• Journal of Veterinary Science
• /
• v.24 no.1
• /
• pp.11.1-11.14
• /
• 2023

Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ⁺CD3⁺ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ⁺ CD4^-CD8⁺ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

• Journal of Korea Entertainment Industry Association
• /
• v.13 no.1
• /
• pp.217-223
• /
• 2019

Colon cancer is the most common form of cancer diagnosis in worldwide. There are growing interests in the health benefits associated with consumption of fruits and vegetables, especially for the prevention of cancer, cardiovascular or other chronic diseases. The objective of the present study was to investigate the antioxidant and anticancer activities of natural product, guarana(GR) and graviola(GV) in human colon carcinoma HCT-116 cells. MTT assay, flow cytometry analysis were employed to investigate the anticancer mechanism and DPPH assay was determined to the antioxidant activity to scavenge free radicals in extract of these. All two extracts showed significantly antioxidant activity at 50mg/ml of concentration. GR and GV reduced HCT-116 cell proliferation in a dose dependent manner. Specially GR treatment(96.65±3.71) also significantly increased the sub-G1 population more than GV(79.58±2.87) treatment in HCT-116 at the concentration of 10mg/ml, as shown by flow cytometry assay. Statistical analyses revealed GR and GV exhibited significantly high (P < 0.05) cytotoxicity in HCT-116. These findings indicate that GN and GV may serve as novel therapeutic agents for colon cancer treatment and future leads for drug development.

• Korean Journal of Radiology
• /
• v.21 no.8
• /
• pp.967-977
• /
• 2020

Objective: To evaluate the effects of tube voltage on image quality in coronary CT angiography (CCTA), the estimated radiation dose, and DNA double-strand breaks (DSBs) in peripheral blood lymphocytes to optimize the use of CCTA in the era of low radiation doses. Materials and Methods: This study included 240 patients who were divided into 2 groups according to the DNA DSB analysis methods, i.e., immunofluorescence microscopy and flow cytometry. Each group was subdivided into 4 subgroups: those receiving CCTA only with different tube voltages of 120, 100, 80, or 70 kVp. Objective and subjective image quality was evaluated by analysis of variance. Radiation dosages were also recorded and compared. Results: There was no significant difference in demographic characteristics between the 2 groups and 4 subgroups in each group (all p > 0.05). As tube voltage decreased, both image quality and radiation dose decreased gradually and significantly. After CCTA, γ-H2AX foci and mean fluorescence intensity in the 120-, 100-, 80-, and 70-kVp groups increased by 0.14, 0.09, 0.07, and 0.06 foci per cell and 21.26, 9.13, 8.10, and 7.13 (all p < 0.05), respectively. The increase in the DNA DSB level in the 120-kVp group was higher than those in the other 3 groups (all p < 0.05), while there was no significant difference in the DSBs levels among these latter groups (all p > 0.05). Conclusion: The 100-kVp tube voltage may be optimal for CCTA when weighing DNA DSBs against the estimated radiation dose and image quality, with further reductions in tube voltage being unnecessary for CCTA.

• Journal of Korean Society on Water Environment
• /
• v.40 no.4
• /
• pp.161-167
• /
• 2024

The demand for bottled water in South Korea is steadily increasing, but there are challenges regarding water sources and violations of water quality standards. Consumers struggle to identify products that do not meet these standards, highlighting the need for improved water management. This study aims to investigate the use of flow cytometry to identify microbial behavior in bottled water. Twelve different bottled water brands were selected for this study. A novel non-culture-based analysis method called total cell count via flow cytometry was utilized, which is not commonly used to assess drinking water quality. This method was compared to conventional culture-based methods for heterotrophic plate count and E. coli experiments, in order to introduce new indicators for hygiene management. Adenosine triphosphate analysis was also conducted to assess cell activity, and total organic carbon was measured to determine the presence of organic matter. The total cell counts varied among the different bottled water brands. The adenosine triphosphate levels ranged from 37.1ng/L to 221.7ng/L, while the total organic carbon ranged from 0.4 to 0.6 mg/L. Furthermore, E. coli was not detected in any of the bottled waters, and with the exception of two cases, the levels of heterotrophic bacteria did not exceed the drinking water standard of 100 CFU/mL. This study demonstrated a correlation between total cell count and heterotrophic plate count, suggesting that non-culture-based analysis could be valuable in promptly assessing microbial contamination, in contrast to the conventional methods that require approximately 48 hours for incubation.

• Journal of Plant Biotechnology
• /
• v.2 no.2
• /
• pp.109-113
• /
• 2000

Somatic hybrids were produced by electrofusion between embryogenic callus protoplasts of 'Syougun' mandarin and leaf protoplasts of grapefruit. Hybridity of the two plants was confirmed by leaf morphological characteristics and random amplified polymorphic DNA (RAPD) analysis. The cpDNA analysis using PCR-RFLP could not distinguish those of both parents. These plants showed normal growth and had chromosome number of 27. These unexpected triploid somatic hybrids might be derived from fused cells between diaploid protoplast of embryogenic calli and diploid protoplast of leaf, because polysomaty, a mixture of haploid cells and diploid cells was observed in the lactose medium-pretreated embryogenic calli of 'Syougun' by flow cytomehy analysis.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.1
• /
• pp.52-55
• /
• 2002

Flow cytometric techniques were used to investigate cell size, protein content and cell cycle behavior of recombinant Saccharomyces cerevisiae strains producing human lysozyme (HLZ). Two different signal sequences, the native yeast $MF\alpha1$ signal sequence and the rat $\alpha-amylase$ signal sequence, were used for secretion of HLZ. The strain containing the rat $\alpha-amylase$ signal sequence showed a higher level of internal lysozyme and lower specific growth rates. Flow cytometric analysis of the total protein content and cell size showed the strain harboring the native yeast signal sequence had a higher total protein content than the strain containing the rat $\alpha-amylase$ signal sequence. Cell cycle analysis indicated that the two lysozyme producing recombinant strains had an increased number of cells in the $G_2+M$ phase of the yeast cell cycle compared with the host strain SEY2102.

• Reproductive and Developmental Biology
• /
• v.29 no.3
• /
• pp.163-168
• /
• 2005

The aim of this study was to investigate the changes in insulin-like growth factor-I (IGF-I) and growth hormone (GH) in serum, the quantitation of spermato-genesis and the comparable relationships among these measurements during pubertal period in New Zealand White male rabbits. To investigate the age-related testicular changes in DNA contents of spermatogenic cells, the fine-needle testicular biopsies from males aged 10 to 28 wks were evaluated by flow cytometry(FCM). Body weight increased significantly between the ages of 12 and 20 wks (P<0.05) and reached 3.4 kg at 28 wks of age. The highest serum IGF-I level (451.3ng/mL) was observed at 20wks of age (P<0.05) and thereafter remained stable at low levels. Serum GH level at 18 wks of age was 183.3 pg/mL which was significantly higher compared to the other ages (P<0.05), and the rising time in serum GH tend to be somewhat earlier than that of IGF-I. The relative percentage of It-cells in testicular cell compartments was $48.2\%$ at the age of 18 wks which significantly increased than those of 16-wk-old (P<0.05) and thereafter increased with the advance of age to $68\%$. The percentage of 2C-cells in testis was $26.8\%$ at 18 wks of age which was significantly lower than $54.3\%$ at 16 wks old (P<0.05). The percentage of 4C-cells was constantly maintained $2\~6\%$ except the $9.9\%$ at 18 wks of age. In conclusion, the results suggest that the puberty onset occurred at about the 18 wks of age and that the IGF-I and GH in serum during the pubertal period showed the age/growth-specific changes and these changes might be related to the spermatogenesis. The DNA FCM combined with fine-needle testicular biopsy could offer a very sensitive method to monitor the quantitative spermatogenic events related to the puberty onset.

• Journal of Korean Society of Forest Science
• /
• v.101 no.2
• /
• pp.291-296
• /
• 2012

Triploids are a useful tool for biomass production and molecular breeding of trees with a long life span. Triploids of the poplar 'Hyunsasi' (Populus alba ${\times}$ P. glandulosa) have been developed by crossing between female diploids and a male tetraploid. The tetraploid was developed around the 1970s at Korea Forest Research Institute by colchicine-induced chromosome doubling. Seedlings of the $F_1$ generation were analyzed using flow cytometry to verify their ploidy status. The mean relative fluorescence index of 3 F1 poplars, labeled as Line- 1, Line-17, Line-18, were approximately 1.5 times higher than those of diploid poplars, and the results clearly indicated that they were triploids. The phenotype of the F1 poplars included larger leaves and thicker stem than diploids, and abnormal leaf morphology, especially in the triploid 'Line-18'. Three triploid lines developed roots more slowly and had less roots than diploid. However, 3 poplar cytotypes (2x, Line-1, Line-17) rooted within 10 days on MS medium. In contrast, compared with the 3 cytotypes, the Line-18 showed about 80% and 70% in the rooting rate and the number of roots. The triploid poplars could be directly utilized for biomass production and with their sterility, they could serve as basic material for genetic transformation. In addition, flow cytometric analysis proved to be an effective and reliable method for screening forest trees for their ploidy level.

• Korean Journal of Clinical Laboratory Science
• /
• v.49 no.4
• /
• pp.382-389
• /
• 2017

Currently, reticulocyte experimental calculation technology used in clinical laboratories are divided two types: manual and automated. Manual reticulocyte counting using a microscopy lacks accuracy due particularly to its low reproducibility, affecting the accuracy of manual reticulocyte count. Moreover, Automatic blood corpuscle analyzer flow cytometry is difficult to be used in underdeveloped countries and small scale laboratories due to relatively high cost. Therefore, this study tried to find a new method to complement these drawbacks. The aim of this study was to compare the stained reticulocytes count by spectrophotometer and also to analyze the statistics of spectrophotometer and flow cytometer. The same 8 EDTA samples were repeated 36 times to compare the agreement between spectrophotometer and flow cytometer. This study measured the specimen diluted 600 times at 700~780 nm by 10 differences. Wavelength between 710 to 730 by absorbance showed a positive correlation between standard data and test data (r=0.967, p<0.01), presenting a correlation between variables. Statistical analyses of regression for test and standard parametric data, the optimal dilution factor was 600 times. Therefore, this study tried to technical utilizes such as contributing economical for the reticulocyte absorbance apply from the auto spectrophotometer, a monitoring system for the reticulocyte relation anemia, etc. Therefore, more extensive studies, including an auto chemical analyzer application, will be needed.

• Korean Journal of Clinical Laboratory Science
• /
• v.54 no.1
• /
• pp.68-72
• /
• 2022

Angioimmunoblastic T-cell lymphoma (AITL) is a lymphoproliferative disorder of mature T follicular helper cells. Atypical lymphoid cells were observed in the bone marrow of an 80-year-old woman, and the flow cytometric determined immunophenotypes of B-cells were unusual, that is, CD19+, CD20-, and CD22- with lambda light chain restriction. Initially, we suspected BM involvement of B-cell lymphoma based on the presence of abnormal B-cells. However, the patient was diagnosed with AITL involving BM. A re-analysis of flow cytometric immunophenotyping revealed a minor, aberrant T-cell population, and the lambda light chain restriction observed by surface staining was considered non-specific binding. This case demonstrates B-cells in patients with EBV-positive T-cell lymphoma may exhibit immunophenotypes resembling those of plasma cells, and that proliferation of abnormal B-cells or plasma cells could also potentially mask underlying T-cell lymphoma. A more integrated approach is required for accurate diagnosis.