Cytokines are protein mediators that possess the ability to assist cell-to-cell communication in immune system-related activities. In general, pathogen endotoxins activate the release of inflammatory mediators, and with time, there is an increase in the cytokine levels in the body. Interleukin (IL)-6 mediates the acute-phase inflammatory response, and elevated IL-6 levels have been reported in peritoneal fluids of women with pelvic inflammation and endometriosis, thereby associating it with oocyte quality and infertility. To overcome subfertility or infertility in humans and animals, the present study was done to examine the effect of recombinant IL-6 on porcine oocytes matured in vitro and subsequently to determine the fertilization rate and embryo development. Porcine oocytes were incubated with varying concentrations of IL-6 (0 - 2 ㎍·mL-1) for 44 h followed by in vitro fertilization and culturing of the oocytes. The oocytes or embryos were fixed with 3.7% paraformaldehyde (PFA) and stained with fluorescence dyes, and the meiotic spindle, chromosome organization, fertilization status and embryo development were subsequently assessed under a fluorescence microscope. We observed induction of an abnormal meiotic spindle alignment in the oocytes incubated with IL-6 compared to the control oocytes incubated without IL-6. Moreover, significantly decreased fertilization rates and embryo development were observed for oocytes incubated with IL-6 (p < 0.05). Thus, an increased IL-6 level during oocyte maturation could be associated with fertilization failure due to an aberrant chromosomal alignment and a disruption of the cortical granules. Taken together, our results indicate that successful assisted reproduction can be achieved by controlling the levels of inflammatory cytokines.
Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.
The physiological and photochemical responses of rice seedling to NaCl stress were investigated through measuring leaf relative water content (RWC), root water uptake and chlorophyll fluorescence. When plants were exposed to increased salinity stress, the visual symptoms of injury were significant at ${\geq}$500 mM NaCl concentration for 4 and 5 day stress periods. The differences in Fv/Fm between control treatment and plants treated with 500 mM and 1,000 mM NaCl were evident after 5 day and 4 day, respectively, whereas in root water uptake its effect was observed at 500 mM and 1,000 mM NaCl at 2 day of salt-stressed periods. Leaf RWC in salt-stressed plants decreased gradually with increasing salinity in exogenous solution and duration of salt stress, and these decrease showed leaf RWC of 58-68% atduration over 2 day stress of 1,000 mM NaCl treatment and 88% at 1 day stress. NaCl stress led to a significant inhibition of the light-induced greening in etiolated rice plants, especially in 4 and 5 day salt-stressed plants, which linearly decreased with NaCl concentration ($R^2$=0.812 and 0.918, respectively). The effects of NaCl stress in rice seedlings indicate that water uptake in root is more sensitive to increasing NaCl concentration and stress duration than Fv /Fm in leaves compared with the same NaCl concentration.
Anatoxin-a is a cyanobacterial neurotoxin with a high toxicity produced by Anabaena, Aphanizomenon and Oscillatoria. Water bloom, formed by Anabaena has been occurring frequently in Lake Yeongchun. It is need to develop a sensitive method for determination of anatoxin-a to control potential hazard in raw water resources. In this study, we developed a highly sensitive analytical method of anatoxin-a using solid phase extraction (SPE) and high performance liquid chromatography (HPLC) with fluorescence detection. Anatoxin-a was converted into a highly fluorescent derivative using 4-fuoro-7-nitro-2,1,3-benzoxadiazole (NBF-F). The method was evaluated in terms of linearity of calibration curve, recovery and repeatability, and the adequate values were obtained. The method detection limit was $0.034\;{\mu}g/g$ and $0.022\;{\mu}g/L$ for algal and water samples, respectively. The concentrations of anatoxin-a were measured in algal and water samples from Lake Andong, Yeongchun and Daechung and ranged from $0.135\;{\mu}g/g$ to $10.979\;{\mu}g/g$ in algal samples and not detected in water samples.
Lee, Ji Young;Kim, Chul Hwan;Park, Jong Hye;Kim, Eun Hea;Wi, Sang Wook;Sung, Yong Joo;Heo, Young Jun;Kim, Yeon Oh;Hwang, In Young
Journal of Korea Technical Association of The Pulp and Paper Industry
/
v.47
no.3
/
pp.34-39
/
2015
Fluorescent whitening agents (FWAs) are widely applied in the papermaking process to improve the CIE whiteness and the ISO brightness of papers. Large amounts of FWAs are used in the paper and paperboard industry, but no standard method exists for quantitative FWA analysis. Therefore, a new method is needed to analyze the presence of FWAs in paper and paperboard to control their additions and applications. In particular, FWA analysis must concentrate on those used for surface treatment of paper because their use in surface applications has increased recently. The purposes of this study were to measure the optical properties of papers surface-treated with oxidized starch containing FWAs and to deduce the regression equations for the quantitative analysis of FWAs used for surface treatments. Surface sizing was carried out to treat the surface of a base paper with T-FWA and H-FWA, and the pickup weight of the surface sized paper was measured to determine the real weight of FWAs transferred onto the paper surface. After surface sizing and preconditioning, the paper was evaluated for CIE whiteness, ISO brightness, fluorescence index, and reflectance at 440 nm using an Elrepho spectrophotometer. The CIE whiteness and the ISO brightness increased linearly as FWA content increased. The fluorescence index and the reflectance showed linear relationships with T-FWA and H-FWA. Linear regression equations were determined for the quantitative analysis of FWAs used for surface treatments.
Seo, Chang-Woo;Lee, Seok-Min;Kang, Sang-Mo;Park, Yeon-Gyeong;Kim, Ah-Yeong;Park, Hyeon-Jin;Kim, Yoonha;Lee, In-Jung
KOREAN JOURNAL OF CROP SCIENCE
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v.62
no.4
/
pp.325-332
/
2017
This research was conducted to evaluate methods of enhancing the waterlogging resistance of soybean plant. Thus, we applied seven types of plant growth regulators (PGRs) to soybean plants and exposed them to waterlogged conditions for a total of 14 days. To evaluate stress resistance, we monitored plant growth characteristics data such as height, chlorophyll content, and chlorophyll fluorescence for 28 days after the initial waterlogging (14 days under waterlogging conditions and 14 days after waterlogging). According to the results, plant height was significantly increased by gibberellin A4 ($GA_4$) treatment compared to the control treatment and waterlogging-only treatment. However, we could not detect plant height owing to plant death when we applied abscisic acid (ABA). Except for $GA_4$ and ABA treatments, plant heights slightly decreased in all treatments compared to the waterlogging-only treatment. The chlorophyll content and chlorophyll fluorescence showed a similar tendency among PGR treatments. The chlorophyll content and chlorophyll fluorescence were significantly increased by ethephon and kinetin treatments 28 days after waterlogging compared to the waterlogging-only treatment. Consequently, kinetin and ethephon treatments induced more resistant phenotypes in soybean plants during or after exposure to waterlogging conditions.
Recently elevated plasma homocysteine concentration is considered an independent risk factor for atherosclerosis and thrombosis with coronary artery disease. Folate and vitamin $B_{12}$ are cofactors and closely related with metabolism of homocysteine. The purpose of this study is to evaluate the correlation between homocysteine and folate and vitamin $B_{12}$ in patients with ischemic heart disease. Twenty-six patients, in whom coronary angiographic finding revealed more than $50\%$ of stenosis at least in one coronary vessel were enrolled as the patient group, and thirty subjects, in whom angiographic finding revealed in not significant stenosis, but complained of chest pain, were selected as the control group. Fasting venous blood was obtained and measured the concentration of plasma total homocysteine, folate and vitamin $B_{12}$ by high performance liquid chromatography and fluorescence detection method. We examined the correlation between homocysteine and folate and/or vitamin $B_{12}$ in the control group and the patient group, respectively. Compared with the control group, the patient group had relatively higher plasma total homocysteine concentration ($10.7\pm4.2\;vs\;9.6\pm3.5$ umol/L), but showed no significant difference. Folate and vitamin $B_{12}$ concentration are low in the patient group, but showed no significant difference between patient and control group. Plasma total homocysteine concentration showed negative correlation with folate and vitamin $B_{12}$ in both the control group and the patient group, and showed significantly negative correlation in patient group {r = -0.550 (p < 0.01) vs r = -0.609 (p < 0.01)}. We knew that the plasma total homocysteine concentration were relatively elevated in patient group compared with the control group. Because plasma total homocysteine concentrations are closely negative correlated with folate and vitamin $B_{12}$ in the patient group, folate and vitamin $B_{12}$ supplement can lower the mortality and morbidity of ischemic heart disease. (Korean J Community Nutrition 10(6) : $963\∼970$, 2005)
Chlorophyll fluorescence, chlorophyll content, graft-taking and growth of grafted cucumber seedlings as affected by photosynthetic photon flux (PPF) of LED lamps were analyzed in this study. Four PPF levels, namely 25, 50, 100, $150{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ were provided to investigate the effect of light intensity on the chlorophyll fluorescence, chlorophyll content, graft-taking and growth of grafted cucumber seedlings. Air temperature, relative humidity, and photoperiod for graft-taking were maintained at $25^{\circ}C$, 90%, $16h{\cdot}d^{-1}$, respectively. Maximum quantum yield (Fv/Fm) of rootstock as affected by PPF was found to be 0.84-0.85 and there was no significant change in Fv/Fm. Even though Fv/Fm of scion measured at 2 days after grafting was lowered to 0.81-0.82, after then it gradually increased with increasing PPF. At 4 days after grafting, the chlorophyll content extracted from scion increased with increasing PPF. Graft-taking ratio of grafted cucumber seedlings was 90-95% as PPF was ranged from $25{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ to $100{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. However, the graft-taking ratio of grafted seedlings healed under PPF of $150{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ was decreased to 80%. Maximum PPF measured required for smooth joining of rootstock and scion was assumed to be $100{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. At healing stage of grafted cucumber seedlings, Fv/Fm of scion decreased and at least two days after grafting were required for rooting of grafted seedlings. Chlorophyll fluorescence response of rootstock and scion was linked to light irradiation. Therefore, it was concluded that physical environment including light and humidity during healing process of grafted seedlings should be controlled more precisely to facilitate root formation and to prevent scion from lowering Fv/Fm. Further studies are required to investigate the effects of root development and joining of vascular bundles of grafted seedlings on the chlorophyll content of scion.
Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.
This study was carried out to quantify the drought stress in grafted watermelon seedlings non-destructively by using chlorophyll fluorescence (CF) imaging technique rather than the visual judgment. Six-day old watermelon seedlings were grown under uniform irrigation for 3 days, and then given drought stress. Afterward, the sensor for the measurement of water content in plug tray cell unit was used to classify the drought-stress level into nine groups from D1 (53.0%, sufficient moisture state) to D9 (15.7%, extremely dry stress), and the 16 CF parameters were measured. In addition, re-irrigation was performed on the drought stressed seedlings(D5 - D9) to determine the growth and photosynthesis recovery level, which was not confirmed by visual judgment. The kinetic curve patterns of CF in three different drought stressed seedling groups were found to be different for the early detection of drought stress. All the 16 CF parameters decreased continuously with exposure to drought stress and drastically decreased from D5 (32.1%) where the visual judgment was possible. The fluorescence decline ratio (Rfd_Lss) started to decrease from the initial drought stress level (D5 - D6), and the Maximum PSII quantum yield (Fv/Fm) was significantly decreased in the later extreme drought stress range (D7 - D9) by re-irrigation recovery test. Thus, Rfd_Lss and Fv/Fm parameters were finally selected as potent indicators of growth and photosynthesis recovery in the initial and later stages of drought stress. Also, to the differences in the numerical values of the individual chlorophyll fluorescence parameters, the drought stress level was intuitively confirmed through the image. These results indicate that Rfd and Fv/Fm can be considered as potential CF parameters for the detection of low and extremely high drought stress, respectively. Furthermore, Fv/Fm can be considered as the best CF parameters for recovery at re-irrigation.
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