• 미생물학회지
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• 제7권1호
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• pp.10-21
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• 1969

The effect of glucose and 2-thiobarbituric acid on the biosynthesis of cell constituents such as protein, carbohydrate, DNA, RNA, phospholipid and PCA-soluble phosphate compounds in Chlorella duing the life cycle was measured, and the changes in the content of these main cellular components of the algal cell were analyzed in connection with the nuclear and cytoplasmic divison. In the normal autotrophic synchronous culture the contents of protein, RNA, and DNA in the cell showed a chracteristic changes according to the progress of cell development, increasing more or less throughout all the life cycle. The synthesis of protein is more prominent in the division period nad that of DNA is more active in the ripening period, while the synthesis of RNA is more rapid in the growing and ripening periods than other developmental stages. The period of division cycle was little affected by glucose in the medium, although the synchrony of the growth and cellular division was disturbed and the n value increased. The cotents of protein, carbohydrate, RNA nad DNA of the cell were increased by the glucose treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment. The synthesis of protein, carbohydrate, DNA, RNA and phospholipid of the cell was also retarded by 2-thiobarbituric acid. In the autotrophic, mixotrophic and 2-thiobarbituric acid-treated cultures, each having different mode cytoplasmic division, a common general schema occurring in the cell during the life cycle may be drawn as follows. The ratio of RNA to protein attains maximum value in the $L_1$-cell stage prior to the nuclear division and thereafter decreases during the periods of ripening and division. The ratio of PCA-soluble phosphate compounds to protein increased from the begining of the culture to $L_4$-cell stage successively and thereafter decreased gradually during the division period, while the ratio of protein to DNA kept almost constant up to the division period and thereafter increased during the division period. Therefore, it is presumed that the increase in the ratio of RNA to protein is to be an inducer of nuclear division and that the cytoplasmic division is induced by the increase in the ratio of protein to DNA.

• 한국양식학회지
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• 제6권3호
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• pp.221-233
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• 1993

범가자미, Verasper variegatus에 대한 유전학적 동정을 위하여 세포 크기, DNA함량, 염색체수 및 핵형분석 등의 세포유전학적 조사와 PCR 기법을 이용한 mtDNA 125 ribosomal RNA gene의 분석을 실시하였다. 본 종의 적혈구와 핵의 평균 부피는 각각 $211.10{\mu}m^3$와 $23.03{\mu}m^3$였으며, haploid DNA content는 0.79 pg/cell로서 잉어의 $46.5\%$, 포유류의 $22.6\%$로 나타났다. 염색체 수는 46개로 모두 acrocentric 염색체로 구성되어 있었으며, heteromorphic한 성 염색체는 관찰되지 않았다. PCR 기법을 이용하여 증폭된 범가자미 mtDNA의 12S rRNA gene segment는 대략 390bp로 나타났고, 12S rRNA gene의 PCR product를 제한 효소로 처리 결과, Ava I, Mae II, Sma I, Xba I는 1개의 restriction site가, Mae I는 2개의 restriction site가 관찰되었다. 범가자미 mtDNA의 12S rRNA gene segment의 염기 서열을 인간과 차넬메기와 비교한 결과, identity가 차넬메기 와는 $81.8\%$, 인간과는 $67.7\%$였다.

• 한국미생물·생명공학회지
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• 제26권4호
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• pp.290-296
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• 1998

Alkaline protease 생산력이 우수한 야생형인 A. oryzae val. oryzae의 핵을 순수 분리하여 영양요구성 변이주인 P. chrysogenum(T$yr^{-}$)의 원형질체에 전달하는 핵전이를 이용하여 종간잡종을 형성시킨 후 이들 형질전환체으로 부터 alkaline protease 고생산 균주를 획득하였다. 핵전이를 위한 원형질체 형성 및 재생 조건에서는 Novozym 234의 농도 1%, 삼투안정제는 0.6 M KCl, 효 소의 처리 시간은 180분 그리고 최적 pH는 5.8로 나타났다. 핵전이에 의한 형질전환의 빈도는 1.3~3.8$\times$$10^{-4}$으로 비교적 낮은 편이었으며, 유전적 안정성, conidia의 크기, DNA함량의 측정 그리고 핵염색의 결과 형질전환체의 핵형은 aneuploid로 추정되었다. 형질전환체의 alkaline protease 활성은 모균주와 비교하여 1.1~2.2배 증가하였다.다.

• Archives of Pharmacal Research
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• 제23권1호
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• pp.79-86
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• 2000

Heterokaryotic nuclear hybrids overcoming the natural barriers of incompatibility have been studied in basidiomycetes. To produce these nuclear hybrids between incompatible mushrooms, which have several potent pharmacological effects, nuclear transfer was performed between Lentinula edodes and Coriolus versicolor. Nuclei from serine auxotrophs of Lentinula edodes, $LE207(Ser^{-})$ were transferred into the protoplasts of arginine auxotrophs of Coriolus versicolor, $CV17(Ser^{-})$using 30% polyethylene glycol 4000 in 10 mM $Cacl_{2}$-glycine solution (pH 8.0). Nulcear transfer progenies were selected by nutritional complementation on minimal media supplemented with 0.6 M sucrose. The progenies were classified based on colony morphology to L. edodes-like, C, versicolor-like and non-parental type. Most of the progenies grew slower than either parent. The number of nuclei per cell was similar but the DNA content varied between progenies. The isozyme patterns of nuclear hybrids resembled either of the parent porfiles or showed a mixed profile.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.23-30
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• 1994

Intergeneric hybrids formed between Aspergillus oryzae var oryzae and Penicillium verruculosum F-3 were obtained by nuclear transfer technique. Several auxotrophic mutants isolated fromconidio-spores of the two strains mutagenized with ultraviolet and N-methyl-N-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of hybrid formation by nuclear transfer were 3$\times $10$^{-5}$ $~$1$\times $10$^{-5}$. From observation of genetic stability, conidial size, DNA content, nuclear stain, it was suggested that their karyptypes are aneuploid. The hybrid posses the 1.3$~$2.2 fold higher amylase activities than those of parental strains. It was also revealed that some hybids had different isozyme patterns compared to those of parental strains by amylase activity assays.

• 한국양식학회지
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• 제21권4호
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• pp.259-264
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• 2008

한국산 누치속(genus Hemibarbus) 어류 3종에 대한 세포유전학적 특성을 구령하기 일해 적혈구 세포 크기, 세포당 DNA 함량 및 염색체 핵형을 분석하였다. 누치 H. labeo, 참마자 H. longirostris 및 어름치 H. mylodon의 적혈구 크기를 비교한 결과, 적혈구 세포 장경, 세포 표면적 및 세포 체적에서 어름치가 다른 2종보다 크게 나타났다(P<0.05). 그러나 DNA 함량은 적혈구 세포 크기와 다른 경향을 보여 2.33-2.51 범위로 3종간에 유의적인 차이를 보이지는 않았다(P>0.05). 누치속 어류 3종의 염색체 수는 모두 2n=50개로 나타났으나 핵형에 있어서는 종별로 서로 달라 누치 16M+16SM+18T,A (Fundamental number: NF, 82), 참마자 18M+16SM+16T,A (NF, 84) 그리고 어름치가 18M+24SM+8T,A (NF, 92)로 나타나 3종은 핵형에서 뚜렷한 차이를 보였다.

• 한국균학회지
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• 제27권6호
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• pp.399-405
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• 1999

${\alpha}-amylase$ 생산력이 우수한 야생형인 S. fiburigera의 핵을 순수분리하여 영양요구성 변이주인 S. cerevisiae(tyr-, ura-)의 원형질체에 전달하는 핵전이를 이용하여 종간잡종을 형성시킨 후 이들 형질전환체로부터 ${\alpha}-amylase$ 고생산 균주를 획득하였다. 핵전이를 위한 원형질체 형성 및 재생조건에서는 Novozym 234의 농도 1%, 삼투안정제는 0.6M KC1, 효소처리 시간은 90분, 그리고 최적 pH는 5.8로 나타났다. 핵전이에 의한 형질전환율은 0.25%로 비교적 낮은 편이었으며, 유전적 안정성, 세포크기의 체적, DNA함량의 측정 그리고 핵염색의 결과 형질전환체의 핵형은 aneuploid로 추정되었다. 형질전환 체의 ${\alpha}-amylase$ 활성은 모균주와 비교하여 약 $1.2{\sim}1.9$배 증가하였다.

• 대한기관식도과학회:학술대회논문집
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• 대한기관식도과학회 1993년도 제27차 학술대회 초록집
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• pp.89-89
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• 1993

후두의 상피세포에서 발생하는 질환중 전암성병변은 이형성(dysplasia)과 상피내암(carcinoma in situ)등이 있으며, 이들은 진행하게되면 침윤성 악성병변으로 전환하게 된다. 따라서 전암성 병변의 정도를 정확히 구분 및 파악함으로써 침윤성 암종으로의 전환여부를 미리 예견한다는 것은 악성 후두질환의 병태파악 및 예방에 중요한 역할을 차지한다. 이에 저자들은 최근 후두경하에 절제생검을 시행한 26례(침윤성 편평상피세포암 14례, 상피증식증 5례, 성대결절 7례)를 대상으로, 22례에서는 생검조직을 touch imprint법으로 도말하여 Feulgen염색한 후 CAS 200 화상분석기로 DNA함량분석을 시행하였고, 전례에 대하여 파라핀 포매조직에서 Ki-67 단크론성 항체(M1B1)를 이용하여 면역효소염색을 시행한 후 화상분석기로 양성표현율을 측정분석하여 다음과 같은 결과를 얻었다. 1) Ki-67 양성표현율은 침윤성 암종에서 31.65$\pm$11.59%, 상피증식증에서는 20.14$\pm$3.38%, 성대결절에서는 11.66$\pm$3.02%이었다. 2) 핵산지수(DNA index)는 침윤성 암종의 경우 비배수성이 10례 중 7례(70%), 상피증식증에서는 5례중 2례(40%), 성대결절에서는 7례 모두 이배수성을 보였다. 3) DNA함량분석에서 5기와 G2/M기를 합한 증식지수(PI)는 침윤성 암종에서 23.42$\pm$11.33%, 상피증식증에서는 13.09$\pm$10.90%, 성대결절에서는 4.50$\pm$1.19%로 침윤성 암종에서 가장 높았다. 이상의 성적에서 성대의 생검조직과 같은 미세조직으로부터도 DNA함량검사와 함께 Ki-67 양성표현율을 측정함으로써 전암성병변의 악성화 가능성 정도를 예견할 수 있었으며, 악성종양 환자의 예후판정에 도움을 얻을 수 있으리라 생각된다.

• Journal of Nutrition and Health
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• 제1권2호
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• pp.107-115
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• 1968

Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo. It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacriflced 24, 48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g. The animals were sacriflced by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin, by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionatin of the homogenate (10% W/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4). For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10% trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue being delipidate by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis. The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utilizing the Caraway's colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser's method, which was followed by Marmur's method of purification for DNA and by Chargaff's method of purification for RNA. The determinations of both DNA and RNA were carried out by the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work. 1. It was observed that the body as well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein. 2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded. 3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the decline as compared to the former. 4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue. 5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein. 6. The possible mechanisms involved during in this acute starvation as bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

• The Korean Journal of Physiology and Pharmacology
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• 제26권3호
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• pp.165-174
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• 2022

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.