In order to measure cellular physiological activity including ion channel activity, protoplasts were isolated from the root tissue of tomato plant. The general methods recommended were not efficient enough to make protoplasts from the root tissue. Among various conditions tested, we found that a two-step treatment of osmosis is very efficient for the isolation of protoplasts. In this procedure, root tissues were preincubated in a solution containing 300 mM sorbitol for 30 min. Then, they moved to the reaction solution containing 700 mM sorbitol as well as cell wall-digesting enzymes. The formation of protoplast was greatly increased by this method. In order to find the optimal condition of the two-step method, various conditions of pH, osmotic pressure, incubation time, and the concentrations of cell wall-digesting enzymes were tested. The yield of protoplast isolation was maximal at pH 5.0 after 2 hr incubation. Mixed enzymes of 3% cellulase, 1 % macerozyme, and 0.1 % pectolyase showed maximal protoplast isolation. The physiological activity of isolated protoplast evaluated by measuring the cellular ATPase activity was as high as that measured from the preparation of root tissue. The protoplasts isolated by this method were remained healthy up to 4 hrs which is enough time to measure the cellular physiological activity. These results show that the two-step treatment of osmotic pressure was successful to obtain high yield of healthy protoplast from tomato root tissue.
Shin, Narae;Xu, Dayuan;Song, Jun Kyung;Park, Jaebum
Korean Journal of Applied Biomechanics
/
v.29
no.3
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pp.157-166
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2019
Objective: This study examined the effects of stimulating fingertip temperature on the patterns of force sharing and stability properties during multi-finger force production tasks. Method: 9 adult subjects (male: 3, female: 6, age: $26.11{\pm}4.01yrs$, height: $169.22{\pm}5.97cm$, weight: $61.44{\pm}11.27kg$) participated in this study. The experiment consisted of three blocks: 1) maximal voluntary contraction (MVC) task, 2) single-finger ramp task to quantify enslaving (i.e., unintended force production by non-task fingers), and 3) 12 trials of multi-finger steady-state force production task at 20% MVC. There were three temperature conditions including body-temperature (i.e., control condition), $40^{\circ}C$, and $43^{\circ}C$, and the stimulation was given to the index finger only for all experimental conditions. Results: There were no significant differences in the MVC forces, enslaving, and the accuracy of performance during the steady-state task between the conditions. However, the share of stimulated index finger force increased with the index fingertip temperature, while the share of middle finger force decreased. Also, the coefficient of variation of both index and middle finger forces over repetitive trials increased with the index fingertip temperature. Under the framework of the uncontrolled manifold (UCM) hypothesis used to quantify indices of multi-finger synergies (i.e., stability property) stabilizing total force during the steady-state task, the two variance components within the UCM analysis increased together with the fingertip temperature, while no changes in the synergy indices between the conditions. Conclusion: The current results showed that fingertip temperature stimulation only to index finger does not affect to muscle force production capability of multi-finger, independence of individual fingers, and force production accuracy by the involvement of all four fingers. The effect of fingertip temperature on the sharing pattern and force variation may be due to diffuse reflex effects of the induced afferent activity on alpha-motoneuronal pools. However, the unchanged stability properties may be the reflection of the active error compensation strategies by non-stimulated finger actions.
This study was performed to investigate the optimum condition of protease production from submerged culture of oak mushroom (Lentinula edodes, Sanlim No. 5) and its enzymatic features. Among several combinations of media, the combination of wheat bran, corn flour, water and corn oil (WB+CF+W+ CO) yielded 84.8 U/g of maximum protease activity. This combination of ingredients, in spite of not being particularly protein-rich in comparison to the other media, allowed for good growth of the fungus and maximal protease production. Comparison of different growth medium liquids indicated that demineralized water afforded the best growth of the fungus and the highest protease activity. Acetate buffer and acidified water negatively affected The protease production peaked around 72 hr of incubation, and decreased thereafter. The molecular weights of produced protease were about 45,000 by Sephadex G-75 chromatography. The pH optimum for protease activity was 4, while maximal activity incubated at 37℃ for 1 hr was observed between pH 4~6. The optimum temperature of this protease was 55℃, and the enzyme was active over a broad temperature range (30~60℃), indicating that this protease would be suitable for a wide range of applications where. different pH and temperature are necessary, such as digestive aids, food industry, beer and tannery industries.
Henry Onyebuchi Okonkwo;Olubunmi Ayokunle Koyejo;Joseph Okechukwu Ariwaodo;Nsien Iniobong Bruno
Journal of Forest and Environmental Science
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v.39
no.2
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pp.73-80
/
2023
Little is known of the life history of Garcinia kola; the objective of this study, therefore, was to assess the fruiting age and tree size of the species in a coastal humid tropical climate plantation condition. A total 103 trees were used in the study viz; 80 ten-year-old trees at reproductive maturity onset and 13 thirty-year-old trees with several cycles of reproduction that constitute two independent variables. Data collected were age of onset of flowering and size at reproductive maturity onset. Relative size at reproductive maturity onset (RSOM) was estimated as size at reproductive maturity onset (SOM) divided by asymptotic maximal size (AMS). Data analysis was conducted using pairwise t-test and principal component analysis (PCA). Reproductive maturity onset (flowering) was recorded in the ten-year-old stand eight (8) years after planting. Mean size at reproductive maturity onset (SOM) was height 5.32±1.7 m, dbh 0.11±0.03 m, total number of branches was 29.6±7.3, crown depth 5.24±1.05 m, crown diameter was 4.78±0.7 m, branch diameter 0.098±0.01 m, leaf length 0.13±0.02 m, leaf breadth 0.37±0.01 m, twig length 0.35±0.11 m and leaf per twig 6±0.84 and asymptotic maximal size (AMS) was height 19.85±0.76 m, dbh 0.95±0.09 m, total number of branches 62±5, crown depth 18.83±0.7 m, crown diameter 12.5±1.64 m, branch diameter 0.5±1.6 m, leaf length 0.16±0.023 m, leaf breadth 0.45±0.12 m, twig length 0.37±0.11 m and leaf per twig 19±7.5. Pairwise t-test analysis showed there was significant differences between SOM and AMS in all growth factors except leaf length, leaf breadth, and twig length. Highest relative size at reproductive maturity onset (RSOM) was recorded in leaf length 0.82, twig length 0.82, and leaf breadth 0.80, while, the lowest was branch diameter 0.11. Four components out of the total of eleven were extracted to explain the relationship in RSOM: Principal component one (PC1) explained 37.23%; PC2 26.4%, PC3 22.73%, and PC4 13.64%.
The purposes of this study were to assess variation of body sway prior to and after submaximal treadmill exercise; to determine the time course of the effects of a fatiguing performed on a treadmill on body sway; and to compare position sense prior to and after exercise in order to assess any variance in proprioception caused by submaximal treadmill exercise. The subjects were twenty-four healthy men in their twenties. They stood barefoot on the Kinesthetic Ability Training Balance Platform to measure body sway. Control trials were performed with eyes alternately open and closed. In the eyes open condition, they were asked to look at a target placed at eye level 1 m in front them. A total of 10 trials, each lasting 20 seconds, were performed. After this series of trials, position sense was measured. Subjects then exercised on the treadmill until 85% of each person's maximal heart rate was reached. The first series of postural sway measurements began immediately after this exercise. The second identical series of postural sway trials was performed at approximately 10 minutes after exercise. The third series was performed approximately 20 minutes after exercise. This allowed approximately 5 minutes of rest between each experimental series. Position sense was measured at approximately 15 and 25 minutes after exercise. The results were as follows: 1) There was a significant increase in body sway after submaximal treadmill exercise compared to pre-exercise values under both visual conditions (p<.05). 2) After submaximal treadmill exercise, under the eyes open condition, the mean value of body sway was significantly increased after both the first and second series (p<.05). Under the eyes closed condition, the mean value of body sway increased significantly after the first series but decreased significantly after the third series (p<.05). 3) Position sense, measured repeatedly after submaximal treadmill exercise, did not change significantly with respect to pre-exercise values (p>.05). These results suggest that fatigue induced by submaximal treadmill exercise produced an increase in body sway in young healthy subjects with or without visual input, but the increase appeared to be lasting less than 15 minutes. No significant change in position sense suggested that proprioception was unaffected by submaximal treadmill exercise-induced fatigue.
Six barley varieties that showed different degree of drought tolerance were grown with and without drought stress treatment (control), and investigated for the temporal changes in growth and several physiological traits after drought treatment. Soil water potential was -0.05 ㎫ at the initial stage of drought treatment and dropped to -0.29 ㎫ at 19 days after withholding irrigation. Soil water potential (SWP) maintained at -0.05 ㎫ in the control. The dry weight (DW) under the drought treatment were reduced compared to the control as follows: Dicktoo-S (short awn), 69% ; Dicktoo-L (long awn), 70%; Dicktoo-T (tetra), 86%; Dongbori-1, 69%; Suwonssalbori-365, 55% and Tapgolbori, ,37%. Dicktoo lines and Dongbori-1 were more tolerant than Suwonssalbori-365 and Tapgolbori. Leaf relative water contents (RWC) and leaf water potential (LWP) decreased obviously under the drought condition, the decrease being greater especially in the less drought-tolerant barley genotypes. Dongbori-1 and Dicktoo-L in drought treatment showed net photosynthesis of 38% and 17% compared to the control, respectively, and the other four genotypes much lower photosynthesis of 1.1% to 7.0%. Stomatal conductance, mesophyll conductance, and the photochemical efficiency (Fv/Fm) of PS II were reduced by drought treatment, the reduction being greater in drought-sensitive genotypes. The drought-tolerant genotypes had greater osmotic adjustment (OA) capacity under water stress. Thus, the decrease of RWC and LWP was lower and the turgor pressure conservation capacity was higher under water stress in drought-tolerant genotypes. Drought-tolerant genotypes showed less decrease of photosynthesis because stomatal conductance, mesophyll conductance and the ratio (Fv/Fm) of the variable to maximal fluorescence of drought-resistant genotype was decreased less in the drought stress condition. In conclusion, the drought-tolerant genotypes had better water conservation capacity through efficient OA, and this led to the lower decrease of photosynthesis and growth in water stress condition.
The Journal of Korean Academy of Orthopedic Manual Physical Therapy
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v.13
no.2
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pp.31-44
/
2007
The purposes of this study were to examine the normal lumbar proprioception and identify the effect of vision and proprioception on lumbar movement accuracy through measuring a reposition error in visual and non-visual conditions and to provide the basic data for use of vision when rehabilitation program is applied. The subjects of this study were 39 healthy university students who have average physical activity level. They were measured the ability to reproduce the target position(50% of maximal range of motion) of flexion, extension, dominant and non-dominant side flexion in visual and non-visual conditions. Movement accuracy was assessed by reposition error(differences between intended and actual positions) that is calculated by the average of absolute value of 3 repeated measures at each directions. The data were analysed by paired samples t-test, independent samples t-test, and repeated measures ANOVA. The results were as follows : 1. Movement accuracy of flexion, extension, dominant side flexion, and non-dominant side flexion was increased in visual condition. 2. There were no differences in the lumbar movement accuracy between sexes in visual and non-visual conditions. 3. In non-visual condition, the movement in coronal plane(dominant and non dominant side flexion) is more accurate than that in sagittal plane(flexion and extension). 4. In non-visual condition, there were no differences in the lumbar movement accuracy between dominant and non-dominant side flexion. In conclusion, this study demonstrates that the movement is more accurate when the visual information input is available than proprioception is only available. When proprioception is decreased by injury or disease, it disturbs the control of posture and movement. In this case, human controls the posture and movement by using visual compensation. However it is impossible to prevent an injury or trauma because most of injuries occur in an unexpected situation. For this reason, it is important to improve the proprioception. Therefore, proprioceptive training or exercise which improve the ability to control of posture and movement is performed an appropriate control of permission or interception of the visual information input to prevent an excessive visual compensation.
Journal of the Korean Society of Food Science and Nutrition
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v.35
no.6
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pp.747-753
/
2006
Tea was known to have different taste and flavor with leaching temperature and time. This study was designed to optimize leaching condition for green tea bag that has been popular to consumers. Soluble solid, total phenolics, and total flavonoid content increased as leaching temperature and time increased. The ranges of maximum leaching condition for these components were $96.1{\sim}99.0^{\circ}C\;and\;5.7{\sim}6.8\;min$. As leaching temperature decreased and leaching time increased, however, electron donating ability (EDA) increased and showed the highest values at $65.3^{\circ}C$ and 7.2 min. The overall acceptability was maximal at $70.2^{\circ}C$ and 3.1min. Based upon the physicochemical and organoleptic properties, it was predicted that the optimal range of leaching conditions for green tea bag was $73{\sim}83^{\circ}C\;for\;5.3{\sim}6.3\;min$.
The purpose of this research was to establish the culture condition for dissociated submandibu -lar gland (SG) cells. After trypsin digestion of SG from 3-4 weeks old mice, dissociated cells were cultured in 1OO/o fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DME) or 0.5-2% low protein serum replacement-DME (LPSR-DME) on plastic surface to form monolayer. The effects of FBS, LPSR and hormones on the growth and function of cultured SG cells were examined. SG cells dissociated by enzyme were successfully cultured and were characterized as epithelial-like cells by light and electron microscope. The maximal DNA synthesis of cultured SG cells was achieved by DME containing 5-10% FBS. The same results were obtained when the effects of LPSR on cell proliferation were examined up to a LPSR concentration of 2%. SG cells cultured in 20/o LPSR-DME expressed a population doubling time of 42.5 hrs and a saturation density of 1.2 $\times$10 5cell/cm$^2$. Dihydrotestosterone (DHT) in medium did not influence on the DNA synthesis of the cultured SG cells, but stimulated protein synthesis of the SG cells. Thyroxine (T4) stimulated protein synthesis of the SCI cells markedly in a dose-dependent fashion. EGF secretion by the cultured SG cells increased significandy by DHT and or T4 trearment. This finding indicated that secretion of EGF by the SG cells was under the control of the hormones such as androgen and thyroid hormones. It seems to be that the culture condition described here can be used as a useful tool for further research on the SCI cells.
When cucumber (Cucumis sativus L.) cotyledon discs were floated on $\delta$-aminolevulinic acid, oxyfluorfen, or rose bengal solution under light condition following 20 h dark incubation, rapid electrolyte leakage from the tissues occurred. The electrolyte leakage from the tissues was dependent on the compounds treated, their concentrations, and the duration of light exposure to the tissues. Dark incubation before exposure to continuous white light enhanced electrolyte leakage from the tissues treated with the compounds and reduced lag period for the activity of the compounds. Electrolyte leakage from the treated tissues was greatly influenced by the light intensity to which they were exposed. Higher light intensities stimulated electrolyte leakage and reduced lag period. Porphyrin biosynthesis inhibitors, gabaculine and 4,6-dioxoheptanoic acid, completely inhibited electrolyte leakage from the oxyfluorfen-treated tissues. Protection against the activity of $\delta$-aminolevulinic acid from electrolyte leakage was complete with 4,6-dioxoheptanoic acid, but not with gabaculine. However, gabaculine and 4,6-dioxoheptanoic acid gave no such protection against rose bengal activity. In summary, our results indicate that $\delta$--aminolevulinic acid, oxyfluorfen, and rose bengal exert their effects by causing electrolyte leakage from the treated tissues in a similar manner, except that oxyfluorfen has an apparent lag period for its action on electrolyte leakage increase. All above compounds require preincubation of treated tissues in darkness and subsequent light exposure with a high intensity for their maximal activities. Our results also support that in the presence of light, $\delta$-aminolevulinic acid and oxyfluorfen cause cellular damage through the indirect generation of singlet oxygen from accumulated tetrapyrroles of porphyrin pathway, whereas rose bengal causes cellular damage through the direct generation of singlet oxygen.
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