The binding properties of oxomemazine to muscarinic receptors using the ability of oxomemazine to inhibit $[^3H]QNB$ binding in membrane fractions of rat cerebrum and guinea pig ventricle and ileum were investigated. $[^3H]QNB$ bound to a single class of muscarinic receptors with a dissociation constant of approximately 60 pM in three tissue preparations. Pirenzepine and oxomemazine inhibited $[^3H]QNB$ binding in cerebrum with a Hill coefficient lower than unity, and the inhibition data were best described by a two-site model. The relative densities of the high $(M_1)\;and\;low\;(M_2)$ affinity sites for pirenzepine were 60 and 40%, with corresponding Ki values of 16 and 431 nM, and those $(O_H\;and\;O_L)$ for oxomemazine 40 and 60%, with corresponding Ki values of 80 and 1350 nM. However, the inhibition data of both drugs vs $[^3H]QNB$ in ventricle and ileum appeared to obey the law of mass-action (Hill coefficient close to 1). The apparent Ki values of pirenzepine were 850 and 250 nM, and those of oxomemazine 1460 and 570 nM in ventricle and ileum, respectively. Thus, oxomemazine like pirenzepine has high affinity for cerebrum, moderate affinity for ileum and low affinity for ventricle. These results suggest that oxomemazine could recognize the muscarinic receptor subtypes with a high affinity for the $M_1$ sites.
The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.
To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the different fusion and activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM + 10% FBS in 5% $CO_2$ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept In frozen. From rabbits treated with FSH in 30% PVP solution and hCG, oocytes were surgically collected from oviducts at 14 h post-hCG injection and stripped off their cumulus cells by re-pipetting in a 300 IU hyaluronidase solution. Oocytes with an extruded first polar body and dense cytoplasm were enucleated by micromanipulation in Ham's F-10 medium+7.5 g/$m\ell$ cytochalasin B. Euncleation was confirmed under a fluorescence microscope after staining with 5 g/$m\ell$ bisbenzimide for 2 min. Each enucleated oocyte was injected with a fetal fibroblast into a perivitelline space. Reconstructed eggs were compared fusion rates either at 2.0 ㎸/cm or 1.6 ㎸/cm(60 sec, double pulses). After fusion, all eggs were activated with the combination of 5 M ionomycin (5 min) and 10 g/$m\ell$ cycloheximide (CHX, 3h), and cultured in CRlaa medium and transferred into TCM199+10% FBS on day 3. Although there was not significantly differ in fusion rate between treatments (60%, 2.0 ㎸/cm vs. 79.4%, 1.6 ㎸/cm), none of them in the eggs fused with 2.0 ㎸/cm developed to blastocyst. In comparison of development and chromosome status between different activation treatments (Group 1; 5 M ionomycin/10 g/$m\ell$ CHX, Group 2; 5 M ionomycin/5 g/$m\ell$ CHX + 2 mM DMAP after fusion with 1.6 ㎸/cm), there were not differ in cleavage and development rates (67.3% and 28.9% in Group 1; 67% and 33% in Group 2). All out of 8 embryos evaluated in Group 1 appeared a normal diploid chromosome sets and mean number of cells (Mean SEM) on day 4.5 of culture was 141.5 23.15 (n=8). It can be concluded that the use of cycloheximide has not happened in chromosome abnormalities, and fetal fibroblasts can be used for cloning in rabbit.
Ann Ji-Young;Sa Soo-Jin;Cao Yang;Lee Sang-Young;Cheon Hee-Tae;Yang Boo-Keun;Park Choon-Keun
Reproductive and Developmental Biology
/
v.30
no.2
/
pp.135-141
/
2006
The present study was conducted to investigate the effects of cumulus cells and porcine follicular fluid (pFF) on plasminogen activator (PA) activity and oocytes maturation in vitro in the pig. The cumulus-oocyte complexes (COCs) and denuded oocytes (DOs) were incubated in NCSU-23 medium with or without 10% pFF for 0, 24, or 48 hr. In the presence of cumulus cells, the proportions of oocytes matured to metaphase-II stage were significantly (P<0.05) higher in medium with pFF than without pFF (69.8 vs. 37.7%, respectively). When COCs and DOs were cultured in the presence of pFF, tissue-type PA (tPA), urokinase-type PA (uPA), and tPA-PA inhibitor (tPA-PAI) were observed in COCs, and PA activities were higher at 48 hr than 24 hr. When COCs and DOs were cultured in the absence of pFF, tPA and tPA-PAI were observed in COCs, and PA activities were increased as duration of culture increased. No PA activities were detected in DOs regardless of pFF supplementation. When porcine oocytes were cultured in the presence of pFF for 24 and 48 hrs, the activities of tPA-PAI, tPA, and uPA were observed in both COCs and DOs. In medium of absence of pFF, PA activities were observed in oocytes with cumulus cells only. On the other hand, three plasminogen-dependent lytic bands (tPA-PAI, tPA, and uPA) were observed in pFF cultures. Particularly uPA activity was higher than the other kinds of PA activity. When oocytes and cumulus cells were separated from porcine COCs at 0 hr of culture, tPA-PAI, tPA, and uPA were detected in cumulus cells at 48 hr of culture, but no PA activities were in DOs. The presence of pFF and cumulus cells in maturation medium stimulated not only nuclear and cytoplasmic maturation in porcine COCs, but also PA production by cumulus cells and COCs. It is possible that PAs produced by cumulus cells migrated through the gap junction between oocyte and cumulus cells. These results suggest that porcine oocytes have no ability to produce PA themselves.
Journal of the Korea Academia-Industrial cooperation Society
/
v.21
no.5
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pp.401-408
/
2020
This study was undertaken among students majoring in Emergency Medical Technology, to comprehend the knowledge, attitudes and preventive behaviors associated with MERS. This study further provides basic data for education related to MERS prevention. A total of 167 students were enrolled in the study. Evaluation of MERS education revealed a higher number of inexperienced subjects: 74 experienced(44.3%) vs. 93 inexperienced(55.7%). Assessing the attitude items for MERS revealed that "have to wear personal protective equipment when dealing with MERS patients" ranked highest (average, 4.54 ± 0.87), whereas "washed my hands more often than usual" ranked highest (average, 3.82 ± 1.06) in the prevention items for MERS. Analyzing the correlation between attitude towards MERS and preventive behavior revealed that "canceled or delayed meeting with friends, eating out, watching sports, shopping, etc." showed that, although negatively correlated, a greater attitude showed a higher level of prevention (r=-0.63, p<0.004). Taken together, results of this study indicate the necessity to develop an education program that improves the level of education and knowledge about MERS, and to impart this education systematically and repeatedly.
Journal of the Korea Academia-Industrial cooperation Society
/
v.21
no.5
/
pp.384-390
/
2020
Purpose: Organophosphate insecticide poisoning can have clinically fatal results. This study aimed to evaluate the relationship between the neutrophil/lymphocyte ratio (NLR) and the occurrence of death in patients with organophosphate insecticide poisoning. Methods: For this retrospective study, data on patients with organophosphate insecticide poisoning who visited the emergency room between January 2008 and November 2018 were collected. The NLR was measured at the time of arrival in the emergency room. The patients were divided into survival and death groups. Results: Overall, 150 patients were enrolled: 15 (10%) in the death group and 135 (90%) in the survival group. In the univariate analysis, the following variables were significantly different between the two groups: age, white blood cell count, amylase level, creatinine level, Acute Physiology And Chronic Health Evaluation (APACHE) II score, and NLR. In the logistic regression analysis of variables with significant differences in the univariate analysis, there were significant differences between the two groups with respect to age, APACHE II score, and NLR. The NLR was significantly higher in the death group than in the survival group (20.83 ± 22.24 vs. 7.38 ± 6.06, p=0.036). Conclusion: High NLR in patients with organophosphate insecticide poisoning may be useful in predicting mortality.
A valid food or dish frequency questionnaire needs to be developed in Korea for accurate dietary assessment because the dietary practices of Koreans are very different from those of other countries. This study was conducted to evaluate the accuracy of the newly developed, semi-quantitative, dish-based frequency questionnaire (Semi-DFQ) with 12-day dietary records (12-DRs) as a gold standard. The study subjects were 115 men and 173 women aged 30-65 years old. We calibrated the frequency, portion size and daily intake of 112 dish items reported in Semi-DFQ with those in 12-DRs by Spearman rank correlation coefficients (SCCs). The consumption frequency and portion size reported in Semi-DFQ were higher than those in 12-DRs. The SCCs for the consumption frequency of various dishes ranged from -0.07 (fried seaweed) to 0.70 (instant coffee), the portion size ranged from -0.09 (cold seaweed soup) to 0.68 (soju), and the daily intake ranged from -0.07 (fried seaweed) to 0.71 (soju). The SCCs were higher for dishes consumed daily, such as steamed rice,milk, coffee and alcohol, than those of foods eaten rarely. The overall agreements between the Semi-DFQ and 12-DRs were low for categories of consumption frequency and portion size, even though some dishes showed high SCCs. The SCCs of the two methods in consumption frequency and amount were higher among the women and younger subject. The results revealed the limitation of the Semi-DFQ for evaluating the status of usual individual intake. Therefore, the Semi-DFQ can be used in addition as dietary records and 24-hour recall depending on the research aims.
Park, Chan Woo;Hwang, Yu Im;Koo, Hwa Seon;Kang, Inn Soo;Yang, Kwang Moon;Song, In Ok
Clinical and Experimental Reproductive Medicine
/
v.41
no.4
/
pp.158-164
/
2014
Objective: To assess whether an early GnRH antagonist start leads to better follicular synchronization and an improved clinical pregnancy rate (CPR). Methods: A retrospective cohort study. A total of 218 infertile women who underwent IVF between January 2011 and February 2013. The initial cohort (Cohort I) that underwent IVF between January 2011 and March 2012 included a total of 68 attempted IVF cycles. Thirty-four cycles were treated with the conventional GnRH antagonist protocol, and 34 cycles with an early GnRH antagonist start protocol. The second cohort (Cohort II) that underwent IVF between June 2012 and February 2013 included a total of 150 embryo-transfer (ET) cycles. Forty-three cycles were treated with the conventional GnRH antagonist protocol, 34 cycles with the modified early GnRH antagonist start protocol using highly purified human menopause gonadotropin and an addition of GnRH agonist to the luteal phase support, and 73 cycles with the GnRH agonist long protocol. Results: The analysis of Cohort I showed that the number of mature oocytes retrieved was significantly higher in the early GnRH antagonist start cycles than in the conventional antagonist cycles (11.9 vs. 8.2, p=0.04). The analysis of Cohort II revealed higher but non-significant CPR/ET in the modified early GnRH antagonist start cycles (41.2%) than in the conventional antagonist cycles (30.2%), which was comparable to that of the GnRH agonist long protocol cycles (39.7%). Conclusion: The modified early antagonist start protocol may improve the mature oocyte yield, possibly via enhanced follicular synchronization, while resulting in superior CPR as compared to the conventional antagonist protocol, which needs to be studied further in prospective randomized controlled trials.
Hayirli, Armagan;Esenbuga, N.;Macit, M.;Lacin, E.;Karaoglu, M.;Karaca, H.;Yildiz, L.
Asian-Australasian Journal of Animal Sciences
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v.18
no.9
/
pp.1310-1319
/
2005
This experiment was conducted to determine the effects of cage density (CD) and humate supplementation (HS) on laying performance, metabolic profile, and egg quality during the peak production period in hens. Lohman layers (n = 180, 46 wks of age) were blocked according to the location of cages and then allocated randomly to two levels of CD (4 or 6 hens per cage or 540 vs. 360 $cm^2$/hen) and three levels of HS (0, 0.15, and 0.30%). Egg production (EP) and feed consumption (FC) were measured daily; egg weight was measured bi-weekly; and BW was measured before and after the experiment. Blood and additional egg samples were obtained at the end of the experiment for determination of metabolic profile and egg quality. The data were analyzed using two-way ANOVA as repeated measures. Except for FC, CD did not affect laying performance parameters. Hens placed in high-density cages had lower FC than hens placed in normal-density cages. Increasing HS level linearly increased FC, EP, and feed conversion ratio (FCR). There was a CD by HS interaction effect on FC and EP. Hens placed in high-density cages had greater serum glucose, total protein, albumin, globulin, Ca, and P concentrations and tended to have greater serum corticosterone concentration than hens placed in normaldensity cages. Increasing HS level linearly increased serum glucose, total protein, albumin, globulin, creatine, and Ca concentrations and linearly decreased serum triglyceride and very low-density lipoprotein concentrations. There was a CD by HS interaction effect on serum glucose and albumin concentrations. There were no alterations in egg quality parameters in response to increasing CD. Albumen index and Haugh unit decreased linearly and other egg quality parameters did not change as HS level increased. In conclusion, increased caging density adversely affected metabolic profile, despite insignificantly deteriorating laying performance. Moreover, benefits from humate supplementation seem to be more noteworthy for hens housed in stressing conditions than for hens housed in standard conditions.
Boyd, R.D.;Touchette, K.J.;Castro, G.C.;Johnston, M.E.;Lee, K.U.;Han, In K.
Asian-Australasian Journal of Animal Sciences
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v.13
no.11
/
pp.1638-1652
/
2000
Prolific females require better nutrition and feeding practice because of larger litter size and the substantial decline in body fat. Life-time pig output will be compromised if body protein and fat are not properly managed. First litter females are especially vulnerable because they can loose ${\geq}15%$ of whole-body protein. Conservation of body protein mass during first lactation minimizes wean to estrus interval and increases second litter size (up to 1.2 pigs). The ability to influence litter-size by amino acid nutrition is a new dimension in our understanding. A P2 fat depth below 12 mm at farrow and below 10 mm at wean compromised wean to estrus interval (>2 d) and next litter size (0.5 to 1.5 pigs) in sows. It is now clear that a 'modest' excess of feed during the first 72 h of pregnancy decreases embryo viability so that the potential for an increased litter size at birth is not realized. The capacity for milk production by prolific young sows is 25% higher than the standard used previously (NRC, 1988). First litter females averaged 9.82 kg milk/d for a 21 d lactation. Second and third litter counterparts averaged 10.35 kg/d. Milk production was 95% of peak by 10 d of lactation and sows were in greatest negative energy and lysine balance during the first 6 d. Nearly 45% of the total loss in body protein occurred within the first 6 d, but this could reduced to 30-35% by using a more aggressive feeding strategy after parturition. There appear to be 2 phases in lactation for lysine need (d 2-12 vs 12-21). Feeding to the higher level alleviates the second litter size decline. The lysine requirement for lactation can be predicted with accuracy, but we are not able to predict the second limiting amino acid. Mammary uptake of valine relative to lysine and recent work with practical diets suggest that the recent NRC (1998) pattern is realistic and that threonine and valine could be co-limiting for corn-soy diets for prolific sows nursing 10-11 pigs. Empirical studies are needed to refine the ideal pattern so that synthetic lysine can be used with more confidence. Milk fat output for the elite sow is extraordinary and poses an unnecessarily high energetic cost. Methods that reduce mammary fat synthesis will benefit the sow and may enhance piglet growth.
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