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Apoptosis-Induced Effects of Extract from Artemisia annua Linné by Modulating Akt/mTOR/GSK-3β Signal Pathway in AGS Human Gastric Carcinoma Cells (AGS 인체 위암 세포에서 Akt/mTOR/GSK-3β 신호경로 조절을 통한 개똥쑥 추출물의 Apoptosis 유도 효과)

  • Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Kim, Sang-Yong;Kim, Young Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.9
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    • pp.1257-1264
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    • 2016
  • Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

An Experimental Study on Time Dependency of Strain for Saturated Clay (포화점토(飽和粘土)의 변형(變形)에 있어서 시간의존성(時間依存性)에 관한 실험적(實驗的) 연구(研究))

  • Park, Byong Kee;Lee, Jin Soo
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.4 no.3
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    • pp.43-52
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    • 1984
  • This paper is concerned with the strain characteristics of the time effect on the remoulded saturated day sampled from the downstream of the Yeongsan river, and the constitutive equation that can generally explain time-dependent behaviors of norma1ly consolidated clay. This paper examines whether or not the afore-said constitutive equation can be applied to the remoulded Mooan-clay. Throughout this study, the conclusions obtained are as follows. 1. Throughout the isotropic consolidation test for 7 days and the isotropic relaxation test, the existence of the static and dynamic yielding surfaces is confirmed respectively. 2. The characteristics of time effect of the deformation, namely, the existence of a unique stress-strain-time relation, is conformed from the experimental result on the Mooan-clay. 3. The prodictions of the stress path and the strain on the Cam-clay theory is not consistent with those observed during the experiments. 4. Constitutive equation(2-3-12) obtained by applying Cam-clay theory to Perzyna's elastic-viscoplasticity theory can explain the behavior of pore water pressure during isotropic stress relaxation, concerned with time dependency under undrained condition. The equation can also explain the results of the undrained triaxial compression test for the clay with different strain rate under the same or different consolidation history. 5. This constitutive equation has eight material parameters which can be determined from triaxial compression tests.

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Hypoglycemic Effect of Culture Broth of Bacillus subtilis S10 Producing 1-Deoxynojirimycin (1-Deoxynojirimycin을 생산하는 Bacillus subtilis S10 배양액의 혈당강하 효과)

  • Cho, Yong-Seok;Park, Young-Shik;Lee, Jae-Yeon;Kang, Kyung-Don;Hwang, Kyo-Yeol;Seong, Su-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1401-1407
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    • 2008
  • 1-Deoxynojirimycin (DNJ) is a strong $\alpha$-glucosidase inhibitor which inhibits hyperglycemia in animals. To select the Bacillus strains highly producing DNJ, 4,000 strains were isolated from soil and grain samples. By the inhibitory activity against $\alpha$-glucosidase, nine Bacillus strains were selected and then identified by 16S rDNA sequencing. B. subtilis S10 was finally selected as the best strain for the production of DNJ. Various carbon sources and nitrogen sources in culture medium were evaluated for the highest production of DNJ. As the results, the optimized concentration of carbon source and nitrogen source was 1.0% galactose and 1.6% polypeptone and the concentration of DNJ produced was 0.75 g/L. The effect of culture supernatant of B. subtilis S10 on lowering blood glucose level was investigated in streptozotocin (STZ)-induced diabetic mice model. Mice were randomly assigned to control group (saline) and three test groups such as acarbose group, silkworm powder group and B. subtilis S10 group. After eight-week oral feeding, blood glucose levels of the B. subtilis S10 and silkworm powder groups were respectively $209.1{\pm}19.6\;mg/dL$ (59.1%) and $208.6{\pm}39.8\;mg/dL$ (59.0%) lower than $510{\pm}10\;mg/dL$ of the control group. These results indicated that the culture supernatant of B. subtilis S10 was able to reduce the blood glucose level in STZ-induced diabetic mice.

Real-Time fMRI-Guided Functional MR Spectroscopy: The Lactate Peaks at the Motor Cortex during Hand-Grasping Tasks (실시간 기능적 자기공명영상 유도 기능적 자기공명분광법: 손운동 수행중 운동피질의 젖산변화 관찰)

  • Choi, Sun-Seob;Oh, Jong-Young;Kang, Myong-Jin;Lee, Jin-Hwa;Yoon, Seong-Kuk;Nam, Kyung-Jin
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.1
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    • pp.33-39
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    • 2008
  • Purpose : There is debate concerning the observation of metabolite changes on MRS at the designated cortex during some tasks. The purpose of this study is to assess the change of the lactate content at the motor cortex during hand-grasping tasks with performing real-time fMRI-guided fMRS. Materials and Methods : Seven healthy volunteers (23-28 years old) underwent realtime fMRI during right hand grasping tasks with using a 1.5 T system. After confirming the activating area, single voxel MRS was preformed at 1) the baseline, 2) during the task and 3) after the task on the activating cortex. The three consecutive spectra were compared for observing the changes of the lactate content by the tasks. The Cho/Cr, NAA/Cr and Lac/Cr ratios were calculated manually from those spectra. Results : MRS during the tasks revealed the lactate peaks at the 1.33 ppm resonance frequency with great conspicuity at the activated area, which was identified on the real-time fMRI. After the task scan, the lactate peaks completely disappeared and the spectra recovered to the values of the baseline scan in all volunteers. At baseline, during the task and after the task, the Cho/Cr ratios were 0.81, 0.76 and 0.77, respectively, and the NAA/Cr ratios were 1.68, 1.65 and 1.72, respectively, and the Lac/Cr ratios were 0.28, 0.41 and 0.30, respectively. During the task, Lac was significantly increased by 46%. Conclusion : We observed prominent lactate peaks on MRS during hand-grasping tasks at the activated area, as was shown on the real-time fMRI. We suggest that fMRS can be used as a sensitive tool for observing the metabolite changes of the functioning brain.

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A Study on the Official Uniform (2nd period) of Meritorious Vassals' Portraits in the Joseon Dynasty (조선시대 공신 초상(功臣肖像)의 관복(제2기) 고찰)

  • Kim, Migyung;Lee, Eunjoo
    • Korean Journal of Heritage: History & Science
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    • v.53 no.3
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    • pp.142-163
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    • 2020
  • In the early Joseon dynasty, the meritorious vassals' portrait of Jeokgae (1467, 1476), Jeongguk (1506)·Jeongnan (1507), and Wisa (1545) were defined as the 2nd period of the official uniform, in which the acheongsaek HeukDanryung appeared and settled. A total of seven portraits were analyzed, including three portraits of Jeokgae (1467, portrait production 1476) Son-so, Oh Jachi, and Jang Malson, and four portraits of Jeongguk (1506)·Jeongnan (1507) Yoo Sunjung, Hong Gyeongju, Lee Woo, and Yoo Hong. In the portrait of the Jeokgae wearing a Yebok HeukDanryung, in the case of the Samo, the height of the Samo, which had been raised at the beginning of King Seongjong, was lowered again, and the parietal part was rounded. The pattern was not expressed on the wide oval side wings of the Samo. In addition, the pattern was not expressed on the acheongsaek Danryung, Dapho, and Cheolrik. The colors of Dapho and Cheolrik were unified in green and red, respectively. The Mu of Danryung and Dapho was expressed as 'in and out wrinkled Mu', and the shoes were expressed in Baekhwa. Another feature is that the Jeokgae's portrait is expressed not as the Pumgye at the time of appointment, but as the Pumgye of the portrait production time. In the portrait of the Jeongguk·Jeongnan wearing a Sibok HeukDanryung, the height of the Samo lowered from the Jeokgae was maintained, while the parietal part was slightly narrowed and bent forward. The side wings of the Samo were expressed as an oval with a uniform width, and still the pattern was not expressed on the side wings. The pattern was not expressed in the costume of the Danghagwan's portrait, but the cloud pattern began to be expressed in the costume of the Dangsanggwan's portrait. And the Danryung was expressed as single Danryung without lining. In the portrait of Dangsang, the red Dapho and green Cheolrik were expressed, and in the portrait of Dangha, the green Dapho and blue Cheolrik were expressed. Therefore, it was found that the color of the undergarment was distinguished according to the Dangsang and Dangha. In Danryung and Dapho, the 'in and out wrinkled Mu' were maintained. The shoes were expressed in Heukhwa, and the rank badge and rank belt reflect the Pumgye at the time of the appointment. On the other hand, the portrait of Wisa could not be confirmed, but through the recording painting of the reign of King Meongjong, it was found that it was similar to the Danryung system of the reign of King Jungjong. Through the 2nd period of the meritorious vassal's portrait of Jeokgae, it was confirmed that the HeukDanryung outfit began to be used as the official uniform of the meritorious vassal's portrait, and that the Pumgye of the portrait production time was reflected in the portrait. After the meritorious vassal's portrait of Jeongguk, the pattern began to be expressed in the costumes, and in particular, it was possible to distinguish Dasang and Dangha by the presence or absence of the HeukDanryung's pattern. In addition, it was confirmed that the expressed aspect of the official uniform was consistent with the practices and regulations at the time.

IL-1 AND TNF-α RELEASE IN HUMAN POLYMORPHONUCLEAR LEUKOCYTES AFTER EXPOSURE TO CALCIUM HYDROXIDE TREATED Porphyromonas endodontalis LIPOPOLYSACCHARIDE (수산화칼슘 처리된 Porphyromonas endodontalis Lipopolysaccharide가 다형핵백혈구의 IL-1과 TNF-α 생성에 미치는 영향에 관한 연구)

  • Park, Chan-Je;Park, Dong-Sung;Yoo, Hyeon-Mee;Oh, Tae-Seok;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.27 no.5
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    • pp.463-472
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    • 2002
  • Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-$\alpha$ from immune cells. Although rnonocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-$\alpha$. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)$_2$ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-$\alpha$, and it was compared with Escherichia coli LPS. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 $\mu\textrm{g}$/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)$_2$ at 37$^{\circ}C$ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4$\times$10$^6$ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10$\mu\textrm{g}$/ml) for 24 hours at 37$^{\circ}C$ in 5% $CO_2$ incubator. The supernatants of cells were collected and the levels of IL-1$\alpha$, IL=1$\beta$ and TNF-$\alpha$ were measured by enzyme-linked immunosorbent assay. The results were as follows ; 1. The levels of IL-1$\alpha$, IL-1$\beta$, TNF-$\alpha$ from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05) 3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with p. endodontalis LPS were significantly lower than those released from PMN stimulated with E coli LPS (p<0.05).

Trans Fatty Acid Isomers of Processed Foods Commonly Consumed in Korea (한국인 상용 가공식품의 trans 지방산 이성체)

  • 노경희;원미숙;송영선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.3
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    • pp.325-337
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    • 2003
  • This study was designed to determine the distribution of trans fatty acids (tFAs) isomers of Processed foods commonly consumed in Korea. The tFAs positional isomers were analyzed using GC/MS spectrometer with HP-23 cis/trans FAME, capillary column (50m $\times$ 0.20 mm, id., 0.2 ${\mu}{\textrm}{m}$ film thickness) for 41 food samples. TFAs isomers were identified by comparing retention time with standards and GC/MS spectrum. In margarines, the content of tFAs ranged from 4.0% to 25.16% and the most abundant positional isomer of tFAs was C18:1 $\Delta$9t. In oils and fats, lards contained higher levels of tFAs (5.70~16.54%) than shortenings (6.77~10.55%). Shortenings contained higher levels of C18:1 $\Delta$9t (3.1~5.1%) than lard (1.6~4.3%), but corn oils had no tFAs. In seasonings, mayonnaise had no C16:1 $\Delta$9t, whereas C18:3t was detected. The content of tFAs in confectioneries was wide (16.20~52.16%). Among them, instant popcorns contained the highest amount of tFAs. Milk and dairy products showed even distribution of tFAS such as C18:1t, C18:2t, and C18:3t. Predominant tFAS isomer of condensed milk and ice cream was C16:1 $\Delta$9t. Frozen french fries and fried chicken contained higher levels of C18:1$\Delta$9t (9.4%), whereas grilled pork (jowl) had no C18:1 $\Delta$9t. The amount of tFAs per serving size was the highest in popcorn, followed by frozen pizza, frozen french fries, fried chicken, and bakeries.

Chemical Properties and Immuno-Stimulating Activities of Crude Polysaccharides from Enzyme Digests of Tea Leaves (녹차 효소 처리 다당의 화학적 특성 및 면역증진 활성)

  • Park, Hye-Ryung;Suh, Hyung Joo;Yu, Kwang-Won;Kim, Tae Young;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.664-672
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    • 2015
  • In order to develop new immuno-stimulating ingredients from mature leaves of green tea, crude polysaccharides were isolated from pectinase digests of tea leaves (green tea enzyme digestion, GTE-0), after which their immuno-stimulating activities and chemical properties were examined. GTE-0 mainly contained neutral sugars (54.9%) such as glucose (14.2%), arabinose (12.2%), rhamnose (11.1%), and galacturonic acid (45.1%), which are characteristic of pectic polysaccharides. The anti-complementary activity of GTE-0 was similar to that of polysaccharide K (used as positive control). Number of morphologically activated macrophages was significantly increased in the GTE-0-treated group. GTE-0 significantly augmented $H_2O_2$ and reactive oxygen species production by murine peritoneal macrophage cells in a dose-dependent manner, whereas production of nitric oxide showed the highest activity at a dose of $100{\mu}g/mL$ among all tested concentrations. Murine peritoneal macrophages stimulated with GTE-0 showed enhanced production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factors-${\alpha}$ in a dose-dependent manner. Further, GTE-0 induced higher phagocytic activity in a dose-dependent manner. In ex vivo assay for cytolytic activity of murine peritoneal macrophages, GTE-0-treated group showed significantly higher activity compared to the untreated group at an effector-to-target cell ratio of 20. The above results lead us to conclude that polysaccharides from leaves of green tea have a potent immuno-stimulating effect on murine peritoneal macrophage cells.

Defence Mechanism of Java Tilapia, Tilapia mossambica, to Clonorchis sinensis (Tilapia mossambica의 간흡충에 대한 방어기전)

  • RHEE Jae-Ku;PAIK Young-Ki;LEE Ho-Il;YANG Hong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.4
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    • pp.317-327
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    • 1987
  • As a series of studies on the suitability as a second intermediate host of Clonorchis sinensis, artificial infection experiments were applied to Tilapia mossambica. And then, in order to elucidate the defence mechanism of the fish to Clonorchis, clonorchicidal substance in the epidermal mucus of the fish was isolated by silica gel column and thin layer chromatography and analyzed for its chemical structure by UV, IR and NMR-spectroscopy. The results obtained are summarized as follows: 1. The cercariae which attempted to contact with the fish in the water were observed under stereomicroscope. After contact, the cercariae began to separate their tail from the body after several minutes and then the number increased to $80\%$ more than 10 minutes after the encounter. But very few cercariae could actually invade the epidermis of the fish. 2. The fish were reared with Parafossarulus manchouricus which were shedding numerous cercariae of Clonorchis in the aquarium for 24 hours. Only a few cercariae could invade the epidermis but most of the invaded cercariae died out before forming their cysts. Very few number of the remaining encysted cercariae were also found to be in a state of suspended animation within 42 hours. 3. In the cases of the control fish, Pseudorasbora parva, numerous cercariae of Clonorchis were found to invade the fish through the epidermis under stereomicroscope. Then many metacercariae of Clonorchis were also found in the fish while they were kept in the aquarium. 4. A sample of the epidermal mucus of Tilapia mossambica was extracted with ethly ether 6 times repeatedly. In silica gel column chromatography, using petroleum ether: chloroform/30:70(v/v) as a first solvent and MeOH as a second solvent, the extract was fractionated into the yellow and brownish red solutions in the first solvent and the clonorchicidal brownish yellow solution in the second sovent. 5. The clonorchicidal brownish yellow solution was added to petroleum ether, and the mixture was stored for 5 days at $5^{\circ}C$ and was, then, separated into supernatant fraction and precipitate. Ten mg/ml of the supernatant fraction killed, in vitro, the excysted metacercariae in 45 minutes but the precipitate in 600 minutes. 6. In silica gel column chromatography, using acetone: benzene/10:90(v/v) as a solvent, the more clonorchicidal supernatant fraction was fractionated into the first fraction with Rf. 0.2966 and the second fraction with Rf. 0.072. In vitro, 10mg/ml of the first fraction killed the excysted metacercariae in 28 minutes, the second fraction in 80 minutes and the first fraction was, therefore, determined to be a final clonorchicidal substance. 7. By this purification procedures, the most clonorchicidal substance from the epidermal mucus of Tilapia mossambica was purified 71-folds with $0.2075\%$ yield. Infra red, nuclear magnetic resonance and ultraviolet spectrometric analysis of the purified substance revealed that the substance is linoleic acid. According to the results of the present studies it seemed that this species can not serve as a proper intermediate host of Clonorchis sinensis, and that defence mechanism to the fluke seems to be correlated with linoleic acid in the epidermal mucus of this species.

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Effect of Brown Seaweed Waste Supplementation on Lactational Performance and Endocrine Physiology in Holstein Lactating Cows (미역부산물의 첨가가 홀스타인 비유우의 비유성적과 내분비생리에 미치는 영향)

  • Lee, H.G.;Hong, Z.S.;Li, Z.H.;Xu, C.X.;Jin, X.;Jin, M.G.;Lee, H.J.;Choi, N.J.;Koh, T.S.;Choi, Yun-Jaie
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.573-582
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    • 2005
  • This study was conducted to investigate effects of the brown seaweed waste(BSW) supplementation on milk production and related endocrine response in serum in Holstein dairy cows. A total of 14 Holstein dairy cows(initial mean live weight 625kg, average lactation days 225, Reproduction 2.4) were randomly allocated into control(basal diet) and treatment groups (4% BSW/basal diet) with 7 replications for 90 days. Dry matter intake was not affected by brown seaweed waste supplementation, but daily milk yield(kg) at the last experiment significantly increased (6.25kg) in treatment group compared with control group(p<0.05) at the last experiment. The plasma insulin-like growth factor(IGF)-1, triiodothyronine($T_3$) and thyroxine($T_4$) levels were significantly increased in treatment group compared with control group(p<0.05), although the concentration of plasma growth hormone(GH) was not significantly different. Milk composition was not significantly different between groups. The somatic cell count(SCC) in milk were significantly reduced in treatment group compared with control group(p<0.05), but antibodies(total IgG, G1, G2) were not significantly different between groups. Therefore we strongly believe that the increased milk yield is related to metabolic hormones as IGF-1, $T_3$ and $T_4$ and the mechanism of reducing SCC in milk must do more study related nonspecific immunsystem in the future.