the each lenses with varying adsorbed time (48 hour) and the pH range (6, 6.8, 7.4, 8.2, 9) of artificial tear. Amount of the proteins absorbed on the contact lenses were measured by using HPLC. Results: Time to reach the equilibrium of protein adsorption for silicone hydrogel lens was taken longer than hydrogel lens. The amount of adsorbed both lysozyme and albumin at equilibrium were greater for the hydrogel lens than the silicone hydrogel lens, and larger for the ionic lens than the non-ionic lens. Lysozyme was more adsorbed on the higher water content of contact lens, whereas albumin was more adsorbed on the lower water content of contact lens. Only lysozyme was adsorbed on the Group IV hydrogel lens of ionic higher water content. The adsorption of protein on contact lens increased with pH of artificial tears as close to the isoelectric point of each protein. Conclusions: The adsorption amount of lysozyme is more affected by the ionic strength of the contact lens surface than the water content of contact lens. Albumin adsorption is more affected by water content than the ionic strength of the contact lens surface. For the adsorption of proteins on the silicone hydrogel lens, the pore size, determined both by the number of Si atoms and the chemical structure of the silicone-containing monomers, as well as the polarity of contact lens should be also considered.
Two different Eelwardsiella tarda isolates, KFE and Edk-2, were obtained from Korea and Japan respectively. On the base of the previous results showing higher pathogenicity of E. tarda KFE compared to that of E. tarda Edk-2 isolate, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Concentration of specific antibody in the serum appeared to be much higher in loach immunized with FKC of E. tarda Edk-2 than those found in loach immunized with FKC of E. tarda KFE. Cross-reaction analysis using agglutination test with normal and antigen-absorbed antisera implied the differences epitopes in the antigens of these E. tarda isolates. For the comparison of bactericidal activity of the produced antibody with different antigens, absorption analysis was performed and confirmed the presence of critical epitopes in the FKC of E. tarda KFE strain. The prophylactic agent, polysaccharide-bound protein (PS-K) injected 1 week before the artificial infection with E. tarda KFE isolates decreased the cumulative mortality in loach and would be on effective method to prevent the occurrence of bacterial infection including E. tarda.
Inefficient rumen microbial fermentation is a major factor limiting intake of low quality roughage in ruminants. In this study, the effect of energy supplementation on rumen microbial fermentation, absorption of balanced digestion products and voluntary feed intake in sheep was investigated. A basal diet of a urea-treated mixture of wheaten chaff and barley straw (3:1 DM) containing 22.2 g N/kg DM was used. Four Merino-cross wethers weighing $45{\pm}4.38\;kg$ and fitted with permanent rumen and abomasal cannulae were allocated to four treatments in a $4{\times}4$ Latin square design. The dietary treatments were basal diet ($E_0$), or basal diet supplemented with sucrose (112.5 g/d) administered to the animals intra-ruminally ($E_R$), abomasally ($E_A$), or through both routes (50:50) ($E_{RA}$). Feed intake (basal and dietary) was increased (p<0.05) by sucrose supplementation through the rumen ($E_R$) or abomasum ($E_A$). However, there was no difference (p>0.05) in intake between animals on the control diet and those supplemented with sucrose through both intraruminal and abomasal routes ($E_{RA}$). The digestibility of DM and OM was highest in $E_R$ and $E_A$ supplemented animals. Although the rumen pH was reduced (p<0.001) in animals supplemented with sucrose entirely intra-ruminally ($E_R$), the in sacco degradation of barley straw in the rumen was not adversely affected (p>0.05). Intra-ruminal sucrose supplementation resulted in a higher concentration of total VFA, acetate and butyrate, while the pattern of fermentation showed a higher propionate: acetate ratio. Intra-ruminal supplementation also increased (p<0.05) the glucogenic potential (G/E) of the absorbed VFA. However, there was no difference (p>0.05) in microbial protein production between the four dietary treatments. Protozoa numbers were increased (p<0.05) by intra-ruminal supplementation of sucrose.
This study was performed to determine effects of the supplementation of Kimchi lactic acid bacterial culture in extruded pellets (EP) on the growth, body composition, blood chemistry and immune response of olive flounder Paralichthys olivaceus. Four hundred eighty individuals averaging 16.1 g were randomly distributed into 12, 180 L flow-through tanks (forty fish per tank). Four concentrations of Kimchi lactic acid bacterial culture (KL) were prepared: Control (0%), 0.1%, 0.2% and 0.5%. Three concentrations (0.1%, 0.2% and 0.5%) of Kimchi lactic acid bacterial culture were each diluted to 10% of EP weight and then fully absorbed by EP for 10 minutes. Each diet was fed to triplicate groups of fish. Fish were hand-fed to apparent satiation twice a day for 8 weeks. At the end of the 8-week feeding trial, the plasma lysozyme and bacterial activities of fish were determined. In addition, the cumulative mortality of fish was monitored for 8 days after their artificial infection with Edwardsiella tarda. The weight gain, specific growth rate, feed efficiency ratio, protein efficiency ratio, protein retention, hepatosomatic index and condition factor of fish were not affected by dietary supplementation with KL. None of the proximate composition, plasma parameters, lysozyme or bactericidal activities of fish was affected by dietary supplementation with KL. However, the cumulative mortalities of fish fed EP containing 0.1% and 0.5% Kimchi lactic acid bacterial culture were relatively low compared to that of fish fed the control diet. In conclusion, dietary supplementation with KL did not effectively improve growth, feed utilization, body composition, plasma chemistry, lysozyme, bactericidal activities or immune response of olive flounder after E. tarda infection under these experimental conditions.
Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.24-24
/
2017
Heavy metals at toxic levels have the capability to interact with several vital cellular biomolecules such as nuclear proteins and DNA, leading to oxidative stress in plants. The present study was performed to explore the metal tolerance mechanism in Sorghum seedling. Morpho-physiological and metal ions uptake changes were observed prominently in the seedlings when the plants were subjected to different concentrations of $CuSO_4$ and $CdCl_2$. The observed morphological changes revealed that the plants treated with Cu and Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cu and Cd was markedly increased by treatment with Cu and Cd, and the amount of interacting ions taken up by the shoots and roots was significantly and directly correlated with the applied level of Cu and Cd. Using the 2-DE method, a total of 24 and 21 differentially expressed protein spots from sorghum leaves and roots respectively, 33 protein spots from sorghum leaves under Cd stress were analyzed using MALDI-TOF/TOF MS. However, the over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. The results obtained from the present study may provide insights into the tolerance mechanism of seedling leaves and roots in Sorghum under heavy metal stress.
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