• 생명과학회지
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• 제15권3호
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• pp.344-351
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• 2005

산화질소(NO)와 칼슘 결합 단백질은 중추신경계의 다양한 세포들에서 나타나며, 이들은 각각 중요 신호전달 분자와 칼슘 완충 분자이다. 본 연구는 햄스터의 시각피질에서 뇌산화질소 합성효소 (nNOS)와 parvalbumin을 포함하는 뉴런들의 분포와 이들의 co-localization 양상을 면역세포화학적 기법을 이용하여 알아보았다. 햄스터 시각피질에서 parvalbumin에 대한 면역 반응성을 나타내는 뉴런들의 전체 수는 nNOS에 대한 면역 반응성을 보이는 뉴런들의 수보다 17배나 많았다. 가장 큰 차이는 시각피질 제5충에서 발견되었으며, 이곳에서 parvalbumin-면역 반응성 뉴런이 nNOS-면역 반응성 뉴런들의 수보다 54.7배나 높았다. nNOS-또는 parvalbumin-면역 반응성 뉴런들은 크기와 형태, 분포 방식이 시각피질에서 유사하게 나타났다. 그러나 이색 면역형광 기법은 햄스터 시각피질에서 nNOS와 parvalbumin을 모두 발현하는 뉴런은 없음을 보여주었다. 본 연구의 결과는 nNOS와 칼슘 결합 단백질 사이의 co-localization양상이 종간에 차이가 존재함을 나타내며 또한 시각피질에 있는 nNOS-면역 반응성 뉴런들의 다양성과 이질성뿐만 아니라 동물 다양성 이해의 중요성을 함께 제시한다고 볼 수 있다.

• 한국잠사곤충학회지
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• 제24권2호
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• pp.55-72
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• 1983

가잠의 휴면성을 난각의 조직학적 특성을 보고자 난각층에 대한 전자현미경상을 관찰하였고 또한 난각의 물리화학적 특성에 대해서는 수분투과성, 염산투과성, 염색성 및 난각 구조단백질을 조사한 결과를 요약하면 다음과 같다. 1. 난각구조를 전자현미경으로 보면 뚜렷이 4층으로 되어 있으며, 또한 휴면난과 비휴면난 및 즉침난 사이에 구조적인 차이가 보인다. 즉 외층 밖에 또 다른 전자밀도가 낮은 얇은 층이 휴면난에는 존재하고 있으나 비휴면 및 즉침난에는 이 층을 볼 수 없다. 2. 산난후 20∼24시간째의 휴면난과 비휴면난 및 즉침난 사이에 난각염산투과성의 차이를 보면 휴면난에 비해 즉침난 및 비휴면난이 모두 투과성이 높았고 또한 난령이 지남에 따라 휴면난은 투과성이 떨어지는데 반하여 즉침난 및 비휴면난은 투과성이 높아졌다. 3. 휴면난의 ether 처리난은 무처리난에 비해 난각의 투과성이 변하는 것으로 보아 난각물질 중 ether 용출물이 존재한다. 4. 난각의 수분투과성도 염산투과성에 있어서와 같이 휴면난이 즉침난 및 비휴면난에 비해 수분투과성이 낮았다. 5. 가잠난각의 조직화학적 특성은 단백질과 당에 대한 염색성이 높아 난각에는 단백질과 당이 풍부하며 특히 지질에 대한 염색성은 거의 음성이나 내층으로 갈수록 진하게 염색되었다. 6. 누에의 난각점착물은 PAS-alcian 반응으로 보아 mucopolysaccharides이다. 7. 난각 구조단백질 중 SH단백질에 대한 전기영동상에 있어서 휴면난과 비휴면난 모두 2개의 band가 인정되나 비휴면난의 경우 이동거리가 빠른 band는 매우 흔적적이며 또한 영동거리도 원점으로부터 멀리 이동하여 두 처리간에 minor component에 차이가 보였다. 8. SH단백질에 대한 아미노산조성에 있어서는 휴면난 및 비휴면난 사이에 큰 차이를 인정할 수 없었고 난각단백질중 SH단백질의 구성아미노산에는 glycine이 가장 많고 cystine함량은 적었다. SI K-2 는 Penicillin, Ampicillin과 Cephalothin에 저항성을 나타냈고 ts-U171, A-N92와 A-N115는 이 세항생제에 민감성을 나타냈다. Temperature - sensitive돌연변이균주를 42$^{\circ}C$에서 24시간 배양후에 28$^{\circ}C$로 옮기어 배양한 결과 ts-U601, -U603, -U604와 -U171은 성장을 더 이상하지 않았다.ta-exotoxin을 생산했는데, 48시간 배양 배지 $m\ell$당 70$\mu\textrm{g}$을 분비한 균주는 BTK-1이고, BTK-37 균주는 $m\ell$당 88$\mu\textrm{g}$(6.1$\times$$10^{8}$ Cells/$m\ell$) BTK-35 균주는 $m\ell$당 81$\mu\textrm{g}$(5.2$\times$$10^{8}$ Cells/$m\ell$)을 생산했고. 그외는 모두 70$\mu\textrm{g}$미만이었다. 3. Beta-exotoxin과 B. thuringiensis 균체을 동시에 per os, interaperitoneal injection, subcuntaneous injection, nasal cavity inoculation, intracerebral injection을 120시간 처리했어도 치사효과를 나타내지 않았다.삼각호마법 and Japan#s 병음한자변환방식 have prospect to be prevalent. The following suggestions can be made from these results, 1) All the

• 한국식품과학회지
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• 제26권3호
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• pp.310-316
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• 1994

한국인의 식생활에서 널리 이용되고 있는 식물성 식품 및 민간요법에서 이용되는 비상용식품 45가지를 선택하여 총페놀함량은 Folin-Denis 법으로, 축합형 탄닌의 함량은 vanillin 법과 modified vanillin 법으로, 단백질 침전성 페놀물질의 함량은 단백질 침전법으로 측정하였다. 총페놀 함량(건물기준)은 $0.1{\sim}5.8%$로 감잎, 밤속껍질, 모과, 호두, 해바라기씨, 칡뿌리에서는 2% 이상의 높은 함량을 보였다. 축합형 탄닌함량(건물기준)은 $0{\sim}48%$로 모과 밤속껍질에서 매우 높이 나타났다. 단백질 침전성 페놀성분의 함량(건물 기준)은 $0.4{\sim}2.2%$로 밤속껍질, 호두, 모과에서 매우 높았고 그들의 단백질 침전능은 트립신보다 펩신이나 알부민에서 더 잘 나타났다. 식품중 페놀성물질 함량간의 상관관계를 보면 총페놀 함량과 단백질 침전성 페놀물질 함량(r=0.65)과 축합형 탄닌함량(r=0.56)간에 높은 상관관계를 보였으나, 축합형 탄닌함량과 단백질 침전성 페놀물질 함량간에는 비교적 낮은 상관관계(r=0.39)를 보였다. 축합형 탄닌의 중합도를 예측하는 vanillin/Folin-Denis 비를 살펴본 결과 모과, 밤속껍질, 수수에서 높은 값을 보여 결국 낮은 중합도를 보였다. 메밀, 도토리, 쑥, 칡 등을 이용한 가공식품에서의 페놀물질 함량은 원료상태에서 보다 낮은 함량을 보여 식품단백질의 이용율에는 큰 장애를 주지 않을 것으로 판단되었다.

• Molecules and Cells
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• 제44권10호
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• pp.736-745
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• 2021

Although various marine ingredients have been exploited for the development of cosmetic products, no previous study has examined the potential of seaweed extracellular vesicles (EV) in such applications. Our results revealed that EV from Codium fragile and Sargassum fusiforme effectively decreased α-MSH-mediated melanin synthesis in MNT-1 human melanoma cells, associated with downregulation of MITF (microphthalmia-associated transcription factor), tyrosinase and TRP1 (tyrosinase-related proteins 1). The most effective inhibitory concentrations of EV were 250 ㎍/ml for S. fusiforme and 25 ㎍/ml for C. fragile, without affecting the viability of MNT-1 cells. Both EV reduced melanin synthesis in the epidermal basal layer of a three-dimensional model of human epidermis. Moreover, the application of the prototype cream containing C. fragile EV (final 5 ㎍/ml) yielded 1.31% improvement in skin brightness in a clinical trial. Together, these results suggest that EV from C. fragile and S. fusiforme reduce melanin synthesis and may be potential therapeutic and/or supplementary whitening agents.

• Journal of Acupuncture Research
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• 제17권2호
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• pp.187-208
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• 2000

By the activation of ovary hormone, many morphological changes occur in the epithelial cell lines and muscle cells in rat uterus. These two cells in uterus are important to the implantation of embryo, maintaining pregnancy and starting parturition. One important change associated with the morphological change of these two cells in uterus is the change on prostaglandin(PG) metabolism. Its presence and synthesis in endometriurn and myometrium in uterus affects estrous cycle and the start of embryo implantation in uterus. It also performs as an important modulator in parturition. So the abnormally weak expression of PG causes difficulty during labor and over-expression causes pre-term labor. PG biosynthesis starts from either free or liberated arachidonic acids from membrane phospholipid by phospholipase. Such arachidonic acids are converted into PG catalyzed by Cyclooxygenase. Under normal physiological condition, Cyclooxygenase-1(COX-1) having 602 units of amino acids controls the synthesis of PG. It acts as a local hormone regulating vasomodulation of blood flow, flexible muscle movement, increasing the blood permeability and contributing the protective role in preserving integrity of the stomach lining and Cyclooxygenase-2 (COX-2) is induced by the inflammation, pregnancy and increased its expression until parturition. Lipid metabolite like PG is located in uterine and expression of COX-2 increased with pregnancy. Increased expression of COX proteins in epithelial cells and myometrial cells are told to increase the muscle contractility in uterus but decreased right after the labor in rat. It is a good sign indicating that COX proteins are deeply related to the start of labor. Currently, Several studies report the use of PG and COX-2 inhibitor as medication for controlled abortion or to prevent pre-term labor but they entail various side-effects. Our study proposed to suggest use of acupuncture as an another mediator to control abortion or pre-term labor without causing unnecessary side-effects by those medicines. Two acupuncture sites, LI-4 & SP-6 were selected due to their known efficacy. From the immunohistochemical staining of COX-2, normal expression of COX-2 protein in nonpregnant SD rat's uterus revealed that COX-2 protein was primarily detected in the lumina epithelial lining and in the epithelial cell lining contacting the stromal cells. High resolution optical microscopic scanning revealed distinguishable staining in the myometrial mucosa. LI-4 acupuncture administered nonpregnant rat's uterus showed strong expression for COX-2 in endometrium contacted with lumina epithelial lining of rat uterus and in myometrial mucosa. Stromal cells showed more staining than untreated nonpregnant rat's uterus and stronger staining in stromal cells contacting myometrial layer compared to untreated nonpregnant rat's uterus. SP-6 acupuncture administered nonpregnant rat's uterus showed weak expression for COX-2 in myometrial layers and stromal cells but no staining was visible in lumina epitheliai and glandular epithelial cells. Few stromal cells and myometrial mucosa were positively stained for COX-2. Pregnant SD rat's uterus was also immunostained for COX-2 expression after 18 days of pregnancy. Unlike to untreated nonpregnant rat's uterus, luminal epithelial cells were not positively stained for COX-2 but stronger staining for COX-2 was revealed in stromal cells. LI-4 acupunctured SD rat's uterus had very strong expression of COX-2 in luminal epithelial lining. Few stromal cells showed stronger positive COX-2 staining and myometrial layers also showed more expression than untreated pregnant rat. SP-6 acupuncture administered pregnant SD rat's uterus showed positive expression of COX-2 in epithelial cells of luminal mucosa layer but weaker than that of LI-4 acupuncture treatment's case. However, strong positive staining was revealed in stromal mucosa and myometrial layers. Virgin SD rat's uterus motility index during LI-4 acupuncture was 66.52 % (Prob〉T = 0.0197) compared to its motility before the acupuncture treatment but the motility index was slighdy elevated up to 79.58 % (Prob〉T = 0.1175) after the acupuncture. During the SP-6 acupuncture treatment for 30 minutes, uterus motility index was 90.52 % (Prob〉T = 0.1832) showing lesser decrement but consequently reached similar motility index decreasal to 79.95 % (Prob〉T = 0.0215) after the acupuncture treatment as LI-4 showed. LI-4 acupuncture tend to be a quick treatment to reducing the uterus motility in a virgin rat but eventually both two acupuncture administration created very similar reduction of uterus motility seeing the index after the both acupunctures. The uterus movement monitored during the LI-4 acupuncture administered for 30 minutes, Pregnant SD rat showed decreased motility down to 77.90 % (Prob〉 T = 0.0076) compared to uterus motility before the acupuncture and it continuously decreased down to 71.81 %(Prob〉T = 0.0214) after the removal of needle. The statistical analysis using paired t-test showed significance difference for both two motility indexs at =0.05. SP-6 acupuncture administered to pregnant SD rat also had similar pattern of decreasing uterus motility index down to 74.70 % (Prob〉T = 0.1730) during the initial 30 minutes acupuncture administration and it was continuously lowered to 71.52 % (Prob〉T = 0.0155) after the acupuncture. The paired t-test resuit for SP-6 suggest prompt response of uterus motility index to the SP-6 acupuncture treatment but consequently reached same level of inducing the motility reduction as LI-4 at =0.05 level.

• Journal of Ecology and Environment
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• 제41권9호
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• pp.265-270
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• 2017

Background: Over the years, pine pollens have been excluded as an allergen due to its relatively large size, low protein content, and waxy hydrophobic layer, despite their abundance. However, recent studies suggest the possibilities of pine pollens being allergens, and it has been reported that allergy symptoms were highly prevalent in areas with considerably large pine forests and high possibility of exposure to the pollen. Therefore, we conducted a comparative analysis of the allergenicities of the pollens from the dominant species of Korean pines, red pine (Pinus densiflora), black pine (Pinus thunbergii), and pitch pine (Pinus rigida), in mice. Methods: The protein composition of the pollens from the three pine species was compared via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The pine pollens and proteins extracted from the pollens were introduced to BALB/c mice by nasal inhalation and application to exposed skin and the IgE produced by the mice were extracted from blood and analyzed via ELISA. Results: SDS-PAGE showed differing protein compositions of the pollens of the three pine species. Analysis of blood IgE compositions showed a similar amount of IgE produced when pollens were applied to skin. In contrast, when mice inhaled the pollens, P. densiflora was shown to induce significantly more IgE production than those of the other two species. Conclusions: The experimental results demonstrate that the pollens of all three South Korean pine species induce IgE production, and this production was more pronounced when the pollens were inhaled than when they were applied to the skin. Of the three species, the pollen of P. densiflora was found to induce the highest level of IgE production.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.756-763
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• 2021

Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

• Journal of Microbiology and Biotechnology
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• 제28권8호
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• pp.1376-1383
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• 2018

The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity ($K_D$) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.

• The Korean Journal of Physiology and Pharmacology
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• 제5권4호
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• pp.287-297
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• 2001

Contraction of smooth muscle is initiated by an increase in cytosolic $Ca^{2+}$ leading to activation of $Ca^{2+}$/ calmodulin-dependnet myosin light chain (MLC) kinase and phosphorylation of MLC. The types of contraction and signaling mechanisms mediating contraction differ depending on the region. The involvement of these different mechanisms varies depending on the source of $Ca^{2+}$ and the kinetic of $Ca^{2+}$ mobilization. $Ca^{2+}$ mobilizing agonists stimulate different phospholipases $(PLC-{\beta},\;PLD\;and\;PLA_2)$ to generate one or more $Ca^{2+}$ mobilizing messengers $(IP_3\;and\;AA),$ and diacylglycerol (DAG), an activator of protein kinase C (PKC). The relative contributions of $PLC-{\beta},\;PLA_2$ and PLD to generate second messengers vary greatly between cells and types of contraction. In smooth muscle cell derived form the circular muscle layer of the intestine, preferential hydrolysis of $PIP_2$ and generation of $IP_3$ and $IP_3-dependent\;Ca^{2+}$ release initiate the contraction. In smooth muscle cells derived from longitudinal muscle layer of the intestine, preferential hydrolysis of PC by PLA2, generation of AA and AA-mediated $Ca^{2+}$ influx, cADP ribose formation and $Ca^{2+}-induced\;Ca^{2+}$ release initiate the contraction. Sustained contraction, however, in both cell types is mediated by $Ca^{2+}-independent$ mechanism involving activation of $PKC-{\varepsilon}$ by DAG derived form PLD. A functional linkage between $G_{13},$ RhoA, ROCK, $PKC-{\varepsilon},$ CPI-17 and MLC phosphorylation in sustained contraction has been implicated. Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to $M_2$ muscarinic receptors activating at least three intracellular phospholipases, i.e. phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD) and the high molecular weight (85 kDa) cytosolic phospholipase $A_2\;(cPLA_2)$ to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic $M_3$ receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the $G_{q/11}$ type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate $(PIP_2),$ producing inositol 1, 4, 5-trisphosphate $(IP_3)$ and DAG. $IP_3$ causes release of intracellular $Ca^{2+}$ and formation of a $Ca^{2+}$-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway.

• 생명과학회지
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• 제30권11호
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• pp.1012-1020
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• 2020

생쥐의 가변부위가 인간의 정상부위에 연결되어 제조된 바이오시밀러 자연항체치료제인 레미케이드는 암괴사인자-알파(TNF-α)에 특이적인 항체로써 카이메릭 단일클론항체이며 류마티스 관절염치료를 위해 개발되었다. 바이오시밀러 레미케이드 항체의 생물학적 기능을 연구하기 위해 우리는 단백질 데이터 은행을 이용한 생물정보학 분석을 수행하여 레미케이드 자연항체와 암괴사인자-알파 항원간의 결합기작특징을 분석하였다. 자연항체를 생산하는 세포의 유전적 불안정성 때문에 레미케이드 항체생산이 제한되므로 우리는 중 사슬 가변부위를 다중펩타이드 링커에 의해 경 사슬 가변부위에 연결된 레미케이드 ScFv항체(Remicade)를 제조하였다. 더욱이 더 높은 생산과 더 높은 항원결합력을 위해 레미케이드 ScFv를 leucine zipper에 융합시켰다. Remicade와 RemicadeScZip ScFv는 대장균에서 발현되었고 Ni⁺-NTA-아가로스 컬럼으로 정제하였다. 정제된 단백질들은 예상한대로 sodium dodecyl sulfate-polyacrylamide electrophoresis에서 28.80 kDa과 33.96 kDa을 나타내었다. Remicade는 ELISA, western blot에서 TNF-α 항원에 대한 결합력이 관찰되지 않았으나 RemicadeScZip은 항원결합력을 나타내었다. 추가적인 BLI분석으로 RemicadeScZip의 TNF-α 항원에 대한 결합력을 재확인시켜주었으며 이 결과는 Leucine zipper가 레미케이드 ScFv의 접힘을 안정화시키고 TNF-α 항원에 대한 결합력을 개선시켰음을 제시해주고 있다.