• Title/Summary/Keyword: Saline Solutions

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Evaluation of time-dependent antimicrobial effect of sodium dichloroisocyanurate (NaDCC) on Enterococcus faecalis in the root canal (이염화이소시아뉼산나트륨 제재의 근관 내 사용 시 시간에 따른 E. faecalis에 대한 항균성 평가)

  • Kim, Hye-Jeong;Park, Se-Hee;Cho, Kyung-Mo;Kim, Jin-Woo
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.121-129
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    • 2007
  • The purpose of this study was to assess the antibacterial effect of sodium dichloroisocyanurate (NaDCC), sodium hypochlorite (NaOCl), and chlorhexidine (CHX) on Enterococcus faecalis and to evaluate and to compare the time-dependant antimicrobial effect of NaDCC with NaOCl and CHX in the root canal in vitro before and after instrumentation. Extracted Human single teeth were prepared by serial instrumentation technique. The samples were autoclaved and contaminated for 3 days with E. faecalis monocultures. The teeth were then divided into 4 groups Each group was irrigated and inserted with 2% NaOCl, 2% NaDCC, 2% CHX and steri)ized saline. After 6, 12, 24, 72h, and 1 week incubation, sterilized paper point was inserted into the root canal. Paper points containing root canal contents were then placed on the agar plate. And then each root cana) was prepared with #4 and #5 GG (Gates-Glidden) drill. The debris were collected in the sterilized microtube and the plates were incubated at $37^{\circ}C$ in an increased $CO_2$ atmosphere. After 24h incubation the growth of bacteria around the paper points were measured. NaOCl and NaDCC solution shows similar antimicrobial effect for E. faecalis at 6, 12, 24, 72h and 1 week. In centrol group, irrigated with sterilized saline, no antimicrobial effect was observed. The results are in agreement with other investigators, who have shown the bactericidal property and possibility of NaDCC as a root canal irrigation solution. Thus it seems that NaDCC solutions can be clinically applied into the root canal within 1 week after dilution.

EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Effects of Green Tea Extract on Intestinal Mucosal Esterification of $^{14}C$-Oleic Acid in Rats (녹차 추출물이 흰쥐 소장세포의 지방 에스테르화 과정에 미치는 영향)

  • Seo, Yun-Jung;Noh, Sang-K.
    • Food Science and Preservation
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    • v.15 no.3
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    • pp.450-455
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    • 2008
  • Previously, we have shown that green tea extract lowers the intestinal absorption of cholesterol, fat, and other fat-soluble compounds. We conducted this study to determine whether green tea extract affects the rate of $^{14}C$-oleic acid esterification into various lipids in the intestinal mucosa of rats. Male Sprague-Dawley ruts were had free access to a nutritionally adequate AIN-93G diet and deionized water. Initially, the rat's mucosal content of total lipids was measured following 1 mL olive oil administration with (green tea group) or without (control group) 100 mg green tea extract powder. At 1 h and 5 h, intestinal segments were extracted for total lipid analysis. Secondly, to measure mucosal esterification rates of lipids, an abdominal incision was made along the midline, and a 10-cm long jejunal segment of the small intestine was ligated in situ. Then, micellar solutions with or without green tea extract were injected into the ligated jejunal segments and incubated for 10 mill. The micellar solution contained $200.0\;{\mu}$ Ci $^{14}C$-oleic acid, $200.1\;{\mu}mol$ unlabelled oleic acid, $66.7\;{\mu}mol$ 2-monooleoylglycerol, $66.7\;{\mu}mol$ palmitoyl-sn-glycero-3-phosphocholine, 2.2 mmol glucose, $50.0\;{\mu}mol$ albumin, and 16.5 mmol Na-taurocholate per L of phosphate buffered saline (pH, 6.3) with or without 8.87 g green tea extract powder. At 10 min, each rat was sacrificed by cervical dislocation under anesthesia and the segment was removed for lipid analysis. Significant differences were observed in mucosal triglyceride content at 1 h and 5 h in ruts given green tea extract. Significant differences in the rate of $^{14}C$-oleic acid esterification into triglycerides and phospholipids fractions were observed between control and green tea groups. However, There were no significant differences in other lipid fractions. These results indicate that the lowered esterification rates of $^{14}C$-oleic acid into triglycerides and phospholipids fractions is attributable to presence of green tea extract. This may be associated with an inhibitory effect of green tea catechin on the mucosal processes of lipids, leading to the inhibition of intestinal absorption of lipids.

On the Applicability of Partially Purified Antigenic Preparations of Paragonimus westermani (부분정제 폐흡충 항원의 유용성 검토)

  • 김석일;강신영조승열
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.257-264
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    • 1983
  • In order to obtain more specific antigenic preparation for the diagnosis of human paragonlmiasis, crude saline extract of whole worm (=PwWWE), secretory.excretory components (PwSEC) and secretion-excretion-free somatic extract (PwSM) of 12 week-old ParagoninBus westermani were filtrated through Sephadex G-200 gel column. The adult Paragonimus worms were obtained from expevimentally infected doge. A total of 11 antigenic solutions was Iyophilised or diluted to adjust protein content of 1mg/ml. To evaluate the antigenicity of crude antigens and fractions, micro-ELISA was done with the sera from P westermani in(ected cases, C. sinensis infected cases and non-infected control cases to detect Paragonimus specific IgG antibody. The results were as follows: 1. When the PwWWE was filtrated through Sephadex G-200 gel, it was separated into three fractions; PwWWE Fr. 1, PwWWE Fr. 2 and PwWWE Fr. 3. The percentage of protein content was 28.0%, 21.6% and 50.4% respectively. The PwSM was also. separated into three fractions; PwSM Fr. 1, PwSM Fr. 2, PwSM Fr. 3. and their percentage of protein content was 41.3%, 38.6% and 20.1%. However, the PwSEC showed different fractionation pattern; i.e. fraction 1 (=PwSEC Fr.1) and 3 (PwSEC Fr. 3) without fraction 2. The percentage of protein content was 14.0% in PwSEC Fr. 1 and 86.0% in PwSEC Fr. 3. 2. When the antigenicity of each Paragonimus crude antigen and fractionated antigen was evaluated for specific IgG aritibody by micro-ELISA in 10 human paragonimiasis sera, PwSEC Fr, 1 was the most potent antigen showing the mean absorbance 1.98. The PwWWE Fr. 1, PwSEC, PwWWE were next to that: their mean absorbance were 1.72, 1.38 and 0.83 respectively. The antigenicity of fractions 2 and 3 was much weaker in binding specific IgG antibody. 3. When the antigens were reacted in micro-ELISA with 10 human clonorchiasis sera, most antigens showed weak reactivity. Each fraction 1 of crude antigens reacted higher than other fractions or crude antigens; the mean absorbance was 0.17 in fraction 1, but in others the absorbances were about 0.06. 4. With non-infected control sera, the result of micro-RLISA revealed almost same pattern with those of the clonorchiasis sera. From the above results, it became apparent that PwWWE Fr. 1, especially PwSEC Fr. 1 was the most potent antigen reacted with Paragonisfaus specific IgG antibody.

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Effects of Various Nucleotides on the Membrane Permeability (Nucleotides가 세포막 투과도에 미치는 영향)

  • Lee, Joong-Woo;Jeong, Seong-Woo
    • The Korean Journal of Physiology
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    • v.23 no.1
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    • pp.13-21
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    • 1989
  • The present study was designed to investigate i) the action of various nucleotides on membrane permeability of rat red blood cell and hepatocyte for $Na^{+}$ and $Rb^{+}$ ii) the characteristics of purinoceptors on these cell membranes. Blood from Sprague-Dawley rats was obtained by carotid arterial cannulation. Red blood cells were then washed 3 times with saline at $4{\circ}C$. Hepatic parenchymal cells were isolated from rat livers by using a modification of the Berry and Friend (1969) method. For the $Na^{+}$ influx studies, isolated RBC and hepatocyte were incubated in incubation medium containing $^{22}Na^{+}0.2\;{\mu}Ci/ml$ at $37^{\circ}C$. After various time intervals samples were removed from the incubation flask and washed out 3 times with ice-cold washing solutions. Cells were destroyed by adding Triton X-100 and TCA solution. After centrifugation, the supernatants were assayed for $^{22}Na^{+}$ by gamma counter. $^{86}Rb^{+}$ was used to simulate $K^{+}$ in these $K^{+}efflux$ studies. Isolated hepatocytes were incubated for 60 min in the loading solution containing $^{86}Rb^{+}\;10\;{\mu}Ci/ml$ at $37^{\circ}C$. After loading, the cells washed out 3 times by centrifugation with washing solution. The cells were incubated in buffer solution at $37^{\circ}C$. At intervals thereafter, samples were removed and centrifuged. The supernatants were analyzed for $^{86}Rb^{+}$ by liquid scintillation counter. The main results of the experiments were: 1) ATP and ATPP increased in both $^{22}Na^{+}$ influx and $^{86}Rb^{+}$ efflux in the red blood cell. Although ADP showed a tendency to increase in RBC membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$, the changes were not significantly different from the control. 2) The Significant changes in $^{22}Na^{+}$ and $^{86}Rb^{+}$ flux by ATP were also demonstrated in hepatocyte. ATPP and ADP showed a tendency to increase in hepatocyte membrane permeability for both ions. 3) Other nucleoside triphosphates-ITP, GTP and CTP-did not change in membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$ in RBC and hepatocyte. In conclusion, not only ATP but also ATPP activate purinoceptors and change in membrane permeability for $Na^{+}$ and $K^{+}$. In order to activate purinoceptors on the cell membrane, the nucleotides have to possess intact adenine moiety and three phosphates or more in its molecule.

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Administration Duration Dependent Effects of Epimedium Herb Extract Solution on the Reproductive Capacities in the Mice (음양곽(淫羊藿)의 수컷 생쥐 생식능력(生殖能力)에 대한 투약기간별(投藥期間別) 효과(效果))

  • Lee, Chang-Hoon;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub;Choi, Jung-Eun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.3
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    • pp.55-68
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    • 2006
  • Purpose : This study was undertaken to evaluate the effects of the different administration duration of Epimedium Herb extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase. Methods : We used the 2-month-old mice and administered the extract solution of Epimedium Herb 0.3 ml/g/day for 30, 60, 90 and 120 days. The control group was administered the normal saline as the same way. We examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase. Also we observed changes of isolated testis before and after administration of Epimedium Herb extract solutions in the mice. And we compared the testicular tissue especially seminiferous tubules with the control and treated group by histochemical methods. Results : The significant differences were observed in the concentration of total sperm and normality of spermatozoa of the Epimedium Herb extract solution administered groups compared to the control group in 60, 90 and 120 days groups. The significant differences were observed the motility of the Epimedium Herb extract solution administered groups compared to the control group in 60 days group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Epimedium Herb extract solution administered groups compared to the control group, respectively. And the activity of hyaluronidase was significantly increased in the Epimedium Herb extract solution administered groups compared to the control group. Conclusion : This study shows that Epimedium Herb has the beneficial effect on the concentration, morphology and motility of sperm, the testicular tissues and the activities of sperm hyaluronidse in 60 days administration group. We can suggest that Epimedium Herb extract solution be useful for the treatment of male sexual dysfunction and infertility.

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MANAGEMENT OF ORAL MUCOSITIS OWING TO CHEMICAL BURN BY INTOXICATION OF AGRICULTURAL CHEMICALS(GRAMOXON) : REPORT OF CASES (농약(Gramoxon)중독에 의한 화상으로 발생된 구강점막염 치험)

  • Yoo, Jae-Ha;Kang, Sang-Hoon;Kim, Hyun-Sil;Baek, Sang-Hum;You, Tae-Min;Lee, Ji-Woong;Chung, Won-Gyun;Kim, Jong-Bae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.2
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    • pp.123-127
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    • 2003
  • Chemical burns onto oral mucosa which are infrequent, may result from contact with a wide variety of chemical agents. The degree of injury depends on the chemical, its concentration, duration of contact, and the natural penetrability and resistance of the tissues involved. Chemicals do not usually "burn" in that they do not cause destruction by hyperthermic activity. Rather, they damage tissue by causing coagulation of protein by one of several processes, reduction, oxidation, desiccation, corrosion, or vesication. Paraquat(Gramoxon) is the most frequently agricultural chemicals that induce the severe toxic reactions onto the organs of human body in Korea. The toxic reaction are composed of pulmonary edema and fibrosis, formation of hyaline membrane, inflammatory reaction and bleeding tendency, owing to the cell damage by the production of superoxide radicals. The contents of essential treatment in paraquat intoxication are commonly airway and breathing maintenance, gastric lavage, much hydration and diuresis, hemoperfusion and medications for the removal of the chemicals and the prevention of various complications. The sedative oral dressings, such as, orabase ointment application, warm saline gargling, lidocaine viscous gargling and oral gargling by the mixed solutions(tetracycline, prednisolone and 10% dextrose water) are important for the improvement of chemical oral mucositis and the comfortable feeding of diet. The authors managed properly two cases of oral chemical mucositis that were occurred by the incorrect use of agricultural chemicals(paraquat) and report the cases with the review of literatures about care of the chemical intoxication and oral mucositis.

Optimization of Nanoencapsulation Process for Azelaic Acid-Milk Nano Powder and Acne Nanocosmetics (Azelaic Acid 함유 밀크 나노분말과 여드름 나노화장품을 위한 나노캡슐의 최적화 공정)

  • Kim, Dong-Myong;Choi, Ji-Eun;Kim, Duck-Hoon;Lee, Jun-Tack
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.1
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    • pp.43-53
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    • 2011
  • The conditions in fluid-bed processor for nanoencapsulation of azelaic acid-milk nano powder for acne nanocosmetics were optimized by response surface methodology (RSM). The maximum value of yield was 70.97 %. The yield was appreciably influenced by inlet air temperature, atomizing pressure, and feeding speed. The particle size increased with an increase in the feeding speed and a decrease in the atomizing pressure. The elution rate in saline solutions was appreciably influenced by inlet air temperature and atomizing pressure. The moisture content increased with higher atomizing pressure, which was demonstrated to be similar to the nanoencapsulation characteristics related to water activity. The Hunter's L and b values increased with an increase in the inlet air temperature. The optimum conditions estimated by RSM for the maximized values of yield, moisture content, particle size and elution rate in skin suitability were $67{\sim}73^{\circ}C$ of inlet air temperature, 0.6 ~ 0.8 mL/min feeding speed and 1.8 ~ 2.0 kg/$cm^2$ of atomizing pressure, respectively. These estimated values were in agreement with those measured by real experiments.

Salinity Tolerance of Blackgram and Mungbean: II. Mineral Ions Accumulation in Different Plant Parts

  • Karim, M.A.;Raptan, P.K.;Hamid, A.;Khaliq, Q.A.;Solaiman, A.R.M.;Ahmed, J.U.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.5
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    • pp.387-394
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    • 2001
  • Blackgram (Vigna mungo) is more salt tolerant than mungbean (Vigna radiata). This study was initiated to know whether the accumulation pattern of mineral ions in different plant parts plays a significant role in the differences in salt tolerance between the two Vigna species. Different mineral ions, viz. N, Cl, Na, K, Mg and Ca in different organs of two varieties of each of blackgram- Barimash-l (susceptible one) and Barimash-2 (tolerant one), and mungbean-Barimung-3 (tolerant one) and Barimung-4(susceptible one), were analyzed after growing with 0, 50, 75 and 100 mM NaCl solutions. The two crops showed a decreased but similar pattern of total N accumulation under saline conditions. The tolerant variety of both the crops showed a less reduction in total N than the susceptible one. Leaves showed the maximum while stem the minimum N, irrespective of levels of salinity. C $l^{[-10]}$ and N $a^{+}$ accumulation increased with the increasing salinity levels. Interestingly, similar to a halophyte, the salt tolerant blackgram exhibited conspicuously higher amount of N $a^{+}$ in the shoot than the salt-susceptible mungbean. However, the tolerant varieties showed less amount of N $a^{+}$ than the susceptible one, especially in blackgram. Seeds of both Vigna spp. accumulated the minimum amount of N $a^{+}$ than other plant parts. $K^{+}$ accumulation decreased by salinity in most of the plant parts, except seeds. Blackgram showed larger reduction in K than mungbean. The $Mg^{++}$ increased in leaves, petioles and stem by salinity while decreased in the roots, podshells and seeds in both the crops. Salinity increased $Ca^{++}$ accumulation in all plant-parts except roots of both Vigna spp. Apparently, the leaves of mungbean accumulated higher concentration of $Ca^{++}$ than blackgram. Varietal differences in the accumulation pattern of $K^{+}$, $Mg^{++}$ and $Ca^{++}$ were not clear. It was concluded that blackgram, presumably, possesses a similar salt tolerance mechanism to halophyte, and the pattern of accumulation of mineral ions in blackgram and mungbean was not fully ascribed to the differences in salinity tolerance between the two Vigna species.gna species.ies.s.ies.

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Contribution of Bulk Flow to Transport Mechanisms of the Membranes Surrounding Amniotic Fluid in the Rabbit

  • Lim, Young-Cheol;Lee, Sang-Jin;Sung, Ho-Kyung
    • The Korean Journal of Physiology
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    • v.28 no.1
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    • pp.79-90
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    • 1994
  • The objective of the present study is to assess the contribution of bulk flow to the regulatory mechanism of amniotic fluid volume and its ionic concentration in the membranes surrounding the amniotic fluid. For quantitative assessment, we prepared 4 kinds of artificial amniotic fIuids (isotonic isovolumetric, hypotonic isovolumetric, isotonic hypervolumetric and hypotonic hypervolumetric ones) by replacing 70% of amniotic fluid of pregnant rabbits with water or normal Tyrode solutions. Isoosmotic saline of 0.5 ml volume containing 0.05% Censored and 15 mM/l LiCl was administered initially into amniotic sacs of all subject animals. Samples of amniotic fluid were collected in after 30 and 90 minute intervals; the concentrations of Censored, $Na^+\;and\;Li^+$ were determined and compared. Followings are the results obtained. 1. from isovolumetric and increased Congcord group, we couldn't find significant change in $Li^+\;and\;Na^+$ concentration in isotonic amniotic fluid. However, $Na^+$ concentration increased significantly as well as a striking increase in Censored concentration in hypotonic amniotic fluid. 2. In isovoIumetric and decreased Censored group, the rate of $[Li^+]$ decrement and the rate of $[Na^+]$ increment were much higher in hypotonic amniotic fluid than in isotonic. 3. In hypervolumetric and increased Censored group, the rate of $Na^+$ efflux increased proportionately with the increment of Censored concentration up to 0.98, which was higher than the rate of $Li^+$ efflux in isotonic amniotic fluid. However, the increment of $Na^+$ concentration was rather related with the initial $Na^+$ concentration in hypotonic amniotic fluid, showing inverse relationship. $Li^+$ concentration increased only when there was a marked increase in Censored concentration and approached near a maximum value or 1. 4. For hypervolumetric and decreased Censored group, the observations were identical to isovolumetric and decreased Censored group. From these results the following conclusions could be made: 1) There is no net movement of water or monovalent cations across the membranes surrounding amniotic fIuid in isotonic isovolumetric condition. In contrast, there is a net efflux of amniotic fluid by osmotic bulk flow, resulting in elevation of $Na^+$ concentration in hypotonic isovolumetric condition. 2) In hypervolumetric conditions, there is a massive efflux of amniotic fluid or solvent drag through the surrounding membranes by fiItrative bulk flow, where the rate of $Na^+$ efflux has a linear relationship with that of water efflux. This is assumed to be carried out through enlarged and newly opened intercellular spaces resulting from increased intraamniotic pressure. 3) Once increasing intraamniotic pressure reaches a point allowing $Li^+$ to pass through during osmotic bulk flow in hypotonic amniotic fIuid, $Na^+$ influx seems to occur by diffusion simultaneously or immediately thereafter, too.

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