Won-Ho Kim;Bo Ram Lee;Hey-Yun Kim;Minji Kim;Jin-Woo Kim
Journal of Korean Dental Science
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v.16
no.2
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pp.182-191
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2023
Purpose: This study investigated the orthodontic tooth movement after weekly parathyroid hormone (PTH) injection in mongrel dogs and analyzes bone formation activity on the tension and pressure sides of the tooth movement in mongrel dogs. Materials and Methods: Three mongrel dogs were used in this study. The first premolar was extracted and orthodontic force using 150 g of closed coil springs between the canine and second premolar was applied. The low-dose PTH group (PTH_1) and high-dose PTH group (PTH_2) received weekly injections of 1.61 ㎍/kg and 3.23 ㎍/kg of PTH, respectively. The control group received weekly injections of 1 ml of saline. Clinical, histomorphometric analysis were carried out. Result: The orthodontic tooth movement was greatest in the PTH_2 group and the lowest in the control group. Fluorescence staining images showed higher bone remodeling on the tension side of the tooth movement in the PTH_1 and PTH_2 groups. PTH_2 group showed a thicker labeling band than the PTH_1 group. PTH_2 group showed the highest mineral apposition rate and bone formation rate, followed by the PTH_1 group and the control group. Conclusion: Weekly intermittent PTH injection, especially in the short-term and at higher doses with orthodontic force, successfully increased orthodontic tooth movement and bone remodeling in mongrel dogs.
In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.
In the past decade, considerable progress has been made in developing the appropriate biotechnology for microalgal mass cultivation aimed at establishing a new agro-industry. This review points out the main biological constraints affecting algal biotechnology outdoors and the requirements for making this biotechnology economically viable. One of them is the availability of a wide variety of algal species and improved strains that favorably respond to varying environmental conditions existing outdoors. It is thus just a matter of time and effort before a new methodology like genetic engineering can and will be applied in this field as well. The study of stress physiology and adaptation of microalgae has also an important application in further development of the biotechnology for mass culturing of microalgae. In outdoor cultures, cells are exposed to severe changes in light and temperature much faster than the time scale re-quired for the cells to acclimate. A better understanding of those parameters and the ability to rapidly monitor those conditions will provide the growers with a better knowledge on how to optimize growth and productivity. Induction of accumulation of high value products is associated with stress conditions. Understanding the physiological response may help in providing a better production system for the desired product and, at a later stage, give an insight of the potential for genetic modification of desired strains. The potential use of microalgae as part of a biological system for bioremediation/detoxification and wastewater treatment is also associated with growing the cells under stress conditions. Important developments in monitoring and feedback control of the culture behavior through application of on-line chlorophyll fluorescence technique are in progress. Understanding the process associated with those unique environmental conditions may help in choosing the right culture conditions as well as selecting strains in order to improve the efficiency of the biological process.
A new concept of "photo" -antisense method has been evaluated, where the inhibition of gene expression by the conventional antisense method is enhanced by photochemical binding between antisense oligonucleotides conjugated with photo-reactive compound and target mRNA or DNA. Fluorescein labeled oligodeoxyribonucleotides (F-DNA) was delivered to cell nuclei in the encapsulated form in multilamellar lecithin liposomes with neutral charge. F-DNA was previously shown to photo-bind to the complementary stranded DNA, and the delivery system using neutral liposome to be effective in normal human keratinocytes. In the present study, we used human kidney cancer G401.2/6TG.1 cell line to be advantageous in reproducible experiments. p53 was adopted as a target gene since antisense sequence information has been accumulated. The nuclear localization ofF-DNA was identified by comparing the fluorescence ofF-DNA with that of Hoechst 33258 under fluorescence microscope. After 7hr incubation to accumulate p53 protein induced by UV -B, p53 protein was quantified by Western blot. After 2hrs from F-DNA application, about 30% of cell population incorporated F-DNA in their nuclei with some morphological change possibly due to liposomal toxicity. Irradiation of visible light longer than 400nm from solar simulator at this time enhanced the inhibitory action of antisense F-DNA. The present results suggest that photo-antisense method is promising to control gene expression in time and space dependent manner. Further improvement of F-DNA delivery to cancer cells in the stability and toxicity is in progress. progress.
Silencing of a specific gene using RNAi (RNA interference) is a valuable tool for functional analysis of a target gene. However, information on RNAi for analysis of gene function in farm animals is relatively nil. In the present study, we have validated the interfering effects of siRNA (small interfering RNA) using both quantitative and qualitative gene silencing in buffalo granulosa cells. Qualitative gene knockdown was validated using a fluorescent vector, enhanced green fluorescence protein (EGFP) and fluorescently labeled siRNA (Cy3) duplex. While quantitatively, siRNA targeted against the luciferase and CYP19 mRNA was used to validate the technique. CYP19 gene, a candidate fertility gene, was selected as a model to demonstrate the technique optimization. However, to sustain the expression of CYP19 gene in culture conditions using serum is difficult because granulosa cells have the tendency to luteinize in presence of serum. Therefore, serum free culture conditions were optimized for transfection and were found to be more suitable for the maintenance of CYP19 gene transcripts in comparison to culture conditions with serum. Decline in fluorescence intensity of green fluorescent protein (EGFP) was observed following co-transfection with plasmid generating siRNA targeted against EGFP gene. Quantitative decrease in luminescence was seen when co-transfected with siRNA against the luciferase gene. A significant suppressive effect on the mRNA levels of CYP19 gene at 100 nM siRNA concentration was observed. Also, measurement of estradiol levels using ELISA (enzyme-linked immunosorbent assay) showed a significant decline in comparison to control. In conclusion, the present study validated gene silencing using RNAi in cultured buffalo granulosa cells which can be used as an effective tool for functional analysis of target genes.
Cai Chun Mei;Van Kyujung;Kim Moon Young;Lee Suk-Ha
KOREAN JOURNAL OF CROP SCIENCE
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v.50
no.5
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pp.361-367
/
2005
Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to determine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety $F_2$-derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on $Victot^3$ microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dyeterminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.
The modern integrated fish-seaweed mariculture has been tested to reduce the environmental impacts of an intensive fed culture. To obtain the best seaweed bioremediation performance, the effects of therapeutants used for fish disease control on the selected seaweed species should be considered. As a selected seaweed, Porphyra yezoensis was tested with six commercial antibiotics including erythromycin thiocyanate_A, erythromycin thiocyanate_B, oxytetracycline, doxycycline, pefloxacin, and amoxicillin trihydrate under the batch incubation at a photon flux density of 10 $\mu$mol ${\cdot}m^{-2}\;{\cdot}\;s^{-1}$ at 15$^{\circ}C$. Among the tested commercial antibiotics, erythromycin thiocyanate_A, erythromycin thiocyanate_B, oxytetracycline, and doxycycline showed decreases in Fv/Fm, the photochemical efficiency of photosystem II, with a dose-dependant and time-dependant manner. From the quenching analysis of chlorophyll fluorescence, three differential patterns were observed in the antibiotics-treated Porphyra: (1) high nonphotochemical quenching (NPQ) and low photochemical quenching (qP) in the cases of Erythromycin thiocyanate_B and amoxicillin trihydrate, (2) high NPQ and high qP in the case of pefloxacin and (3) low NPQ and low qP in the case of oxytetracycline. These results indicated that antibiotics affected in various ways on the photosynthetic apparatus, reflecting differential lesion sites of antibiotics. In addition, the rates of ammonium uptake also decreased with a decrease of Fv/Fm in P. yezoensis thalli treated with erythromycin thiocyanate_B and oxytetracycline. Therefore, the four antibiotics mentioned could affect the bioremediation capacity of the selected seaweed species in the integrated fish-seaweed mariculture system due to the decrease of photosynthetic activity and the simultaneous decrease of ammonium uptake.
Tarakhovskaya, Elena R.;Kang, Eun-Ju;Kim, Kwang-Young;Garbary, David J.
ALGAE
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v.27
no.2
/
pp.125-134
/
2012
Germanium dioxide ($GeO_2$) has been used for many years in the cultivation of red and green algae as a means of controlling the growth of diatoms. Brown algae are sensitive to $GeO_2$, however, the basis of this sensitivity has not been characterized. Here we use embryos of $Fucus$$vesiculosus$ to investigate morphological and physiological impacts of $GeO_2$ toxicity. Morphometric features of embryos were measured microscopically, and physiological features were determined using pulse amplitude modulated (PAM) fluorometry. At 5 mg $L^{-1}$$GeO_2$, embryos grew slower than controls and developed growth abnormalities. After 24 h, initial zygote divisions were often oblique rather than transverse. Rhizoids had inflated tips in $GeO_2$ and were less branched, and apical hairs were deformed, with irregularly aligned, spheroidal cells. Minimum fluorescence ($F_0$) showed minor differences over the 10 days experiment, and pigment levels (chlorophylls $a$, $c$ and total carotenoids) showed no difference after 10 days. Optimum quantum yield increased from ca. 0.52 at 24 h to 0.67 at 5 days, and $GeO_2$-treated embryos had higher mean values (significant at 3 and 5 days). Optimum quantum yield of photosystem II (${\Phi}_{PSII}$) was stable in control thalli after 5 days, but declined significantly in $GeO_2$. Addition of silica (as $SiO_2$) did not reverse the effects of $GeO_2$. These results suggest that $GeO_2$ toxicity in brown algae is associated with negative impacts at the cytological level rather than metabolic impacts associated with photosynthesis.
Seo, Mi-Rae;Nam, Mi-Yeon;Kim, Beom-Sik;Nam, Seung-Eun;Kim, In-Chul;Park, You-In
Membrane Journal
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v.21
no.2
/
pp.163-170
/
2011
Membrane fouling control in water treatment may be the main obstacle for wider implementation and lower cost. A novel fluorescent spectroscope sensor device for membrane fouling integrity monitoring has been developed and evaluated in this study. PSf membranes for water treatment has been fabricated with three types of organic fluorescent materials, OB, FP, KCB. The fluorescent signal from membrane surface was analyzed throughout the filtration process. It was found that the fluorescent signal due to the membrane fouling decreased and the developed device is reliable for membrane fouling monitoring.
Effect of silicon (Si) nutrition on growth and temperature stress tolerance of Nephrolepis exaltata 'Corditas' grown in a soilless substrate was examined. In vitro-grown acclimatized plantlets were transplanted into the pots containing a coir-based substrate. A nutrient solution containing 0, 50, or $100mg{\cdot}L^{-1}$ Si was supplied through a drip-irrigation system. After 5 months of cultiv ation, S i-treated and -untreated p lants were grown at 10, 25, or $40{\pm}1^{\circ}C$ under a 12 h photoperiod with $530{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD and 60% RH. After 7 days, chlorophyll content and chlorophyll fluorescence parameters were measured. Silicon nutrition had a negative effect on growth characteristics of N. exaltata 'Corditas'. However, Si-treated plants had more tolerance to temperature stress than the control plants. The Fv/Fm value was not significantly different when the plants were exposed to $25^{\circ}C$. However, significant difference in Fv/Fm was recorded when plants were exposed to 10 or $40^{\circ}C$. Thus, Fv/Fm could be used as an indicator of low and high temperature tolerance in ferns. The present study also suggests that application of Si may be used to enhance temperature tolerance of ferns.
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