• 대한수의학회지
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• 제63권1호
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• pp.3.1-3.9
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• 2023

The efficiency of somatic cell nuclear transfer (NT) in pigs is low and requires enhancement. We identified the most efficient method for zona pellucida (ZP) removal and blastomere aggregation in pigs and investigated whether the aggregation of NT and parthenogenetic activation (PA) of blastomeres could reduce embryonic apoptosis and improve the quality of NT-derived embryos by investigating. Embryonic developmental competence after ZP removal using acid Tyrode's solution or protease (pronase E). The embryonic developmental potential of NT-derived blastomeres was also investigated using well-of-the-well or phytohemagglutinin-L. We analyzed apoptosis in aggregate-derived blastocysts. The aggregation rate of protease-treated embryos was lower than that of Tyrode's solution-treated embryos (69.2% vs. 88.3%). No significant difference was observed between phytohemagglutinin-L and well-of-the-well (35.7%-38.5%). However, 2P1N showed a higher number of blastocysts compared to 3N (73.8% vs. 24.3%) and an increased blastocyst diameter compared to the control and 1P2N (274 ㎛ vs. 230-234 ㎛). In blastomeres aggregated using phytohemagglutinin-L, the apoptotic cell ratio was significantly higher in 1P2N and 3N than in 3P (5.91%-6.46% vs. 2.94%, respectively). Our results indicate that aggregation of one NT embryo with two PA embryos improved the rate of blastocysts with increased blastocyst diameter.

• Asian-Australasian Journal of Animal Sciences
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• 제28권1호
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• pp.135-142
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• 2015

This study was conducted to evaluate the effect of inclusion of propolis extraction residue in the feed of broilers from 1 to 21 d of age on phagocytic activity of macrophages, cutaneous basophil hypersensitivity response to phytohemagglutinin, antibody production against Newcastle disease, lymphoid organ weight and hematological profile and to determine the optimal level of inclusion. 120 chicks, reared in metabolism cages until 21 days of age, were distributed in a completely randomized design, with five treatments (0%, 1%, 2%, 3%, and 4% of propolis residue) and six replications. The relative weight of thymus and monocyte percentage were affected by propolis residue, with a quadratic response (p<0.05) and lowest values estimated at 2.38% and 2.49%, respectively. Changes in relative weight of cloacal bursa and spleen, percentage of lymphocyte, heterophil, basophil, eosinophil, and heterophil:lymphocyte ratio, antibody production against Newcastle disease, phagocytic activity of macrophages and the average number of phagocytosed erythrocytes were not observed. The nitric oxide production with regard to positive control (macrophages+erythrocytes) decreased linearly (p<0.05) with increased doses of propolis residue. The remaining variables of nitric oxide production (negative control - macrophages, and difference between the controls) were not affected by propolis residue. The cutaneous basophil hypersensitivity response to phytohemagglutinin as determined by the increase in interdigital skin thickness exhibited a quadratic response (p<0.05), which predicted a lower reaction response at a dose of 2.60% of propolis residue and highest reaction response after 43.05 hours of phytohemagglutinin injection. The inclusion of 1% to 4% of propolis extraction residue in broiler diets from 1 to 21 days of age was not able to improve the immune parameters, despite the modest changes in the relative weight in thymus, blood monocyte percentage, nitric oxide concentration, and interdigital reaction to phytohemagglutinin.

• BMB Reports
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• 제35권5호
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• pp.472-475
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• 2002

Phaseolus vulgaris phytohemagglutinin L is a homotetrameric-leucoagglutinating seed lectin. Its three-dimensional structure shows similarity with other members of the legume lectin family. The tetrameric form of this lectin is pH dependent. Gel filtration results showed that the protein exists in its dimeric state at pH 2.5 and as a tetramer at pH 7.2. Contrary to earlier reports on legume lectins that possess canonical dimers, thermal denaturation studies show that the refolding of phytohemagglutinin L at neutral pH is irreversible. Differential scanning calorimetry (DSC) was used to study the denaturation of this lectin as a function of pH that ranged from 2.0 to 3.0. The lectin was found to be extremely thermostable with a transition temperature around $82^{\circ}C$ and above $100^{\circ}C$ at pH 2.5 and 7.2, respectively. The ratio of calorimetric to vant Hoff enthalpy could not be calculated because of its irreversible-folding behavior. However, from the DSC data, it was discovered that the protein remains in its compact-folded state, even at pH 2.3, with the onset of denaturation occurring at $60^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.891-900
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• 2004

UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1164-1169
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• 2001

A five-week trial was conducted to evaluate the effect of organic chromium from different sources on growth performance, immune response and serum parameters of weaned pigs. One hundred and eighty Tianjin white pigs weaned at $35{\pm}1$ days of age, were allotted to three treatments with six replicates and10 pigs per pen. Pigs were fed corn-soybean-whey-fishmeal basal diets with either no supplemental Cr, $200{\mu}g/kg$ Cr as chromium picolinate (CrPi), or $200{\mu}g/kg$ Cr as chromium yeast (Cr-yeast). To assess humoral immune response, all pigs were immunized with swine fever virus on day 21 and two pigs from each pen were immunized with pure albumin on day 14. Cell-mediated immunity was measured by determining the double skinfold thickness (DST) of two pigs from each pen before and 24h after stimulation with phytohemagglutinin (PHA) on day 28. The results indicated that: (1) diets with Cr-yeast increased average daily gain (ADG, p<0.05) and tended to increase average daily feed intake (ADFI, p<0.10). Diets with CrPi did not increase ADG and ADFI (p>0.05). (2) Dietary CrPi or Cr-yeast supplementation did not affect blood urea nitrogen, glucose, or cholesterol (p>0.05), but blood urea nitrogen in CrPi and Cr-yeast supplemented groups and blood glucose in the Cr-yeast supplemented group were significantly influenced by sampling days (p<0.05). (3) Serum proteins (TP, ALB, and GLB) were influenced by sampling days (p<0.05), but not by dietary Cr treatment (p>0.10). (4) There were no significant differences among treatments in the titers of albumin antibody and swine fever virus antibody (p>0.05) or DST before and after PHA stimulation (p>0.05), indicating that organic chromium has no significant effect on the immune function of weaning pigs. Therefore, these results agree with other research that the effects of supplemental Cr are variable in weanling pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제10권5호
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• pp.471-477
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• 1997

A study detennined whether certain biochemical and physiological variables were altered during marginal Zn deficiency. Ten weaned crossbred Hereford Angus heifer calves, weighing $163{\pm}2kg$, were utilized. Five calves were fed a Zn - deficient (- Zn) brome-alfalfa hay diet containing 17 mg Zn/kg diet DM, and five calves were fed a Zn-adequate (+Zn) diet with 23 mg Zn/kg diet DM from $ZnSO_4$ added to the - Zn diet (total diet, 40 mg Zn/kg diet DM), for 32 d. At 21 d the - Zn calves had a reduction (p < .05) in feed efficiency. By 25 d, plasma Zn and alkaline phosphatase concentrations were reduced (p < .05) in the - Zn calves. Blood urea nitrogen, glucose, insulin, IGF-I, Cu plasma concentration and Zn and Cu concentrations of red blood cell (RBC) and liver were not altered (p > .05) by the - Zn diet through 25 d. In response to a single i. m. injection of dexamethasone (20 mg) on d 25, calves fed the two dietary Zn amounts showed no changes (p > .05) in plasma or RBC Zn and Cu concentrations, serum IGF-I, insulin, and glucose when measured at 6, 12, 24, 48, 72, and 96 h after injection. In response to an intradermal injection of phytohemagglutinin on d 30, cell mediated immune (CMI) response was reduced (p < .05) in the - Zn calves. These observations indicate that during a marginal Zn deficiency in calves, there was a decrease in feed efficiency, plasma Zn, serum alkaline phosphatase, and CMI response.

• 한국축산식품학회지
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• 제25권2호
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• pp.250-256
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• 2005

To investigate the effect of bovine colostral whey fractions on in vitro proliferation of mouse splenocytes, polypeptide fractions were separated from acid whey into 3 fractions depending on molecular weight by ultrafiltration: Fraction I, which contains the polypeptide larger than 10,000 Da, Fraction n, which contains the polypeptide ranging from 1,000 Da to 10,000 Da and Fraction III, which contains the polypeptide smaller than 1,000 Da. Fraction II showed the highest proliferative effect of mouse splenocytes among the colostral whey fractions and this proliferative activity increased in dose dependent manner. Unheated Fraction II and Fraction III showed significantly (p<0.01) higher proliferative effects than others but heated Fraction II showed the highest enhancing effect of mouse splenocyte among heated whey fractions (p<0.01). The supplementation of Fraction II and Fraction m showed greater proliferative effect of mouse splenocytes stimulated by concanavalin A (Con A) than that of whole whey or Fraction L Proliferative effect of mouse splenocytes stimulated by phytohemagglutinin (PHA) was the highest when Fraction II was supplemented Proliferative effect of the colostral whey fractions on mouse splenocytes by stimulation of lipopolysaccharide (LPS) was markedly enhanced by supplementation of Fraction II and Fraction m compared with whole whey and Fraction L It was estimated that colostral whey fraction containing IGF-I positively affected proliferation of mouse splenocyte.

• 한국식품영양과학회지
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• 제31권1호
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• pp.1-6
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• 2002

강낭콩을 가열한 후 B. subtilis ATCC 51189로 발효시켜 새로운 활성 단백질의 생성여부를 확인하였다. 생강낭콩의 수용성 단백질은 열처리에 의하여 감소하였으며, 발효과정에서 단백질의 양은 변화하지 않았으나, 새로운 아미노산이 합성되었다. 생강낭콩의 수용성 단백질(raw protein, RP)은 분자량 118 kDa의 PHA(phytohemagglutinin)의 전형적인 모습을 보였으나, 열처리한 단백질(heated protein, HP)의 경우, RP의 band는 사라지고, 저분자 peptide로 변화되었으며, 발효과정에 의하여 분자량 18.0 kDa의 새로운 단백질(fermented protein, FP)이 생성된 것을 확인하였다. 또한, RP는 128 HU, HP는 4HU, FP는 32HU의 적혈구응집효과를 보였으며, 위암세포(SNU-1)에 대하여 RP 및 FP는 비교적 높은 농도($IC_{50}$/ $\mu\textrm{g}$/mL)에서 항암활성을 보였다. 그리고, RP 및 FP의 경우 1 $\mu\textrm{g}$/mL에서 림프구 증식효과를 보였고, IFN-${\gamma}$, IL-12의 분비를 촉진하였다. 그러나 HP는 항암효과, 림프구 증식효과, cytokine의 분비촉진효과를 보이지 않았다. 따라서, 강낭콩 lectin은 가열에 의하여 활성물질이 파괴되나, B. subtilis에 의한 발효과정에서 새로운 cytokine 분비촉진물질이 생성되는 것을 확인할 수 있었다.

• Journal of Community Nutrition
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• 제7권1호
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• pp.42-48
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• 2005

This study was done to investigate effects of antioxidant supplementation on phytohemagglutinin (PHA) -stimulated interleukin-2 (IL-2) production by peripheral blood mononuclear cells (PBMCs) in elderly women. This study was designed as a placebo-controlled, single-blinded, randomized intervention trial. Twenty four elderly women aged over 60 years, visitings social welfare center in Seoul were divided into 3 groups, placebo (n = 8), vitamin C supplemented (n = 8) , and vitamin E supplemented (n = 8) groups. Experimental groups were given either 1000mg of L-ascorbic acid or 400 ill of d- $\alpha$-tocopherol for 4 weeks. There was no significant difference in antioxidant vitamins intakes and their plasma levels among pre-intervention groups. Plasma vitamin C or E levels was significantly increased after vitamin C or E sup-plementations. The increases of plasma thiobarbituric acid-reactive substance (TBARS) levels in the placebo group were significantly higher than those of the supplemented 2 groups. There were no significant differences in the changes of plasma IL-2 level between pre- and post-intervention among the 3 groups. However there was a significant increase in PHA­stimulated IL-2 production by PBMCs after 4-week vitamin E or vitamin C supplementation. Particularly, vitamin E supplemented group showed a higher PHA-stimulated IL-2 production than vitamin C supplemented group. These results indicate that vitamin E or vitamin C supplementation might enhance mitogen-stimulated cytokine production by immune cells, which could be one of the factors to improve health status in the elderly.

• 한국가축번식학회지
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• 제27권3호
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• pp.233-240
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• 2003

본 연구는 난자의 성숙시간, PHA-P 처리 또는 활성화 방법이 소 미수정란의 탈핵, 재구축란의 융합, 활성화 또는 체외발육에 미치는 영향을 검토하였다. 미수정란은 성숙 후 16∼24시간에 탈핵을 실시하고, PHA-P 처리 또는 무처리된 귀 피부세포를 이식 후 전기융합을 실시하였다. 후자의 경우는 융합 전에 PHA-P로 15분간 배양하였다. 융합란은 A23187과 CHXM 혹은 DMAP의 병용처리에 의해 활성화를 유기하고, 7∼9일간 체외배양하였다. 탈핵율은 성숙 후 16∼18시간에 실시한 경우(70.2∼92.3%)가 성숙 후 20∼24시간(44.3∼3.4%)에 비하여 유의적으로 높았다(P<0.05). M-II기 염색체의 위치는 성숙배양 시간이 길어짐에 따라 제 1 극체와의 간격이 멀어졌다. Donor 세포 혹은 재구축란에 PHA-P를 처리한 경우는 무처리구에 비하여 융합율이 향상되었다(P<0.05). 핵이식배의 분할율 및 배반포 발달율은 A23187+DMAP 처리구에서 78.6%와 32.9%로, A23187+CHXM 처리구에 비하여 유의적으로 높았다(P<0.05). 본 실험 결과는 성숙후 18시간에 탈핵을 실시하는 것이 효과적이며, donor세포 또는 융합 전 재구축란의 PHA-P 처리가 융합율 향상시킬 수 있고, 또한, 융합란을 A23187과 DMAP으로 병용처리 함으로써 난자의 활성화 및 배반포 발육율을 향상시켜, 결과적으로 핵이식기술의 효율성을 증진시킬 수 있을 것으로 사료된다.