• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1363-1367
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• 2014

This research was conducted to compare differences in colon cancer lymphatic vessel invasion (LVI) with D2-40 antibody labeling and regular HE staining, blood vessel invasion (BVI) with CD34 antibody labeling and HE staining and to assess the possibility of using D2-40-LVI/CD34-BVI in combination for predicting stage II colon cancer prognosis and guiding adjuvant chemotherapy.Anti-D2-40 and anti-CD34 antibodies were applied to tissue samples of 220 cases of stage II colon cancer to label lymphatic vessels and small blood vessels, respectively. LVI and BVI were assessed and multivariate COX regression analysis was performed for associations with colon cancer prognosis. Regular HE staining proved unable to differentiate lymphatic vessels from blood vessels, while D2-40 selectively labeled lymphatic endothelial cell cytosol and CD34 was widely expressed in large and small blood vessels of tumors as well as normal tissues. Compared to regular HE staining, D2-40-labeling for LVI and CD34-labeling for BVI significantly increased positive rate (22.3% vs 10.0% for LVI, and 19.1% vs 9.1% for BVI). Multivariate analysis indicated that TNM stage, pathology tissue type, post-surgery adjuvant chemotherapy, D2-40-LVI, and CD34-BVI were independent factors affecting whole group colon cancer prognosis, while HE staining-BVI, HE staining-LVI were not significantly related. When CD34-BVI/D2-40-LVI were used in combination for detection, the risk of death for patients with two or one positive results was 5.003 times that in the LVI(-)&BVI(-) group (95% CI 2.365 - 9.679). D2-40 antibody LVI labeling and CD34 antibody BVI labeling have higher specificity and accuracy than regular HE staining and can be used as molecular biological indicators for prognosis prediction and guidance of adjuvant chemotherapy for stage II colon cancer.

• International Journal of Oral Biology
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• 제40권4호
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• 대한미생물학회지
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• 제3권1호
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• pp.51-65
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• 1968

The site of the synthesis of Japanese encephalitis virus(JEV) in the actinomycin-treated and infecter PS Y15 cells(a porcine kidney stable cell line) was observed by the immunofluorescent antibody technique, acridine orange staining, and the autoradiographic analysis. In the parallel studies by immunofluorescent technique and acridine orange staining it the infected cells, Viral protein(as an antigen) and viral RNA were detected at the same site of cytoplasm. In the autoradiographic analysis, the cytoplasmic labeling of $^3H$-uridine was due to the synthesis of JEV-RNA, while the nucleolus and nucleus were not involved. In the autoradiographic studies on the secton of infected cells, the $^3H$-uridine was frequently incorporated around the cytoplasmic vacuoles. This localization of labeling agreed with the site of acridine orange positive granules. The results suggest that the syntheses of the viral RNA and viral protein occurred in the similar site of cytoplasm of the infected cells, and also the virus particles seem to be assembled in the sites of the viral RNA and protein syntheses.

• Journal of Korean Neurosurgical Society
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• 제39권6호
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• pp.432-437
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• 2006

Objective : In order to establish the index of degeneration, the authors performed a histochemical study with Safranin-O staining and investigated the occurrence of apoptosis in the human intervertebral disc. Methods : Eighteen intervertebral disc specimens surgically extracted from the patients and two additional specimens from the autopsied cases were stained with Safranin-O for proteoglycan according to a standard protocol. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate- biotin nick end labeling[TUNEL] was used to detect the fragmented DNA known to be associated with apoptotic cell death and classification scheme was formulated for categorization of the degree of Safranin-O staining [normal, moderate reduction, faint] by modification of Makin's histological-histochemical grading. The Kruskal-Wallis H test and Chi-square test were used for statistical analysis. Results : The statistical results showed a significant difference in the mean age between "normal" Safranin-O staining group and the others [19.3 versus 55, 43.4, p=0021]. However, there was no statistically significant correlation between Safranin-O staining and MR grading of disc degeneration. Only six of eighteen surgical specimens and none in autopsies showed positive apoptotic cells in TUNEL staining. Conclusion : The determination of the degree of degeneration in surgically obtained disc tissue per se by histochemical staining or by the degree of apoptosis that corresponds to its morphologic change was not feasible.

• Animal cells and systems
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• 제1권1호
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• pp.93-98
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• 1997

The present study investigated the cellular localization of a-subunit of Gq (Gaq) protein in developing L8E63, rat skeletal muscle cell line. The colocalization of Gaq with actin cytoskeleton was demonstrated by double-labeling experiments. In mononucleated myoblasts, the immuno-fluorescence staining pattern of Gaq was almost identical with that of F-actin visualized with rhodamine-conjugated phalloidin. However, this colocalization of Gaq with cytoskeleton was not maintained in multinucleated myotubes. The staining pattern of Gaq in myotubes did not match with any specific subcellular structure, but appeared as a uniformly distributed diffuse staining throughout the whole cell surface. Interestingly, change in the expression level of Gaq was not detected during myoblast differentiation, suggesting that actin-associated Gaq protein might dissociate from the cytoskeleton as cells differentiate. Immunocytochemical experiments using specific antibodies directed against several G proteins indicated that the subcellular localizations of Gai1, Gai2, Gai3, and Gao were different from those obtained with Gaq.

• 대한의생명과학회지
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• 제12권2호
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• pp.105-112
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• 2006

This study was undertaken to investigate the morphological changes between normal and atretic follicle after gamma irradiation and treatment of follicle stimulating hormone (FSH). The ovaries of each group of treated immature mice were prepared the paraffin sections after 0, 6, 12, and 24 hours (hrs) of those treatment. Hematoxylin-eosin (HE) stain, reticulin stain, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) immunohistochemical stain were performed on the each paraffin sections. As the results of HE staining, the condensed nuclei of oocytes were observed in the atretic primordial follicles, on the other hand the condensations of granulosa cell nuclei were prominent in the atretic primary, preantral, and antral follicles. Only the granulosa cells of atretic follicle were stained specifically with TUNEL staining but not stained in the theca cells, which suggested granulosa cells degenerated through apoptosis. In the reticulin staining, the basement membranes of atretic follicle which was stained weakly showed irregular structure and detachment from the follicles. The ratio of normal to atretic follicle in control and FSH treated group was about 33% but this ratio increased rapidly over 90% in the 6, 12, and 24 hrs group after the irradiation. It could be suggested that the gamma irradiation is the useful tool far the induction of follicle atresia and immunohistochemical staining methods are essential in the study of follicle atresia.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 2010년도 하계학술대회 논문집
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• pp.280-280
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• 2010

Alzheimer disease is a progressive neurodegenerative disease of the aged, characterized by memory loss and dementia. For diagnosis of Alzheimer disease we have simply modified macrophage with amyloid beta bonded with different molecules. Modified Macrophage was observed with microscope for co-localization of amyloid beta molecule. For this experiment we used fluoroscene labeling substances. The macrophage was modified also with cell staining method. For cell staining method was used avidin-biotin reaction principles. All experiments were carried out on poly-L-lysine coated and sterilized glass substrates. In the presentation we will show the further investigations and applications with modified macrophage.

• Journal of Oral Medicine and Pain
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• 제43권4호
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• pp.99-111
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• 2018

$Sj{\ddot{o}}gren^{\prime}s$ syndrome (SjS) is a common autoimmune disorder characterized by lymphocytic infiltration in the salivary and lacrimal glands, resulting in severe dry mouth or eyes. As a result, most of SjS patients suffer from oral dryness and can visit the department of oral medicine with or without diagnosis of SjS. Therefore, oral medicine specialists should know clues, which may indicate the diagnosis of SjS from the clinical and laboratory investigations. By the recent SjS criteria, SjS can be diagnosed by focus score, ocular staining, Schirmer's test, unstimulated whole saliva flow rate, and anti-SSA/Ro antibodies. The aim of this article is to review the diagnostic criteria, clinical investigation, and basic aspect related to SjS and to make oral medicine specialists play an important role in the detection of emerging SjS.

• Journal of Animal Science and Technology
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• 제63권1호
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• pp.58-68
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• 2021

Several herbs including Artemisia are known to possess conceptive property. In the present study, mouse spermatozoa were incubated with ethanol extract of Artemisia vulgaris leaves. The effect of extract on acrosome exocytosis was studied by labeling spermatozoa with fluorescein isothiocyanate (FITC) peanut agglutinin and by staining with Coomassie blue. Viability and membrane integrity were studied by Trypan-blue staining and hypo-osmotic swelling test. Artemisia extract at very low concentration caused precocious acrosome reaction and loss of sperm viability. Acrosome reaction increased remarkably from 22.63% to 88.42% with increasing extract concentration from 0 to 2,000 ㎍/mL. However, the viability loss of spermatozoa was increased from 11.71% in control to 63.73% in samples treated, evaluated by Trypan-blue staining method. Membrane damage caused by the extract, evaluated by hypo-osmotic swelling test was even low, ranging from 2.27% to only 24.23%. These results indicate that Artemisia extract might block fertilization by causing precocious acrosome exocytosis in spermatozoa. A direct contraceptive effect was tested by injecting the plant extract into the vagina of female mice and then allowing them to mate with normal males. The treated female mice delivered significantly fewer litters in comparison to the control.

• Animal Bioscience
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• 제35권1호
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• pp.96-104
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• 2022

Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P_2b, P₂₁, and P₂₂) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P₂₁) and positive association with free water (P₂₂). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.