• Journal of Microbiology and Biotechnology
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• 제31권1호
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• pp.144-153
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• 2021

Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrids, using a flexible linker to develop more promising catalytic bioscavengers against a broad range of OPNAs. These GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNA analogs (kcat/Km (106 M-1 min-1): 59.8 ± 0.06 (paraoxon), 5.2 ± 0.02 (PNPDPP) and 47.0 ± 6.0 (DFP)) and V-type OPNA analogs (kcat/Km (M-1 min-1): 504.3 ± 48.5 (DSM) and 1324.0 ± 47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.

• Korean Journal of Acupuncture
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• 제38권4호
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• pp.197-208
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• 2021

Objectives : The aim of this study was to conduct a scoping review focusing on acupuncture points information applied in experimental studies using electroacupuncture for stroke. Methods : First, we set the research question of this study to identify which acupuncture points were studied for various symptoms of post-stroke sequelae in clinical and animal experiments. For this purpose, among the records searched through EMBASE, experimental studies including acupuncture points used in electroacupuncture treatment for stroke were selected as inclusion criteria for this study. The acupoints information used in each experiment was extracted from the included studies according to the type of study design and symptoms of stroke sequelae, and quantitative analysis was performed. Results : A total of 973 studies using acupuncture treatment, of which 429 papers including electroacupuncture were analyzed. Most of the animal experiments were conducted on general conditions instead of specific symptoms of stroke, and in clinical studies, electroacupuncture studies were conducted on symptoms such as hemiplegia and spasticity, cognitive and mental problems, dysphagia, shoulder problems, and depression that occur after stroke. Acupuncture points such as GV20, ST36, LI11, and LI4 were mainly used regardless of stroke symptoms. Except for these acupuncture points, GV26, GV14, GV16, and GV24 were frequently used in animal experiments, and TE5, LI15, LI10, and LR3 were widely used in clinical studies. Conclusions : Although this study extracted and analyzed only the frequent acupuncture points in the electroacupuncture study for stroke. However, in the future, it is necessary to be able to secure the evidence for acupuncture treatment elements through comparative studies between different acupuncture points or other elements constituting electroacupuncture.

• Molecules and Cells
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• 제42권11호
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• pp.783-793
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• 2019

When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.

• 대한한의진단학회지
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• 제19권2호
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• pp.101-108
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• 2015

Objectives This paper focused on developing the acupuncture-moxibustion device for head. Disadvantages of the existing devices for head moxibustion are its hume and inconvenience in operating because of the hair. Methods The proposed acupuncture-moxibustion device for GV20(Baihui) is 10mm long as needle body and 6mm long as needle root whose feature are 1~2 turns spiral coil. The spiral coil contains the moxa in order not to drop the moxa while operating Results The direct temperature on the burning moxa were measured by TK-305 infrared thermometer. But the direct temperature on the needle of acupuncture-moxibustion device were not able to measure by TK-305 infrared thermometer. By definition equation, the indirect temperatures on the needle of acupuncture-moxibustion device were able to be calculated. Conclusion A very safe, small and cheap acupuncture-moxibustion device for GV20(Baihui) of head is proposed. It will be useful to generate the heat shock protein and to operate in preventing and curing the Alzheimer's disease.

• Journal of Acupuncture Research
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• 제19권6호
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• pp.24-34
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• 2002

Objective : To report seborrheic dermatitis of the scalp which is treated by Bee-venom Acupuncture therapy. Methods : Under the assumption that Be-venom Therapy may be effective for treatment seborrheic dermatitis, Bee-venom acupuncture was injected subcutaneously into the lesion including Gv24(神庭), Gv23(上星), Gv22, Gv20(白會), B4(曲差), B5(五處), B6(承光), S8(頭維), etc. Results : The symptoms including erythematous plaque, scale, itching were resolved clearly except a little scale by Bee-venom acupuncture Therapy. Conclusions : Bee-venom acupuncture Therapy is effective in the treatment of seborrheic dermatitis. if more clinical trials like this are proved to be effective in seborrheic dermatitis, we can expect that Bee-venom acupuncture Therapy is used to treatment of seborrheic dermatitis as good method.

• Journal of Acupuncture Research
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• 제32권4호
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• pp.11-16
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• 2015

Objectives : The purpose of this study is to determine the safe needling depth of Pungbu($GV_{16}$) retrospectively by using magnetic resonance imaging (MRI). Methods : We chose 114 Brain or C-spine MRI images from the Sang-Ji hospital picture archiving communication system. We measured the shortest distance from skin to cerebral dura mater passing by posterior edge of the foramen magnum on the sagittal view for the depth of Pungbu. We analyzed the differences between male and female measured values by using a student t-test. Results : The average depth of male insertion was $49.71{\pm}6.32mm$ and the shortest depth of insertion was 36.29 mm. The average depth of female insertion was $39.84{\pm}5.25mm$ and the shortest depth of insertion was 30.02 mm. The results showed a significant difference according to gender (p=0.00). Conclusions : The depth of male insertion is deeper than that of female, and the safe needling depth in the case of males is 36.29-67.35 mm, while the safe needling depth in the of females is 30.02-52.18 mm.

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.223-227
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• 2005

In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture $43.8\%$ of oocytes cultured in only TCM 199 remained at GV stage whereas $56.2\%$ of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than $90\%$ of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.357-366
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• 1996

The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1695-1701
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• 2002

The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.

• 한국임상수의학회지
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• 제23권1호
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• pp.65-68
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• 2006

개 추간판 디스크의 2증례를 수침치료를 하였다. 증례 1은 경부 디스크(C3-C4)의 증례이었으며, 1-2차 치료에서는 덱사메타손(1 mg/ml), 3-5차 치료에서는 염산치아민(25mg/ml)을 각각 수침하였다. 사용혈위는 GV-16, GB-20, BL-10, LU-7, LI-4 및 SI-6이었다. 2-5차 치료에는 GV-6, GV-20 및 SP-6의 혈위를 추가하였다. 3-5차 치료에는 2% 염산리도카인(0.2ml)를 이용하여 견갑아래근 및 상완세갈래근의 압통점에 각각 수침하였다. 5차 치료 후 마비증상은 소실되어 치료를 중단하였으며, 5개월 후 증상의 재발은 인정되지 않았다. 증례 2는 요부디스크의 증례(L1-L2)로 1-2차 치료에는 덱사메타손 수침을 3-4차치료에는 염산치아민을 각각 수침하였다. 사용혈위는 GV-6를 주혈로, ST-36, ST-40, GB-34, ST-41 및 BL-40을 부혈로 각각 사용하였다. 또한 허리장골 늑골근 및 대퇴네갈래근의 압통점에 2% 염산리도카인(0.2ml)를 각각 수침하였다. 4차 치료 후 마비증상이 호전되어 치료를 중단하였고, 5개월 후 현재 까지 증상의 재발은 인정되지 않았다. 이상의 증례를 통하여, 덱사메타손과 염산치아민의 수침 및 리도카인을 이용한 압통점치료의 병용은 개의 경부 및 요부디스크의 증상을 효과적으로 콘트롤 할 수 있는 것으로 판단되었다.