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Isolation and Genetic Transformation of Primordial Germ Cell (PGC)-Derived Cells from Cattle, Goats, Rabbits and Rats

  • Lee, C.K.;Moore, K.;Scales, N.;Westhusin, M.;Newton, G.;Im, K.S.;Piedrahita, J.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.5
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    • pp.587-594
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    • 2000
  • At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

Prediction of response by FDG PET early during concurrent chemoradiotherapy for locally advanced non-small cell lung cancer

  • Kim, Suzy;Oh, So Won;Kim, Jin Soo;Kim, Ki Hwan;Kim, Yu Kyeong
    • Radiation Oncology Journal
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    • v.32 no.4
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    • pp.231-237
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    • 2014
  • Purpose: To evaluate the predictive value of the early response of $^{18}F$-flurodeoxyglucose positron emission tomography (FDG PET) during concurrent chemoradiotherapy (CCRT) for locally advanced non-small cell lung cancer (NSCLC). Materials and Methods: FDG PET was performed before and during CCRT for 13 NSCLC patients. Maximum standardized uptake value ($SUV_{max}$), mean standardized uptake value ($SUV_{mean}$), metabolic tumor volume (MTV), and total lesion glycolysis (TLG) were measured and the changes were calculated. These early metabolic changes were compared with the standard tumor response by computed tomograms (CT) one month after CCRT. Results: One month after the completion of CCRT, 9 patients had partial response (PR) of tumor and 4 patients had stable disease. The percent changes of $SUV_{max}$ ($%{\Delta}SUV_{max}$) were larger in responder group than in non-responder group ($55.7%{\pm}15.6%$ vs. $23.1%{\pm}19.0%$, p = 0.01). The percent changes of $SUV_{mean}$ ($%{\Delta}SUV_{mean}$) were also larger in responder group than in non-responder group ($54.4%{\pm}15.9%$ vs. $22.3%{\pm}23.0%$, p = 0.01). The percent changes of MTV ($%{\Delta}MTV$) or TLG ($%{\Delta}TLG$) had no correlation with the tumor response after treatment. All the 7 patients (100%) with $%{\Delta}SUV_{max}{\geq}50%$ had PR, but only 2 out of 6 patients (33%) with $%{\Delta}SUV_{max}$ < 50% had PR after CCRT (p = 0.009). Likewise, all the 6 patients (100%) with $%{\Delta}SUV_{mean}{\geq}50%$ had PR, but only 3 out of 7 patients (43%) with $%{\Delta}SUV_{mean}$ < 50% had PR after CCRT (p = 0.026). Conclusion: The degree of metabolic changes measured by PET-CT during CCRT was predictive for NSCLC tumor response after CCRT.

Comparison of Indigenous Browses and Sunflower Seed Cake Supplementation on Intake and Growth Performance of Dual-purpose Goats Fed Buffel Grass (Cenchrus ciliaris) Hay

  • Komwihangilo, D.M.;Chenyambuga, S.W.;Lekule, F.P.;Mtenga, L.A.;Muhikambele, V.R.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.7
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    • pp.966-972
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    • 2005
  • A study to compare the effects of supplementing Delonix elata, Grewia similis, Tamarindus indica and sunflower seed cake on intake and growth rate of dual-purpose goats fed low quality Buffel grass (Cenchrus ciliaris) hay was carried out. Twenty-eight male goats aged five to seven months (mean weight 12.93${\pm}$3.94 kg) were randomly allocated to four dietary groups in a completely randomised design. The diets were hay plus Grewia similis, hay plus Delonix elata, hay plus Tamarindus indica and hay plus sunflower seed cake. All diets were supplemented with maize bran. The experimental period was 90 days. Voluntary dry matter intake of the supplements was higher for Tamarindus indica (275.5 g/day) and Grewia similis (201.8 g/day) and lowest for sunflower seed cake (81g/day). Goats supplemented with Grewia similis had the highest hay intake (183.8 g/day) while those supplemented with sunflower seed cake had the lowest hay intake (98.9 g/day). Animals fed browse supplements gained significantly more weight (p<0.001) than those with sunflower seed cake. There were no significant differences in live weight change between goats fed the different browses. However, those fed Tamarindus indica gained an average of 20.79 g/d which was slightly higher than the gains for those on Grewia similis and Delonix elata while those fed sunflower seed cake lost weight. Correspondingly, goats supplemented with browse leaf meals had higher feed conversion ratios than those supplemented with sunflower seed cake and required 23.91 to 35.06 g DM of feed to produce one g of weight gain per day. In a separate study, the DM disappearance pattern indicated that Grewia similis and Delonix elata were highly degradable compared to Tamarindus indica. At 24 h of incubation, DM degradability was 627, 588 and 345 g/kg DM for Grewia similis, Delonix elata and Tamarindus indica, respectively. In another study in vivo DM digestibility ranged from 46.1% (for hay alone) to 56.2% (for hay plus Grewia similis). It was concluded that the addition of Tamarindus indica, Grewia similis and Delonix elata leaf meals to Cenchrus ciliaris hay resulted in increased total DM intake, in vivo digestibility and growth rate. Therefore, leaf meals of indigenous browses particularly Tamarindus indica and Grewia similis could be used as supplementary feeds for small ruminants grazing on poor quality roughages during the dry season rather than use of expensive, less effective and intermittently available sunflower seed cake.

Gas Exchanges and Dehydration in Different Intensities of Conditioning in Tifton 85 Bermudagrass: Nutritional Value during Hay Storage

  • Pasqualotto, M.;Neres, M.A.;Guimaraes, V.F.;Klein, J.;Inagaki, A.M.;Ducati, C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.807-815
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    • 2015
  • The present study aimed at evaluating the intensity of Tifton 85 conditioning using a mower conditioner with free-swinging flail fingers and storage times on dehydration curve, fungi presence, nutritional value and in vitro digestibility of Tifton 85 bermudagrass hay dry matter (DM). The dehydration curve was determined in the whole plant for ten times until the baling. The zero time corresponded to the plant before cutting, which occurred at 11:00 and the other collections were carried out at 8:00, 10:00, 14:00, and 16:00. The experimental design was randomised blocks with two intensities of conditioning (high and low) and ten sampling times, with five replications. The high and low intensities related to adjusting the deflector plate of the free iron fingers (8 and 18 cm). In order to determine gas exchanges during Tifton 85 bermudagrass dehydration, there were evaluations of mature leaves, which were placed in the upper middle third of each branch before the cutting, at every hour for 4 hours. A portable gas analyser was used by an infrared IRGA (6400xt). The analysed variables were photosynthesis (A), stomatal conductance (gs), internal $CO_2$ concentration (Ci), transpiration (T), water use efficiency (WUE), and intrinsic water use efficiency (WUEi). In the second part of this study, the nutritional value of Tifton 85 hay was evaluated, so randomised blocks were designed in a split plot through time, with two treatments placed in the following plots: high and low intensity of cutting and five different time points as subplots: cutting (additional treatment), baling and after 30, 60, and 90 days of storage. Subsequently, fungi that were in green plants as well as hay were determined and samples were collected from the grass at the cutting period, during baling, and after 30, 60, and 90 days of storage. It was observed that Tifton 85 bermudagrass dehydration occurred within 49 hours, so this was considered the best time for drying hay. Gas exchanges were more intense before cutting, although after cutting they decreased until ceasing within 4 hours. The lowest values of acid detergent insoluble nitrogen were obtained with low conditioning intensity after 30 days of storage, 64.8 g/kg DM. The in vitro dry matter of Tifton 85 bermudagrass did not differ among the storage times or the conditioning intensities. There was no fungi present in the samples collected during the storage period up to 90 days after dehydration, with less than 30 colony forming units found on plate counting. The use of mower conditioners in different intensities of injury did not speed up the dehydration time of Tifton 85.

Estimation of Genetic Parameters and Trends for Weaning-to-first Service Interval and Litter Traits in a Commercial Landrace-Large White Swine Population in Northern Thailand

  • Chansomboon, C.;Elzo, M.A.;Suwanasopee, T.;Koonawootrittriron, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.5
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    • pp.543-555
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    • 2010
  • The objectives of this research were the estimation of genetic parameters and trends for weaning-to-first service interval (WSI), and litter traits in a commercial swine population composed of Landrace (L), Large White (T), LT, and TL animals in Chiang Mai, Northern Thailand. The dataset contained 4,399 records of WSI, number of piglets born alive (NBA), litter weight of live piglets at birth (LBW), number of piglets at weaning (NPW), and litter weight at weaning (LWW). Variance and covariance components were estimated with REML using 2-trait analyses. An animal model was used for WSI and a sire-dam model for litter traits. Fixed effects were farrowing year-season, breed group of sow, breed group of boar (litter traits), parity, heterosis (litter traits), sow age, and lactation length (NPW and LWW). Random effects were boar (litter traits), sow, permanent environment, and residual. Heritabilities for direct genetic effects were low for WSI (0.04${\pm}$0.02) and litter traits (0.05${\pm}$0.02 to 0.06${\pm}$0.02). Most heritabilities for maternal litter trait effects were 20% to 50% lower than their direct counterparts. Repeatability for WSI was similar to its heritability. Repeatabilities for litter traits ranged from 0.15${\pm}$0.02 to 0.18${\pm}$F0.02. Direct genetic, permanent environment, and phenotypic correlations between WSI and litter traits were near zero. Direct genetic correlations among litter traits ranged from 0.56${\pm}$0.20 to 0.95${\pm}$0.05, except for near zero estimates between NBA and LWW, and LBW and LWW. Maternal, permanent environment, and phenotypic correlations among litter traits had similar patterns of values to direct genetic correlations. Boar genetic trends were small and significant only for NBA (-0.015${\pm}$0.005 piglets/yr, p<0.004). Sow genetic trends were small, negative, and significant (-0.036${\pm}$0.013 d/yr, p<0.01 for WSI; -0.017${\pm}$0.005 piglets/yr, p<0.007, for NBA; -0.015${\pm}$0.005 kg/yr, p<0.01, for LBW; -0.019${\pm}$0.008 piglets/yr, p<0.02, for NPW; and -0.022${\pm}$0.006 kg/yr, p<0.003, for LWW). Permanent environmental correlations were small, negative, and significant only for WSI (-0.028${\pm}$0.011 d/yr, p<0.02). Environmental trends were positive and significant only for litter traits (p<0.01 to p<0.0003). Selection based on predicted genetic values rather than phenotypes could be advantageous in this population. A single trait analysis could be used for WSI and a multiple trait analysis could be implemented for litter traits.

A Study on Nursing Students의 Attitudes toward the Elderly (노인에 대한 간호대학생의 태도조사 연구)

  • 김은일
    • Journal of Korean Academy of Nursing
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    • v.12 no.1
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    • pp.1-12
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    • 1982
  • Presently, there is increasing demand for geriatric nursing care due to increasing proportion of elderly population and extended age. And, of particular importance in deforming the effectiveness of nursing care given to the elderly is the attitude of nursing personnel toward the elderly patient. This study was intendent to investigate the attitudes of baccalaureate nursing students toward the elderly, and to investigate any possible correlation between these attitudes and the following variables: the students' levels (freshman, sophomore, junior, senior), their preferences for var-ious fields of specialization in nursing after graduation, their clinical experiences with geriatric-patients, their personal experience with elderly family members, their educational experience of the elderly's health problems and geriatric nursing, and their experience with relatives and acquaintances over 60 years of age. Therefore, the purpose of this study was to compile basic data about the attitudes of nursing students toward the elderly in order to contribute to the improvement of geriatric nursing. The subjects for this study were 259 nursing students: 163 in the College of Nursing at E-womans University., and 96 in the Department of Nursing in K. College. Data was collected from the 11th of Sep., to the 8th of Oct., 1981, using the Tuckman-Lorge Attitude Questionnaire which contained 131 items excepting 6 items. The attitudes toward the elderly was scored by the use of mean and standard deviation. The relations between attitudes toward the elderly and six independent vairables were analysed by T-test. The results which were obtained are as follows : 1. The attitude score of nursing students was M=71.67 S.D= 16.18. Nursing students agreed with 54.8% of the stereotypic statements, and they had stereotypic attitudes toward the elderly relatively. (P<0.01). 2. There was a significant difference between the attitudes of nursing students toward the elderly, and sophomores had a less stereotypic attitudes than juniors and seniors (P<0.01). 3. There was significant difference between the attitudes toward the elderly and the preferences for various fields of specialization in nursing after graduation, and nursing students those who wanted maternity fields had a more stereotypic attitudes than others (P<0.05, p<0.01). 4. There was no significant difference between the attitudes toward the elderly and the clinical experience with the geriatric patients (P>0.05). 5. There was no significant difference between the attitude3 toward the elderly and the personal experience with elderly family members (F>0.05). 6. There was no significant difference between the attitudes toward the elderly and the educational experience of the elderly's health problems and geriatric nursing (P>0.05). 7. There was no significant difference between the attitudes toward the elderly and the experience with relatives and acquaintances over 60 years of age (P>0.05).

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Distribution of Major Deciduous Tree Species in Relation to the Characteristics of Topography in Mt. Joongwang, Gangwon Province(I) (강원도 중왕산 지역의 지형 특성에 따른 주요 활엽수종의 분포(I))

  • Um, Tae Won;Lee, Don Koo
    • Journal of Korean Society of Forest Science
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    • v.95 no.1
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    • pp.91-101
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    • 2006
  • This study was conducted to analyze the relationships between distributions of major hardwood species and characteristics of topography in natural hardwood forest at Mt. Joongwang, Pyongchang-gun, Gangwon-do which belongs to mid-northern part of the temperate forest. Acer pictum var. mono, Acer manshuricum and Quercus mongolica were affected by elevation, while Ulmus davidiana var. japonica and Fraxinus rhynchophylla were affected by the change of slope. Importance values of A. pictum, Betula costata, Ulmus laciniata and Cornus controversa in valley were 13.41%, 5.68% and 3.29%, respectively, which were higher than in any other topography. Species diversity of valley was the highest with 0.98. Ratio of A. pictum and Q. mongolica in dominant species were 24% and 14%, respectively in valley, while were 19% and 23% in slop. Q. mongolica ratio in ridgeline was higher than any other species. Importance values of A. pictum increased continuously until elevation 1,200 m. importance values of F. rhynchophylla was 6.6% which was the highest value found on elevation 800 m. Distributions of major hardwood species were different according to the elevation. The ratio of Q. mongolica and A. pictum were 6.4%~17.5% and 6.0%~10.7% respectively, which were the highest ratio. Species diversity increased according to the increase of the elevation.

Usefulness of serum cystatin C to determine the dose of vancomycin in neonate

  • Shin, Jeong Eun;Lee, Soon Min;Eun, Ho Seon;Park, Min Soo;Park, Kook In;Namgung, Ran
    • Clinical and Experimental Pediatrics
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    • v.58 no.11
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    • pp.421-426
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    • 2015
  • Purpose: The vancomycin dosage regimen is regularly modified according to the patient's glomerular filtration rate (GFR). In the present study, we aimed to assess the usefulness of serum cystatin C (Cys-C) concentration, compared with serum creatinine (SCr) concentration, for predicting vancomycin clearance (CLvcm) in neonates. Methods: We retrospectively analyzed the laboratory data of 50 term neonates who were admitted to the neonatal intensive care unit and received intravenous vancomycin, and assessed the pharmacokinetic profiles. Creatinine clearance (CLcr) and GFR based on Cys-C (GFRcys-c) were estimated using the Schwartz and Larsson formulas, respectively. Results: The mean CLvcm (${\pm}$standard deviation) was $74.52{\pm}31.17L/hr$, the volume of distribution of vancomycin was $0.67{\pm}0.14L$, and vancomycin half-life was $9.16{\pm}17.42hours$. The SCr was $0.46{\pm}0.25mg/dL$ and serum Cys-C was $1.43{\pm}0.34mg/L$. The peak and trough concentrations of vancomycin were $24.65{\pm}14.84$ and $8.10{\pm}5.35mcg/mL$, respectively. The calculated GFR based on serum creatinine concentration (GFR-Cr) and GFRcys-c were $70.2{\pm}9.45$ and $63.6{\pm}30.18mL/min$, respectively. The correlation constant for CLvcm and the reciprocal of Cys-C (0.479, P=0.001) was significantly higher than that for CLvcm and the reciprocal of SCr (0.286, P=0.044). GFRcys-c was strongly correlated with CLvcm (P=0.001), and the correlation constant was significantly higher than that for CLvcm and CLcr (0.496, P=0.001). Linear regression analysis showed that only GFRcys-c was independently and positively correlated with CLvcm (F=41.9, P<0.001). Conclusion: The use of serum Cys-C as a marker of CLvcm could be beneficial for more reliable predictions of serum vancomycin concentrations, particularly in neonates.

Usefulness of FDG-PET/CT as a Diagnostic Tool for Routine Post Therapy Evaluation in Endometrial Cancer (자궁내막암의 치료 후 루틴 추적검사 방법으로서 FDG-PET/CT의 유용성)

  • Lee, Shin-Jae;Jeon, Tae-Joo;Kim, Seung-Jo;Kim, Hee-Jin;An, Hee-Jung
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.301-308
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    • 2009
  • Purpose: The aim of this study was to evaluate the usefulness of FDG-PET/CT as follow up imaging tool in patients with endometrial cancer after therapy. Material and Methods: One hundred one patients with endometrial cancer who underwent FDG PET/CT after the treatment of this disease were included in this study population (25-79 yr old, Mean age 50.6 yr old) and all these patients also performed various laboratory and imaging studies such as serum tumor marker, CT or MRI. The lesions having increased focal FDG uptake were classified into benign, equivocal, and malignant one according to their pattern and activity. Tumor recurrence was confirmed by histopathological results and other clinical and imaging data. Results: Among the 19 patients with 30 malignant or equivocal hot uptakes, 11 of 14 patients supposed to be malignant finding in PET/CT were proved to be tumor recurrence, while one of 5 patients with equivocal lesions were recurred malignancy, Two false negative cases were turned out to be peritoneal carcinomatosis, Estimated sensitivity, specificity and accuracy of PET/CT for diagnosis of recurrence in endometrial carcinoma after treatment were 86 %, 92 % and 91 %, respectively. Positive and negative predictive values in the same issue were 63% and 98%, respectively. Conclusion: FDG-PET/CT is useful for regular work up of endometrial carcinoma after the treatment because of its high negative predictive value as well as high sensitivity and specificity.

Studies on Biological Activities of the Polysaccharides and Oligosaccharides of Orostachys japonicus (와송 다당체 및 올리고당류의 생리 활성)

  • Kim Ki Hoon;Kim Eun Young;Kim Yea Oon;Baek Geum Ok;Kim Han Bok;Lee Dong Seok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.334-341
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    • 2004
  • Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.