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Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts (체외생산된 소 완전탈출 배반포기배의 동결보존)

  • Lee, K.S.;Kim, E.Y.;Yi, B.K.;Nam, H.K.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.141-148
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    • 1999
  • This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

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Effect of OPU (Ovum Pick-Up) Duration on the Rate of Collected Ova and In Vitro Produced Blastocyst Formation (OPU(Ovum Pick-Up) 채란기간이 난자 및 수정란 생산에 미치는 영향)

  • Jin, Jong-In;Kwon, Tae-Hyeon;Choi, Byeong-Hyun;Kim, Sung-Soo;Jo, Hyun-Tae;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.25 no.1
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    • pp.15-20
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    • 2010
  • This study was performed to identify the optimal timing for oocyte donor replacement during OPU procedure. OPU was carried out to collect oocytes from every donor at an interval of $3{\sim}4$ days (2 times a week). The collected oocytes were matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml of FSH and 1 mg/ml of estradiol for 24 h. After 24 h of exposure to sperm, the presumptive zygotes were cultured in CR1aa medium supplemented with 4 mg/ml of BSA for 3 days before being changed to CR1aa medium with 10% of FBS for another $3{\sim}4$ days. The mean numbers of retrieved oocytes were remained constantly up to 3 months ($6.0{\pm}0.5$, $6.2{\pm}0.7$, $5.2{\pm}0.6$), but significantly decreased at over 4 to 6 months ($3.7{\pm}0.5$, $2.8{\pm}0.4$, $1.2{\pm}0.2$) (p<0.05). The blastocyst development potential was also very similar rate from 1 to 3 months (37.2%, 40.4% and 44.6%), but significantly decreased from 4 to 6 months (24.8%, 29.3% and 28.6%, respectively) (p<0.05). The production of OPU derived embryos in periods of 1 to 3 months ($2.2{\pm}0.3$, $2.5{\pm}0.3$ and $2.3{\pm}0.4$) were significantly higher than those in 4 to 6 months ($0.9{\pm}0.2$, $0.8{\pm}0.2$ and $0.3{\pm}0.2$, respectively) (p<0.05). In conclusion, the efficient periods for the production of OPU derived embryos was until 4 months, twice per week to produce over 64 transferable embryos and then replace new donor after 3 months use. The best replacement time is 3 months and could be maximized production of OPU derived embryos.

Combined Chemoradiotherapy vs Radiotherapy Alone for Locally Advanced Squamous Cell Carcinoma of the Head and Neck (국소적으로 진행된 두경부 편평상피세포종양의 방사선- 항암화학 병용요법과 방사선단독치료의 비교)

  • Jeong, Hyeon-Ju;Suh, Hyun-Suk;Kim, Chul-Soo;Kim, Re-Hwe;Kim, Sung-Rok
    • Radiation Oncology Journal
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    • v.14 no.1
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    • pp.9-15
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    • 1996
  • Purpose: The traditional approach with surgery and/or radiotherapy(RT) for advanced head and neck cancer Provides anticipated cure rates of $10-65\%$ depending on stages and sites. Recently, combined modality with chemotherapy have been extensively investigated in attempts to improve survival and local control. We retrospectively analysed our experience of 31 patients with advanced head and neck cancer. Materials and Methods : November 1983 to October 1994. 31 Patients with Stage III and IV squamous cell head and neck cancer were treated with RT. Sixteen patients were treated with RT alone, and IS patients were treated with combined RT plus chemotherapy. All patients were treated with 4-MV LINAC and radiation dose ranged from 5000 cGy to 7760 cGy (median 7010 cGy). In combined group, 7 patients were treated with cisplatin plus 5-FU 2 patients were treated with methotrexate plus leucovorin plus 5-FU plus cisplatin or carboplatin, and 6 patients were treated with cisplatin as a radiosensitizer. Results : Median follow up period was 16 months (range 4-134 months). The major responses (CR+PR) were noted in 10 patient ($66.6\%$) of the RT alone group and 14 patient ($93.3\%$) of the chemoradiation group. There was no statistical difference in CR rate between the two groups The overall survival rates at 5 years were $23.4\%$ in the radiation alone group, $23.5\%$ in the chemoradiation group Disease-free survival rates at 3 years were $44.5\%$ in the radiation alone group, $40\%$ in the chemoradiation group. There was no statistical differences in overall survival rates and disease-free survival rates between the two groups. Local recurrences occurred in $71.5\%$ of the radiation alone group, $72.7\%$ of the chemoradiation group and distant metastasis occurred in $14.4\%$ of radiation alone group, $9.1\%$ of the chemoradiation group. The frequencies of complications were comparable in both groups except hematologic toxicity Conclusion : Total response rates in the combined chemotherapy and radiotherapy was relatively higher than radiotherapy alone. But our result failed to show any survival benefit of the combined chemotherapy and radiotherapy. The accrual of large number of patients and long term follow-un may be necessary to confirm the present result of combined chemotherapy and radiotherapy.

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The Effects of Preoperative Radiation Therapy in Resectable Rectal Cancer - in view of pathologic aspects - (절제 가능한 직장암에서 수술전 방사선 치료의 효과 -병리 조직학적인 연구를 중심으로-)

  • Choi, Ihl-Bong;Jang-Ji-Young;Kim, In-Ah;Shinn-Kyung-Sub;Lee, Jong-Suh;Chang-Suk-Kyun;Choi, Kyu-Young;Kim, Young-Ha;Kim, Jun-Gi;Chun-Chung-Soo;Kay-Chul-Seung
    • Radiation Oncology Journal
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    • v.15 no.1
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    • pp.49-56
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    • 1997
  • Purpose : To evaluate the pathologic effects of preoperative radiotherapy o the resectable distal rectal cancer, we analyzed the results of postoperative pathologic findings for the patients with preoperative radiotherapy ant surgery Materials and Methods: From July 1995 to April 1996, we treated sixteen patients of resectable rectal cancer with preoperative radiation therapy and curative surgery At diagnosis, Thomas Jefferson (TJ) system was used for the clinical stage of the Patients. We treated the patients with conventional radiation therapy of 4500~5000cGy before surgery. The surgery was carried out 4 weeks after completion of radiation therapy. Modified Astler Coller (MAC) system was used for the postoperative pathologic stage. We analyzed the pathologic stages and findings according to preoperative clinical stage and compared with those of the control group in similar clinical stages. Result : All patients were treated with sphincter preservation surgery after Preoperative radiation therapy. Pathoiogic complete response (CR) was shown in 1 case $(6.3\%)$. We compared the results between preoperative radiation therapy group (Preop.RT group) and surgery only group (control group). In TJ stage II, among nine patients of Preop.RT group, 8 patients $(88.9\%)$ were in MAC stage 8 except 1 CR patient, but among 17 patients of control group. 11 patients$(64.7\%)$ were in MAC stage B and 6 Patients $(35.3\%)$ in MAC stage C. In TJ stage III, among 7 patients of Preop.RT group, 4 patients $(57.1\%)$ were in MAC stage B and 3 patients$(42.9\%)$ in MAC stage C. Among 14 Patients of control group, 4 patients $(28.6\%)$ were in MAC stage B and 10 Patients $(71.4\%)$ in MAC stage C. Above results showed that postoperative Pathologic stage was decreased in Preop.RT group with statistical significance (P=0.049). The postoperative Pathologic findings (blood vessel invasion. Iymphatic vessel invasion, perineural invasion) were decreased in the Preop.RT group compared with those of control group. But statistical significance was found only in Iymphatic vessel invasion (p=0.019). Conclusion : The Postoperative pathologic stages and adverse Prognostic pathologic findings were decreased in preoperative radiation therapy group. The Iymphatic vessel invasion and MAC stage C findings were abruptly decreased in Preoperative radiation therapy group. The preoperative radiation therapy was found to be effective in resectable rectal cancer. The patients group in our study was very small and long term follow up was not done. Therefore, further study about this issues is needed.

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The Effect of Nasal BiPAP Ventilation in Acute Exacerbation of Chronic Obstructive Airway Disease (만성 기도폐쇄환자에서 급성 호흡 부전시 BiPAP 환기법의 치료 효과)

  • Cho, Young-Bok;Kim, Ki-Beom;Lee, Hak-Jun;Chung, Jin-Hong;Lee, Kwan-Ho;Lee, Hyun-Woo
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.190-200
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    • 1996
  • Background : Mechanical ventilation constitutes the last therapeutic method for acute respiratory failure when oxygen therapy and medical treatment fail to improve the respiratory status of the patient. This invasive ventilation, classically administered by endotracheal intubation or by tracheostomy, is associated with significant mortality and morbidity. Consequently, any less invasive method able to avoid the use of endotracheal ventilation would appear to be useful in high risk patient. Over recent years, the efficacy of nasal mask ventilation has been demonstrated in the treatment of chronic restrictive respiratory failure, particularly in patients with neuromuscular diseases. More recently, this method has been successfully used in the treatment of acute respiratory failure due to parenchymal disease. Method : We assessed the efficacy of Bilevel positive airway pressure(BiPAP) in the treatment of acute exacerbation of chronic obstructive pulmonary disease(COPD). This study prospectively evaluated the clinical effectiveness of a treatment schedule with positive pressure ventilation via nasal mask(Respironics BiPAP device) in 22 patients with acute exacerbations of COPD. Eleven patients with acute exacerbations of COPD were treated with nasal pressure support ventilation delivered via a nasal ventilatory support system plus standard treatment for 3 consecutive days. An additional 11 control patients were treated only with standard treatment. The standard treatment consisted of medical and oxygen therapy. The nasal BiPAP was delivered by a pressure support ventilator in spontaneous timed mode and at an inspiratory positive airway pressure $6-8cmH_2O$ and an expiratory positive airway pressure $3-4cmH_2O$. Patients were evaluated with physical examination(respiratory rate), modified Borg scale and arterial blood gas before and after the acute therapeutic intervention. Results : Pretreatment and after 3 days of treatment, mean $PaO_2$ was 56.3mmHg and 79.1mmHg (p<0.05) in BiPAP group and 56.9mmHg and 70.2mmHg (p<0.05) in conventional treatment (CT) group and $PaCO_2$ was 63.9mmHg and 56.9mmHg (p<0.05) in BiPAP group and 53mmHg and 52.8mmHg in CT group respectively. pH was 7.36 and 7.41 (p<0.05) in BiPAP group and 7.37 and 7.38 in cr group respectively. Pretreatment and after treatment, mean respiratory rate was 28 and 23 beats/min in BiPAP group and 25 and 20 beats/min in CT group respectively. Borg scale was 7.6 and 4.7 in BiPAP group and 6.4 and 3.8 in CT group respectively. There were significant differences between the two groups in changes of mean $PaO_2$, $PaCO_2$ and pH respectively. Conclusion: We conclude that short-term nasal pressure-support ventilation delivered via nasal BiPAP in the treatment of acute exacerbation of COPD, is an efficient mode of assisted ventilation for improving blood gas values and dyspnea sensation and may reduce the need for endotracheal intubation with mechanical ventilation.

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Ultrasonographic study on the masseter muscle thickness of adult Korean (한국인 성인의 교근 두께에 관한 초음파검사적 연구)

  • Cha, Bong-Kuen;Park, In-Woo;Lee, Yeun-Hee
    • The korean journal of orthodontics
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    • v.31 no.2 s.85
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    • pp.225-236
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    • 2001
  • It is widely accepted that the shape and structure of bone are closely related to the activity of attached muscle. Numerous clinical and animal experimental studies indicated the significant effects of masticatory muscle function on maxillofacial morphology. Recently, the development of ultrasonography has spread throughout different fields of medicine. In the clinical examinations, ultrasonography is a convenient, inexpensive technique to apply with accurate and reliable results. The aim of this study is to assess the thickness of the masseter muscle and its correlation to maxillofacial skeleton by examining 35 male and 15 female dental students at Kangnung National University. The masseter muscle thickness of the subjects were measured by ultrasonographic scanning with a 7.5MHz linear probe, and their maxillofacial morphology were investigated by lateral cephalometric radiographs. The relationship between the masseter muscle thickness and maxillofacial morphology of normal adult was statistically analyzed, and the following results were obtained. 1. The average thickness of male masseter muscle was 13.8${\pm}$1.71mm in the relaxed state and 14.8${\pm}$1.77mm at maximal clenching state, while that of female was 11.6${\pm}$1.58mm and 12.4${\pm}$1.47mm, respectively. Ethnic difference in thickness of the masseter muscle and maxillofacial skeleton was found when the results of many researchers were compared with those of this study. 2. The thickness of the masseter muscle in both sexes increased significantly at maximal clenching state than in relaxed state(P<0.05). 3. The masseter muscle thickness of male was greater than that of female both in the relaxed state and maximal clenching states(P<0.05). 4. In males, the thickness of the masseter muscle was negatively correlated with the mandibular plane angle and positively correlated with the mandibular ramus height and anterior cranial base length(P<0.05). It may suggest that the male with thicker masseter muscle has smaller facial divergence. 5. No significant correlation was found between the masseter muscle thickness and maxillofacial morphology in females(P<0.05). Therefore, these data suggest that ultrasonography can add valuable information to the conventional examinations of masseter muscle function.

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Investigation of Relationships between Soil Physico-chemical Properties and Topography in Jeonbuk Upland Fields (전북지역 밭 토양의 지형별 물리화학적 특성)

  • Ahn, Byung-Koo;Lee, Jae-Hyoung;Kim, Kab-Cheol;Choi, Dong-Chil;Lee, Jin-Ho;Han, Seong-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.3
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    • pp.268-274
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    • 2010
  • The properties of upland soils are much more dependent upon topography than those of paddy soils, and they give us very useful information to manage the upland fields. Therefore, we investigated the selected physical and chemical properties of upland soils at 84 and 150 topographic sampling sites, respectively. The topographic sites included 34.7% of local valley and fans, 18.7% of hilly and mountains, 20.0% of mountain foot slopes, 14.0% of alluvial plains, 8.0% of diluvium, and 4.6% of fluvio-marine deposits. Based on the investigation, soil textures in Jeonbuk upland fields were mostly sandy loam, sandy clay loam, clay loam, and clay soils, especially sandy clay loam soils were evenly distributed in all of the topographic sites. Soil slopes in the sites ranged from 0 to 15%, which showed an optimal condition for farm land. Soil bulk density and compaction values were from 1.19 to 1.24 g $cm^{-3}$ and from 12.1 to 13.9 mm, respectively. As comparing with the optimal conditions of soil chemical properties for upland soils proposed by National Institute of Agricultural Science and Technology, Korea, 37%, 42.7%, 93.0% of the sites were within optimum levels with soil pH, content of soil organic matter, and electrical conductivity, respectively. However, 64.0%, 47.3%, 48.7%, and 42.7% of the upland soils contained excess levels of exchangeable K, Ca, and Mg, and available phosphorus, respectively. In addition, the contents of heavy metals, As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn, in the Jeonbuk upland soils were much less than threshold levels.

Effect of Resistant Starch on the Large Bowel Environment and Plasma Lipid in Rats with Loperamide-Induced Constipation (저항전분 투여가 Loperamide 유도 변비 쥐의 대장환경과 혈청지질에 미치는 효과)

  • Sin, Hyun-Ju;Kim, Kwang-Ok;Kim, Sung-Hong;Kim, Young-Ah;Lee, Hye-Sung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.684-691
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    • 2010
  • The present study was conducted to evaluate the effect of resistant starch (RS) on the large bowel function and plasma lipids in rats with constipation induced by Loperamide. Animals were divided into six groups: normal control-5% cellulose, constipation-5% cellulose, constipation-5% pectin, constipation-5% RS-type 2 (RS2), constipation-8% RS2 and constipation-5% RS type 3 (RS3) groups, and fed experimental diets for five weeks. The results from RS groups were compared with those from other dietary fiber groups. The groups supplemented with RS3 or high level of RS2 showed significantly increased counts of bifidobacteria in the cecum than the other groups. The production of total short chain fatty acids in the cecal contents was significantly high in pectin, RS3 and high RS2 groups. The pH in the cecal contents of the RS supplemented groups was significantly decreased compared with the cellulose supplemented groups. The production of prostaglandin E2 in the colon mucus of the RS groups was higher than the normal group; however, it was significantly decreased compared to the cellulose or pectin supplemented constipated groups. The thickness of the mucus layer and the production of mucus from epithelial cells were significantly increased in RS3 group compared to the constipated cellulose group. Supplementation of resistant starch significantly elevated the ratio of HDL-cholesterol to total cholesterol and significantly lowered plasma atherogenic index compared with cellulose or pectin supplementation in constipated rats. The results of the present study demonstrated that resistant starch supplementation may help in improving the large bowel environment by stimulation of bifidobacterial proliferation, reduction of pH and inflammation factor and by increased production of mucus. It has also been found that an additional health benefit is improvement in lipid levels of serum.

Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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STUDIES ON THE UTILIZATION OF ANTARCTIC KRILL 1. Compositional Characteristics of Fresh Frozen and Preboiled Frozen Krill (남대양산 크릴의 이용에 관한 연구 1. 크릴의 식품원료학적인 성상)

  • PARK Yeung-Ho;LEE Eung-Ho;LEE Kang-Ho;PYEUN Jae-Hyeung;RYU Hong-Soo;CHOI Su-An;KIM Seun-Bong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.191-200
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    • 1979
  • For the use of antarctic krill as a fond protein source its compositional characteristics were investigated as the first part of the work includes other subjects such as processing of drill paste, concentrates, and fermented or seasoned product. In general composition of fresh frozen and preboiled frozen krill on board, the contents of crude fat and free amino nitrogen were higher in the former than in the latter which contained a high amount of ash. VBN was rather high as much as 37.6 and $26.4\;mg\%$ in both fresh frozen and preboiled krill. The pH of drill homogenates was 7.1 to 7.2 in both cases. Such a low pH might be attributed to a long term storage and temperature fluctuations during frequent transshipping. The amino acid competition of fresh frozen krill meat showed relatively high amount of glutamic acid, aspartic acid, lysine, proline, and leucine while methionine, histidine, serine, tyrosine, and phenylalanine were lower. Among the essential amino acids lysine and leucine were higher and methionine was lower. In tile composition of free amino acid proline, lysing, arginine, and alanine were higher comparatively to the contents of histidine, aspartic acid, serine, and threonine. It is noteworthy for nutritional qualification that tile essential amino acids particularly as lysine were abundant similarly to that of fishes. Heavy metal contents of krill meat 0.039 to 0.048 ppm as Hg, 0.06 to 0.11 ppm as Pb, less than 0.32 ppm as Zn, 0.008 to 0.012 ppm as Cd, 0.61 to 0.68 ppm as Fe, 0.87 to 1.37 ppm as Cu, and nondetective as Cr. A high Cu content seems to be resulted by tile blood pigment of crustacea. The ratio,1 of edible portion to non-edible portion were 37:63 in fresh frozen and 42:58 in preboiled frozen krill respectively. Release of drip after thawing was more in fresh frozen than in preboiled frozen drill marking $36\%$ and $24\%$ of both respectively.

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