Background: Anemia is quite common in lung cancer patients and known to decrease the quality of life. Darbepoetin alfa is an erythropoiesis-stimulating protein approved for administration to cancer patients. This study examined the efficacy and safety of darbepoetin alfa in lung cancer patients with a hemoglobin concentration <10 g/dl during chemotherapy. Methods: Lung cancer patients (n=178) received darbepoetin alfa at doses of 1.91 ${\mu}g/kg$ per week until the hemoglobin concentration increased to >10 g/dl. The efficacy and safety were measured by comparing the hemoglobin concentration and assessing the adverse events. Results: After chemotherapy, the hemoglobin concentration decreased to 9.03${\pm}$0.64 g/dl. With the darbepoetin alfa treatment, the hemoglobin concentration increased to 10.09${\pm}$1.17 g/dl after 4 weeks reaching a peak hemoglobin concentration of 10.45${\pm}$1.18 g/dl. The changes in hemoglobin after 4 and 8 weeks with treatment were 1.08${\pm}$1.24 g/dl and 1.38${\pm}$1.59 g/dl (p<0.01). At least a 1 g/dl or more increase in hemoglobin was observed in 62.4% of patients. There were no serious adverse effects except for some mild reactions. Conclusion: Darbepoetin alfa administered to lung cancer patients appears to be an effective, well-tolerated treatment for chemotherapy induced anemia.
ATP is the energy source synthesized at the electron transferase that consist of complex I, II, III, IV and V in mitochondrial cristae. The complex V functions as ATPase which composed of sub-complex $F_0$ and $F_1$. Porin or VDAC (voltagedependent anion-selective channel), is a family of small pore-forming proteins of the mitochondrial outer membrane, and play important roles in the regulated flux of anion, proton and metabolites between the cytosolic and mitochondrial compartments. The channel allows the diffusion of negatively charged solutes such as succinate, malate, and ATP in the fully open state, but of positively charged ions in subconducting state. In this study, in order to investigate the relationship of the function and localization between porin and ATPase we observed the distribution of porin and ATPase in the mitochondria of the bovine heart. Monoclonal antibodies against porin and ATPase ${\beta}$-subunit were used to detect porin and ATPase using light microscope with immunohistochemistry and immunofluorescence, and using electron microscope with immunogold-labeling. ATPase were stained in longitudinal section region in cardiac muscle, porin were stained in longitudinal section region in cardiac muscle. We viewed more specific pattern of localization and distribution of these proteins using immunofluorescence method. There were some region which were labeled with porin or ATPase respectively, and others which were labeled both proteins in cardiac muscle. The electron microscope results showed that immunogold labeled porin were labeled locally at mitochondrial outer membrane and ATPase were labeled evenly at mitochondrial cristae. But ATPase was not labeled at mitochondria cristae. These results confirmed the subcellular localizations of porin and ATPase in mitochondrial outer membrane and cristae. Also, we assumed that ATP synthesis always does not activation in all mitochondria exist in the bovine cardiac muscle.
Park, Jong-Sug;Kim, Jong-Bum;Kim, Kyung-Hwan;Ha, Sun-Hwa;Han, Bum-Soo;Kim, Yong-Hwan
Journal of Plant Biotechnology
/
v.29
no.4
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pp.265-275
/
2002
Flavonoid biosynthesis is one of the most extensively studied areas in the secondary metabolism. Due to the study of flavonoid metabolism in diverse plant system, the pathways become the best characterized secondary metabolites and can be excellent targets for metabolic engineering. These flavonoid-derived secondary metabolites have been considerably divergent functional roles: floral pigment, anticancer, antiviral, antitoxin, and hepatoprotective. Three species have been significant for elucidating the flavonoid metabolism and isolating the genes controlling the flavonoid genes: maize (Zea mays), snapdragon (Antirrhinum majus) and petunia (Prtunia hybrida). Recently, many genes involved in biosynthesis of flavonoid have been isolated and characterized using mutation and recombinant DNA technologies including transposon tagging and T-DNA tagging which are novel approaches for the discovery of uncharacterized genes. Metabolic engineering of flavonoid biosynthesis was approached by sense or antisense manipulation of the genes related with flavonoid pathway, or by modified expression of regulatory genes. So, the use of a variety of experimental tools and metabolic engineering facilitated the characterization of the flavonoid metabolism. Here we review recent progresses in flavonoid metabolism: confirmation of genes, metabolic engineering, and applications in the industrial use.
Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.
The soybean stay green mutant genotype (SSG) derived from the nuclear gene, d1d2, and cytoplasmic gene, cytG, inhibit the breakdown of chloroplast in the leaves, pod walls, seed coats, and embryos during maturity. Soybean seed with black seed coat and green cotyledon (SBG) are preferred than black seed coat with yellow cotyledon (SBY) especially for cooking with rice and as source of traditional food in Korea. The researchers evaluated the seed's chlorophyll content of SSG and introduced SSG to the SBG variety breeding program. The seed chlorophyll content of SSG with d1d2 was $39.93{\sim}60.80\;{\mu}g/g$ and SSG with cytG $38.08{\sim}39.89\;{\mu}g/g$. The Korean SBG variety which was derived from SSG with cytG, contains $16.35{\sim}37.73\;{\mu}g/g$. The composition of seed chlorophyll differs according to the genetic background of SSG genotype. Inheritance study showed that cotyledon color was segregated 15:1 (yellow:green) at $F_2$ seed indicating two recessive genes control green cotyledon as revealed by previous study. Only less than 3% soybean lines showed black seed coat with green cotyledon among crosses SBY and SSG (d1d2). Results showed that SSG with d1d2 can be used as a good source for SBG with high chlorophyll content in the seed cotyledon, but due to the complex genetic behavior, breeding resource of SBG with d1d2 should be prepared to improve the breeding efficiency for development SBG variety.
Kim, Myo-Kyung;Lee, Sun-Hee;Choi, Su-Jin;Choi, Hye-Won;Park, Dong-Wook;Lim, Chun-Kyu;Song, In-Ok;Lee, Hyoung-Song
Clinical and Experimental Reproductive Medicine
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v.37
no.4
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pp.329-338
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2010
Objective: This study was carried out to know whether cryopreservation of sibling 2PN zygotes could increase the cumulative delivery rates in the patients who had less than 10 fertilized zygotes. Methods: A retrospective analysis was performed in 138 in vitro fertilization-embryo transfer (IVF-ET) cycles with less than 10 fertilized zygotes during January 2003 to December 2007 in Cheil General Hospital. These cycles were divided into two groups. In Group I (n=86), all fertilized embryos were cultured to transfer on day 3 without cryopreserved embryos at the 2PN stage. In Group II (n=52), among fertilized zygotes, some sibling zygotes were frozen at the 2PN stage, the remainder were cultured to transfer. Clinical outcomes in fresh ET cycles and cumulative ongoing pregnancy rates after subsequent frozen-thawed (FT)-ET cycles were compared. Results: There were no significant differences in female mean age, number of retrieved oocytes and total fertilized embryos between two groups, Number of cultured embryos was significantly lower in Group II ($5.2{\pm}0.5$) than in Group I ($8.4{\pm}0.7$) (p<0.01). Also, number of transferred embryos was significantly lower in Group II ($3.3{\pm}0.6$) compared with Group I ($3.6{\pm}0.6$) (p<0.01). ${\beta}$-hCG positive rates and delivery rates (51.2 vs. 46.2 % and 41.9 vs. 34.6 %, respectively) after fresh ET were slightly higher in Group I than in Group II. However, the differences were not statistically significant. Also, the cumulative delivery rates after subsequent FT-ET cycles were not significantly different between Group I (48.8%) and Group II (50.0%). Conclusion: This study showed that cryopreservation of sibling 2PN zygotes from patients who had less than 10 zygotes in the fresh ET cycles did not increase cumulative delivery outcomes. But, it could provide an alternative choice for patients due to offering more chance for embryo transfers if pregnancy was failed in fresh IVF-ET cycles.
The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.
Journal of the Korean Society of Food Science and Nutrition
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v.37
no.1
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pp.109-116
/
2008
In this study, the breads with medicinal herbs (MH) composites showing immunostimulating activity were prepared and their characteristics were examined. Fourteen kinds of medicinal herbs were extracted with hot water and divided into 3 groups (MH-1, MH-2, MH-3) based on their contents. All groups showed immunostimulating activity in terms of macrophage phagocytosis, nitrite production, cytostatic activity and cytokine production. In the preparation of breads containing MH extracts of various contents (0, 30, 50, 70, and 100%), there was no significant difference among dough pHs of all groups after first fermentation, but loaf volume was significantly (p<0.05) increased in 70% added group while decreased in 30%, 50%, and 100% added groups compared to the control. The "a" and "b" values of bread crumb increased with the content of MH extracts while "L" value decreased, but these values of bread crust were similar to the control group. Most improvements in hardness, adhesiveness, gumminess and chewiness of bread were noticed by the addition of 70% MH extracts, but those of springiness, cohesiveness and resilience were mostly by the 50% addition ones. Through the sensory evaluation, it was revealed that mouth feeling, taste and overall preference decreased at breads containing 70% and 100% extracts, although appearance and crumb texture were not significantly (p<0.05) different among all groups.
Previously, we have shown that green tea extract lowers the intestinal absorption of cholesterol, fat, and other fat-soluble compounds. We conducted this study to determine whether green tea extract affects the rate of $^{14}C$-oleic acid esterification into various lipids in the intestinal mucosa of rats. Male Sprague-Dawley ruts were had free access to a nutritionally adequate AIN-93G diet and deionized water. Initially, the rat's mucosal content of total lipids was measured following 1 mL olive oil administration with (green tea group) or without (control group) 100 mg green tea extract powder. At 1 h and 5 h, intestinal segments were extracted for total lipid analysis. Secondly, to measure mucosal esterification rates of lipids, an abdominal incision was made along the midline, and a 10-cm long jejunal segment of the small intestine was ligated in situ. Then, micellar solutions with or without green tea extract were injected into the ligated jejunal segments and incubated for 10 mill. The micellar solution contained $200.0\;{\mu}$ Ci $^{14}C$-oleic acid, $200.1\;{\mu}mol$ unlabelled oleic acid, $66.7\;{\mu}mol$ 2-monooleoylglycerol, $66.7\;{\mu}mol$ palmitoyl-sn-glycero-3-phosphocholine, 2.2 mmol glucose, $50.0\;{\mu}mol$ albumin, and 16.5 mmol Na-taurocholate per L of phosphate buffered saline (pH, 6.3) with or without 8.87 g green tea extract powder. At 10 min, each rat was sacrificed by cervical dislocation under anesthesia and the segment was removed for lipid analysis. Significant differences were observed in mucosal triglyceride content at 1 h and 5 h in ruts given green tea extract. Significant differences in the rate of $^{14}C$-oleic acid esterification into triglycerides and phospholipids fractions were observed between control and green tea groups. However, There were no significant differences in other lipid fractions. These results indicate that the lowered esterification rates of $^{14}C$-oleic acid into triglycerides and phospholipids fractions is attributable to presence of green tea extract. This may be associated with an inhibitory effect of green tea catechin on the mucosal processes of lipids, leading to the inhibition of intestinal absorption of lipids.
Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.
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