Even though titanium(Ti) and its alloys are the most used dental implant materials, there are some problems that Ti wears easily and interferes normal osteogenesis due to the metal ions. Ti coated with bioactive ceramics such as hydroxyapatite has also such problems as the exfoliation or resorption of the coated layer, Recent studies on implant materials have been proceeding to improve physical properties of the implant substrate and biocompatibility of the implant surfaces. The purpose of the present study was to examine the physical property and bone tissue compatibility of bioinert nitrides ion plated Ti, Button type specimens(14mm in diameter, 2.32rrun in height) for the abrasion test and cytotoxicity test and thread type implants(3.75mm in diameter, 6mm in length) for the animal experiments were made from Ti(grade 2) and 316LVM stainless steel. Ti specimens were ion plated with TiN, ZrN by the low temperature arc vapor deposition, and the depth profile of the TiN/Ti, ZrN/Ti ion plated surface was examined by Auger Electron Spectroscopy. Three kind of button type specimens .of TiN/Ti, ZrN/Ti and Ti were used for abrasion test, and HEPAlClC7 cells and CCD cells were cultivated for 4 days with the specimens for cytotoxicity test. Thread type implants of TiN/Ti, ZrN/Ti, Ti, 316LVM were implanted on the femur of 6 adult dogs weighing 10kg-13kg. Two dogs were sacrified for histological examination after 45 days and 90 days, and four dogs were sacrified for the removal torque test of the implant') after 90 days. The removal torque force was measured by Autograph (Shimadzu Co., AGS-1000D series, Japan). Abrasion resistance of TiN/Ti was the highest, and that of ZrN/Ti and Ti were followed. The bioinert nitride ion plated Ti had much better abrasion resistance, compared with Ti, In the cytotoxicity test, the number of both cells were increased in all specimens, and there were no significant difference in cytotoxic reaction among all groups (p>0.1), In histological examination, 316LVM showed the soft tissue engagement in interface between the implant and bone, but the other materials after 45 days noted immature new bone formation in the medullary portion along the implant surface, and those after 90 days showed implant support by new bone formation in both the cortical and the medullary portion, The removal torque force of Tilv/Ti showed significantly higher than that of Ti(p(O,05). The difference in removal torque force between TiN/Ti and ZrN/Ti was not significant(p>0.05), and that of 316LVM was lowest among all groups(p<0.05). These results suggest that bioinert nitrides ion plated Ti can resolve the existing problems of Ti and bioactive ceramics, and it may be clinically applicable to human.
The present experiments were focussed to modify a short slow-cooling protocol used for freezing of early stage embryo(Testart et al., 1986) and also to apply the modified method for the cryopreservation of hamster oocytes with Zona or without. The protocol was modified by changing the 4-step equilibration into 1-step and the 1-step thawing into 2-step. The oocytes were added in 1.5M PROH and 0.1M Sucrose, seeded at $-7^{\circ}C$, slow cooled($0.3^{\circ}C$/min) to $-30^{\circ}C$ before plunging to $-196^{\circ}C$. The oocytes were thawed at $23-25^{\circ}C$ air(20sec/150sec) and/or 33-35 water(10sec). The survival of the frozen-thawed oocytes was determined by morphologic criteria and their fertilizing ability was also estimated by Sperm Penetration Assay(SPA) system(Chang et al, 1990) using fertile men semen sample. One-step equilibration showed slightly higher survival rate(83.9% vs. 71.0%) and fertilization rate(83.9% vs. 71.0%) compared with four-step(p>0.05). And two-step thawing(air & water exposing) of oocytes frozen after 1-step equilibration showed significantly higher survival rate(96.3%) than one-step thawing at air(85.2%) or water(65.0%) only(p<0.05). Therefore, by the modified method(l-step equilibration & 2-step thawing), Zona-intact(ZI) and Zona-free(ZF) oocytes were frozen and thawed. ZI-oocytes showed significantly higher survival rate(95.4%, 308/323 vs. 67.6%, 240/355) than ZF-oocytes(P<0.01). But the survival of ZF-oocytes was as high as ZI-oocytes in fourteen of twenty-four replicates. ZI-oocytes was also significantly higher fertilization rate($92.4{\pm}8.9%$ vs. $63.7{\pm}18.5%$) and higher mean number of penetrated sperm($6.2{\pm}4.2$ vs. $3.9{\\pm}3.3$) than ZF-oocytes, but not higher than control(fresh oocytes;$99.3{\pm}2.4%$, $8.4{\pm}4.2$)(P<0.001). Conclusively, this modified method will contribute to freeze effectively the hamster oocytes for simplifing of the logical consideration of performing SPA and also to freeze the human and other animal oocytes.
Choi, Chang Soon;Hong, Minha;Kim, Ki Chan;Kim, Ji-Woon;Yang, Sung Min;Seung, Hana;Ko, Mee Jung;Choi, Dong-Hee;You, Jueng Soo;Shin, Chan Young;Bahn, Geon Ho
Biomolecules & Therapeutics
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v.22
no.5
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pp.406-413
/
2014
to valproic acid (VPA) during pregnancy produces ASD-like core behavioral phenotypes as well as hyperactivity in offspring both in human and experimental animals, which makes it a plausible model to study ASD-related neurobiological processes. In this study, we examined the effects of two of currently available attention defecit hyperactivity disorder (ADHD) medications, methylphenidate (MPH) and atomoxetine (ATX) targeting dopamine and norepinephrine transporters (DAT and NET), respectively, on hyperactive behavior of prenatally VPA-exposed rat offspring. In the prefrontal cortex of VPA exposed rat offspring, both mRNA and protein expression of DAT was increased as compared with control. VPA function as a histone deacetylase inhibitor (HDACi) and chromatin immunoprecipitation experiments demonstrated that the acetylation of histone bound to DAT gene promoter was increased in VPA-exposed rat offspring suggesting epigenetic mechanism of DAT regulation. Similarly, the expression of NET was increased, possibly via increased histone acetylation in prefrontal cortex of VPA-exposed rat offspring. When we treated the VPA-exposed rat offspring with ATX, a NET selective inhibitor, hyperactivity was reversed to control level. In contrast, MPH that inhibits both DAT and NET, did not produce inhibitory effects against hyperactivity. The results suggest that NET abnormalities may underlie the hyperactive phenotype in VPA animal model of ASD. Profiling the pharmacological responsiveness as well as investigating underlying mechanism in multiple models of ASD and ADHD may provide more insights into the neurobiological correlates regulating the behavioral abnormalities.
Oh, Jang-Hoon;Kim, Hyug-Gi;Woo, Dong-Cheol;Rhee, Sun Jung;Lee, Soo Yeol;Jahng, Geon-Ho
Progress in Medical Physics
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v.29
no.1
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pp.29-41
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2018
The objective of this study was to evaluate the chemical exchange saturation transfer (CEST) effect of amino acids and neurotransmitters, which exist in the human brain, depending on the concentration, pH, and amplitude of the saturation radiofrequency field. Phantoms were developed with asparagine (Asn), ${\gamma}-aminobutyric$ acid (GABA), glutamate (Glu), glycine (Gly), and myoinositol (MI). Each chemical had three different concentrations of 10, 30, and 50 mM and three different pH values of 5.6, 6.2, and 7.4. Full Z-spectrum CEST images for each phantom were acquired with a continuous-wave radiofrequency (RF) saturation pulse with two different $B_1$ amplitudes of $2{\mu}T$ and $4{\mu}T$ using an animal 9.4T MRI system. A voxel-based CEST asymmetry was mapped to evaluate exchangeable protons based on amide (-NH), amine ($-NH_2$), and hydroxyl (-OH) groups for the five target molecules. For all target molecules, the CEST effect was increased with increasing concentration and B1 amplitude; however, the CEST effect with varying pH displayed a different trend depending on the characteristics of the molecule. On CEST asymmetric maps, Glu and MI were well visualized around 3.0 and 0.9 ppm, respectively, and were well separated macroscopically at a pH of 7.4. The exchange rates of Asn, Glu, BABA, and Gly usually decreased with increasing pH. The CEST effect was dependent on the concentration, acidity of the target molecules, and B1 amplitude of the saturation RF pulse. The CEST effect for Asn can be observed in a 9.4T MRI system. The results of this study are based on applying the CEST technique in patients with neurodegenerative diseases when proteins in the brain are increased with disease progression.
The Artemisia capillaris THUNB is a perennial herb that belongs to the family Compositae spp and probably the most common plant among the various herbal folk remedies being used in the treatment of abdominal pain, hepatitis, chronic liver disease, jaundice and coughing in Korea. Recently the biological and pharmacological actions of herb have been studied well such as antibacterial, antidiabetic and antitumor activities. This experiment was conducted to investigate antitumor and immunomodulatory effects of Artemisia capillaris extracts against Hepa-1c1c7 and Sarcoma 180 cancer cells in in vivo experimental tests. In in vivo experimental tests using 210 ICR mice, based on flow cytometry, CD3+, CD4+, CD8+ and TNF-${\alpha}+$ splenocytes were significantly (p<0.05) reduced in the Hepa-1c1c7 and Sarcoma 180 inoculated vehicle controls, HP and SP, compared to those of the intact vehicle control on both the $28^{th}$ day and the $42^{nd}$ day, respectively. These decreases of CD3+, CD4+, CD8+ and TNF-${\alpha}+$ cells induced by tumor inoculations were significantly (p<0.05) inhibited by mACH treatment regardless of the type of experiments and tumor cells inoculated. The results suggest that Artemisia capillaris methanol extracts have prominent antitumor effects on the cancer cell lines Hepa-1c1c7 and Sarcoma 180.
Objectives : The use of natural products with therapeutic properties is as ancient as human civilisation and, for a long time, mineral, plant and animal products were the main sources of drugs. Catalposide, the major iridoid glycoside isolated from the stem bark of Catalpa ovata G. Don (Bignoniceae) has been shown to possess anti-microbial and anti-tumoral properties. Heme oxygenase-1 (HO-1) is a stress response protein and is known to play a protective role against the oxidative injury. In this study, we examined whether catalposide could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 protein expression and HO activity. We also examined the effects of catalposide on the productions of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and nitric oxide (NO) on RAW 264.7 macrophages activated with the endotoxin lipopolysaccharide. Methods : HO-1 expression in Neuro 2A cells was measured by Western blotting analysis. NO and $TNF--{\alpha}$ produced by RAW 264.7 macrophage were measured by Griess reagent and enzyme-linked immunosorbent assay, respectively. Results : The treatment of the cells with catalposide resulted in dose- and time-dependent up-regulations of both HO-1 protein expression and HO activity. Catalposide protected the cells from hydrogen peroxide-induced cell death. The protective effect of catalposide on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX, a HO inhibitor. Additional experiments revealed the involvement of CO in the cytoprotective effect of catalposide-induced HO-1. In addition, catalposide inhibited the productions of $TNF--{\alpha}$ and NO with significant decreases in mRNA levels of $TNF--{\alpha}$ and inducible NO synthase. Conclusions : Our results indicate that catalposide is a potent inducer of HO-1 and HO-1 induction is responsible for the catalposide-mediated cytoprotection against oxidative damage and that catalposide may have therapeutic potential in the control of inflammatory disorders.
The Artemisia capillaris THUNB is a perennial herb that belongs to the family Compositae spp. and probably the most common plant among the various herbal folk remedies being used in the treatment of abdominal pain, hepatitis, chronic liver disease, jaundice and coughing in Korea. This experiment was conducted to investigate the effects of Artemisia capillaris extracts on the amounts of splenocytes-derived cytokine ($TNF-{\alpha},\;IL-1{\beta}$ and IL-10). In in vivo experimental tests using 210 ICR mice with Hepa-1c1c7 or sarcoma 180 cancer line, splenocytes derived cytokine contents were significantly (p < 0.05) reduced in the Hepa-1c1c7 and Sarcoma 180 inoculated vehicle controls, HP and SP, compared to those of the intact vehicle control on both the $28^{th}$ day and the $42^{nd}$ day, respectively. However, these decreases of $TNF-{\alpha},\;IL-1{\beta}$ and IL-10 levels induced by tumor inoculations were significantly (p < 0.01, p < 0.05) inhibited by mACH (Artemisia capillaris methanol extracts) treatment regardless of the type of experiments and tumor cells inoculated. The results suggest that Artemisia capillaris methanol extracts have prominent anti-inflammation effects on the cancer cell lines Hepa-1c1c7 and Sarcoma 180.
Hypertermia for the treatment of cancer has been introduced for a long time and the biological effect for the use of hyperthermia to treat malignant tumors has been well established and encouraging clinical results have been obserbed. Unfortunately, however, the engineering or technical aspects of hyperthermia for the deep seated tumors has not been satisfactory. We developed the radiofrequency capactive hyperthermia device (Greenytherm-GY8) in cooperation with Yonsei Cancer Center and Green Cross Medical Corporation. It was composed with $8{\sim}10MHz$ RF generator, capacitive electrode, matching system, cooling system, temperature measuring system and control PC computer. The thermal profile was investigated in agar phantom, animals and in human tumors, heated with capactivie RF device. Deep and homogeneous heating could be achieved in a large phantom of 25cm diameter and 19cm thick when heated with a pair of 23cm diameter electrodes, coupled to both bases of the phantom, when the size of the two electrodes was not the same, the region near the smaller electrode was preferentially heated. It was, therefore, possible to control the depth of heating by choosing proper size of electrodes. Therapeutic temperature $(42^{\circ}C{\sim}43^{\circ}C)$ could be obtained in the living animal experiments. Indications are that deep heating of humn tumors might be achieved with the capacitive method, provided that subcutaenous fat layer is cooled by temperature controlled bolus and large size of electrodes.
Purpose : By using the micro-imaging unit modified from NMR spectrometer, the high resolution MRI protocols of finer than 100 micron in 5 minutes, is sought for mouse, which plays a central role in animal studies Materials and Methods : C57BL/6 mouse, lighter than 50 gram, is used for the experiments. The superconducting magnet is vertical type with 89 mm inner diameter at 4.9 Tesla. The diameter of rf-coil is 30 mm. Mostly used techniques are the fast spin echo and the gradient echo pulse sequence. Results : For 2D images, proton density and T2 weighted images are obtained and their optimum experimental variables were sought. Minute structure of mouse brain can be recognized and 3D brain image is also obtained additionally. 3D image will be useful particularly for the dynamic contrast study using various contrast agents. Conclusion : Like the case of human and other small animals, the high resolution of mouse brain is enough to recognize the minute structure of it. Recently, similar studies are reported domestically, but it seems only a beginning stage. Due to easiness of breeding/control, mouse MRI study will soon play a vital part in brain study.
Kim, Yong-Hoon;Park, Jun-Young;Cha, Mi-Kyong;Lee, Sang-Moo;Kim, Hyeon-Tae;Uh, Soo-Taek;Chung, Yeon-Tae;Park, Choon-Sik
Tuberculosis and Respiratory Diseases
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v.40
no.1
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pp.16-22
/
1993
Background: The Microbicidal and cytotoxic activities of neutrophils are to a large extent dependent on a burst of oxidative metabolism which generates superoxide anion, hydrogen peroxide, and other reactive products of oxygen. The respiratory burst of PMN is initiated by intracellular calcium mobilization that follows immune or particular stimulation and is very sensitive to modulation by c-AMP or adenosine. Despite its antagonism against adenosine, earlier study has demonstrated potent theophylline inhibition of the PMN respiratory burst at variable ranges of blood concentrations of theophylline in the healthy normal volunteers and in the septic animals pretreated or early post-treated with aminophylline (AMPH) or pentoxifylline. However it is unclear whether theophylline inhibits the superoxide generation or not in the established human sepsis caused by acute pneumonia, as taking into consideration of the fact that full activation of neutrophils have occurred within minutes after the septic insult in the animal experiments. Methods: We measured the $O_2$ generation of peripheral arterial neutrophils obtained from 11 human septic subjects caused by acute pneumonia before and 1 hour after completion of continuous AMPH infusion. Patients were identified and studied within 48 hour of admission. All subjects were administered an intravenous loading and maintenance dose of AMPH. The generation of $O_2$ was measured at a discrete time point (60 min) by the reduction of ferricytochrome c.PMA (10 ${\mu}g/ml$) was used as a stimulating agent. PMNs were isolated at a concentration of $2{\times}10^6$ cells/ml. The arterial oxygen tension, blood pressure and heart rates were also checked to evaluate the systemic effects of AMPH in the acute pneumonia. Results: The mean serum concentration of AMPH at 60 minutes was $8.8{\pm}0.6{\mu}g/ml$. Sixty minutes after AMPH infusion the generatition of $O_2$ was decreased from $0.076{\pm}0.034$ to $0.013{\pm}0.004$(OD) (p<0.05) and from $0.177{\pm}0.044$ to $0.095{\pm}0.042$(OD) (p<0.01) in the resting and stimulated PMNs respectively. $PaO_2$ was not changed after AMPH infusion. Conclusion: AMPH may compromise host defense by significant inhibition of neutrophil release of superoxide anion and it had no effect on improving $PaO_2$ in the acute pneumonia.
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