• BMB Reports
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• 제52권8호
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• pp.514-519
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• 2019

Osteoclasts are multinucleated giant cells derived from myeloid progenitors. Excessive bone resorption by osteoclasts can result in serious clinical outcomes for which better treatment options are needed. Here, we identified fibronectin leucine-rich transmembrane protein 2 (Flrt2), a ligand of the Unc5 receptor family for neurons, as a novel target associated with the late/maturation stage of osteoclast differentiation. Flrt2 expression is induced by stimulation with receptor activator of nuclear factor-kB ligand (RANKL). Flrt2 deficiency in osteoclasts results in reduced hyper-multinucleation, which could be restored by RNAi-mediated knockdown of Unc5b. Treatment with Netrin1, another ligand of Unc5b which negatively controls osteoclast multinucleation through down regulation of RANKL-induced Rac1 activation, showed no inhibitory effects on Flrt2-deficient cells. In addition, RANKL-induced Rac1 activation was attenuated in Flrt2-deficient cells. Taken together, these results suggest that Flrt2 regulates osteoclast multinucleation by interfering with Netrin 1-Unc5b interaction and may be a suitable therapeutic target for diseases associated with bone remodeling.

• Molecules and Cells
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• 제45권3호
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• pp.112-121
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• 2022

Calorie restriction (CR) and the activation of autophagy extend healthspan by delaying the onset of age-associated diseases in most living organisms. Because protein kinase CK2 (CK2) downregulation induces cellular senescence and nematode aging, we investigated CK2's role in CR and autophagy. This study indicated that CR upregulated CK2's expression, thereby causing SIRT1 and AMP-activated protein kinase (AMPK) activation. CK2α overexpression, including antisense inhibitors of miR-186, miR-216b, miR-337-3p, and miR-760, stimulated autophagy initiation and nucleation markers (increase in ATG5, ATG7, LC3BII, beclin-1, and Ulk1, and decrease in SQSTM1/p62). The SIRT1 deacetylase, AKT, mammalian target of rapamycin (mTOR), AMPK, and forkhead homeobox type O (FoxO) 3a were involved in CK2-mediated autophagy. The treatment with the AKT inhibitor triciribine, the AMPK activator AICAR, or the SIRT1 activator resveratrol rescued a reduction in the expression of lgg-1 (the Caenorhabditis elegans ortholog of LC3B), bec1 (the C. elegans ortholog of beclin-1), and unc-51 (the C. elegans ortholog of Ulk1), mediated by kin-10 (the C. elegans ortholog of CK2β) knockdown in nematodes. Thus, this study indicated that CK2 acted as a positive regulator in CR and autophagy, thereby suggesting that these four miRs' antisense inhibitors can be used as CR mimetics or autophagy inducers.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.615-629
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• 2021

An active compound, triterpene saponin, astersaponin I (AKNS-2) was isolated from Aster koraiensis Nakai (AKNS) and the autophagy activation and neuroprotective effect was investigated on in vitro and in vivo Parkinson's disease (PD) models. The autophagy-regulating effect of AKNS-2 was monitored by analyzing the expression of autophagy-related protein markers in SH-SY5Y cells using Western blot and fluorescent protein quenching assays. The neuroprotection of AKNS-2 was tested by using a 1-methyl-4-phenyl-2,3-dihydropyridium ion (MPP⁺)-induced in vitro PD model in SH-SY5Y cells and an MPTP-induced in vivo PD model in mice. The compound-treated SH-SY5Y cells not only showed enhanced microtubule-associated protein 1A/1B-light chain 3-II (LC3-II) and decreased sequestosome 1 (p62) expression but also showed increased phosphorylated extracellular signal-regulated kinases (p-Erk), phosphorylated AMP-activated protein kinase (p-AMPK) and phosphorylated unc-51-like kinase (p-ULK) and decreased phosphorylated mammalian target of rapamycin (p-mTOR) expression. AKNS-2-activated autophagy could be inhibited by the Erk inhibitor U0126 and by AMPK siRNA. In the MPP⁺-induced in vitro PD model, AKNS-2 reversed the reduced cell viability and tyrosine hydroxylase (TH) levels and reduced the induced α-synuclein level. In an MPTP-induced in vivo PD model, AKNS-2 improved mice behavioral performance, and it restored dopamine synthesis and TH and α-synuclein expression in mouse brain tissues. Consistently, AKNS-2 also modulated the expressions of autophagy related markers in mouse brain tissue. Thus, AKNS-2 upregulates autophagy by activating the Erk/mTOR and AMPK/mTOR pathways. AKNS-2 exerts its neuroprotective effect through autophagy activation and may serve as a potential candidate for PD therapy.

• 대한의생명과학회지
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• 제15권3호
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• pp.261-263
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• 2009

Methylprednisolone is a synthetic glucocorticoid which is usually taken intravenously for many neurosurgical diseases which cause edema including brain tumor, and trauma including spinal cord injury. Methylprednisolone reduces swelling and decreases the body's immune response. It is also used to treat many immune and allergic disorders, such as arthritis, lupus, psoriasis, asthma, ulcerative colitis, and Crohn's disease. To identify genes expressed during methylprednisolone treatment against neurons of rats (PC12 cells), DNA microarray method was used. We have isolated 2 gene groups (up- or down-regulated genes) which are methylprednisolone differentially expressed in neurons. Lipocalin 3 is the gene most significantly increased among 772 up-regulated genes (more than 2 fold over-expression) and Aristaless 3 is the gene most dramatically decreased among 959 down-regulated genes (more than 2 fold down-expression). The gene increased expression of Fgb, Thbd, Cfi, F3, Kngl, Serpinel, C3, Tnfrsf4 and Il8rb are involved stress-response gene, and Nfkbia, Casp7, Pik3rl, I11b, Unc5a, Tgfb2, Kitl and Fgf15 are strongly associated with development. Cell cycle associated genes (Mcm6, Ccnb2, Plk1, Ccnd1, E2f1, Cdc2a, Tgfa, Dusp6, Id3) and cell proliferation associated genes (Ccl2, Tnfsf13, Csf2, Kit, Pim1, Nr3c1, Chrm4, Fosl1, Spp1) are down-regulated more than 2 times by methylprednisolone treatment. Among the genes described above, 4 up-regulated genes are confirmed those expression by RT-PCR. We found that methylprednisolone is related to expression of many genes associated with stress response, development, cell cycle, and cell proliferation by DNA microarray analysis. However, We think further experimental molecular studies will be needed to figure out the exact biological function of various genes described above and the physiological change of neuronal cells by methylprednisolone. The resulting data will give the one of the good clues for understanding of methylprednisolone under molecular level in the neurons.

• 한국지능시스템학회:학술대회논문집
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• 한국퍼지및지능시스템학회 1993년도 Fifth International Fuzzy Systems Association World Congress 93
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• pp.975-976
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• 1993

This talk presents the overview of the author's research and development activities on fuzzy inference hardware. We involved it with two distinct approaches. The first approach is to use application specific integrated circuits (ASIC) technology. The fuzzy inference method is directly implemented in silicon. The second approach, which is in its preliminary stage, is to use more conventional microprocessor architecture. Here, we use a quantitative technique used by designer of reduced instruction set computer (RISC) to modify an architecture of a microprocessor. In the ASIC approach, we implemented the most widely used fuzzy inference mechanism directly on silicon. The mechanism is beaded on a max-min compositional rule of inference, and Mandami's method of fuzzy implication. The two VLSI fuzzy inference chips are designed, fabricated, and fully tested. Both used a full-custom CMOS technology. The second and more claborate chip was designed at the University of North Carolina(U C) in cooperation with MCNC. Both VLSI chips had muliple datapaths for rule digital fuzzy inference chips had multiple datapaths for rule evaluation, and they executed multiple fuzzy if-then rules in parallel. The AT & T chip is the first digital fuzzy inference chip in the world. It ran with a 20 MHz clock cycle and achieved an approximately 80.000 Fuzzy Logical inferences Per Second (FLIPS). It stored and executed 16 fuzzy if-then rules. Since it was designed as a proof of concept prototype chip, it had minimal amount of peripheral logic for system integration. UNC/MCNC chip consists of 688,131 transistors of which 476,160 are used for RAM memory. It ran with a 10 MHz clock cycle. The chip has a 3-staged pipeline and initiates a computation of new inference every 64 cycle. This chip achieved an approximately 160,000 FLIPS. The new architecture have the following important improvements from the AT & T chip: Programmable rule set memory (RAM). On-chip fuzzification operation by a table lookup method. On-chip defuzzification operation by a centroid method. Reconfigurable architecture for processing two rule formats. RAM/datapath redundancy for higher yield It can store and execute 51 if-then rule of the following format: IF A and B and C and D Then Do E, and Then Do F. With this format, the chip takes four inputs and produces two outputs. By software reconfiguration, it can store and execute 102 if-then rules of the following simpler format using the same datapath: IF A and B Then Do E. With this format the chip takes two inputs and produces one outputs. We have built two VME-bus board systems based on this chip for Oak Ridge National Laboratory (ORNL). The board is now installed in a robot at ORNL. Researchers uses this board for experiment in autonomous robot navigation. The Fuzzy Logic system board places the Fuzzy chip into a VMEbus environment. High level C language functions hide the operational details of the board from the applications programme . The programmer treats rule memories and fuzzification function memories as local structures passed as parameters to the C functions. ASIC fuzzy inference hardware is extremely fast, but they are limited in generality. Many aspects of the design are limited or fixed. We have proposed to designing a are limited or fixed. We have proposed to designing a fuzzy information processor as an application specific processor using a quantitative approach. The quantitative approach was developed by RISC designers. In effect, we are interested in evaluating the effectiveness of a specialized RISC processor for fuzzy information processing. As the first step, we measured the possible speed-up of a fuzzy inference program based on if-then rules by an introduction of specialized instructions, i.e., min and max instructions. The minimum and maximum operations are heavily used in fuzzy logic applications as fuzzy intersection and union. We performed measurements using a MIPS R3000 as a base micropro essor. The initial result is encouraging. We can achieve as high as a 2.5 increase in inference speed if the R3000 had min and max instructions. Also, they are useful for speeding up other fuzzy operations such as bounded product and bounded sum. The embedded processor's main task is to control some device or process. It usually runs a single or a embedded processer to create an embedded processor for fuzzy control is very effective. Table I shows the measured speed of the inference by a MIPS R3000 microprocessor, a fictitious MIPS R3000 microprocessor with min and max instructions, and a UNC/MCNC ASIC fuzzy inference chip. The software that used on microprocessors is a simulator of the ASIC chip. The first row is the computation time in seconds of 6000 inferences using 51 rules where each fuzzy set is represented by an array of 64 elements. The second row is the time required to perform a single inference. The last row is the fuzzy logical inferences per second (FLIPS) measured for ach device. There is a large gap in run time between the ASIC and software approaches even if we resort to a specialized fuzzy microprocessor. As for design time and cost, these two approaches represent two extremes. An ASIC approach is extremely expensive. It is, therefore, an important research topic to design a specialized computing architecture for fuzzy applications that falls between these two extremes both in run time and design time/cost. TABLEI INFERENCE TIME BY 51 RULES {{{{Time }}{{MIPS R3000 }}{{ASIC }}{{Regular }}{{With min/mix }}{{6000 inference 1 inference FLIPS }}{{125s 20.8ms 48 }}{{49s 8.2ms 122 }}{{0.0038s 6.4㎲ 156,250 }} }}

• Applied Biological Chemistry
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• 제10권
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• pp.69-100
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• 1968

1 : 탁주양조(濁酒釀造)에 가장 큰 미생물원(微生物源)이 되는 누룩중(中)의 미생물군(微生物群)과 효소(酵素)를 조사하기 위(爲)하여 충남대학교(忠南大學校) 농과대학(農科大學)에서 제조(製造)한 누룩(S)와 시판(市販) 누룩(T)을 공시(供試)하여 사상균(絲狀菌), 호기성세균(好氣性細菌), 유산균(乳酸菌) 및 효모(酵母)를 검색(檢索) 계수(計數)하고 특(特)히 효모(酵母)는 TTC-agar 처리(處理)로서 그 정색별(呈色別)에 따라 유별(類別)하였으며 또한 Amylase 및 Protease 역가(力價)를 측정(測定)하여 얻은 결과(結果)는 다음과 같다. (a) S 누룩 1gm. 중(中)에는 Aspergilles eryzae group: $240{\times}10^5$, Black aspergilli: $163{\times}10^5$, Rhizopus: $20{\times}10^5$, Penicillia: $134{\times}10^5$, 호기성(好氣性) 세균(細菌): $9{\times}10^6{\sim}2{\times}10^7$, 유산균(乳酸菌): $3{\times}10^4$개(個)이였으며 T 누룩 1gm.중(中)에는 Aspergilles oryzae group: $836{\times}10^5$, Black aspergilli: $268{\times}10^5$, Rhizopus: $623{\times}10^5$, Penicillia: $264{\times}10^5$, 호기성(好氣性) 세균(細菌): $5{\times}10^6{\sim}9{\times}10^6$, 유산균(乳酸菌): $3{\times}10^4$개(個)이였다. (b) S 누룩중(中) 호기성세균(好氣性細菌)은 $80{\sim}90%$가 Bacillus subtilis 계(系)인것 같았으며 T 누룩의 것은 70%내외(內外)가 구균(球菌)이었고 유산균(乳酸菌)은 양(兩)누룩이 다 약(約) 80%가 구상유산균(球狀乳酸菌)이였다. (c) S 누룩 1gm.중(中)의 효모(酵母)는 약(約) $6{\times}10^4$개(個)로서 이 중(中) TTC Pink 효모(酵母)가 56.5%, Red pink 효모(酵母): 16%, Red 효모(酵母): 8%, White 효모(酵母): 19.5%이었으며 T 누룩 1gm.중(中)의 효모(酵母)는 약(約) 개(個)로서 이 중(中) Pink: 42%, Red Pink: 21%, Red: 28%, White: 9%이였다. (d) S 누룩 1gm.중(中)의 효소력(酵素力)은 액화형(液化型) amylase: $D^{40}^{\circ}_{30}^'=32\;W.V.$, 당화형(糖化型) amylase: 43.32 A.U., Acid protease 181 C.F.U., Alkaline pretease: 240 C.F.U.이였으며 T 누룩 1gm.의 효소력(酵素力)은 액화형(液化型) amylase: $D^{40}^{\circ}_{30}^'=32\;W.V.$, 당화형(糖化型) amylase: 34.92 A.U., Acid protease 138 C.F.U., Alkaline pretease: 31 C.F.U.이였다. 2 : S, T 누룩으로서 탁주(濁酒)를 담금하여 전양조(全釀造) 기간중(期間中) 매(每) 12시간(時間)의 미생물군(微生物群) 및 효소력(酵素力)의 소장(消長)을 측정(測定)하고 주효모(主酵母)라고 인정(認定)한 Pink와 Red 효모(酵母)를 분리배양(分離培養)하여 유산첨가(乳酸添加)로 pH를 4.2로 조절(調節)한 S, T 각구(各區)술덧에 $1{\times}10^6ml$개(個)를 첨가(添加) 양조(釀造)한 각(各)술덧중(中)의 미생물군(微生物群)의 소장(消長)을 측정(測定)하여 얻은 결과(結果)는 다음과 같으며 이 실험에서도 효모(酵母)는 TTC 정색별(呈色別)로 유별(類別) 계수(計數)하였다. (a) 각시료구(各試料區)에서 사상균(絲狀菌)은 담금후(後) $2{\sim}3$일경(日頃)부터 검출(檢出)되지 않았으며 호기성(好氣性) 세균(細菌)은 담금 직후(直後) 술덧 매(每) ml 당(當) S 누룩 사용(使用)의 3구(區)에서 $15{\times}10^7{\sim}35{\times}10^7$개(個), T 누룩 사용(使用)의 3 구(區)에서 $8.2{\times}10^7{\sim}12{\times}10^7$개(個)가 검출(檢出)되어 이중(中) 구균(球菌)은 유산(乳酸) 무첨가(無添加)로 담금한 S 및 T구(區)에서는 36시간경(時間頃)까지 많은 증식(增殖)을 하다가 기후(其後) 급속(急速)히 감소(減少)되고 유산(乳酸) 첨가(添加)로 담금한 SP, SR, TP, 및 TR 구(區)에서는 처음부터 거의 검출(檢出)되지 않고 Bacillus는 각구(各區)에서 다 같이 많은 기복(起伏)이 있는 소장(消長)을 하였으나 말기(末期)에는 초기(初期)의 약(約) $1/5{\sim}1/10$ 수(數)로 감소(減少)되었다. (b) 유산균(乳酸菌)은 담금후(後) 24시간경(時間頃)에 S 구(區)에서는 술덧 매(每) ml 당(當) 약(約) $7.4{\times}10^7$ 개(個)가 검출(檢出)되어 $3{\sim}4$일경(日頃)까지 약(約) $2{\times}10^8$ 개(個)로 증식(增殖)하고 그후(後) 급속(急速)히 감소(減少)되어 말경(末頃)에는 약(約) $4{\times}10^5$ 개(個)가 존재(存在)하였으며 T 구(區)에서는 24시간경(時間頃)에 검출(檢出)되었으며 3일경(日頃)에는 약(約) $3{\times}10^7$ 개(個), 말경(末頃)에는 약(約) $2{\times}10^5$ 개(個)가 존재(存在)하였다. 한편 SP, SR, TP, 및 TR 구(區)에서는 각구(各區)마다 24시간경(時間頃)에 약(約) $4{\times}10^5$ 개(個)로 현저(顯著)히 적은 수(數)가 검출(檢出)되었으며 기후(其後) 별(別) 변동(變動)없는 소장(消長)을 하거나 사멸(死滅)되어 거의 검출(檢出)되지 않았다. (c) 각구(各區)술덧에서 검출(檢出)된 유산균(乳酸菌)은 대부분(大部分) 구상균(球狀菌)이였으며 또한 유산균(乳酸菌)의 소장(消長)은 술덧중(中)에 생성(生成)되는 유산(乳酸) 및 Alcohol 함량(含量)과 부합(符合)되는 경향(傾向)이였다. (d) 효모(酵母)는 24시간경(時間頃)부터 증식(增殖)이 뚜렷하여 술덧 매(每) ml당(當) S 구(區)에서는 약(約) $2{\times}10^8$ 개(個)가 되고 48시간경(時間頃)에는 약(約) $4{\times}10^8$ 개(個)가 되어 계속(繼續)하다가 후기(後期)에 다시 증가(增加)되어 $5{\sim}7{\times}10^8$ 개(個)가 되었으며 T 구(區)에서는 24시간경(時間頃)에 $4{\times}10^8$ 개(個)가 되었고 기후(其後) 기복(起伏)을 보이면서 $2{\sim}5{\times}10^8$ 개(個)로 계속(繼續)하였다. (e) S, T 양구(兩區)술덧중(中)에서 소장(消長)한 효모(酵母)는 TTC Pink 효모(酵母)가 90%이상(以上)을 차지하고 Red pink 및 Red 효모(酵母)는 전양조기간(全釀造期間)을 통(通)하여 $2{\times}10^6{\sim}3{\times}10^7$ 개(個) 사이에서 소장(消長)하였다. (f) SP 에서는 효모(酵母)는 24시간경(時間頃)에 S 구(區)보다 2배(倍)가 되는 약(約) $5{\times}10^8$ 개(個)의 Pink 효모(酵母)가 검출(檢出)되었으며 중기말경(中期末頃)까지는 S 구(區)보다 많은 수(數)이였으나 후기(後期)에는 별차(別差)없는 수(數)가 되었다. (g) SR 구(區)에서 소장(消長)하는 총효모수(總酵母數)는 SP 구(區)와 대차(大差)는 없었으나, 첨가(添加)해준 Red 효모(酵母)는 초기(初期)에 다소(多少) 많이 검출(檢出)되고 3일후(日後)부터는 S 구(區)와 별차(別差)없는 수(數)로 소장(消長)하였으며 TR구(區)에서도 Red 효모(酵母)가 초기(初期)에는 T 구(區)에 비(比)해서 많았으나 중기이후(中期以後)부터는 T 구(區)와 별차(別差)없는 경향(傾向)이어서 Red 효모(酵母)를 가(加)한 구(區)에서도 Pink 효모(酵母)가 훨씬 우세(優勢)함을 보여 본실험(本實驗)에서 검출(檢出)된 Red 효모(酵母)는 탁주양조(濁酒釀造)에서 후기(後期)까지 계속(繼續) 생육(生育)은 하나 많은 증식(增殖)은 하지 않았다. (h) TP 구(區)에서 Pink 효모(酵母)는 2일경(日頃)에 약(約) $5{\times}10^8$ 개(個)가 되어 T 구(區)에 비(比)해서 많고 기후(其後) 감소(減少)되는 경향(傾向)이나 T 구(區)보다는 많으며 후기(後期)에서는 역시(亦是) T 구(區)와 별차(別差)없는 수(數)가 되었다. (i) 술덧중(中)의 효모(酵母) 소장(消長)과 동시(同時)에 Alcohol 생성량(生成量)을 측정(測定)한 결과(結果) Pink 효모(酵母)를 첨가(添加)한 구(區)에서 초기(初期)에 다소(多少) 많은 생성량(生成量)을 보였으나 중기(中期) 이후(以後)는 효모(酵母) 무첨가구(無添加區)와의 차이(差異)를 인정(認定)할 수 없었고 또 Alcohol생성(生成)은 효모(酵母) 총수(總數)와 비례(比例)하지 않았다. (j) 액화형(液化型) Amylase는 담금 후(後) 12시간경(時間頃)까지 가장 강(强)하고 24시간경(時間頃)에 일단(一旦) 감소(減少)되었다가 시간경(時間頃)에 최고(最高)로 증가(增加)된 후(後) 74시간경(時間頃)까지 서서(徐徐)히 감소(減少)되고 기후(其後)는 급(急)히 감소(減少)를 보였다. (k) 술덧발효중(醱酵中) Alkaline protease는 불규칙적(不規則的)이기는 하나 계속(繼續) 감소(減少)되는 소장(消長)이었고 Acid protease는 24시간경(時間頃)에 최고(最高)로 증가(增加)하였다가 급(急)히 감소(減少)된 후(後) 다시 증가(增加)하는 불규칙적(不規則的)인 소장(消長)을 보였으나 Alkaline protease 보다는 계속(繼續) 강(强)하였었다. 3 : 본실험(本實驗)에서 가장 많이 검출(檢出)된 TTC Pink 효모(酵母)와 계속(繼續) 나타난 2주(株)의 Red Pink 효모(酵母) 및 1주(株)의 를 동정(同定)하고 이들의 생리적성질(生理的性質)을 검사(檢査)한 결과(結果)는 다음과 같다. (a) TTC Pink 효모(酵母)(B-50P)와 2주(株)의 Red Pink 효모(酵母)(B-54RP 및 B-60RP)는 Saccharomyces cerevisiae 형(型)이였고 Red 효모(酵母)(B-53P)는 Hansenula subpelliculosa 형(型)이였다. (b) 분리동정(分離同定)한 발효력(醱酵力)을 측정(測定)한 결과(結果) 2주(株)의 TTC Red Pink 효모(酵母)가 가장 강(强)하고 Pink 효모(酵母)가 다소(多少) 떨어지며 Red효모(酵母)는 현저(顯著)히 약(弱)함을 보였고 특(特)히 Pink 및 2주(株)의 Red Pink 효모(酵母)는 초발효력(初醱酵力)이 강(强)하여 탁주양조(濁酒釀造)에 적합(適合)함을 알았으며 Red효모(酵母)는 발효력(醱酵力)은 약(弱)하나 Ester생성력(生成力)이 강(强)하여 탁주양조(濁酒釀造)에 중요(重要)한 역할(役割)를 하는것을 알았다. 따라 탁주양조(濁酒釀造)에는 TTC Red Pink 및 Pink로 정색(呈色)되는 Saccharomyces cerevisiae 형(型)이 우량(優良)함을 추정(推定) 할 수 있었다. (c) 분리동정(分離同定)한 4주(株)효모(酵母) 아초산내성(亞硝酸耐性)은 강(强)하였으며 유산내성(乳酸耐性)은 국즙(麴汁) 배지(培地)에서 3%정도(程度)이었으나 Red효모(酵母)는 더 강(强)하였고 Alcohol 내성(耐性)은 Hayduck배지(培地)에서 Pink 및 Red Pink 효모(酵母)는 3%정도(程度)이고 맥아즙(麥芽汁) 배지(培地)에서는 13%정도(程度)이었으나 Red 효모(酵母)는 이들보다 훨씬 약(弱)하였으며 Gelatin 액화(液化)는 2주(株)가 다 40일(日)까지 (-)이였다. 4 : 탁주양조중(濁酒釀造中)의 발효도(醱酵度)는 2일경(日頃)에 총발효율(總醱酵率)의 $70{\sim}80%$가 이루어지고 $3{\sim}4$일경(日頃)까지 90%내외(內外)가 진행(進行)되어 주발효(主醱酵)는 이 시기(時期)에 종료(終了)됨을 보였으며 또한 탁주양조(濁酒釀造)에 있어서 담금한 총전분량(總澱粉量)에 대(對)한 주정(酒精) 발효율(醱酵率)은 65%내외(內外)가 됨을 알 수 있었다. 5 : 제등(齊藤)가 탁주(濁酒)술덧에서 분리(分離)한 Saccharomyces coreanus가 본실험(本實驗)에서 전연(全然) 검출(檢出)되지 않은 이유(理由)는 1930년경(年頃)부터 누룩제조(製造)에 국균(麴菌)을 종균(種菌)으로 접종(接種)하였으며 또한 탁주양조(濁酒釀造)에 일본국(日本麴)도 혼합사용(混合使用)하여서 탁주양조(濁酒釀造)에 있어서 미생물상(微生物相)이 완전(完全)히 달라진 탓이 아닌가 생각되며 이것을 뒷받침 하는 것으로 과거의 약탁주(藥濁酒) 고유미(固有味)가 달라진 것을 들 수 있다.