Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.
Under a typical Mediterranean environment in southern Australia, the evaporation rate increases significantly in hot summers, resulting in highly saline drinking water for grazing animals. Also in the cropping areas, dryland salinity is a problem. Grazing animals under these environments can ingest excessive amount of salt from feed, drinking water and soil, which can lead to a reduction in growth rate. To understand the impact of high salt intake on grazing deer, two experiments were conducted to assess the effect of salt levels in drinking water on feed intake and growth rate of red and fallow weaner deer. The results revealed that fallow deer did not show any abnormal behaviour or sickness when salt level in drinking water was increased from 0% to 2.5%. Feed intake was not affected until the salt content in water exceeded 1.5%. Body weight gain was not affected by 1.2% salt in drinking water, but was reduced as salt content in water increased. Compared with deer on fresh water, the feed intake of red deer on saline water was 11-13% lower when salt level in drinking water was 0.4-0.8%. An increase in salt level in water up to 1% resulted in about a 30% reduction in feed intake (p<0.01). Body weight gain was significantly (p=0.004) reduced when salt level reached 1.2%. The deer on 1% salt tended to have a higher (p=0.052) osmotic pressure in serum. The concentration of P, K, Mg and S in serum was affected when salt level in water was over 1.0%. The results suggested that the salt level in drinking water should be lower than 1.2% for fallow weaner deer and 0.8% for red weaner deer to avoid any reduction in feed intake. Deer farmers need to regularly test the salt levels in drinking water on their farms to ensure that the salt intake of grazing deer is not over the levels that deer can tolerate.
The objective of the current study was to determine acute plasma stress responses in two size groups of juvenile Epinephelus akaara (average body weight: $8.4{\pm}2.1$ and $3.3{\pm}0.6g$; 150 and 120 days after hatch, respectively) exposed to abrupt salinity drops (from 34 practical salinity unit, PSU seawater to 18, 10 PSU (experiment 1) or 26, 18, 10 PSU (experiment 2), respectively). Plasma glucose, glutamic oxalate transaminase, glutamic pyruvate transaminase, red blood cell counts, and gill histology were determined during 72 h exposure. Significantly increased plasma glucose, glutamic oxalate transaminase levels, and red blood cell counts were observed in fish exposed to 18 or 10 PSU. Histological changes, such as hyperplasia and lifting of epithelium in the gill secondary lamellae, were also observed in fish exposed to 18 or 10 PSU at 72 h post-drop. E. akaara exposed to sudden salinity drops to 18 or 10 PSU still seems to undergo the primary adjustment phase before fish reaches a new homeostasis, whereas fish exposed to 26 PSU seems to mount osmotic changes. Therefore, the no observed adverse effect levels for 72 h acute salinity challenge was 26 PSU in our study, and salinity drop to 18 PSU and below can possibly cause acute adverse effect, in which fish could be vulnerable to additional stresses such as a temperature changes or handling stress.
Kim, Ha Na;Seo, Hyun Won;Kim, Bong Seok;Lim, Hyun Ju;Lee, Ha Na;Park, Jin Suck;Yoon, Young Jin;Oh, Jong Woo;Oh, Mi Jin;Kwon, Jin;Oh, Chan Ho;Cha, Dong Seok;Jeon, Hoon
Natural Product Sciences
/
v.21
no.2
/
pp.128-133
/
2015
Lindera obtusiloba has been widely used as a traditional medicine for the treatment of lots of diseases, including abdominal pain, bruise, and hepatocirrhosis. Here in this study, we elucidated the lifespan-extending effect of methanolic extract of Lindera obtusiloba (MLO) using Caenorhabditis elegans model system. We found that MLO has potent lifespan extension activities under normal culture condition. Then, we determined the protective effects of MLO on the stress conditions such as osmotic, thermal and oxidative stress. To reveal possible mechanism of MLO-mediated lifespan, we further investigated the effect of MLO on the antioxidant enzyme activities and intracellular ROS levels. Our results demonstrated that superoxide dismutase and catalase activities were significantly up-regulated by MLO treatment, resulted in reduced intracellular ROS levels. In this work, we also tested whether MLO-mediated longevity activity was associated with aging-related factors such as food intake and growth. Our data revealed that both of pharyngeal pumping rate and body length were significantly shifted by MLO treatment, indicating these factors were involved in MLO's lifespan-extension effects. Although MLO induces reduction in food intake, the body movement of MLO-fed aged worms was not decreased, compared to untreated control worms, indicating MLO might extend lifespan without affecting healthspan.
The studies aimed to distinguish between initial (ionic or osmotic) effects of salinisation on growth and the longer-term consequences of excessive salt accumulation. Tall and dwarf varieties of rice were used to provide different growth rates. There was no significant effect upon the day-to-day pattern of growth, upon the ultimate length of leaves that were developing at the time of, or shortly after, salinisation with 50 mM NaCl. Leaves that developed after prolonged exposure of the plants to salinity were shorter. Addition of NaCl, KCl or mannitol to the root medium brought about a cessation of leaf elongation within one minute. Growth at a reduced rate restarted abruptly after a lag period that depended upon the external concentration. Elongation rate recovered to its original value within 24 hours after exposure to 50 mM NaCl, though not at higher concentrations. Addition of NaCl at concentrations up to 100 mM elicited no short-term effect upon photsynthetic gas exchange. No change in turgor pressure was detectable in the growing zone with the resolution of the miniature pressure probe used (about 70 kPa). It is concluded that the initial growth reduction in rice caused by salinisation is due to a limitation of water supply. A clear distinction is made between the initial effects of salt which are recoverable, and the long-term effects which result from the accumulation of salt within expanded leaves.
Kim, Hong-Sung;Park, Sang-Min;Yoon, Sang-Jun;Hwang, Dae-Youn;Jung, Young-Jin
Polymer(Korea)
/
v.33
no.5
/
pp.509-513
/
2009
For examining an effect of lithium bromide on structure and property of chitosan/fibroin blend, we investigated the structural characteristic of chitosan/fibroin blend films using solution with lithium bromide which was removed during a casting. The chitosan/fibroin blend formed a complex with the dissolved bromine/lithium ions. The crystalline phase of the complex was found in the blend film at LiBr concentration of 0.6 mol/L. The degree of crystallization was decreased with increasing the concentration of LiBr. The hydrated crystalline phase of chitosan was formed in the blend film that lithium bromide was removed in the process of casting by neutralization and osmotic action. The crystallinity of this film was increased largely as compared with that of the film without lithium bromide. The complexed blend film formed hydrogel absorbing plenty of water.
In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.
Kim, Sang Hyun;Im, Jun Sang;Kim, Bong Seok;Lim, Hyun Ju;Oh, Jong Woo;Park, Jin Suck;Yoon, Young Jin;Lee, Ha Na;Cha, Dong Seok;Jeon, Hoon
Korean Journal of Pharmacognosy
/
v.46
no.1
/
pp.38-43
/
2015
Ligularia stenocephala has a wide range of types of constituents with various pharmacological properties. Here in this study, we examined the effect of methanolic extract of L. stenocephala (MLS) on the lifespan and stress tolerance using Caenorhabditis elegans model system. We found that lifespan of wild-type worms was significantly lengthened in the presence of MLS in a dose dependent manner. MLS also elevated the tolerance of worms against osmotic, heat shock, and oxidative stress. We also demonstrated in vivo antioxidant capacity of MLS by checking intracellular reactive oxygen species levels as well as antioxidant enzyme activities such as catalase and superoxide dismutase. We further investigated several aging-related factors, including pharyngeal pumping rate and body length. Here, we showed that MLS exerts longevity effect independent of both factors. In addition, body movement of aged worms was significantly elevated, suggesting MLS could enhance healthspan as well as lifespan.
Cloud droplet activation process is well described by $K{\ddot{o}}hler$ theory and several parameterizations based on $K{\ddot{o}}hler$ theory are used in a wide range of models to represent this process. Here, we test the two different method of calculating the solute effect in the $K{\ddot{o}}hler$ equation, i.e., osmotic coefficient method (OSM) and ${\kappa}-K{\ddot{o}}hler$ method (KK). To do that, each method is implemented in the cloud droplet activation parameterization module of WRF-CHEM (Weather Research and Forecasting model coupled with Chemistry) model. It is assumed that aerosols are composed of five major components (i.e., sulfate, organic matter, black carbon, mineral dust, and sea salt). Both methods calculate similar representative hygroscopicity parameter values of 0.2~0.3 over the land, and 0.6~0.7 over the ocean, which are close to estimated values in previous studies. Simulated precipitation, and meteorological variables (i.e., specific heat and temperature) show good agreement with reanalysis. Spatial patterns of precipitation and liquid water path from model results and satellite data show similarity in general, but on regional scale spatial patterns and intensity show some discrepancy. However, meteorological variables, precipitation, and liquid water path do not show significant differences between OSM and KK simulations. So we suggest that the relatively simple KK method can be a good alternative to the OSM method that requires various information of density, molecular weight and dissociation number of each individual species in calculating the solute effect.
We investigated some important factors which affect the transdermal flux of ketoprofen, a nonsteroidal anti-inflammatory agent, as a first step to provide some basic knowledge for the development of a iontophoretic transdermal patch system. Factors such as current density, polarity, buffer (HEPES) and electrolyte concentration and pH were studied using hairless mouse skin. The effect of poly(L-lysin), which is known to affect the electro-osmotic flow through skin, on flux was also studied. Passive flux was about $20\;{\mu}g/cm^2hr$ at pH 4.0, but was negligible at pH 7.4 where all ketoprofen molecules dissolved are ionized (ketoprofen pKa=5.94). At pH 4.0, application of anodal current increased the flux further above the passive level, however anodal flux at pH 7.4 was much smaller than passive flux at pH 4.0. The application of cathodal current at pH 4.0 increased the average flux to $30-40\;{\mu}g/cm^2hr$, depending on the current density applied. At pH 7.4, cathodal flux was only about $5\;{\mu}g/cm^2hr$. Decrease in buffer and electrolyte concentration increased this cathodal flux about 10 fold. However decrease in HEPES buffer concentration 100 fold did not affect the flux. Anodal flux of acetaminophen was much larger than cathodal flux, indicating that electroosmotic flow can be playing an important role in the flux. Poly(L-lysin) increased the cathodal flux at pH 7.4. These results provide some important insights into the mechanism of transdermal flux of ketoprofen and the role of electroosmotic flow.
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