• Title/Summary/Keyword: tissue samples

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Necrotrophic Fungus Pyrenophora tritici-repentis Triggers Expression of Multiple Resistance Components in Resistant and Susceptible Wheat Cultivars

  • Andersen, Ethan J.;Nepal, Madhav P.;Ali, Shaukat
    • The Plant Pathology Journal
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    • v.37 no.2
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    • pp.99-114
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    • 2021
  • Tan spot of wheat, caused by Pyrenophora tritici-repentis (Ptr), results in a yield loss through chlorosis and necrosis of healthy leaf tissue. The major objective of this study was to compare gene expression in resistant and susceptible wheat cultivars after infection with Ptr ToxA-producing race 2 and direct infiltration with Ptr ToxA proteins. Greenhouse experiments included exposure of the wheat cultivars to pathogen inoculum or direct infiltration of leaf tissue with Ptr-ToxA protein isolate. Samples from the experiments were subjected to RNA sequencing. Results showed that ToxA RNA sequences were first detected in samples collected eight hours after treatments indicating that upon Ptr contact with wheat tissue, Ptr started expressing ToxA. The resistant wheat cultivar, in response to Ptr inoculum, expressed genes associated with plant resistance responses that were not expressed in the susceptible cultivar; genes of interest included five chitinases, eight transporters, five pathogen-detecting receptors, and multiple classes of signaling factors. Resistant and susceptible wheat cultivars therefore differed in their response in the expression of genes that encode chitinases, transporters, wall-associated kinases, permeases, and wound-induced proteins, among others. Plants exposed to Ptr inoculum expressed transcription factors, kinases, receptors, and peroxidases, which are not expressed as highly in the control samples or samples infiltrated with ToxA. Several of the differentially expressed genes between cultivars were found in the Ptr resistance QTLs on chromosomes 1A, 2D, 3B, and 5A. Future studies should elucidate the specific roles these genes play in the wheat response to Ptr.

Multiresidue matrix solid phase dispersion (MSPD) extraction and HPLC determination of tetracyclines in animal muscle tissue (시료고체상분산(matrix solid phase dispersion)전처리법을 이용한 식육중 테트라사이클린계 항생물질 동시정량분석)

  • Kang, Hwan-goo;Son, Seong-wan;Cho, Byung-hoon;Lee, Hye-sook;Park, Shin-ja;Kim, Jae-hak;Cho, Myung-haing
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.541-550
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    • 1996
  • Tetracycline antibiotics have been widely used not only therapeutics but feed additives. There are many methods for the isolation and determination of tetracycline antibiotics in animal muscle tissue. But those methods take much time and labor, so it is difficult to analyse many samples simultaneously. A rapid isolation method and liquid chromatographic determination of tetracycline antibiotics in animal muscle tissue (bovine, porcine, chicken) is presented. Blank control and tetracyclines fortified samples (0.5g) were blended with $C_{18}$ containing 0.05g each of oxalic acid and disodium ethylenediaminetetraacetate. After homogenize, homogenate was transferred to glass column made from 10ml glass syringe and compressed to 4~4.5ml volume. A column made from the $C_{18}$/meat matrix was washed with hexane (8ml) and dichloromethane (8ml, if needed), following which the tetracyclines were eluted,vith methanol or 0.01M methanolic oxalic acid (8ml). The eluates containing tetracyclines analytes were free from interfering compounds when analysed by HPLC with UV detection (photodiode array at 360nm). Standard curve for each tetracycline showed a linear response at the range of $0.05{\sim}1.0{\mu}g/ml$ and tetracycline antibiotics were eluted within 4ml of eluted volume. All tetracycline antibiotics except tetracycline were stable during the concentration process at $40^{\circ}C$ and time required for concentration was 3~4 hours. Fortified samples containing oxalic aicd and EDTA represented more good recoveries than those of not-contained sample. Recoveries were 91.8~110.1% (oxytetracycline; OTC), 57.7~79.5% (tetracycline; TC), 78.1~88.6% (chlortetracyclines; CTC) and 88.4~100.6% (doxycycline; DC) in pork tissue, 101.1~126.8% (OTC), 66.4~75.4% (TC), 79.2~88.1% (CTC) and 69.3~86.7% (DC) in beef tissue, and 90.8~95.6% (OTC), 66.2~84.4% (TC), 75.7~77.2% (CTC) and 55.6~80.7% (DC) in chicken muscle tissue. The detection limits validated in muscle tissue by this method were $0.05{\mu}g/g$ for OTC and TC, and $0.1{\mu}g/g$ for CTC and DC.

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Difference of Microbiology according to Tissue Sampling in Diabetic Ulcers (만성 당뇨발에서 표재조직 및 심부조직 세균배양검사의 비교)

  • Rhee, Sung-Mi;Han, Seung-Kyu;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.37 no.1
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    • pp.1-6
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    • 2010
  • Purpose: Diabetic foot infection is one of the most common and severe complications of diabetes mellitus that delays healing of the wound. Deep tissue biopsy is considered to be the gold standard method for antibiotic selection in treating infected chronic diabetic ulcers. However, swab culture or superficial tissue biopsy is often performed for a microbiologic test since deep tissue biopsy has limitations in application. The purpose of this study is to find out whether microbiologic results of swab culture or superficial tissue biopsy could be used for selection of antibiotics in treating diabetic ulcers. Methods: This study involved 42 patients with diabetic foot ulcers with negative results in bone probing test. Tissue samples for microbiologic tests were collected from all the patients by using superficial cotton swab, superficial tissue, and deep tissue. The microbiologic results of deep tissue biopsy were compared with swab culture and superficial tissue biopsy statistically. Results: Microbiology of the deep tissue showed the same results with those of the swab culture and superficial tissue in 67% and 71%, respectively. Statistical analysis demonstrated that the microbiology of the swab culture and superficial tissue does not coincide with that of the deep tissue. Conclusion: These results suggest that the microbiology of the swab culture and superficial tissue is not concordant with that of the deep tissue in infected chronic diabetic ulcers. To select appropriate antibiotic regimen, the speci specimen for the microbiologic test should be obtained from deep tissue.

Endophilin A2: A Potential Link to Adiposity and Beyond

  • Alfadda, Assim A.;Sallam, Reem M.;Gul, Rukhsana;Hwang, Injae;Ka, Sojeong
    • Molecules and Cells
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    • v.40 no.11
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    • pp.855-863
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    • 2017
  • Adipose tissue plays a central role in regulating dynamic cross-talk between tissues and organs. A detailed description of molecules that are differentially expressed upon changes in adipose tissue mass is expected to increase our understanding of the molecular mechanisms that underlie obesity and related metabolic co-morbidities. Our previous studies suggest a possible link between endophilins (SH3Grb2 proteins) and changes in body weight. To explore this further, we sought to assess the distribution of endophilin A2 (EA2) in human adipose tissue and experimental animals. Human paired adipose tissue samples (subcutaneous and visceral) were collected from subjects undergoing elective abdominal surgery and abdominal liposuction. We observed elevated EA2 gene expression in the subcutaneous compared to that in the visceral human adipose tissue. EA2 gene expression negatively correlated with adiponectin and chemerin in visceral adipose tissue, and positively correlated with $TNF-{\alpha}$ in subcutaneous adipose tissue. EA2 gene expression was significantly downregulated during differentiation of preadipocytes in vitro. In conclusion, this study provides a description of EA2 distribution and emphasizes a need to study the roles of this protein during the progression of obesity.

Elevated Aurora Kinase A Protein Expression in Diabetic Skin Tissue

  • Cho, Moon Kyun;An, Je Min;Kim, Chul Han;Kang, Sang Gue
    • Archives of Plastic Surgery
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    • v.41 no.1
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    • pp.35-39
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    • 2014
  • Background Aurora kinase A (Aurora-A) plays an important role in the regulation of mitosis and cytokinesis. Dysregulated Aurora-A leads to mitotic faults and results in pathological conditions. No studies on Aurora-A expression in human diabetic skin tissue have been reported. In light of this, we explored the expression of Aurora-A in human diabetic skin tissue. Methods Aurora-A protein was evaluated by western blotting in 6 human diabetic skin tissue and 6 normal skin specimens. Results Increased expression of Aurora-A protein was detected in all diabetic skin tissue samples in both western blot analysis and immunohistochemical staining. However, in the case of the normal skin tissue, no bands of Aurora-A protein were detected in either the western blotting analysis or the immunohistochemical staining. Conclusions Thus far, there have been no studies on the expression of Aurora-A in diabetic skin tissue. However, we believe that oxidative DNA damage related to the expression of Aurora-A protein and Aurora-A could be involved inhuman diabetic skin tissue.

Tissue Level Based Deep Learning Framework for Early Detection of Dysplasia in Oral Squamous Epithelium

  • Gupta, Rachit Kumar;Kaur, Mandeep;Manhas, Jatinder
    • Journal of Multimedia Information System
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    • v.6 no.2
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    • pp.81-86
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    • 2019
  • Deep learning is emerging as one of the best tool in processing data related to medical imaging. In our research work, we have proposed a deep learning based framework CNN (Convolutional Neural Network) for the classification of dysplastic tissue images. The CNN has classified the given images into 4 different classes namely normal tissue, mild dysplastic tissue, moderate dysplastic tissue and severe dysplastic tissue. The dataset under taken for the study consists of 672 tissue images of epithelial squamous layer of oral cavity captured out of the biopsy samples of 52 patients. After applying the data pre-processing and augmentation on the given dataset, 2688 images were created. Further, these 2688 images were classified into 4 categories with the help of expert Oral Pathologist. The classified data was supplied to the convolutional neural network for training and testing of the proposed framework. It has been observed that training data shows 91.65% accuracy whereas the testing data achieves 89.3% accuracy. The results produced by our proposed framework are also tested and validated by comparing the manual results produced by the medical experts working in this area.

Tritium Distribution in Some Environmental Samples-Rices, Chinese Cabbages and Pine Needles in Korea (국내 환경시료(쌀, 채소, 솔잎) 중 삼중수소의 분포)

  • Kim, Chang-Kyu;Cho, Yong-Woo;Han, Man-Jung;Pak, Chan-Kirl
    • Journal of Radiation Protection and Research
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    • v.17 no.2
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    • pp.25-35
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    • 1992
  • To evaluate tritium level in some environmental samples, tissue-free water tritium (TFWT) and tissue-bound tritium (TBT) were analyzed in rices, chinese cabbages and pine needles collected at 12 locations in Korea. The TFWT was recovered by freeze-drying of the samples and the TBT was obtained in the form of water by combustion of the dried samples. Tritium was measured by liquid scintillation counter. The concentrations of TFWT were in the range of $0.96{\sim}3.96 Bq/1,\; 0.83{\sim}3.40 Bq/1\;and\;1.02{\sim}3.01 Bq/1$ in rices, chinese cabbages and pine needles, respectively. The mean specific activity ratios (TBT/TFWT) were 0.94, 1.71 and 1.39 in rices, chinese cabbages and pine needles, respectiviely. This excess TBT in the samples may be attributed to the fact that the residence time of TBT in the plant is longer than that of TFWT. The specific activity ratio depends on the plant species, the exposed time to tritiated atmosphere, atmospheric moisture, temperature and diffusion factor.

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Application of MALDI Tissue Imaging of Drugs and Metabolites: A New Frontier for Molecular Histology

  • Shanta, Selina Rahman;Kim, Young-Jun;Kim, Young-Hwan;Kim, Kwang-Pyo
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.149-154
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    • 2011
  • Matrix assisted laser desorption ionization (MALDI) mass spectrometry is commonly used to analyze biological molecules such as proteins, peptides and lipids from cells or tissue. Recently MALDI Imaging mass spectrometry (IMS) has been widely applied for the identification of different drugs and their metabolites in tissue. This special feature has made MALDI-MS a common choice for investigation of the molecular histology of pathological samples as well as an important alternative to other conventional imaging methods. The basic advantages of MALDI-IMS are its simple technique, rapid acquisition, increased sensitivity and most prominently, its capacity for direct tissue analysis without prior sample preparation. Moreover, with ms/ms analysis, it is possible to acquire structural information of known or unknown analytes directly from tissue sections. In recent years, MALDI-IMS has made enormous advances in the pathological field. Indeed, it is now possible to identify various changes in biological components due to disease states directly on tissue as well as to analyze the effect of treated drugs. In this review, we focus on the advantages of MALDI tissue imaging over traditional methods and highlight some motivating findings that are significant in pathological studies.

Is optimal cutting temperature compound essential embedding solution treatment to cryo-sectioning of brain tissue?

  • Baek, Hye Kyung;Song, Ji Ae;Yi, Sun Shin
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.85-89
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    • 2016
  • We tested a set of conditions for obtaining optimal tissue quality in preparation for histology in samples of mouse brain. C57BL/6J mice were sacrificed and perfused with 4% paraformaldehyde, after which the brains were removed and dehydrated in 30% sucrose solution. The brains were then divided into four groups according to freezing temperature and usage of optimal cutting temperature (OCT) compound. Next, we stained the sectioned brain tissues with Harris hematoxylin and eosin Y and immunohistochemistry was performed for doublecortin. The best quality tissue was obtained at $-25^{\circ}C$ and by not embedding with the OCT compound. When frozen at $-25^{\circ}C$, the embedded tissue was significantly damaged by crystals, while at $-80^{\circ}C$ there were no meaningful differences between qualities of embedded- and non-embedded tissues. Overall, we identified a set of conditions to obtain quality frozen brain sections. Our developed protocol will help resolve matters associated with damage caused to sectioned brain tissue by crystal formation during freezing.

Cross-Linked Collagen Scaffold from Fish Skin as an Ideal Biopolymer for Tissue Engineering

  • Biazar, Esmaeil;Kamalvand, Mahshad;Keshel, Saeed Heidari;Pourjabbar, Bahareh;Rezaei-Tavirani, Mustafa
    • Korean Journal of Materials Research
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    • v.32 no.4
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    • pp.186-192
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    • 2022
  • Collagen is one of the most widely used biological materials in medical design. Collagen extracted from marine organisms can be a good biomaterial for tissue engineering applications due to its suitable properties. In this study, collagen is extracted from fish skin of Ctenopharyngodon Idella; then, the freeze drying method is used to design a porous scaffold. The scaffolds are modified with the chemical crosslinker N-(3-Dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) to improve some of the overall properties. The extracted collagen samples are evaluated by various analyzes including cytotoxicity test, SDS-PAGE, FTIR, DSC, SEM, biodegradability and cell culture. The results of the SDS-PAGE study demonstrate well the protein patterns of the extracted collagen. The results show that cross-linking of collagen scaffold increases denaturation temperature and degradation time. The results of cytotoxicity show that the modified scaffolds have no toxicity. The cell adhesion study also shows that epithelial cells adhere well to the scaffold. Therefore, this method of chemical modification of collagen scaffold can improve the physical and biological properties. Overall, the modified collagen scaffold can be a promising candidate for tissue engineering applications.