• Title/Summary/Keyword: R peptide

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Identification of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Golpayegani Cattle by DNA Test

  • Mosafer, J.;Nassiry, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1691-1695
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    • 2005
  • The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

A Comparison of Ammonia and Preformed Protein as a Source of Nitrogen for Microbial Growth in the Rumen of Sheep Given Oaten Chaff

  • Kanjanapruthipong, J.;Leng, R.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.4
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    • pp.351-362
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    • 1998
  • Microbial growth efficiency in the rumen was studied in sheep given hourly, 31.25 g oaten chaff with either 0.31 and 0.88 g urea or 1.88 and 5.63 g casein (exp. 1) and 33.33 g oaten chaff with 1.04 casein or 0.3, 0.6 and 0.9 g urea or the mixture of the casein and urea (exp. 2). Concentrations of ruminal fluid ammonia increased with increasing nitrogenous supplements. Organic matter digestibility in sacco in the rumen was not different irrespective of N sources. Isoacids and valeric acid increased with increasing ingested casein but decreased with increasing urea intake. Peptide and amino acid pools in ruminal fluid increased with increasing ammonia concentrations (exp. 2) suggesting that proteolytic activity and transportation of peptides and amino acids across microbial membrane of rumen microbes may be regulated by the metabolite mechanism (intracellular amino acids and $NH_4{^+}$, respectively). Densities of total viable and cellulolytic bacteria in ruminal fluid increased with increasing ammonia levels but that of small Entodinia decreased. The density of fungal sporangia growth on oat leaf blades decreased with increasing ammonia concentrations but appeared to remain constant in the presence of casein. Efficiency of net microbial cell synthesis was 15-28% higher when ammonia concentrations increased from 100 to above 200 mg N/l regardless of N sources. In conclusion, supplementation of preformed protein had no effect on rumen digestion and microbial growth efficiency. This could not be accounted for its effect on ruminal fluid ammonia. Increased microbial growth efficiency with increasing ammonia levels may be due to a reduction in the turnover of microbial cells within the rumen.

Regulation of m-Calpain Activity by α-Synuclein and Its C-terminal Fragment (α-syn61-140)

  • Lee, In-Hwan;Kim, Hyun-Jin;Lee, Choong-Hwan;Paik, Seung R.
    • Bulletin of the Korean Chemical Society
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    • v.27 no.7
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    • pp.1001-1004
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    • 2006
  • The m-calpain activity hydrolyzing a fluorogenic substrate of N-Succinyl-Leu-Leu-Val-Tyr-7-amino-4-methylcourmarin (LLVY-AMC) was significantly stimulated by more than two-fold in the presence of 5$\mu$M $\alpha$synuclein at $15{^{\circ}C}$. The stimulation was also confirmed with azocasein. The stimulation of the peptide hydrolyzing activity required structural intactness of $\alpha$-synuclein since the C-terminally or N-terminally modified proteins such as $\beta$-synuclein, $\alpha$-syn1-97, and $\alpha$-syn61-140 did not increase the proteolytic activity. Instead, however, the N-terminally truncated $\alpha$-syn61-140 was shown to drastically suppress the calpain activity. Since the N-terminal truncation was known to be the primary cleaving event of calpain-mediated proteolysis of $\alpha$-synuclein and the $\alpha$-syn61-140 has been demonstrated to be resistant against the calpain digestion, it has been proposed that the intracellular calpain activity could be regulated in a reciprocal manner by $\alpha$-synuclein and its proteolyzed C-terminal fragment. Based on the results, a possible physiological function of $\alpha$-synuclein has been suggested as a calpain regulator which contains both stimulatory and inhibitory activities.

Molecular Simulations for Anti-amyloidogenic Effect of Flavonoid Myricetin Exerted against Alzheimer’s β-Amyloid Fibrils Formation

  • Choi, Young-Jin;Kim, Thomas Donghyun;Paik, Seung R.;Jeong, Karp-Joo;Jung, Seun-Ho
    • Bulletin of the Korean Chemical Society
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    • v.29 no.8
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    • pp.1505-1509
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    • 2008
  • Comparative molecular simulations were performed to establish molecular interaction and inhibitory effect of flavonoid myricetin on formation of amyloid fibris. For computational comparison, the conformational stability of myricetin with amyloid $\beta$ -peptide (A$\beta$ ) and $\beta$ -amyloid fibrils (fA$\beta$) were traced with multiple molecular dynamics simulations (MD) using the CHARMM program from Monte Carlo docked structures. Simulations showed that the inhibition by myricetin involves binding of the flavonoid to fA$\beta$ rather than A$\beta$ . Even in MD simulations over 5 ns at 300 K, myricetin/fA$\beta$ complex remained stable in compact conformation for multiple trajectories. In contrast, myricetin/A$\beta$ complex mostly turned into the dissociated conformation during the MD simulations at 300 K. These multiple MD simulations provide a theoretical basis for the higher inhibitory effect of myricetin on fibrillogenesis of fA$\beta$ relative to A$\beta$ . Significant binding between myricetin and fA$\beta$ observed from the computational simulations clearly reflects the previous experimental results in which only fA$\beta$ had bound to the myricetin molecules.

The Effect of GCSB-5 a New Herbal Medicine on Changes in Pain Behavior and Neuroglial Activation in a Rat Model of Lumbar Disc Herniation

  • Cho, Hee Kyung;Kim, So-Yeon;Choi, Mi Jung;Baek, Seung Ok;Kwak, Sang Gyu;Ahn, Sang Ho
    • Journal of Korean Neurosurgical Society
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    • v.59 no.2
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    • pp.98-105
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    • 2016
  • Objective : Lumbar disc herniation can induce sciatica by mechanical compression and/or chemical irritation. The aim of this study was to compare the effects of GCSB-5 (Shinbaro$^{(R)}$) and NSAIDs on pain-related behavior and on the expressions of microglia, astrocytes, CGRP, TRPV1, IL-6, and CX3CL1 in a rat model of lumbar disc herniation. Methods : 112 male Sprague-Dawley rats underwent implantation of nucleus pulposus to a dorsal root ganglion (DRG). Rats were divided into five groups as follows; a saline group (the vehicle control group) (n=27), a 10 mg/kg aceclofenac group (the aceclofenac group) (n=22), and 100, 300 or 600 mg/kg GCSB-5 groups (the GCSB-5 100, 300, or 600 groups) (n=21 for each group). Rats were tested for mechanical allodynia at 3 days after surgery and at 1 day, 3 days, 7 days, 14 days, 21 days, 28 days, 35 days, 42 days, 49 days, and 56 days after treatment commencement. Immunohistochemical staining of microglia (Iba1), astrocytes (GFAP), CGRP, and TRPV1, and PCR for IL-6 and CX3CL1 were performed on spinal dorsal horns and DRGs at 56 days after medication commencement. Results : After 56 days of GCSB-5 300 administration, mechanical withdrawal thresholds were significantly increased (p<0.05), and immunohistochemical expressions of Iba1, GFAP, CGRP, and TRPV1 were reduced than other groups, but this difference was not statistically significant. Conclusion : These results indicate GCSB-5 reduces mechanical allodynia and downregulates neuroglial activity and the expressions of CGRP and TRPV1 in the spinal segments of a rat model of lumbar disc herniation.

Correlation of PD-L1 Expression Tested by 22C3 and SP263 in Non-Small Cell Lung Cancer and Its Prognostic Effect on EGFR Mutation-Positive Lung Adenocarcinoma

  • Kim, Taehee;Cha, Yoon Jin;Chang, Yoon Soo
    • Tuberculosis and Respiratory Diseases
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    • v.83 no.1
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    • pp.51-60
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    • 2020
  • Background: Programmed death-ligand 1 (PD-L1) expression is tested by immunohistochemistry (IHC)-22C3, SP263, and SP142. The aim of this study is to evaluate the correlation among the three methods of PD-L1 IHC in non-small cell lung cancer (NSCLC) and clinical significance of PD-L1 expression in lung adenocarcinoma with an epidermal growth factor receptor (EGFR)-tyrosine kinase domain mutation. Methods: The results of 230 patients who were pathologically confirmed as having NSCLC; tested using PD-L1 IHC 22C3, SP263, and SP142 methods; and evaluated via the peptide nucleic acid clamping method to confirm EGFR mutation, were analyzed in this study. Results: 164 patients underwent both the SP263 and 22C3 tests. There was a significant positive correlation between the outcomes of the two tests (Spearman correlation coefficient=0.912, p<0.001), with a derived regression equation as follows: 22C3=15.2+0.884×SP263 (R2=0.792, p<0.001). There was no relationship between the expression of PD-L1 and clinical parameters, including EGFR-tyrosine kinase inhibitor (TKI) mutation. The PD-L1 expression in patients treated with EGFR-TKI yielded a 2-month-shorter progression period than that in the PD-L1-negative group. However, this did not reach statistical significance (PD-L1<1% vs. PD-L1≥1%, 10 months vs. 8 months). Conclusion: The results of the 22C3 and those of SP263 methods were in good correlation with one another. Since the PD-L1 expression is not influenced by the EGFR mutation, it is necessary to perform a PD-L1 test to set the treatment direction in the patients with EGFR-mutant NSCLC.

Comparative antigen analysis of yrichomonus vaginulis by enzyme-linked immunoelectrotransfer blot technique (효소면역 전기영동 이적법을 이용한 질트리코모나스 항원의 비교 분석)

  • 민득영;임미혜
    • Parasites, Hosts and Diseases
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    • v.30 no.4
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    • pp.323-328
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    • 1992
  • Analysis of siR isolates of Trichemenes veginalis was carried out with sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12~170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with difFerent mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum(HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 9:k kDa on EITB may connote the important significance on immune response in T. vaginalis infection.

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Bioinspired Polymers that Control Intracellular Drug Delivery

  • Allan S. Hoffman;Patrick S. Stayton;Oliver-Press;Niren-Murthy;Chantal A. Lackey;Charles-Cheung;Fiona-Black;Jean Campbell;Nelson Fausto;Themis R. Kyriakides;Paul-Bornstein
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.205-212
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    • 2001
  • One of the important characteristics of biological systems os their ability to change im-portant properties in response to small environmental signals. The molecular mechanisms that biological molecules utilize to sense and respond provide interesting models for the development of "smart" polymeric biomaterials with biomimetic properties. An important example of this is the protein coat of viruses, which contains peptide units that facilitate the trafficking of the virus into the cell via endocytosis, then out of the endosome into the cytoplasm, and from there into the nucleus, We have designed a family of synthetic polymers whose compositions have been de-signed to mimic specific peptides on viral coats that facilitate endosomal escape. Our biomimetic polymers are responsive to the lowered pH whinin endosomes, leading to distruption of the en-dosomal membrane and release of important biomolecular druges such as DNA, RNA, peptides and proteins to the cytoplasm before they are trafficked to lysosomes and degraded by lysosomal en-zymes. In this article, we review our work on the design, synthesis and action of such smart, pH-sensitive polymers.

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Bee-Venom theraphy -Method of Clinical Approach- (봉독요법 -임상활용방법을 중심으로-)

  • 이재동
    • The Journal of Korean Medicine
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    • v.21 no.3
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    • pp.3-8
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    • 2000
  • 1. Definition : Bee-venom therapy does not involve actual bee-stings: it is a treatment method with acquired bee-venom extract through the electric stimulus on the bee, It is injected subcutaneusly on the acupuncture point after refining, according to the diagnosis of constitution and disease. 2. History : Around 2000 B.C., records that Bee-venom was used for therapy were written in the medical book of Babylonia and Papirus of ancient Egypt. Hippocrates, who is called the father of Medicine, said that Bee-venom is Arcanum, which means mysteric medicine. In Oriental medicine, B.C. 200, there was a clinical record that the meat suspended in front of the bee house on the tree in order to get bee-venom, was attached on the lesion. 3. Mechanism of Action : There are two aspects: 1) The effect of stimulating acupunture point It is similar to the chemical moxa. I think that there are several methods of stimulating the acupuncture point: For example, a simple needle is a mechanical stimulus, Moxa is a heating stimulus and electric and Raser acupunture etc. And another stimulus: in the ancient orient, a chemical stimulus called Chungu(Tianjiu), is attached to the lesions by using grinded insects (ex. Mylaris phalerate PALL.) which have toxin. So Bee venom therapy is similar to this. 2) The effect of biochemical ingredients Bee venom consists of 40 kinds of ingredients. For example, me Iii tin, Apamin, Pospholipase A2, MCD peptide, Adolapin and so on. They have effects which have been proven through experimentation l) tonifying mechanism of the body through increasing hormon secretion 2) tonifying immune system through proliferation of WBC, lymphocytes, macrophage 3) anti-inflammatory reaction Therefore Bee venom therapy is the representative 3rd Medicine, which combined East & West medicine. 4. Application of disease : L.B.P and HIVD, O.A, R.A, degenerative arthritis, shoulder pain and other pain diseases. 5. Therapic methods : According to constitution and disease, proper concentration of bee venom is injected on acupunture point, 2 times a week. Generally one term is consisted of 15times. 6. Contraindication : Heart disease, TBc, DM, kidney disease(nephritis), pregnancy, woman in menstruation 3-4 persons per 100,000 persons may have severe allergic reaction.

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Description of Nearly Completed Mitochondrial Genome Sequences of the Garden Chafer Polyphylla laticollis manchurica, Endangered in Korea (Insecta: Coleoptera)

  • Kim, Min Jee;Kim, Ki-Gyoung;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.1
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    • pp.185-202
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    • 2013
  • In this study, we present the nearly complete mitogenome sequences of the garden chafer, Polyphylla laticollis manchurica, which is listed as an endangered species in Korea. The P. l. manchurica mitogenome, which includes unfinished whole A+T-rich region and a partial srRNA was 14,473-bp long, possessing typical sets of genes (13 PCGs, 22 tRNA genes, and 2 rRNA genes). Gene arrangement of the P. l. manchurica mitogenome was identical to the common one found in the majority of insects. The 5 bp-long motif sequence (TAGTA) that has been suggested to be the possible binding site for the transcription termination peptide for the major-strand was also found in the P. l. manchurica mitogenome between $tRNA^{Ser}$(UCN) and ND1. The start codon for COI gene and ATPase8 was designated as a typical TTG. All tRNAs of the P. l. manchurica showed a stable canonical clover-leaf structure of other mt tRNAs, except for $tRNA^{Ser}$(AGN), DHU arm of which could not form stable stemloop structure. As has been previously determined, the high A/T content was unanimously observed in P. l. manchurica in terms of A/T bias in the third codon position (73.5%) compared with the first (66.4%) and second codon position (66.2%). The PCGs encoded in major-strands are slightly T-skewed, whereas those of the minor-strand are A-skewed, indicating strand asymmetry in nucleotide composition in the Coleoptera including P. l. manchurica.