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Effect of TNF-$\alpha$ Gene Transfer to Respiratory Cancer Cell Lines on Sensitivity to Anticancer drugs (호흡기계암세포주에서 TNF-$\alpha$ 유전자의 이입이 항암제 감수성에 미치는 효과)

  • Mo, Eun-Kyung;Lee, Jae-Ho;Lee, Kye-Young;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Choi, Hyung-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.3
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    • pp.302-313
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    • 1995
  • Background: Tumor necrosis factor(TNF) showed antitumor cytolytic effects on sensitive tumor cells in numerous in vivo and in vitro studies. But it could not be administered systemically to human because of severe systemic adverse effects at effective concentrations against tumor cells. Many studies showed that a high concentrations of TNF in the local milieu may evoke in vivo TNF-responsive mechanisms sufficient to suppress tumor growth. Recently developed technique of TNF gene transfer to tumor cells using retrovirus vector could be a good candidate for local TNF administration. TNF is also known to synergistically enhance in vitro cytotoxicity of chemotherapeutic drugs targeted to DNA topoisomerase II against TNF-sensitive tumor cell lines. In this study the in vitro chemosensitivity against DNA topoisomerase II targeted chemotherapeutic drugs was evaluated using some respiratory cancer cell lines to which TNF gene had been transferred. Method: NCI-H2058, a human mesothelioma cell line, A549, a human lung adenocarcinoma cell line and WEHI 164 cell line, a murine fibrosarcoma cell line were treated with etoposide and doxorubicin, which are typical topoisomerase II - targeted chemotherapeutic agents, at different concentration. The resultant cytotoxicity was measured by MIT assay. Then the cytotoxicity of the same chemotherapeutic agents was measured after TNF-$\alpha$ gene-transfer and the two results were compared. Results: The cytotoxicity was not increased significantly in WEHI164 cell line and A549 cell line but statistically significant increase was observed in H2058 cell line when TNF-$\alpha$ gene was transferred(p<0.05). Conclusion: These findings show that TNF-$\alpha$ gene transfer to respiratory cancer cell lines results in variable effects on chemosensitivity against topoisomerase II inhibitor among different cell lines in vitro and can be additively cytotoxic in certain selective tumor cell lines.

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Optimum Condition of Soil Dispersion for Remediating Heavy Metal-Contaminated Soils using Wet Magnetic Separation (중금속 오염 토양 정화를 위한 습식자력선별법 사용 시 최적 토양분산 조건)

  • Chon, Chul-Min;Park, Jeong-Sik;Park, Sook-Hyun;Kim, Jae-Gon;Nam, In-Hyun
    • Economic and Environmental Geology
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    • v.45 no.2
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    • pp.121-135
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    • 2012
  • Soil dispersion and heavy metal leaching with two heavy metal-contaminated soils were studied to derive the optimal dispersion condition in the course of developing the remedial technology using magnetic separation. The dispersion solutions of pyrophosphate, hexametaphosphate, orthophosphate and sodium dodecylsulfate (SDS) at 1 - 200 mM and the pH of solutions was adjusted to be 9 - 12 with NaOH. The clay content of suspension as an indicator of dispersion rate and the heavy metal concentration of the solution were tested at the different pHs and concentrations of the dispersion solution during the experiment. The dispersion rate increased with increasing the pH and dispersion agent concentration of the solution. The dispersion efficiency of the agents showed as follows: pyrophosphate > hexametaphosphate > SDS > orthophosphate. Arsenic leaching was sharply increased at 50 mM of phosphates and 100 mM of SDS. The adsorption of $OH^-$, phosphates and dodecysulfate on the surface of Fe- and Mn-oxides and soil organic matter and the broken edge of clay mineral might decrease the surface charge and might increase the repulsion force among soil particles. The competition between arsenic and $OH^-$, phosphates and dodecylsulfate for the adsorption site of soil particles might induce the arsenic leaching. The dispersion and heavy metal leaching data indicate that pH 11 and 10 mM pyrophosphate is the optimum dispersion solution for maximizing dispersion and minimizing heavy metal leaching.

Comparative Evaluation of Colon Cancer Stemness and Chemoresistance in Optimally Constituted HCT-8 cell-based Spheroids (적정 구성 배양 HCT-8 기반 대장암 스페로이드의 암 줄기세포능 및 항암제 내성 평가의 비교 평가 연구)

  • Lee, Seung Joon;Kim, Hyoung-Kab;Lee, Hyang Burm;Moon, Yuseok
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1313-1319
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    • 2016
  • Cancer is a complex disease heterogeneously composed of various types of cells including cancer stem-like cells responsible for relapse and chemoresistance in the tumor microenvironment. The conventional two-dimensional cell culture-based platform has critical limitations for representing the heterogeneity of cancer cells in the three-dimensional tumor niche in vivo. To overcome this insufficiency, three-dimensional cell culture methods in a scaffold-dependent or -free physical environment have been developed. In this study, we improved and simplified the HCT-8 colon cancer cell-based spheroid culture protocol and evaluated the relationship between cancer stemness and responses of chemosensitivity to 5- Fluorouracil (5-FU), a representative anticancer agent against colon cancer. Supplementation with defined growth factors in the medium and the culture dish of the regular surface with low attachment were required for the formation of constant-sized spheroids containing $CD44^+$ and $CD133^+$ colon cancer stem cells. The chemo-sensitivities of $CD44^+$ cancer stem cells in the spheroids were much lower than those of $CD44^-$ non-stem-like cancer cells, indicating that the chemoresistance to 5-FU is due to the stemness of colon cancer cells. Taken together, the inflammation and oncogenic gut environment-sensitive HCT-8 cell-based colon cancer spheroid culture and comparative evaluation using the simplified model would be an efficient and applicable way to estimate colon cancer stemness and pharmaceutical response to anticancer drugs in the realistic tumor niche.

Survey the Occurrences and Establishment of Environment-friendly Control System of Ricania shanthungensis in Jeonnam Province (전남지역 갈색날개매미충 발생현황과 친환경 방제)

  • Choi, Duck-Soo;Ma, Kyung-Cheol;Kim, Hyo-Jeong;Lee, Jin-Hee;Oh, Sang-A;Kim, Seon-Gon
    • Korean Journal of Organic Agriculture
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    • v.26 no.3
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    • pp.439-452
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    • 2018
  • This study was carried out to establish of environment-friendly control system and to survey the occurrence status of Ricania shanthungensis in Jeonnam province from 2016 to 2017. R. shanthungensis occurred at 1,344 ha in 61 towns of 15 cities in Gwangju and Jeonnam province. At four sites in the stationary area, the densities of eggmass and pupae were lower in 2017 than in 2016, the hatching rate decreased, and the first hatching time, emergence of adults, and the start of spawning tended to be slower. Telenomus sp., an egg parasitoid of R. shanthungensis, was found in most areas. Sunchon had the highest rate of 46.1%, followed by Muan 40.6, Goksung 29.2, Gurye 25.8, and Gwangju 17, respectively. The optimum spraying time for the control of R. shanthungensis was early March. The spraying material and the dilution multiples are 10 times of the machine oil at environmental friendly cultivation and 500 times of the chlorpriphosphate wp. at normal cultivation. When sprayed these materials, 95% of egg prohibited to hatch. There was more than 80% of the insecticidal effects on the organic materials of the machine oil emulsion, the Sophora root extrect + microbial extract agent, and the castor emulsion. For adults, Sophora root extrect + microbial extract, neem extract, and machine oil were better. Four kinds of chemical pesticides such as dinotefuran wp were effective for nymph and adult control. We have developed an adult catching device using the most preferred daylight color and behavioral habits of R. shanthungensis. The capture device consisted of two daylight compact lamps (30W and 20W), a yellow plate, and a catcher using water, and caught about 700 individuals a day. Based on the above results, we have established a system for controlling and life cycle of R. shanthungensis in Jeonnam province.

Toxicity Assessment and Establishment Acceptable Daily Intake of Penthiopyrad (펜티오피라드(Penthiopyrad)의 독성평가와 일일섭취허용량 설정)

  • Jeong, Mi-Hye;Hong, Soon-Sung;Park, Kyung-Hun;Kim, Chan-Sub;Park, Jae-Eup;Hong, Moo-Ki;Im, Moo-Hyeog;Kim, Young-Bum;Han, Bum-Seok;Han, Jeung-Sul
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.478-489
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    • 2010
  • Penthiopyrad is a fungicide agent in types of pyrazole which is showing the effect of prevention in fungal disease and powdery mildew. In order to register this new pesticide, reports of acute toxicity and chronic toxicity by animal study were examined to set acceptable daily intake to evaluate hazards of consumers. Acute toxicity was low in toxic, and it did not have the effect of acute dermal toxicity, acute eye irritation, or skin sensitization. As the result of the study in chronic toxicity, the common effect of chemical appeared in the liver and thyroid which was proven as a toxic effect. Two-generation reproduction toxicity, genotoxicity, and prenatal development toxicity were not proven. As the result of carcinogenic study, increase of thyiroid follicular adenoma in the rat and the frequency of liver hepatocellar adenoma in mice were also increased. However, it was decided that the threshold value on the effect in chemicals could be controlled through study liver enzyme induction. Therefore, the ADI for penthiopyrad is 0.081 mg/kg/ bw/day, based on the NOAEL of 8.10 mg/kg bw/day of twelve-months dogs study and applying an uncertainty factor of 100.

Phytic Acid Protects the Formation of Colonic Aberrant Crypt Foci Induced by Azoxymethane in Male F344 Rats (랫드에서 azoxymethane으로 유도된 대장 전암병변에 대한 피티산의 방어 효과)

  • Hue, Jin-Joo;Lee, Yea-Eun;Lee, Ki-Nam;Nam, Sang-Yoon;Ahn, Byeong-Woo;Yun, Young-Won;Lee, Beom-Jun
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.264-270
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    • 2008
  • Phytic acid(PA) (Inositol hexaphosphate, $IP_6$) is a naturally occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plants and mammalian cells. Recently PA has received much attention for its role in anticancer activity. In the present study, the preventive effects of PA on colon carcinogenesis were investigated. Six-week old Fisher 344 male rats were fed a AIN-93G purified diet and PA(0.5% or 2% PA in water) for 8 weeks. The animals received two ($1^{st}\;and\;2^{nd}$ week) injections of azoxymethane(AOM, 15 mg/kg b.w.) to induce colonic aberrant crypt foci(ACF). After sacrifice, the total numbers of aberrant crypts(AC) and ACF in colonic mucosa were examined after staining with methylene blue. Blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. AOM induced the total numbers of $142.3{\pm}22.3$ ACF/colon and $336.6{\pm}55.1$ AC/colon. PA at the doses of 0.5 and 2% decreased the numbers of ACF and AC/colon in a dose-dependent manner. The numbers of ACF/colon and AC/colon by PA at the dose of 0.5% were $124.4{\pm}28.5\;and\;302.7{\pm}67.3$, respectively. PA at the dose of 2% significantly decreased the ACF and AC numbers to $109{\pm}18.1\;and\;254.8{\pm}50.6$, respectively(p<0.01). Especially, 2% PA significantly reduced the number of large ACF(${\geq}4$ AC/ACF) from $26.8{\pm}6.2$ ACF/colon to $15{\pm}6.7$ ACF/colon(p<0.01). Although some parameters in blood counts and serum chemistry were changed compared with the control, no specific toxicity was found. These findings suggest that phytic acid can be a chemopreventive agent for colon carcinogenesis resulting from inhibition of the development of ACF in the F344 rat.

Detection of Human Papillomavirus and Expression of p53, c-erbB-2 Protein in Inverted Papilloma of the Nasal Cavity and Paranasal Sinuses (비강 및 부비동의 반전성 유두종에서 인유두종바이러스검출과 p53및 c-erbB-2의 발현)

  • Cho Jae-Shik;Baik Joon;Lim Sang-Chul;Cho Yeon;Yoon Je-Hwan;Seo Duk-Jung;Park Chang-Soo
    • Korean Journal of Head & Neck Oncology
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    • v.17 no.2
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    • pp.162-168
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    • 2001
  • Background: Inverted papilloma(IP) of the nasal cavity and paranasal sinuses is a benign neoplastic condition that can be associated with squamous cell carcinoma (SCC). Several studies have indicated an etiologic role for viruses in the development of inverted papilloma. And it is necessary to find out the significance of a biologic markers such as p53, c-erbB-2 to predict the malignant potential. The purposes of this study are to detect HPV in inverted papilloma of the nasal cavity and paranasal sinus, to examine role of HPV as an etiological agent, to examine the relationship between HPV subtype and malignant transformation of inverted papilloma, and to investigate the relation between expression rate of p53, c-erbB-2 and HPV in recurrent or malignant transformation cases. Material and Methods: Thirty two cases of inverted papilloma(IP) in the nasal cavity and paranasal sinuses were reviewed and classified into 3 groups; simple IP, IP with dysplasia group, IP with squamous cell carcinoma group. Paraffin embedded achival tissue was used in this study. The HPV was detected by in situ hybridzation (ISH) using HPV type 6/11, 16/18, 31/33/35 DNA probes. Expression of p53 and c-erbB-2 was examined by immunohistochemical staining. Results: 1) The HPV was detected in 6(19%) out of 32 cases. 2) The HPV 6/11 was dectected in 4 out of 21 cases of simple IP, HPV 16/18 in 1, HPV 31/33/35 in lout of 8 cases of IP with dysplasia respectively. 3) The positive expression of p53 was 13 cases out of 32 cases; 2 out of 21 cases of simple IP, all of 8 cases of IP with dysplasia and 3 cases of IP with squamous cell carcinoma 4) The positive expression of c-erbB-2 was in 24 out of 32 cases; 16 out of 21 cases of simple IP, 6 out of 8 cases of IP with dysplasia, 2 out of 3 cases of IP with squamous cell ca. 5) The recurrence of IP occurred in lout of 6 cases of positive for HPV, in 4 out of 26 cases negative for HPV. 6) The recurrence of IP occurred only in positive cases for p53. 7) The recurrence of IP occurred in 4(17%) out of 24 cases positive for c-erbB-2, in 1(13%) out of 8 cases negative for c-erbB-2. Conclusion: The p53 expression was associated with Inverted papillomas exhibiting evidence of malignant transformation. Also, there was a correlation between the p53 expression and recurrence.

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The Therapeutic Effect of Lactobacillus reuteri in Acute Diarrhea in Infants and Toddlers (영유아의 급성 설사에서 Lactobacillus reuteri의 치료 효과)

  • Eom, Tae-Hun;Oh, Eun-Young;Kim, Young-Hoon;Lee, Hyun-Seung;Jang, Pil Sang;Kim, Dong-un;Kim, Jin-Tack;Lee, Byung-Churl
    • Clinical and Experimental Pediatrics
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    • v.48 no.9
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    • pp.986-990
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    • 2005
  • Purpose : Certain strains of lactobacilli are known to accelerate recovery from acute diarrhea. Lactobacillus reuteri is isolated from human breast milk and a commonly occurring Lactobacillus species with therapeutic potential in acute diarrhea. The purpose of the present study was to investigate the therapeutic effect of L. reuteri in acute diarrhea in young children. Methods : Fifty patients between 6 and 36 months of age hospitalized with acute diarrhea (rotavirus in 40 percent) were randomized into two groups to receive either $10^8$ colony-forming units of L. reuteri or a matching placebo, twice a day for their length of hospitalization, or for up to 5 days. Antidiarrheal drugs were not prescribed to either group. The clinical outcome of diarrhea was evaluated. Results : The mean duration of watery diarrhea after initiation of treatment was 2.3 days for the L. leuteri group(n=25) vs. 2.9 days for the placebo group(n=25)(P=0.072). By the second day of treatment, watery diarrhea persisted in 64 percent of patients receiving L. reuteri, compared to 84 percent of those receiving placebo(P=0.006). On the second day, the mean frequency of watery diarrhea was 1.9 in the L. leuteri group and 3.4 in the placebo(P=0.046). Also, vomiting continued to the second day in 16 percent of patients receiving L. reuteri and 40 percent of those recieving placebo(P=0.031). Conclusion : L. reuteri is effective as a therapeutic agent in acute diarrhea in children.

The Effects of Dimethyl Sulfoxide and Sodium Thiosulfate for the Prevention of Tissue Necrosis due to Extravasaion of Mitomycin-C (혈관외로 유출된 Mitomycin-C에 의한 조직괴사 예방을 위한 Dimethyl Sulfoxide와 Sodium Thiosulfate의 효과)

  • Woo, Sang-Hyun;Choi, Byung-Cheol;Kim, Ki-Hyung;Seul, Jung-Hyun;Jung, Tae-Eun
    • Journal of Yeungnam Medical Science
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    • v.13 no.2
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    • pp.243-250
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    • 1996
  • Extravasation of toxic chemotherapeutic agents cause severe skin ulceration and necrosis which often need secondary surgical intervention. Still, there were not established antidote agent in case of extravasation with mitomycin-c. Dimethyl sulfoxide is known as an effective chemical scavenger of toxic hydroxyl free radical and sodium thiosulfate also was demonstrated significant protector from mitomycin-c induced ulceration by a few experimental studies. Author investigated necrotic area of mitomycin-c injected site and compare to the effectiveness of topical treatment with dimethyl sulfoxide and intradermal injection of sodium thiosulfate according to starting times, forty five mice were divided into 3 groups. Control group(n=5) had no treatment after subcutaneous injection of mitomycin-c. Experimental group I and II were 20 mice treated dimethyl sulfoxide and sodium thiosulfate, respectively. Depending on the starting time of treatment, group I and II were subdivided into 1, 2, 3 and 4 as immediate, 6 hours, 12 hours and 24 hours after mitomycin-c injection. Histologic studies of the necrotic area and survival area after treatment were performed using hematoxylin-eosin staining. The mean necrotic area of group I was significantly decreased depending on the starting time of treatment compared with control group(p<0.01). The results means there was no necrosis area which was treated with topical sodium thiosulfate within 6 hours, and it showed also significant decrease of necrosis area within 24 hours. There was also no necrosis area in group II-1 and significant decrease of necrosis area II-2 and III-3. But, effctiveness of intradermal injection of sodium thiosulfate was not found in group II-4 which was started after 24 hours. Hisotolgic findings showed a bland coagulative necrosis without inflammatory changes and no granulation tissue. The significant difference that cytoplasmic loss of subcutaneous fat and decrease number of hair follicles between two groups resulted from the methods of treatment by topical application and intradermal injection. In conclusion, immediate treatments with topical dimethyl sulfoxide or intradermal injection of sodium thiosulfate signifcantly prevents necrosis by extravasation of mitomycin-c.

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Studies on Adrenoceptors Involved in Regulation of Sodium Transport in Frog Skin (개구리 피부에 있어서 Na 수송을 조절하는 Adrenoceptors에 관한 연구)

  • Choi Bong-Kyu;Kim Kyung-Keun;Kim Heung-Kyu;Kook Young-Johng
    • The Korean Journal of Pharmacology
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    • v.22 no.1 s.38
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    • pp.24-33
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    • 1986
  • To ascertain the existence of various adrenoceptors involved in active transport of sodium in the frog skin and to delineate their physiological roles, the influence of various adrenergic agonists and antagonists on the potential difference (PD), short-circuit current (SCC) and total skin conductance (TSC) of the isolated frog skin of Rana nigromaculata were investigated. PD and SCC were determined with Ussing's technique. Drugs were administered to the serosal side of the skin. Experimental results were summarized as follows: 1. The responses to norepinephrine (NE, $6{\times}10^{-8}-6{\times}10^{-5})M$), phenylephrine (PE, $5{\times}10^{-6}-5{\times}10^{-4}M$) and epinephrine (Epi, $5.5{\times}10^{-7}-5.5{\times}10^{-5}M$) were characterized by marked elevation of PD & SCC in dose-related fashion, but the maximal effect attained by Epi was less than those of NE and PE. 2. These increments of PD & SCC were significantly inhibited by prazosin $(2{\times}10^{-6}M)$, a speciflc ${\alpha}_1$-adrenoceptor blocker. The stimulatory effect on PD & SCC were completely abolished by phenoxybenzamine (PBZ, $3.3{\times}10^{-5}M$), an irreversible ${\alpha}$-adrenoceptor blocking agent. Furthermore, with a larger doses of Epi produced marked decline of PD & SCC after the PBZ pretreatment. 3. Isoproterenol (ISP), a ${\beta}$-adrenoceptor agonist, in concentrations ranging from $5{\times}10^{-7}$ to $5{\times}10^{-6}M$ produced dose-related decrease in PD & SCC, which could be abolished by pretreatment with propranolol $(4{\times}10^{-6}M)$, a specific ${\beta}$-adrenoceptor blocker. It was further noted that the effects of Epi on PD & SCC were markedly potentiated by Propranolol pretreatment. 4. Clonidine as well as guanabenz produced increases in PD & SCC and these effects were inhibited more specifically by prazosin pretreatment than by yohimbine. These results indicated that there exist in the frog skin two distinctive types of adrenoceptors, ${\alpha}$ and ${\beta}$, which roughly corresponds to those in mammals, and that the ${\alpha}$ type of adrenoceptors mediate the stimulation of PD & SCC, whereas ${\beta}$-adrenoceptors mediate the inhibition. However, based on evidence at hand, no conclusion could be drawn on the subtype of ${\alpha}$-adrenoceptors which is involved in the stimulation of sodium transport in the frog skin.

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