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Effects of FBS(Fetal Bovine Serum) and pFF(Porcine Follicular Fluid) on In Vitro Maturation and Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

  • Moon, Hyo-Jin;Shim, Joo-Hyun;Hwang, In-Sun;Park, Mi-Rung;Kim, Dong-Hoon;Ko, Yeoung-Gyu;Park, Choon-Keun;Im, Gi-Sun
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.85-91
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    • 2009
  • In this study, in vitro maturation system using fetal bovine serum (FBS) or porcine follicular fluid (pFF) was investigated to produce comparable oocytes to those derived from in vivo. Control group of oocytes was cultured in TCM 199 supplemented with 0.1% polyvinyl alcohol (PVA). Other three groups of oocytes were cultured in TCM 199 supplemented with 10% FBS, 10% pFF or 5% FBS + 5% pFF, respectively. After 44 h maturation, oocytes with the first polar body were activated with two electric pulses (DC) of 1.2 kv/cm for 30 ${\mu}sec$. Also, matured oocytes of four groups were reconstructed and fused. Reconstructed embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. The oocytes matured in the medium supplemented with FBS or/and pFF showed significantly higher maturation rates (64.0 vs. 73.9 to 85.2%). In PA embryos, cleavage rates (89.7 vs. 77.1 to 86.6%) and blastocysts rates (30.0 vs. 16.2 to 26.2%) were significantly higher in pFF group (p<0.05). In NT embryos, there was no difference among treatments in cleavage rate, but the blastocyst rates (28.5 vs. 15.5 to 24.6%) were significantly higher in pFF group (p<0.05). The apoptosis rate was significantly higher (p<0.05) in the control than other groups (10.8 vs. 4.9 to 8.2% for PA, 3.1 vs. 0.5 to 1.3% for NT). In order to select the comparable oocyte to in vivo oocytes, each group of oocytes was stained with Brilliant cresyl blue (BCB) after 42h maturation. The matured oocytes were separated according to color of cytoplasm; stained group (BCB+) and unstained group (BCB-). The oocytes matured in the presence of FBS or/and pFF showed significantly higher staining rates (70.3 to 72.7 vs. 35.1%) (p<0.05). To verify the fact that the supplementation of FBS or/and pFF can increase the maturation rates, cdc2 kinase activity, the catalytic subunit of MPF, was determined. The cdc2 kinase activity of the oocytes matured in the medium supplemented with FBS or/and pFF was significantly higher than control group (6.7 to 9.3 vs. 3.8). In conclusion, the supplementation of FBS or/and pFF can support in vitro maturation rate of porcine oocytes through the increment of cdc2 kinase activity level in the cytoplasm.

Total Dose Effect on Normal Skin of Hybrid Mice by Conventional Fractionated Irradiation (고식적 분할조사시 방사선의 전조사량이 잡종 마우스의 정상피부에 미치는 병리조직학적 변화)

  • Jung, Kyung-Hee;Kim, Myung-Se;Choi, Won-Hee
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.261-267
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    • 1986
  • Development of supervoltage treatment machine may minimize skin reaction by skin-sparing effect, but skin damage is still one of "the dose limiting factor" in radiation therapy. In spite of these importance, systemic histopathologic studies of skin in similar conditions which used in clinical treatment has not been performed so far. 60mice were irradiated with conventional fraction ($200{\times}5/wk$) and whole abdominal field ($2{\times}3cm$, from symphysis pubis to xyphoid process). Used machine was 250KV, 24mA, orthovoltage x-ray machine. Histopathological changes of acute skin reaction at the level of total irradiation dose were analyzed and the possible mechanism of later chronic changes were investigated. Obtained results are as follows: 1. In 1,000 rad irradiated group, only mild epidermal edema is noted. 2. In 2,000 rad irradiated group, slightly decreased number and size of hair follicles and appendages, dermal edema and scanty infiltration of inflammatory cells are visible. 3. In 3,000 rad irradiated group, marked increased capillary congestion and prominant infiltration of inflammatory cells are observed. 4. In 4,000 rad irradiated group, vascular wall thickening with proliferation of endothelial cells are prominant. Dermal thinning and hyalinization are newly developed. 5. In 5,000 rad irradiated group, complete desquamation of epidermis is not seen, despite of acceleration of all above mentioned changes.

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Sterilization of Yakju(Rice Wine) Using a Batch-type High Voltage Pulsed Electric Field System (고전장펄스를 이용한 약주의 회분식 살균)

  • Kim, Su-Yeon;Park, Young-Seo;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1247-1253
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    • 1999
  • Yakju(rice wine) was sterilized with high-voltage pulses of short time on a batch pulsed electric field(PEF) system. The initial microbial counts of Yakju were 7.52 X $10^4\;CFU/mL$ for total aerobes, 2.20 X $10^4\;CFU/mL$ for lactic acid bacteria and 7.08 X $10^4\;CFU/mL$ for yeasts. The pH, acidity and electric conductivity of Yakju were 3.36, 0.462% and 1.24 mS/cm, respectively. Yakju was treated with 2-250 of pulses exponential-wave formed electric pulses under the field strength of 12.5-25 kV/cm. The critical strengths of the electrical field for the sterilization of Yakju were 7.5 kV/cm for total aerobes, 8.5 kV/cm for lactic acid bacteria and 6.5 kV/cm for yeasts. Logarithmic survival rates decreased linearly at low pulse number, but curvilinearly at high pulse number. The PEF sterilization kinetics of Yakju could be analysed by In s = In A-k In (n) and the sterilization rate constant increased with electric field strength and the size of target microorganisms. No changed in pH, acidity, and the growth of microorganisms were found in the PEF treated Yakju during the storage for 6 weeks at both $4^{\circ}C$ And $30^{\circ}C$.

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GIS Database and Google Map of the Population at Risk of Cholangiocarcinoma in Mueang Yang District, Nakhon Ratchasima Province of Thailand

  • Kaewpitoon, Soraya J;Rujirakul, Ratana;Joosiri, Apinya;Jantakate, Sirinun;Sangkudloa, Amnat;Kaewthani, Sarochinee;Chimplee, Kanokporn;Khemplila, Kritsakorn;Kaewpitoon, Natthawut
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1293-1297
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    • 2016
  • Cholangiocarcinoma (CCA) is a serious problem in Thailand, particularly in the northeastern and northern regions. Database of population at risk are need required for monitoring, surveillance, home health care, and home visit. Therefore, this study aimed to develop a geographic information system (GIS) database and Google map of the population at risk of CCA in Mueang Yang district, Nakhon Ratchasima province, northeastern Thailand during June to October 2015. Populations at risk were screened using the Korat CCA verbal screening test (KCVST). Software included Microsoft Excel, ArcGIS, and Google Maps. The secondary data included the point of villages, sub-district boundaries, district boundaries, point of hospital in Mueang Yang district, used for created the spatial databese. The populations at risk for CCA and opisthorchiasis were used to create an arttribute database. Data were tranfered to WGS84 UTM ZONE 48. After the conversion, all of the data were imported into Google Earth using online web pages www.earthpoint.us. Some 222 from a 4,800 population at risk for CCA constituted a high risk group. Geo-visual display available at following www.google.com/maps/d/u/0/edit?mid=zPxtcHv_iDLo.kvPpxl5mAs90&hl=th. Geo-visual display 5 layers including: layer 1, village location and number of the population at risk for CCA; layer 2, sub-district health promotion hospital in Mueang Yang district and number of opisthorchiasis; layer 3, sub-district district and the number of population at risk for CCA; layer 4, district hospital and the number of population at risk for CCA and number of opisthorchiasis; and layer 5, district and the number of population at risk for CCA and number of opisthorchiasis. This GIS database and Google map production process is suitable for further monitoring, surveillance, and home health care for CCA sufferers.

Feasibility Study of Deep Inspiration Breath-Hold Based Volumetric Modulated Arc Therapy for Locally Advanced Left Sided Breast Cancer Patients

  • Swamy, Shanmugam Thirumalai;Radha, Chandrasekaran Anu;Kathirvel, Murugesan;Arun, Gandhi;Subramanian, Shanmuga
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.9033-9038
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    • 2014
  • Background: The purpose of this study was to assess the feasibility of deep inspiration breath-hold (DIBH) based volumetric modulated arc therapy (VMAT) for locally advanced left sided breast cancer patients undergoing radical mastectomy. DIBH immobilizes the tumor bed providing dosimetric benefits over free breathing (FB). Materials and Methods: Ten left sided post mastectomy patients were immobilized in a supine position with both the arms lifted above the head on a hemi-body vaclock. Two thermoplastic masks were prepared for each patient, one for normal free breathing and a second made with breath-hold to maintain reproducibility. DIBH CT scans were performed in the prospective mode of the Varian real time position management (RPM) system. The planning target volume (PTV) included the left chest wall and supraclavicular nodes and PTV prescription dose was 5000cGy in 25 fractions. DIBH-3DCRT planning was performed with the single iso-centre technique using a 6MV photon beam and the field-in-field technique. VMAT plans for FB and DIBH contained two partial arcs ($179^{\circ}-300^{\circ}CCW/CW$). Dose volume histograms of PTV and OAR's were analyzed for DIBH-VMAT, FB-VMAT and DIBH-3DCRT. In DIBH mode daily orthogonal ($0^{\circ}$ and $90^{\circ}$) KV images were taken to determine the setup variability and weekly twice CBCT to verify gating threshold level reproducibility. Results: DIBH-VMAT reduced the lung and heart dose compared to FB-VMAT, while maintaining similar PTV coverage. The mean heart $V_{30Gy}$ was $2.3%{\pm}2.7$, $5.1%{\pm}3.2$ and $3.3%{\pm}7.2$ and for left lung $V_{20Gy}$ was $18.57%{\pm}2.9$, $21.7%{\pm}3.9$ and $23.5%{\pm}5.1$ for DIBH-VMAT, FB-VMAT and DIBH-3DCRT respectively. Conclusions: DIBH-VMAT significantly reduced the heart and lung dose for left side chest wall patients compared to FB-VMAT. PTV conformity index, homogeneity index, ipsilateral lung dose and heart dose were better for DIBH-VMAT compared to DIBH-3DCRT. However, contralateral lung and breast volumes exposed to low doses were increased with DIBH-VMAT.

MORPHOLOGICAL CHANGES OF DENTIN SURFACE TREATED WITH VARIOUS DENTIN SURFACE CONDITIONERS (수종(數種) 표면처리제(表面處理劑)에 의(依)한 상아질(象牙質) 표면(表面)의 형태(形態) 변화(變化)에 관(關)한 연구(硏究))

  • Cho, Jin-Ho;Choi, Ho-Young;Min, Byung-Soon;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.13 no.2
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    • pp.323-334
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    • 1988
  • The purpose of this study was to observe the effect of dentin surface conditioners on the dentin surfaces. Freshly extracted human molars were used in this study. They were stored at $4^{\circ}C$ saline solution before experiment. The crown portions of the teeth were cut in various directions by means of wet diamond point to expose dentin which include transverse, vertical oblique, horizontal and oblique cut to the long axis (Fig. 1). Each tooth was then mounted with self curing acrylic resin in brass ring to expose the flattened dentin surfaces. Final finish was accomplished by grinding the dentin specimens with wet No. 180 and No. 600 grit silicon carbide abrasive paper until a 6.0mm in diameter on a dentin surface was exposed without pulp exposure. The specimens were divided into 9 groups according to the modes of dentin treatment procedure. The following surface treatments were applied on these preparation surfaces; Group 1: unetched (control group) after finish with No. 600 silicon carbide abrasive paper. Group 2: etched with 30% phosphoric acid for 60s Group 3: etched with 10-3 solution for 60s Group 4: Cleaned with 5% NaOCl for 30s Group 5: applied Dentin Adhesit Group 6: cleaned with 5% NaOCl followed by applying the Dentin Adhesit$^{(R)}$ Group 7: applied Photo Bond on the unetched dentin followed by applying the Photo Clearfil Bright Group 8: Etched with 30% phosphoric acid followed by applying Photo Bond and Photo Clearfil Bright Group 9: etched with 10-3 solution followed by applying Photo Bond and Photo Clearfil Bright All the specimens were stored in $37^{\circ}C$ under 50% relative humidity for 24 hours before observations. The specimens in 7, 8, and 9 group, omitting the group 1 to 6, were demineralized in 10% HCl for 10s in order to observe the resin tags. All the specimens in each group were then dried at room temperature. The dried specimens were ion coated with Eiko ion coater (Eiko-engineering Co.), and observed in Hitachi S-430 Scanning electron microscope (Hitachi, Co. Tokyo) at 15KV. The following results were obtained as follows; 1. The smear layers were still remained in group 1,2,4,5, and 6. 2. There is no effect of 5% NaOCl and 30% phosphoric acid on the changes of dentin morphology 3. The dentin treated with 10-3 solution, indicating the tubules opened when the smear layer and the dental plug dissolved. 4. In case of applying the bonding agents the resin tag was not formed at the deep area of dentinal tubules, but in case of applying the Dentin Adhesit$^{(R)}$ that was not.

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Non-thermal Pasteurization of Carrot Juice by High Voltage Pulsed Electric Fields with Exponential Decay Pulse (고전압 Exponential Decay Pulse를 이용한 당근주스의 비열(非熱) 살균)

  • Ha, Koo-Yong;Shin, Jung-Kue;Lee, Seok-Hoon;Cho, Hyung-Yong;Pyun, Yu-Ryang
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1577-1582
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    • 1999
  • Carrot juice inoculated with $2\;{\times}\;10^8\;cfu/mL$ of Escherichia coli was treated with pulsed electric fields(PEF) for the purpose of a development of new cold pasteurization processes. Inactivation of E. coli in carrot juice increased with increase in intensity of the electric field strength and treatment time. The cells were suspended at concentration of ca. $2\;{\times}\;10^8$ cells per ml. A reduction of 4D was obtained at 40 kv/cm and 256 exponential decay pulses at room temperature. Critical electric field strength(Ec) and treatment time(tc) needed for inactivation of E. coli were 11.74 kV/cm and $3.6\;{\mu}s$ at room temperature, respectively. The combination of PEF and thermal treatment inactivated E. coli more effectively. The reductions of up to 5.5D were observed when the carrot juice was treated with PEF of 22.5 kV/cm and $205\;{\mu}s$ at $50^{\circ}C$. PFF treatment did not effect in color, pH, $^{\circ}Brix$, titratable acidity and ${\alpha}-,\;{\beta}-carotene$ contents of carrot juice.

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Fabrication of Electrospun Juniperus Chinensis Extracts loaded PVA Nanofibers (향나무 추출물을 함유하는 PVA 나노섬유 제조)

  • Kim, Jeong-Hwa;Lee, Jung-Soon
    • Science of Emotion and Sensibility
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    • v.19 no.2
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    • pp.35-42
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    • 2016
  • Electrospinning is a simple and effective process for producing nanofiber with diameter range from nanometers to micrometers which have high specific surface area. Hence, medicated nanofibers can be readily fabricated using a solution containing a mixture of a plant-extracts and a polymer. It has proved that Juniperus Chinensis can be effectively used for the prevention of UV and SLS-induced advers skin reaction such as radical production, inflammation and skin cell damage. It also found that Juniperus Chinensis has efficient ingredient of antifungal activity and house dust mite repellent effect. The fabrication of PVA nanofibers containing Juniperus Chinensis extracts by electrospinning has been studied. PVA/Juniperus Chinensis extracts composite nanofibers were produced at different Juniperus Chinensis concentrations (0.25, 0.5, 1.5 wt. %). The parameters of electrospinning including polymer contents, voltage and tip-to-collector distance (TCD) were optimized for fabrication process. The study show that 12 wt. % PVA, 10kV applied voltage and TCD 10~20 cm are the best condition to obtain uniform PVA/Juniperus Chinensis extracts composite nanofibers. Morphologies of the electrospun composite nanofiber were observed by using a field emission scanning electron microscope. It has been found that the average diameters of fibers increased by the adding of Juniperus Chinensis extracts. As the results, PVA/Juniperus Chinensis extracts composite nanofibers having a diameter in the range from 310~360 nm were successfully prepared via an electrospinning.

The agonistic action of URO-K10 on Kv7.4 and 7.5 channels is attenuated by co-expression of KCNE4 ancillary subunit

  • Lee, Jung Eun;Park, Christine Haewon;Kang, Hana;Ko, Juyeon;Cho, Suhan;Woo, JooHan;Chae, Mee Ree;Lee, Sung Won;Kim, Sung Joon;Kim, Jinsung;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.6
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    • pp.503-516
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    • 2020
  • KCNQ family constitutes slowly-activating potassium channels among voltage-gated potassium channel superfamily. Recent studies suggested that KCNQ4 and 5 channels are abundantly expressed in smooth muscle cells, especially in lower urinary tract including corpus cavernosum and that both channels can exert membrane stabilizing effect in the tissues. In this article, we examined the electrophysiological characteristics of overexpressed KCNQ4, 5 channels in HEK293 cells with recently developed KCNQ-specific agonist. With submicromolar EC50, the drug not only increased the open probability of KCNQ4 channel but also increased slope conductance of the channel. The overall effect of the drug in whole-cell configuration was to increase maximal whole-cell conductance, to prolongate the activation process, and left-shift of the activation curve. The agonistic action of the drug, however, was highly attenuated by the co-expression of one of the β ancillary subunits of KCNQ family, KCNE4. Strong in vitro interactions between KCNQ4, 5 and KCNE4 were found through Foster Resonance Energy Transfer and co-immunoprecipitation. Although the expression levels of both KCNQ4 and KCNE4 are high in mesenteric arterial smooth muscle cells, we found that 1 μM of the agonist was sufficient to almost completely relax phenylephrine-induced contraction of the muscle strip. Significant expression of KCNQ4 and KCNE4 in corpus cavernosum together with high tonic contractility of the tissue grants highly promising relaxational effect of the KCNQ-specific agonist in the tissue.

SCANNING ELECTRON MICROSCOPIC STUDY ON RESIN INFILTRATIONS AFTER DENTIN TREATMENTS (수종 상아질접착제의 상아질침투양상에 관한 주사현미경적 연구)

  • Kwon, Gyun-Won;Park, Sang-Jin;Min, Byung-Soon;Choi, Ho-Young;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.18 no.1
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    • pp.38-53
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    • 1993
  • The purpose of this study was to observe the resin infiltration pattern into dentin by various dentin bonding agents. Freshley extracted 36 sound human molars were used in this study. They were stored at $4^{\circ}C$ physiologic saline solution before experiment. All the teeth were cross-sectioned to expose dentin below about 3.0mm at the cusp tip and above 2.0mm at the cemento-enamel junction with Crystal Cutter (MC411 D, Maruto Co., Japan), and were made into specimens for this study (Fig. 1). The specimen experimental groups were divided into 9 groups by dentin surface treatment as following procedures: Group I: Treated with Gluma Cleanser followed by Gluma Primer and Sealer Group 2. Treated with Gluma Cleanser followed by Scotch bond 2 Adhesive Group 3: Treated with Gluma Cleanser followed by Tenure Solution A, Band Visar Seal Group 4: Treated with Scotchprep followed by Scotch bond 2 Adhesive Group 5: Treated with Scotchprep followed by Gluma Primer and Sealer Group 6: Treated with Scotch prep followed by Tenure Solution A, Band Visar Seal Group 7: Treated with Tenure Conditioner followed by Tenure Solution A, Band Visar Seal Group 8: Treated with Tenure Conditioner followed by Scotchbond 2 Adhesive Group 9: Treated with Tenure Conditioner followed by Gluma Primer and Sealer 27 specimens of 36 specimens were divided into 9 groups (Group 1-9), and were used for observation of resin tags. Remaining 9 specimens were divided into 3 groups (Group 1,4 and 7), and were used for observation of fractured dentin surfaces. Specimens to observe the resin tag were demineralized with 20% HCl for 14 hours, specimens to observe the fractured dentin surfaces were demineralized with 10% HCl for 3 minutes. All the specimens were gold-coated with Eiko ion coater (Eiko-engineering Co.), and observed under Scanning electron microscope (Hitachi S-2300) at 20 KV. The following results were obtained: 1. In group 1 treated with Gluma Cleanser, Gluma Primer, and Sealer, most resin tags were more than $100{\mu}m$. 2. In group 4 treated with Scotch prep and Scotchbond 2 Adhesive, most resin tags were about $10{\mu}m$. 3. In group 7 treated with Tenure conditioner, Tenure Solution A, B, and Visar Seal, most resin tags were about $10{\mu}m$ but occasionally resin tags were more than $100{\mu}m$. 4. In groups 2,3,5,6,8 and 9, the lengths of resin tags were inconsistent and the amount of resin tags were reduced.

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