• Title/Summary/Keyword: silver staining

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Comparative Analysis of Muscle Proteome from Porcine White and Red Muscles by Two-dimensional Electrophoresis (이차원전기영동법을 이용한 white muscle과 red muscle간의 단백질 발현양상의 비교분석)

  • Kim, N.K.;Joh, J.H.;Chu, K.S.;Park, H.R.;Park, B.Y.;Kim, O.H.;Lee, C.S.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.731-738
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    • 2003
  • The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

Effect of Post-treatment Using Acidic Amino Acids during Hair Coloring on Hair Condition (산성 아미노산 후처리가 헤어컬러링 시 모발에 미치는 영향)

  • Lee, Jin Young;Lee, Sang Hyun
    • Journal of the Korea Convergence Society
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    • v.12 no.11
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    • pp.427-434
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    • 2021
  • Modern people express their beauty through hair coloring, but hair can be damaged by repeated chemical treatments. In order to increase the durability of dyeing and minimize the hair damage, in this study, the acidic amino acids including aspartic acid (Asp) and glutamic acid (Glu) were used to post-treat hair during hair coloring. The post-treatment with 0.75% Asp and Glu solution was carried out at room temperature for 20 minutes after dyeing bleached hair with cherry red and blue silver colors. After repeated shampooing of 1, 5, 10, 15, 20, and 25 times, L*a*b* value of dyed hair was measured to confirm the dyeing durability, and the changes in tensile strength, porosity, and surface properties of the hair were also analyzed to determine the condition of the hair. In the case of cherry red and blue silver staining, the Asp and Glu experimental group showed higher color persistence than the control group, and the Asp and Glu experimental group showed higher tensile strength, lower porosity and smooth surface properties than the control group. In particular, the Asp test group showed superior color persistence and lower hair damage than the Glu test group. This study, therefore, if damaged in dyeing and bleaching in the field of hair after treatment with asp glu a combination of hair cosmetics in the development of basic data look forward to be.

Acupuncture Treatment at HT8 Protects Hippocampal Cells in Dentate Gyrus on Kainic Acid-Induced Epilepsy Mice Model (소부혈(少府穴) 자침(刺鍼)이 Kainic Acid로 유도(誘導)된 간질(癎疾) 동물(動物) 모델의 해마(海馬) 치상회(齒狀回)에 미치는 영향(影響))

  • Kim, Seung-Tae;Chung, Joo-Ho;Jeong, Wu-Byung;Kim, Jang-Hyun;Kang, Min-Jung;Hong, Mee-Sook;Park, Hae-Jeong;Kim, Yeon-Jung;Park, Hi-Joon;Lee, Hye-Jeong
    • Korean Journal of Acupuncture
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    • v.24 no.4
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    • pp.99-110
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    • 2007
  • Objectives : Epilepsy is one of the most common serious brain disorders that affect people of all ages, and it is characterized by recurrent unprovoked seizures. We examined whether acupuncture can reduce both the incidence of seizures and hippocampal cell death in dentate gyrus (DG) using a mouse model of kainic acid (KA)-induced epilepsy. Methods : ICR mice ($20{\sim}25$ g) were given acupuncture once a day at acupoint HT8 (sobu) bilaterally during 2 days before KA injection. After an intracerebroventricular injection of 0.1${\mu}g$ of KA, acupuncture treatment was subsequently administered once more (total 3 times), and the degree of seizure was observed for 20 min. Three hours after injection, we confirmed the neural cell death using cresyl violet staining and silver impregnation staining, and determined the expressions of c-Fos and glutamate decarboxylase (GAD)-67 using immunohistochemistry techniques in the DG. Results : KA induced epileptic seizure, neural cell death, increased c-Fos expression and decreased GAD-67 expression in the DG. Acupuncture treatment at HT8 reduced the severity of the epileptic seizure and inhibited neural cell death from KA. In addition, acupuncture normalized the expressions of c-Fos and GAD-67 in the same areas. Conclusions : These results demonstrated that acupuncture treatment at HT8 may reduce the KA-induced epileptic seizure and neural cell death in the DG possibly by normalizing c-Fos expressions and the gamma-aminobutyric acid neurons.

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Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene

  • Pruthviraj, D.R.;Usha, A.P.;Venkatachalapathy, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.315-320
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    • 2016
  • Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5'-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.

Potential Vaccine Targets against Rabbit Coccidiosis by Immunoproteomic Analysis

  • Song, Hongyan;Dong, Ronglian;Qiu, Baofeng;Jing, Jin;Zhu, Shunxing;Liu, Chun;Jiang, Yingmei;Wu, Liucheng;Wang, Shengcun;Miao, Jin;Shao, Yixiang
    • Parasites, Hosts and Diseases
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    • v.55 no.1
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    • pp.15-20
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    • 2017
  • The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.

Molecular Characterization of Hallikar Breed of Cattle Using Microsatellite Markers

  • Kumar, S. Naveen;Jayashankar, M.R.;Nagaraja, C.S.;Govindaiah, M.G.;Saravanan, R.;Karthickeyan, S.M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.622-626
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    • 2006
  • Molecular characterization of Hallikar, the native cattle breed of Karnataka, was undertaken using 19 cattle specific, highly polymorphic microsatellite markers recommended by FAO. The genomic DNA was subjected to PCR amplification and alleles were resolved through six per cent denaturing PAGE with a 10 bp DNA ladder followed by silver staining. Genotyping of animals was done based on allele size. The number of alleles ranged from three to nine with allele sizes ranging from 102 bp to 294 bp. These alleles were distributed in the frequency range between 0.0306 and 0.8673 in the population. The mean observed number of alleles was $6.368{\pm}1.4225$. The mean observed and expected heterozygosities were $0.7515{\pm}0.1734$ and $0.7850{\pm}0.1381$, respectively. The high heterozygosity observed implies presence of higher genetic variability within Hallikar breed. The PIC (Polymorphism Information Content) values ranged from 0.2322 (ETH152) to 0.8654 (ETH225). The percentage of polymorphic loci obtained was 100 as all the 19 microsatellite markers were found to be polymorphic. Except for ETH152, all the other loci had high PIC values, indicating that these markers are highly informative for characterization of Hallikar breed. The population was tested for Hardy-Weinberg equilibrium at 19 microsatellite loci, and at 74 per cent of the loci the population was found to be in disequilibrium.

Immunologic effects of somatic antigens of house dust mite (Dermatophagoides pteronyssinus) against canine sarcoptic mite (Sarcoptes scabiei var. canis) infestation (집먼지진드기 체항원을 이용한 개 옴 감염증에 대한 면역효과)

  • Yoon, In-Soo;Kim, Jae-Won;Jee, Cha-Ho
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.689-696
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    • 2003
  • Canine sarcoptic mite (Sarcoptes scabiei var. canis) burrow usually in the stratum corneum of the skin of dogs and rabbits. Antigens from the burrowing mites induce cutaneous inflammatory reaction and humoral and cell-mediated immune response in the host. The effect of immunization induced by somatic antigens of house dust mite (Dermatophagoides spp.) has been evaluated to control the canine sarcoptic mite in this experiment. Twelve common antigens (187, 142, 126, 120, 109, 92, 80, 68, 51, 30, 25, 17 kDa) were found using SDS-PAGE with silver staining and Western blot between canine sarcoptic mite and house dust mite. In order to evaluate the immunologic effect of these common antigens 10 New Zealand white rabbits were divided as 4 groups such as negative control (group I), positive challenged control (group II), vaccinated (group III), and vaccinated-challenged (group IV) groups. Group II was artificially infested with about 1,000 canine sarcoptic mites and group III and IV were immunized with somatic antigens of house dust mite. In addition group IV was artificially infested with about 1,000 canine sarcoptic mites and group II, IV were treated with ivermectin. At the 8 weeks of the vaccination with common antigen, the antibody titers of all groups of II, III and IV had been increased. Both infestation score and live canine sarcoptic mite counts of group IV were lower than group III. Infestation score of group II become 0 by 2 weeks and group IV by 4 weeks after infestation. These results suggest that house dust mite, which is easy to culture in vitro, can be a vaccine candidate for protection of canine sarcoptic mite infestation.

Prognostic Significance of Two Dimensional AgNOR Evaluation in Local Advanced Rectal Cancer Treated with Chemoradiotherapy

  • Gundog, Mete;Yildiz, Oguz G;Imamoglu, Nalan;Aslan, Dicle;Aytekin, Aynur;Soyuer, Isin;Soyuer, Serdar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8155-8161
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    • 2016
  • The prognostic significance of AgNOR proteins in stage II-III rectal cancers treated with chemoradiotherapy was evaluated. Silver staining was applied to the $3{\mu}m$ sections of parafin blocked tissues from 30 rectal cancer patients who received 5-FU based chemoradiotherapy from May 2003 to June 2006. The microscopic displays of the cells were transferred into the computer via a video camera. AgNOR area (nucleolus organizer region area) and nucleus area values were determined as a nucleolus organizer regions area/total nucleus area (NORa/TNa). The mean NORa/TNa value was found to be $9.02{\pm}3.68$. The overall survival and disease free survival in the high NORa/TNa (>9.02) patients were 52.2 months and 39.4 months respectively, as compared to 100.7 months and 98.4 months in the low NORa/TNa (<9.02) cases. (p<0.001 and p<0.001 respectively). In addition, the prognosis in the high NORa/TNa patients was worse than low NORa/TNa patients (p<0.05). In terms of overall survival and disease-free survival, a statistically significant negative correlation was found with the value of NORa/TNa in the correlations tests. Cox regression analyses demostrated that overall survival and disease-free survival were associated with lymph node status (negative or positive) and the NORa/TNa value. We suggest that two-dimensional AgNOR evaluation may be a safe and usable parameter for prognosis and an indicator of cell proliferation instead of AgNOR dots.

Pneumocystis jiroveci Pneumonia Mimicking Miliary Tuberculosis in a Kidney Transplanted Patient

  • Jung, Ju Young;Rhee, Kyoung Hoon;Koo, Dong Hoe;Park, I-Nae;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.2
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    • pp.127-130
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    • 2009
  • Bilateral interstitial infiltration in chest radiography, which may be fine granular, reticular or of ground glass opacity, is the typical radiographic findings of Pneumocystis jiroveci pneumonia. Recently, atypical radiographic features, including cystic lung disease, spontaneous pneumothorax or nodular opacity, have been reported intermittently in patients with P. jiroveci pneumonia. We report the case of a 29-year-old woman with a transplanted kidney whose simple chest radiography and HRCT scan showed numerous miliary nodules in both lungs, mimicking miliary tuberculosis (TB). Under the presumptive diagnosis of miliary TB, empirical anti-TB medication was started. However, Grocott methenamine silver nitrate staining of a transbronchial lung biopsy tissue revealed P. jiroveci infection without evidence of TB. These findings suggest that even in TB-endemic area other etiology such as P. jiroveci as well as M. tuberculosis should be considered as an etiology of miliary lung nodules in mmunocompromised patients.

Characterization of Hemolytic Aeromonas sp. MH-8 Responding to the Exposure of Green Tea Catechin, EGCG (녹차 카테킨 EGCG의 노출에 따른 식중독 세균인 용혈성 Aeromonas sp. MH-8의 특성조사)

  • Kim, Dong-Min;Oh, Kye-Heon
    • KSBB Journal
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    • v.31 no.4
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    • pp.228-236
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    • 2016
  • The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.