The nutritional components, antioxidant, and neuroprotective effects of water and a 50% methanol extract from litchi fruit pericarp were investigated. The most abundant mineral, amino acid, and fatty acid were K, proline, and palmitic acid, respectively. In addition, the total water phenolics and 50% methanol extracts were 8.02 and 12.28 mg/g, respectively. The DPPH, ABTS radical scavenging activities and ferric reducing antioxidant power of the water and 50% methanol extracts showed dose-dependent antioxidant activity. In a cell viability assay using MTT, almost all extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase leakage was also inhibited by the pericarp extracts. In particular, the 50% methanol extract showed a higher cell membrane protective effect than the water extract at the highest concentration. Consequently, these data suggest that litchi fruit pericarp can be utilized as an effective and safe functional food substances for natural antioxidants and may reduce the risk of neurodegenerative disorders.
The Journal of the Korean Society for Microbiology
/
v.21
no.1
/
pp.151-161
/
1986
In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.
This study was conducted to study the properties of the water-soluble natural chelating agents from garbage compost and activated sewage sludge responsible for Fe chelation, which is closely associated with the effectiveness in correcting iron chlorosis in plant. The water-soluble fraction of these materials was fractionated by menas of Sephadex gel filtration and the fractions of Fe chehates were traced by radioactive $^{59}Fe$. The fractions were examined by ultraviolet and infrared. spectroscopy and stability constants for Fe. The water-soluble fraction from garbage compost was separated by Sephadex G-25 into approximately four fractions. Most of the added $^{59}Fe$ was associated with fraction I, which appeared at the void volume. Further fractionation by Sephadex G-50 indicated that the molecular weight of water-soluble chelating agents is in the approximate range of 5000 to 10,000. The water-soluble fraction from activated sewage sludge gave six fractions by Sephadex G-25. Most of the added $^{59}Fe$ was found in the fraction I,II, and III, The molecular weights of most chelating agents associated with $^{59}Fe$ appeared to be less than 5,000 and those of fraction I that appeared at the void volume was in the range of 5,000 to 1,000. Discrepancy between radio activity count and UV absorption indicated the heterogeneity of the fractions obtained by Sephadex gel filtration. Ultraviolet absorption spectra of all fractions separated by Sephadex G-25 and containing chelating agents showed no differences. Fraction IV and V of sewage extract showed absorption maxima and shifting similar to nucleic acid components suggesting the presence of decomposition products of nucleic acid. Similarity fraction VI contained phenolic type amino acid groups. Fraction I of compost extract contained most of the added $^{59}Fe$ and showed weak but extra definite absorption in the 1230, and $1270cm^{-1}$ region, suggesting that extra oxygen groups in polyphenolic structure were probably involved in Fe chelation. In sewage extract, fraction I,II, and III in which most of the $^{59}Fe$ was found, showed strong definite polypeptide absorption in the region of $1540cm^{-1}$ due to NH deformation and C-N stretching of amide groups in the peptidebond. These extra functional groups in fraction I, II, and III appeared to be associated with Fe chelation. The other fractions, not associated with $^{59}Fe$, still have carboxyl and hydroxyl groups, suggesting that these functional groups in these water extracts may not independently form the Fe chelates. Precipitation of ferric hydroxide precluded measuring the stability constants for Fe-chelates. However, the formation constants for Zn chelates as log K values for compost extract and sewage extract at pH 4.0 from which the strength of chelation with Fe could be presumed, were 8.23, and 9.75, respectively, indicating strong complexation with metals. The chelating capacity of compost extract containing 6.5 g organic matter per liter was 0.82 mM, and that of sewage extract containing 5.3 g per liter was 0. 64 mM.
For the study of method for salt elimination aimed at reforming tidal land into normal paddy fields in a short period with reduction of periods requiring for elimination of saline, CHP (a kind of Ca-hum ate), a soil conditioner made of peat as a main material was tried. In the pot experiment, effect on elimination of salt, improvement of physical-chemical characteristics and rice cultivation test were studied. The results of these tests are as follows: 1, CHP treatment somewhat improves aggregation state with some effect on aggregation. 2. CHP treatment is remarkably effective in permeability which increases with 1.0 percent treatment by three times in percolation rate, and by 4.5 times in volume of leached water respectively. 3. With the increase of CHP amounts, salt was eliminated in short period. When 80% of the total Na was leached in 1.0% CHP-A treated pot, control pot begins permeable. 4. CEC and phosphorous absorption capacity are not influenced by CHP treatment. 5. Growing state of rice is greatly influenced by rainfalls. Growth of rice in tidal land however are almost similar to those in normal paddy fields with layer amounts of CHP treatment. With salt content in the soils, saline hazard and numbers of ineffective stems, amounts of unmatured grain are increased. 6. With the treatment of CHP yields of rough rice were increased. With 0.5% CHP treatment the yields were similar to those of the normal paddy fields. With 1.0% CHP-A treatment, the yields were increased by 15 times more than those of none treated soil and by 25 percent more than normal paddy soils.
Many studies have shown that the development of mouse early 2-cell embryos in vitro is related with the intracellular $Ca^{2+}$ changes. In ICR strain mouse, the development of embryos arrests at early 2-cell stage, but the arrested early 2-cell embryos can be rescued by the addition of $Ca^{2+}$-related materials. Acetylcholine (ACh) increases intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) via the mAChR-PLC-IP3 pathway in mouse oocytes. We examined whether ACh rescues 2-cell block in mouse. In early 2-cell embryos, ACh increased [$Ca^{2+}$]i in a dose-dependent manner (p<0.001), and had an effect on rescue of 2-cell block and embryonic development. To identify the signal pathway involved in ACh-induced rescue of 2-cell block, we first applied an agonist of ACh receptor (AChR). Like ACh, carbachol increased intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) and atropine, an antagonist of ACh receptor, blocked the ACh-induced $Ca^{2+}$ increase. In $Ca^{2+}$-free medium, ACh also increased [$Ca^{2+}$]i, indicating that $Ca^{2+}$ increased by ACh is mainly released from the intracellular $Ca^{2+}$ store. The ACh-induced $Ca^{2+}$ increase was blocked by PLC inhibitor (U73122), ryanodine receptor (RyR) antagonist (dantrolene), and CaM KII inhibitor (KN-93), but not by IP3R antagonists (xestospongin C). These results show that ACh increases intracellular $Ca^{2+}$ concentration via mAChR/PLC/RyR, and this contributes to the rescue of 2-cell block.
Because main barley straw management is changing these days from off-fields to burning that may relate to air quality concerning the global warming, this study was conducted to investigate the effects of barley-straw management practices on greenhouse gas emissions during rice cultivation in rice-barley double cropping system. The treatments were barley straw burning, off-field usage of barley straw and incorporation of barley straw in paddy fields. Laboratory experiment showed that burning of barley straw at the rate of $4.5Mg\;ha^{-1}$ emitted GHGs in the amounts of 4,607, 19.5, and $0.9kg\;ha^{-1}$ of $CO_2$, $CH_4$, and $N_2O$, respectively. During the rice cultivation of the rice-barley double cropping system, the highest GHG emission by evaluated close-static chamber method was observed from the soil incorporation of barley straw with 387 and $1.0kg\;ha^{-1}$ of $CH_4$ and $N_2O$, respectively. The GHGs emissions from the barley straw burning and off-field usage treatments were 233 and $160kg\;ha^{-1}$ for $CH_4$ and 0.80 and $0.79kg\;ha^{-1}$ for $N_2O$, respectively. The barley straw burning treatment showed the greatest GHGs emission among barley straw management practices in rice-barley double cropping system when considering GHGs emissions both during burning and from paddy fields during the cropping seasons. As a result, the GHGs emissions recorded in the barley straw incorporation to soil and off-field usage treatments were 22.4 and 66.8%, respectively, less than sum of GHGs emissions from the burning of barley straw and from paddy fields during rice cultivation.
Purpose : To elucidate the effects of pentoxifylline and diltiazem on the late response of the salivary glands of the rat after irradiation. Materials and Methods : Sixteen Sprague-Dawley rats were divided into 4 groups : (a) irradiation alone (b) irradiation with pentixifylline (PTX) (c) irradiation with diltiazem (DTZ) (d) irradiation with both PTX and DTZ. Irradiation was given in a single fraction of 16 Gy using 4 MV photon energy through an anterior port encompassing the left side of the salivary gland leaving the right side of salivary gland as a control. PTX, 20 mg/kg and/or DTZ, 50 mg/kg were infused intraperitoneally before irradiation, Two rats from each group were sacrificed on the 10th week and the rest was sacrificed on the 16th week after irradiation. Histopathologic examinations were undertaken for each section and the proportion of vacuolated cells out of the total number of cells under light microscopic fields was calculated. The statistical significance in the difference of the proportion of the vacuolated cells among the experimental groups was evaluated by a $x^2$-test. Results : Irradiated salivary glands of the 10th week group revealed markedly increased number of vacuolated cells compared to those of unirradiated control. The proportion of vacuolated cells was significantly reduced in both the PTX group (p value=0.001) and the combined PTX and DTX group compared to those of irradiation alone group. The DTZ alone group did not reveal the significant reduction of vacuolated cells compared to those of irradiation alone group (p value, >0.05). The 16th week groups revealed similar findings to those of the 10th week group, but the degree of chronic inflammatory cell infiltrates and interstitial fibrosis was increased and the number of acinar cells was reduced compared to those of the 10th week group. Conclusions : PTX significantly reduced the late radiation response of salivary glands, but DTZ did not reduce the same degree as PTX did. Taking the positive results of this study into consideration, it seems reasonable to apply PTX into the clinical trial for the head and neck irradiation to reduce the late radiation sequelae of salivary glands in the near future. At the same time the further experiment to clarify the subcellar mechni는 involved in PTX should be preceded.
Dans le contentieux m$\acute{e}$dical, il est difficile de prouver l'existence d'un lien de causalit$\acute{e}$ entre la faute m$\acute{e}$dicale et les pr$\acute{e}$judices subis par la victime. R$\acute{e}$partir bien de façon raisonnable la charge de la preuve du lien causal est alors une des questions pr$\acute{e}$occup$\acute{e}$es par la doctrine cor$\acute{e}$enne. La Cour supr$\hat{e}$me cor$\acute{e}$enne semble toutefois facilliter l'indemnisation des victimes dans les cas o$\grave{u}$ la responsabilit$\acute{e}$ du m$\acute{e}$decin a ${\acute{e}}t{\acute{e}}$ mise en cause, et cela en admettant des fois une solution op$\acute{e}$rant un renversement de la charge de la preuve du lien causal. Une telle attitude a ${\acute{e}}t{\acute{e}}$ m$\hat{e}$me affirm$\acute{e}$e dans un arr$\hat{e}$t rendu r$\acute{e}$cemment en cas de dommage caus$\acute{e}$ par le fait du produit de sant$\acute{e}$, notamment pour le cas de contamination virale par voie de transfusion. La Cour a $\acute{e}$galement reconnu que l'action se pr$\acute{e}$scrit $\grave{a}$ partir du moment de la consolidation du pr$\acute{e}$judice. Aux termes de cette $\acute{e}$tude, on pourra constater que le juge français reconna$\hat{i}$t aussi l'assouplissement de la charge de la preuve du lien de causalit$\acute{e}$ en mati$\acute{e}$re d'action m$\acute{e}$dicale. Il faudra toutefois souligner que le ph$\acute{e}$nom$\acute{e}$ne ne soit pas g$\acute{e}$n$\acute{e}$ralis$\acute{e}$ en droit français, d'autant plus que la pr$\acute{e}$somption de l'existence de la causalit$\acute{e}$ en la mati$\grave{e}$re a $\acute{e}$t$\acute{e}$ admise de mani$\grave{e}$re restrictive par la l$\acute{e}$gislation sp$\acute{e}$cifique. Tel $\acute{e}$tait notamment le cas pour les accidents de la contamination par le virus du sida ou de l'h$\acute{e}$patite C survenus apr$\grave{e}$s la transfusion. En d$\acute{e}$finitive, on peut dire qu'en droit français, le principe est maintenu en cas de manquement $\grave{a}$ une obligation de r$\acute{e}$sultat n$\acute{e}$e du contrat m$\acute{e}$dical, tandis que la Cour de cassation admet parfois en mati$\grave{e}$re de droit commun de la responsabilit$\acute{e}$ contractuelle la pr$\acute{e}$somption de causalit$\acute{e}$ en cas d'inex$\acute{e}$cution des obligations de r$\acute{e}$sultat. En fait, la pr$\acute{e}$somption de causalit$\acute{e}$ dans le contentieux m$\acute{e}$dical pourra mener les m$\acute{e}$decins $\grave{a}$ se diriger vers les traitements d$\acute{e}$fensifs. Cette situation peut m$\hat{e}$me conduire $\grave{a}$ emp$\hat{e}$cher le d$\acute{e}$veloppement de la science m$\acute{e}$dicale, enfin $\grave{a}$ une situation d$\acute{e}$savantageuse aux patients. Il y a alors lieu de se m$\acute{e}$fier des int$\acute{e}$r$\hat{e}$ts d$\acute{e}$s$\acute{e}$quilibr$\acute{e}$s entre le m$\acute{e}$decin et le patient. De ce point de vue, on peut estimer que le droit français donne des suggestions aux juristes cor$\acute{e}$ens dans la recherche des solutions plus ad$\acute{e}$quates en ce qui concerne la charge de la preuve du lien causal en mati$\acute{e}$re de responsabilit$\acute{e}$ m$\acute{e}$dicale.
Red rice (local name "Sore") as a weed has been a serious threat to rice production in direct-seeded rice culture in Ganghwagun, Gyeonggi province. In the Ganghwagun 508 ha out of 1,420 ha in the Samsanmyeon area is infested with red rice. The average lowland rice yield is about 4,300 kg/ha in the Ganghwagun, but the average upland rice yield is about 2,000 kg/ha in infested area. This study was carried out in order to clarify the ecological characteristics of red rice and factors affecting the competitive ability of five red rice varieties, collected from Samsanmyeon in 1981, with rice cultivar. Five varieties-Monggeunsare, Salsare, Ginkaragsare, Galsaegsalsare, Galsaegkaragsare-showed the same morphological characteristics of cultivated Japonica type, Chuncheongbyeo, but red rice tillers more profusely, is taller and lodges more easily than Chucheongbyeo. It shatters easily about 10-15 days after heading date, and at this time the hull is discolored in yellow or dark brown. There are many types of red rice with short or long owns on the spikelet, occasionally with or without own on the spikelet in the same hill, and the grains are short or long. In red reice leaf blast occurs more severely than in cultivated Indica/Japonica type, Teabaegbyeo, particulary serious in Monggeunsare. When red rice invaded in direct-seeded rice, number of panicles of rice became reduced more than other yield components.
This experiment was carried out to produce herbicide resistant potatoes hawing only chimeric phosphinothricin acetyltransferase (PAT) genes without using antibiotic selectable marker. The pDY502 vector having only PAT gene was reconstructed for transformation of potato. The reconstructed vector was introduced to Agrobacterium tumefaciens MP90 disarmed, and they were used for potato transformation. Hormonal requirement for plant regeneration from leaves and stem explants of potato was investigated. From this experiment, MS medium treated with IBA 0.1 mg/L + BA 0.5 mg/L was the best for potato regeneration, and the ratio of shoot regeneration was 54% for leaf and 46% for stem in that condition. For transformation, explants of potato leaves and stems were cocultured with A. tumefaciens MP90 containing reconstructed vector harvoring only PAT gene. When the potato explants were placed on various concentrations of bialaphos and all the potato explants were dead on medium with over 5.0mg/L bialaphos. By this selection methods, the explants cocultured with Agrobacterium produced the putative transgenic shoots on medium with 5mg/L bialaphos treatment after 3-4 weeks. Second selection was performed by transferring the shoot tips of putative transgenic to medium containing 20mg/L of bialaphos. The shoot tips grew well on the second selection medium, indicating the production of successful transgenic plants. But normal shoots were dead in same cytotoxic medium. Incorporation of the PAT gene into transgenic potatos were confirmed by PCR analysis of DNA and Southern hybridization. These results show that the PAT gene can serve as a selectable marker and herbicide resistant genes for transformation of potato.
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