This experiment was conducted to identify the enzyme treatment time, calcium ion effect, enzyme concentration and leaf position for protoplast isolation. It was also performed to determine the adequate molarity on protoplasts, and to investigate the incubation time, pH, PEG concentration and DMSO effect for protoplast fusion. The results obtained were summarized as follows ; The optimal time of incubation in enzyme solution was 4 hours. And the protoplast releasing time was delayed by $CaCl_2{\cdot}2H_2O$ addition to the enzyme solution compared with no added one. The viability had kept up to above 95% until the 4 hours after digestion. The high viability of the protoplast was preserved more than 16 hours by adding $CaCl_2{\cdot}2H_2O$ to digestion solution. The enzyme concentration had no effect on protoplast yield in range from 1% to 5% and the first or second leaf from the top of the plant produced the highest protoplast yield among the leaf position tested. The purity of healthy protoplast was better in 0.4M and 0.5M sucrose than in others, and the percentage of protoplast aggregation was more 20% to 50% in PEG 6,000 compared with 4,000 and PEG 1,500. Even though the percentage of protoplast aggregation was less increased by 3% to 7% than without DMSO, its treatment was effective to induce binucleated protoplasts.
Purpose: This study assessed the food intake and nutritional status of the elderly in long-term care facilities in order to provide adequate food services and improve the nutritional status. Methods: The survey was carried out from August 2019 to October 2019 for the elderly in long-term care facilities located in Gwangju Metropolitan City. The survey was conducted to collect data from 199 elderly persons (34 males and 165 females) aged over 65 years old. The food intake was assessed using a 1-day 24-hour recall method. Results: More than 90% of the subjects were over 75 years old. Forty five percent of the subjects were active, 44.2% of the subjects perceived themselves as not being healthy. Dementia and Parkinson's disease were the most common diseases, followed by hypertension, musculo-skeletal disease, diabetes, and stroke. Only 25.6% of the subjects had most of their teeth intact, and 44.7% of the subjects had difficulty in chewing and swallowing. The total food intake was 1,127 g in males and 1,078 g in females. The most frequently consumed foods were kimchi, cooked rice with multi-grains, soybean soup, cooked rice with white rice, yogurt, pumpkin porridge, soy milk, and duck soup. The average energy intake of the subjects was 1,564.9 kcal in males and 1,535.5 kcal in females. The overall nutritional status of the elderly in the long-term care facilities was poor. In particular, the intake of vitamin D and calcium, vitamin C, riboflavin, and potassium were very low. The intake of vitamin D was 5 ㎍, and 86.4% of the elderly were below the estimated average requirement, while the intake of sodium was high. Conclusion: The results of this study can be used to understand the health and nutritional status and to improve the food services and nutrition management for the elderly in longterm care facilities.
The objective of this study was to investigate the effects of nutrition education based on ubiquitous healthcare (u-health) service on changes in dietary habits, nutrition intake, and risk factors for metabolic syndrome in male workers. In total, 72 male office workers with at least three risk factors of the National Cholesterol Education Program-Adult Treatment Panel III were recruited as subjects. Anthropometric measurements and biochemical analyses were conducted on all subjects. Dietary habits and nutrient intake were determined by a questionnaire using the 24-hour dietary recall method before and after nutrition education. Subjects measured their body composition, blood pressure, and physical activity more than once per week during the 12 weeks using the u-health care equipment and sent these data to a central database system using a personal computer. Individual nutrition counseling was provided four times on the first, fourth, eight, and twelfth weeks. The results showed significant decreases in abdominal circumference, body fat (%), diastolic blood pressure, serum triglycerides, and serum total cholesterol during the 12 weeks. Subjects with more than 12 measurements showed significant reductions in abdominal circumference, body fat (%), serum triglyceride, and serum total cholesterol. Mean intake of dietary fiber, animal calcium, potassium, vitamin C, and folic acid after nutrition education were higher than those before nutrition education. Participants showed significant increases in the frequencies of consuming protein foods (meat, fish, eggs, beans, tofu, etc.) and vegetables. In conclusion, nutrition education through the u-health service resulted in positive effects on the risk factors for metabolic syndrome, nutrient intake, and dietary habits.
The objective of this study was to establish a good methodology to isolate single smooth muscle cells that are alive and respond properly to pharmacological agents. Canine urinary bladders were employed as the source of single cells, and acetylcholine, atropine and imipramine were used as indicators of pharmacological responsiveness. Imipramine, an antidepressant drug exhibited the anticholinergic and calcium antagonizing properties on rat detrusor muscle. To establish a control value for a further experiment to elucidate the mechanism of action of imipramine on detrusor muscle, we measured the concentration-response of single cells to acetylcholine in the presesnce of imipramine by length of the cells and compared the result with the response in the presence of atropine. Tiny chops of smooth muscle taken from anesthetized canine urinary bladder were incubated in collagenase solution at $36^{\circ}C$ for 17-20 minutes. The collagenase solution included collagenase 1.2 mg/ml, soybean tryspin inhibitor 0.08 mg/ml, bovine serum albumin 2% in 10 ml Krebs-Henseleit buffer solution aerated with a consistent breeze of 95/5% $O_2/CO_2$, to maintain the pH at 7.4. After washing with plain K-H solution on 450 mesh, cells were dissociated from the digested tissue for 12-15 minutes. Cell suspension was transfered in 5 ml test tubes and acetylcholine was added for the final concentration to be $10^{-14}M{\sim}10^{-9}M$. To find the optimal time to fix the cells to determine the contractile responses, 1% acrolein was added 5, 10, 20, 30, 60 and 120 seconds after the administration of ACh. The length of cells fixed by acrolein were measured by microscaler via CCTV camera on phaes-contrast microscope. The average length of 50 cells from a slide glass was taken as the value of a sample at the very concentration point. Single cells were isolated from canine detrusor. The length of untreated cells varied from 82 ${\mu}m$ to 94 ${\mu}m$. The maximal response to actylcholine $10^{-9}M$ was accomplished within 5 seconds of exposure, and the shortening was $19{\pm}3$%. Atropine reduced the contraction of the cells concentration-dependently. Imipramine which exerts a cholinergic blocking action on some smooth muscles also reduced the contraction concentration-dependently and by a similar pattern as atropine. These findings document that imipramine may exerts a cholinergic blocking activity in the single smooth muscle cells isolated from canine urinary bladder.
Kang, Hyeok Joong;Piao, Min Yu;Lee, In Kyu;Kim, Hyun Jin;Gu, Min Jeong;Yun, Cheol-Heui;Seo, Jagyeom;Baik, Myunggi
Asian-Australasian Journal of Animal Sciences
/
v.30
no.4
/
pp.505-513
/
2017
Objective: This study was performed to evaluate whether ambient temperature and dietary glycerol addition affect growth performance, and blood metabolic and immunological parameters, in beef cattle. Methods: Twenty Korean cattle steers ($405.1{\pm}7.11kg$ of body weight [BW], $14.2{\pm}0.15$ months of age) were divided into a conventional control diet group (n = 10) and a 2% glycerol- added group (n = 10). Steers were fed 1.6% BW of a concentrate diet and 0.75% BW of a timothy hay diet for 8 weeks (4 weeks from July 28th to August 26th and 4 weeks from August 27th to September 26th). Blood was collected four times on July 28th, August 11th, August 27th, and September 26th. Results: The maximum indoor ambient temperature-humidity index in August (75.8) was higher (p<0.001) than that in September (70.0), and in August was within the mild heat stress (HS) category range previously reported for dairy cattle. The average daily gain (ADG; p = 0.03) and feed efficiency (p<0.001) were higher in hotter August than in September. Glycerol addition did not affect ADG and feed efficiency. Neither month nor glycerol addition affected blood concentrations of cortisol, triglyceride, or non-esterified fatty acid. Blood concentrations of cholesterol, low-density lipoprotein, high-density lipoprotein, glucose, and albumin were lower (p<0.05) on August 27th than on September 26 th, and blood phosphorus, calcium and magnesium concentrations were also lower on August 27th than on September 27th. Glycerol addition did not affect these blood parameters. Percentages of $CD4^+$ T cells and $CD8^+$ T cells were higher (p<0.05) on July 28th than on August 27th and September 26th. The blood $CD8^+$ T cell population was lower in the glycerol supplemented-group compared to the control group on July 28th and August 27th. Conclusion: Korean cattle may not be significantly affected by mild HS, considering that growth performance of cattle was better in hotter conditions, although some changes in blood metabolic and mineral parameters were observed.
Kim, Sun Young;Yi, Ho Keun;Lee, Jung Chang;Hwang, Dong Jin;Hwang, Pyoung Han;Lee, Dae Yeol;Cho, Soo Chul
Clinical and Experimental Pediatrics
/
v.46
no.6
/
pp.541-547
/
2003
Purpose : Gap junction intercellular communication(GJIC) is an important mechanism of the bystander effect in herpes simplex thymidine kinase/ganciclovir(HSVtk/GCV) gene therapy Therefore, we attempted to enhance the bystander effect in vitro by exogenous overexpressing connexin 37(Cx37) in cells to increase GJIC. Methods : NIH3T3 cells were transfected with the Cx37 and HSVtk gene or the HSVtk gene alone by the calcium phosphate method, and we detected their expression from these cells by RT-PCR. GCV-mediated cytotoxicity and the bystander effect of each transfectant was then assessed and compared. Results : Cells transfected with HSVtk became sensitive to low concentration of GCV. We found significantly increased cytotoxicity in HSVtk/GCV gene therapy after introduction of the HSVtk and Cx37 genes together compared with the cytotoxicity seen after introduction of the HSVtk gene in vitro. Co-expression of the HSVtk and Cx37 genes potentiates HSVtk/GCV gene therapy through the bystander effect. Conclusion : These results indicated that the increase of GJIC using Cx37 have potentiated the bystander effect of HSVtk/GCV therapy, and may be a new approach to improve response in suicidal cancer gene therapy.
Purpose:The pathophysiology of magnesium, the second highest common compound in humans, is still unclear, especially in preterm babies. We accessed the association between total magnesium (tMg), ionized Mg (iMg), and gestational age (GA) and that between serum magnesium (sMg) and intraventricular hemorrhage (IVH) in preterm babies. Methods:In all, 119 inborn preterm infants admitted between July 2006 and February 2008 were divided into the IVH group (19) and the control group (100) and were prospectively analyzed. TMg, iMg, pH, total calcium (tCa), and ionized Ca (iCa) levels were determined immediately after delivery or within 3 hours after birth, and their correlation with GA were investigated. Results:TMg was not correlated with GA, tCa, iCa, and pH. IMg was correlated with tMg (r=0.288, P=0.002) and iCa (r=0.212, P=0.021); however, it was not correlated with GA and pH. Mean GA and birth weight were significantly lower (P=0.002) and smaller (P=0.030) in the IVH group. Mean sMg was higher in the IVH group ($2.5{\pm}0.9mg/dL$) than in the control group ($2.1{\pm}0.6mg/dL$) (P=0.021). SMg was a risk factor even after logistic regression analysis (OR, 2.798; 95% C.I., 1.265-6.192; P=0.011). Conclusion:In less than 37-week-old preterm babies, tMg and iMg were similar, regardless of GA. High sMg may be a risk factor for IVH in premature babies, regardless of their exposure to antenatal magnesium.
Glutamate and aspartate may evoke an increase in membrane permeability to monovalent cations and $Ca^{++}$. However, it is uncertain whether $Ca^{++}$ influx is mediated by voltage dependent $Ca^{++}$ channels or by excitatory amino acid activated channels. In addition, the influences of excitatory amino acids on $Ca^{++}$ uptake by neuronal tissues as well as the responses of their actions to extracellular $Mg^{++}$ concentration are different. $K^{+}$ induced $Ca^{++}$ uptake by synaptosomes was dependent on extracellular $Mg^{++}$ up to 5 mM and at concentration of 10 mM, $Ca^{++}$ influx was rather reduced. In $Na^{+}$ rich media, glutamate-and aspartate-induced $Ca^{++}$ uptake was increased by $Mg^{++}$ in a dose independent manner. However, the response for NMDA was inhibited by $Mg^{++}$ at concentrations above 2 mM. $K^+$-and glutamate-induced $Ca^{++}$ influx s were inhibited by 2,4-dinitrophenol, chlorprom-azine and verapamil but not by tetraethylammonium chloride. Tetrodotoxin effectively inhibited the action of glutamate but did not affect that of $K^+$. The response for MNDA was inhibited by 2, 4-dinitrophenol and tetrodotoxin, slightly inhibited by verapamil, and not affected by tetraethylammonium chloride. In $Na^{++}$ rich medium, depolarizing action of glutamate, aspartate and MNDA on synaptosomes was not demonstrated, whereas these agents stimulated $Ca^{++}$ uptake and caused $Ca^{++}$ influx induced depolarization at mitochondria. On the other hand, the activities of synaptosomal ATPases were not affected by excitatory amino acids at 5 mM. The results suggest that glutamate or NMDA induced $Ca^{++}$ influx at synaptosomes exhibits different responses for extracellular $Mg^{++}$ Ex citatory amino acids induced $Ca^{++}$ influx at synaptosomes may be associated with increased permeability of membrane for $Na^{++}$ and $Ca^{++}$ except $K^{++}$ and membrane depolarization due to increased ionic permeability.
The effects of external $Ca^{2+}$ and $Ca^{2+}-antagonists$ on the spontaneous contractions and electrical activities were investigated in guinea-pig stomach in order to clarify the mechanism for the generation of slow waves. Electrical responses of circular smooth muscle cells were recorded using glass capillary microelectrodes filled with 3 M KCl. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows: 1) The amplitude of spontaneous contractions was maximal at around 2-4 mM $Ca^{2+}$, whereas their frequency was inversely related with external $Ca^{2+}$ within the range of 0.5 to 16 mM $Ca^{2+}$. 2) Verapamil suppressed the amplitude of spontaneous contraction in a dose-dependent manner, while the frequency of spontaneous contractions was almost not changed over the whole concentration of verapamil $(0.01{\sim}5\;mg/l)$. 3) Manganese increased both the amplitude and the frequency of spontaneous contractions dose-dependently in low $Mn^{2+}$ (below 0.05 mM $Mn^{2+}$), while their amplitude and frequency were decreased in high $Mn^{2+}$ (above 0.1 mM $Mn^{2+}$). 4) The ampltude and maximum rate of rise of slow waves were incrased in high $Ca^{2+}$ solution. In $Ca^{2+}-free$ solution, the spontaneous contractions recorded simultaneously with slow waves ceased and tonic contraction ($Ca^{2+}-free$ contracture) was developed in parallel with membrane depolarization and the disappearance of slow waves. 5) Verapamil (1 mg/1) decreased the amplitude and maximum rate of rise of slow waves and it depolarized the membrane by about 6 mV, whereas the frequency of slow waves was not affected by verapamil. 6) Manganese showed different characteristic effects between low and high $Mn^{2+}$ on the slow waves: In low $Mn^{2+}$ (0.05 mM $Mn^{2+}$), the initial rapid increases and the subsequent gradual decreases in three parameters of slow waves (amplitude, rate of rise, and frequency of slow waves) till a new steady state were observed. However, in high $Mn^{2+}$ (0.5 mM $Mn^{2+}$) slow waves disappeared and membrane was depolarized. From the above results, the following conclusions could be made: 1) $Ca^{2+}$ is necessary for a generation of the slow waves, even though it is small amount. 2) Verapamil suppresses the spontaneous contractions of gastric antral strip by the decreases in amplitude and maximum rate of rise of slow waves, while this drug does not block the $Ca^{2+}-channel$ involved in the generation of slow waves. 3) Manganese has dual actions on the $Ca^{2+}-channels$; the $Ca^{2+}-channel$ involved in the generation of slow waves (or Na-Ca exchange system) or the channel for the generation of spike potentials are stimulated by a low concentration of $Mn^{2+}$, while both the $Ca^{2+}$. Channels are blocked by high concentration of $Mn^{2+}$.
Purpose: The aim of this study was to explore the relationship of ready-to-eat cereal (RTEC) consumption with nutrition and health status. Examination of health status for this project included obesity, abdominal obesity, hypertension, hypertriglyceridemia, hypercholesterolemia, low-HDL-cholesterolemia, diabetes, anemia, and metabolic syndrome. Methods: Two groups, RTEC consumers and those who did not consume RTEC, were identified using 24-hour dietary recall data from the 2012 Korea National Health and Nutrition Examination Survey (KNHANES). Nutritional intakes and risk factors of the two groups were compared using covariates-adjusted statistical procedures. Statistical analyses were performed using SAS survey procedures, and strata, cluster, and weight were considered. Subjects of analysis of nutritional intake were between the ages of 1 and 75, and those considered in the risk factor analysis were between the ages of 19 and 75. Results: Results showed that 3.8% of the Korean population was RTEC consumers. Compared to the subjects who did not intake RTEC, RTEC consumers exhibited significantly higher intakes of calcium, thiamin, riboflavin, and vitamin C. It was also discovered that the percentage of people whose intakes were less than EAR decreased with RTEC consumption. RTEC consumption showed significant association with decreased systolic blood pressure, diastolic blood pressure, serum triglyceride, and serum total cholesterol. Consequently, prevalence of hypertension among RTEC consumers was significantly lower than that among non-consumers, and the odds ratio for hypertension was 0.19 after adjusting the models for covariates. Conclusion: Results of this study clearly suggest an association of RTEC consumption with improved nutritional status and cardiometabolic risk profile in Korean adults. Conduct of additional studies will be necessary in order to determine the nature of these relationships.
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