• Journal of Radiopharmaceuticals and Molecular Probes
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• v.6 no.1
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• pp.3-9
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• 2020

Tetraiodothyroacetic acid (tetrac) is a derivative of thyroid hormone T₄ and causes anti-angiogenesis by blocking T₄ binding to integrin α_vβ₃. In this study, we synthesized [^99mTc]Tc-Cys-Asp-Gly(CDG)-tetrac and evaluated it in vitro as a tumor angiogenesis imaging ligand. The CDG was conjugated to tetrac as a chelator for technetium-99m labeling. The cold vial containing CDG-tetrac, sodium glucoheptonate, and reducing agent was completed under nitrogen-filled atmospheric glove bag. [^99mTc]Tc-CDG-tetrac was synthesized in quantitative yield by heating the cold vial with [^99mTc]TcO₄^- at 100℃ for 30 min. In vitro serum stability of [^99mTc]Tc-CDG-tetrac was measured by incubating the radioligand in 50% fetal bovine serum at 37℃ and analyzing the incubation mixture by radio-TLC, which showed high stability over 6 h (≥ 98%). Cell binding study was carried out by incubating [^99mTc]Tc-CDG-tetrac with human umbilical vein endothelial (HUVE) cells at 37℃ for 6 h. The cell binding of the radioligand increased from 100% at 0.5 h to 293.7% at 6 h in a time-dependent manner. For blocking study, the cells were incubated with the radioligand in the presence of either tetrac (20 μM) or cRGDyK (20 μM) at 37℃ for 4 h. The results demonstrated that the cell binding of the radioligand was inhibited by tetrac (19.1%) or cRGDyK (35.6%), indicating specific binding of the radioligand to integrin α_vβ₃. Thus, this study suggests that [^99mTc]Tc-CDG-tetrac may be a potential radioligand for tumor angiogenesis imaging.

• Applied Biological Chemistry
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• v.30 no.4
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• pp.345-350
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• 1987

Three lots of mejues were prepared with three different strains of Aspergillus oryzae, Bacillus natto, Bacillus subtilis and one was made by conventional method. The four different soy bean pastes were analyzed for compositions of free amino acids, sugars and organic acids during the period of fermentation. Amino nitrogen contents in the samples of A. oryzae were higher than others throughout the aging period. The amounts of each amino acids were varied markedly among the samples after 20-days, while glutamic acid, aspartic acid, lysine and phenylalanine were dominant in all samples after 90-days. Glucose contents were found to be in the range of $0.46{\sim}2.66%$ and other sugars of fructose, sucrose, rhamnose and maltose were less than 0.35%. The levels of total free sugar were relatively higher in the samples prepared with B. natto than others. Citric, lactic, malic, acetic and oxalic acids were identified, and the content of lactic acid was higher in the samples of A. oryzae, whereas citric acid was higher in conventional method.

• Journal of Mushroom
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• v.18 no.2
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• pp.164-173
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• 2020

The extracts of four species of ectomycorrhizal mushrooms-Cantharellus cinnabarinus (OK1247), Lactarius parallelus (OK1264), Tricholoma matsutake (OK1282), and Ramaria botrytis (OK1283)-were prepared to determine their antioxidant activities and nutritional properties. R. botrytis extract displayed the highest 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (33.8%), ferric reducing antioxidant power (0.38), reducing power (0.35), total polyphenol (13.83 mg gallic acid equivalent/g), and flavonoid contents (2.56 mg quercetin equivalent/g). L. parallelus extract displayed the highest nitrite scavenging activity. Analysis of amino acid contents revealed that C. cinnabarinus extract had the highest total amino acid (1,046.1 mg/kg) and essential amino acid (404.9 mg/kg) contents, while R. botrytis extract had the lowest total amino acid (708.3 mg/kg) and essential amino acid (247.3 mg/kg) contents. Among the amino acid components detected in the four ectomycorrhizal mushrooms, cysteine was the most abundant, accounting for 14.3~20.7%, followed by phenylalanine, which accounted for 9.5~13.4% of all amino acids. In summary, the antioxidant activities were the highest in R. botrytis extract, and the amino acid content was the highest in C. cinnabarinus extract, among the four ectomycorrhizal mushrooms.

• Journal of Microbiology
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• v.45 no.2
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• pp.158-167
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• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.459-469
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• 1995

As S-nitrosothiols were proposed as nitrergic carriers in vascular and nonvascular smooth muscle, we have investigated the relaxant properties of several S-nitrosothiols in the porcine retractor penis(PRP) muscle and compared them with the effects of exogenously added NO, electrical field stimulation(EFS) of NANC nerves and sodium nitroprusside(SNP). Also the influences of oxyhemoglobin and hydroquinone on the relaxant responses were investigated. In addition, effects of NO on membrane potentials and its involvement in the generation of inhibitory junction potential(IJP) were investigated with conventional intracellular microelectrode technique. The results were summerized as follows. 1. Frequency-dependent relaxations of PRP muscle were induced by EFS to NANC nerve. Tetrodotoxin($1{\times}10^{-6}M$) abolished the relaxations of PRP muscle induced by EFS, and L-NAME(($2{\times}10^{-5}M$) and methylene blue($4{\times}10^{-5}M$) inhibited the relaxations. L-NAME-induced inhibition of the relaxations was reversed by L-arginine($1{\times}10^{-3}M$), but not by D-arginine. 2. Exogenous NO($1{\times}10^{-5}-1{\times}10^{-4}M$), sodium nitroprusside(($1{\times}10^{-7}-1{\times}10^{-4}M$) induced dose-dependent relaxations of PRP muscle. All S-nitrosothiols($1{\times}10^{-7}-1{\times}10^{-4}M$) tested relaxed the PRP muscle in dose-dependent manner and the potency order was SNAP>GSNO>CysNO>SNAC. 3. Oxyhemoglobin($5{\times}10^{-5}M$) blocked the relaxation induced by exogenous NO and inhibited EFS-, S-nitrosothiols-, and SNP-induced relaxation. 4. Hydroquinone($1{\times}10^{-4}M$) also abolished the relaxations induced by exogenous NO, and reduced the relaxations induced by S-nitrosothiols, but did not affect EFS- and SNP-induced relaxations. 5. SNP($2{\times}10^{-6}-5{\times}10^{-6}M$) relaxed muscle strips but the membrane potentials were not affected. 6. EFS with several pulses(1ms, 2Hz, 80V) produced an inhibitory junction potential(IJP) with muscle relaxation. They were abolished by TTX($2{\times}10^{-6}M$). $N^G$-nitro-$_{\small{L}}$-arginine(L-NNA, $2{\times}10^{-5}M$) abolished the muscle relaxation, but had no effect on IJP.

• Korean Journal of Food Science and Technology
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• v.36 no.3
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• pp.448-455
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• 2004

Changes in quality characteristics of Bijijang (fermented soybean curd residus) prepared at $35^{\circ}C\;and\;40^{\circ}C$ for 0, 12, 24, 36, and 48 hr were investigated. Acidity of Bijijang increased, whereas pH and Hunter's color values decreased during fermentation. Immediately after Bijijang preparation, ${\alpha}-and\;{\beta}-amylase$ activities were very low, ${\beta}-Amylase$ activity during fermentation increased rapidly, with those fermented at $40^{\circ}C$ higher than at $35^{\circ}C$. Neutral pretense activity was significantly higher than acidic pretense activity, and increased gradually after 12 hr. Change in total nitrogen content in Bijijang was insignificant, whereas contents of amino-type and water-soluble nitrogens increased significantly during fermentation. Major free amino acids of Bijijang were Arg, Pro, Glu, Thr, Ser, and Lys at initial fermenting stage, and, as fermentation progressed, contents of Cys, Met Glu, Ile, Leu, and Phe increased. Reducing sugar contents of Bijijang fermented at $40^{\circ}C$ were higher than those fermented at $35^{\circ}C$. Sucrose content decreased and glucose content increased. Glucoside (genistin and daidzin) contents decreased and aglycone (genistein and daidzein) contents increased during preparation of Biji and fermentation of Bijijang. Contents of free sugars and isoflavones were higher in Bijijang fermented at $40^{\circ}C$ than at $35^{\circ}C$. Based on these results, fermentation at $40^{\circ}C$ for 48 hr was determined to be optimum fermentation condition for Bijijang.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.1
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• pp.62-71
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• 1997

The apparent nutrient digestibilities were examined by using chromic oxide indicator according to the various fecal collection methods in juvenile and adult Korean rockfish (Sebastes schlegeli). Feces were collected from three replicate groups of fish by dissection, stripping or decantation using fecal collector attached to fish rearing tank, respectively. The digestibilities of dry matter, protein, lipid, and energy were affected by fecal collection methods (P<0.01), but not affected by fish size. The digestibilities of nutrient determined by stripping or decantation methods were significantly higher than those determined by dissection method (P<0.01). No significant differences in the digestibilities of protein, lipid and energy were found between the stripping and decantation methods in adult fish (P>0.01). The digestibilities of dry matter, protein, lipid, energy, nitrogen-free extract, and total amino acids in juvenile and adult fish were 58, 93, 94, 79, 32, and $93\%$, and 61, 94, 96, 80, 29, and $94\%$, respectively, when they were measured by decantation method. Methionine, cystine and valine digestibilities were significantly lower than those of other amino acids in both juvenile and adult fish (P<0.01). Results indicate that stripping or decantation with fecal collector could be a reliable digestibility procedure for measuring the nutrient digestibilities in Korean rockfish.