Intracellular calcium concentration ($[Ca^{2+}]_i$) may play a crucial role in a variety of neuronal functions. Here we report that in primary culture of mouse cerebellar granule cells nicotinic acetylcholine receptors (nAChRs) are expressed in a specific developmental stage and involved in the regulation of intracellular calcium homeostasis. Nicotine-mediated calcium responses were measured using $^{45}Ca^{2+}$ or fluorometrically using the calcium-sensitive fluorescent dye fura-2. Maximal uptake of $^{45}Ca^{2+}$ evoked by nicotine in mouse cerebellar granule cells were revealed $8{\sim}12$ days in culture. In contrast, nicotine did not alter the basal $^{45}Ca^{2+}$ uptake in cultured glial cells. In cerebellar granule cells nicotine-evoked $^{45}Ca^{2+}$ uptake was largely blocked by the NMDA receptor antagonists. Glutamate pyruvate transaminase (GPT). which removes endogenous glutamate, also prevented nicotine effects, implying the indirect involvement of glutamate in nicotine-mediated calcium responses. Fluorometric studies using fura-2 showed two phases of nicotine-evoked $[Ca^{2+}]_i$ rises: the initial rising phase and the later plateau phase. Interestingly, the NMDA receptor antagonists and GPT appeared to inhibit only the later plateau phase of nicotine-evoked $[Ca^{2+}]_i$ rises. The present results imply that nicotine mediated $^{45}Ca^{2+}$ uptake and $[Ca^{2+}]_i$ rises are attributed to the calcium fluxes through both nAchRs and NMDA receptors in a time-dependent manner. Consequently, nAChRs may play an important role in neuronal development by being expressed in a specific developmental stage and regulating the intracellular calcium homeostasis.
In this study, the antioxidant and cytotoxic effects and the flavonoid contents of leaf extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz. were compared. The flavonoid contents of the acetone + methylene chloride (A+M) and methanol (MeOH) extracts of L. lucidus Turcz. leaves were 55.7 and 233.2 mg/g, respectively. In a DPPH assay, A+M and MeOH extracts from L. lucidus Turcz leaves had a greater scavenging effect than those of S. sieboldii Miq. leaves (p<0.05). In an ABTS assay, MeOH extracts from S. sieboldii Miq. and L. lucidus Turcz (0.5 mg/ml concentration) leaves had scavenging effects of 85% and 91%, respectively (p<0.05), suggesting that both of the MeOH extracts had greater scavenging effects than both A+M extracts. In a 120 min ROS production assay, all tested extracts decreased the cellular ROS production induced by H2O2 compared to that produced by exposure to the extract-free control. The MeOH extract from L. lucidus Turcz leaves had a greater inhibitory effect on cellular ROS production (p<0.05). Treatment with A+M and MeOH extracts from both S. sieboldii Miq. and L. lucidus Turcz. leaves showed a dose-dependent increased cytotoxicity against the growth of AGS, HT-29 cancer cells, and HT-1080 (p<0.05). Both A+M extracts had a greater inhibitory effect on the growth of all cancer cells than both MeOH extracts. These results suggest that the MeOH extract of L. lucidus Turcz. leaves is effective in scavenging free radicals and inhibiting cellular oxidation, while the A+M extract inhibits proliferation of three types of cancer cell.
The massive, fractured and porous-type of glauconite, which is subdivided by surface morphology, occur in subtidal sand and semiconsolidated intertidal sand/mud from continental shelf of the southeastern Yellow Sea. This area is presumed to be a part of Holocene transgressive tidal systems tract. The glauconite, pellet-like grains with diameter of 0.1 to 1 mm, is scattered in surface sand sediments. Results of X-ray diffraction data of the minerals are monoclinic with $a=5.242{\AA}$, $b=9.059{\AA}$, $c=10.163{\AA}$, ${\beta}=100.5^{\circ}$, $V=474.53{\AA}^3$. Thermal treatments on the oriented glauconite increase the X-ray diffraction intensity near $10{\AA}$ (001), suggesting the presence of some expandable layers. Specific gravity of the glauconite is $2.60{\pm}0.45gm/cc$ on the basis of chemical composition and unit-cell dimensions. Based on $O_{10}(OH)_2$, chemical composition of glauconites, octahedral Fe content ranges from 1.19 to 2.06 atoms, corresponding octahedral AI is 0.18 to 0.76 atoms, which progressively substitute Fe for AI with increasing from porous to massive-type. The Mg content ranges from 0.35 to 0.54 atoms, and shows higher with increasing Al contents. A systematic increase of interlayer K from 0.34 to 0.71 is also observed with apparent increases from porous to massive-type, and related to a proportion of expandable layers. The clay preserved in glauconite, which is recognized as ordered/disordered (massive to fractured-type). The interstratified illite/smectite (porous-type), contains 7 to 27 % expandable layers. The glauconite seems to originate from post depositional authigenic growth in reducing environments promoted by the dissolution of clay minerals and biogenic debris.
Kim, Tae-Won;Lee, Geon-Hee;Jeon, Byeong-Gyun;Lee, Sung-Ho
Journal of Life Science
/
v.28
no.10
/
pp.1163-1169
/
2018
In order to utilize the residue that is thrown away after an onion harvest, we analyzed the physiological activity and cytotoxicity of fermented and hot water extracts of the residue. The pH of the extracts were all acidic, and organic matter content was 0.75% in the fermented extract and four times more than 0.19% in the hot water extract. The contents of nitrogen, phosphoric acid, calcium, and magnesium components, except for the potassium component among macroelements, were higher in the fermented extract than in the hot water extract. The content of iron and silicon among the micro-elements was also higher in the fermented extract than in the hot water extract. In addition, the content of boron was higher in the hot water extract than in the fermented extract. The total polyphenol contents of the fermented and hot water extracts were $16.2{\pm}3.3mg{\cdot}g^{-1}$ and $14.6{\pm}1.4mg{\cdot}g^{-1}$, respectively, which was $1.6mg{\cdot}g^{-1}$ higher in the fermented extract than in the hot water extract. However, the total flavonoid contents of the fermented and hot water extracts were $0.1{\pm}0.1mg{\cdot}g^{-1}$ and $4.8{\pm}0.7mg{\cdot}g^{-1}$, respectively, which was $4.7mg{\cdot}g^{-1}$ higher in the hot water extract than in the fermented extract. DPPH and ABTS radical scavenging ability for antioxidant activity were higher in the hot water extract than the fermented extract. The cytotoxicity of the extract using MTT assay showed cell viability of 101.6% and 97.9% in the fermented and hot water extracts, respectively. It was confirmed that there was no cytotoxicity in either extract.
Kim, Sang Tae;Lee, Jae-Jung;Park, Dae-Hak;Yang, In;Han, Gyu-Seong;Ahn, Byoung Jun
Journal of the Korean Wood Science and Technology
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v.43
no.1
/
pp.122-134
/
2015
This study was conducted to investigate the potential of torrefied larch wood as a raw material of pellets. First of all, larch chip was torrefied at the temperatures of 230, 250 and $270^{\circ}C$ for 30, 50 and 70 min. Secondly, moisture content, moisture absorption, higher heating value and ash content of the torrefied chip were measured to examine the effects of torrefaction conditions on the fuel characteristics of larch. Thirdly, surfaces of the torrefied chip were observed by light microscope (LM), field emission scanning microscope (FE-SEM) and SEM-energy dispersive spectroscopy (EDXS). With the increases of torrefied temperature and time, contents of lignin increased and those of hemicellulose reduced. Moisture content of torrefied larch chip was greatly lower than that of non-torrefied chip. Moisture absorption of the torrefied chip decreased as torrefaction temperature increased. As torrefaction temperature increased, higher heating value and ash content of larch chip increased. However, durability of torrefied-larch pellets was remarkably lower comparing to non-torrefied-larch pellets. When surface of larch chip was observed by LM and FE-SEM, surface color and cell wall of the chip was getting darker and more collapsed with the increases of torrefaction conditions. Through the analysis of SEM-EDXS, distribution and quantity of lignin existing on the surface of larch chip increased with the increases of torrefied conditions. In conclusion, $270^{\circ}C$/50 min might be an optimal condition for the torrefaction of larch with the aspect of fuel characteristics, but torrefaction condition of $230^{\circ}C$/30 min should be considered according to the durability of torrefied-larch pellets.
The bacteriological and physiochemical analysis of seawater in Tongyeong harbor was conducted to evaluate sanitary conditions, The samples were collected at 6 stations established once a month from January to December, 2000. During the study period, the ranges of temperature, transparency, chemical oxygen demand, dissolved oxygen, dissolved nitrogen, phosphate and chlorophyll-a were $6.8\sim25.2^{\circ}C,\;1.0\sim2.5\;m,\;1.79\sim2.41\;mg/L,\;5.7\sim10.1\;mg/L,\;6.59\sim10.53{\mu}g-at/L,\;0.56\sim1.01{\mu}g-at/L\;and\;1.21\sim9.54\;mg/m^3$, respectively, The viable cell counts of seawater in Tongyeong harbor ranged from $3.0\times10^4CFU/mL\;to\;6.9\times10^6CFU/mL$. The coliform group and fecal coliform MPN's of the samples were ranged $23\~4,600\;MPN/100\;mL$ (means 540 MPN/100 mL) and $11\~1,600\;MPN/100\;mL$ (means 210 MPN/100 mL), respectively, The coliform group was classified with IMViC reactions and pathogenic vibrios were analyzed. Two hundred eighteen strains that were obtained from seawater samples in Tongyeong harbor represented Escherichia coli group, $66.1\%$; Citrobacter freundii group, $11.0\%$; Enterobacter aerogenes, $9.6\%$; and unknown, $13.3\%$, respectively. During the study period, infectious bacteria such as Vibrio cholerae O1, Salmonella sp. and Shigella sp. were not detected from the samples, but detection ratios of V. parahaemolyticus, V cholerae non-O1 and V. vulnificus were $10.0\sim30.1\%$.
To utilize canned oyster processing waste water effectively, this study was carried out to prepare instant powdered soup using the waste water (IPSW), Instant powdered souu from oyster hot-water extracts (IPSE) was prepared by mixing hot-water extracts powder (15 g) with table salt (5 g), cream powder (19 g), milk replacer (12 g), wheat flour (20 g), corn flour (15 g), starch (5 g), glucose (7.5 g) and onion powder (1.5 g). In preparing IPSW, mixed powder from wash water and boiling liquid waste, instead of powder from hot-water extracts and table salt, was added (powder from boiling liquid waste: powder from wash water= 12: 8) and other additives were added in proportion to those in the IPSE, The IPSW consists mainly of carbohydrates (about $72\%$). It was not different from the IPSE. The volatile basic nitrogen, viable cell counts, coliform group of the IPSW contains 33.4 mg/100g, $2.2\times10^4CFU/g$, <180 MPN/100g, respectively, and its water activity has 0.257. So it was a hygienically safe and conservable instant food. The main fatty acids of IPSW were 16: 0 and 18: 1n-9. Its chemical score of protein was $61.4\%$ and its main inorganic matter was iron. According to a sensual evaluation, in contrast to the IPSE, the IPSW had a bit lower aroma but better taste, It was concluded from the above chemical and sensory evaluation that even the boiling liquid waste which had been mostly abandoned because of its high table salt content can be used as a good material for instant powdered soup if it's powdered and mixed adequately with powder from wash water, and its table salt content is properly adjusted.
Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.
The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.
The seasonal variations of nitrifying bacterial population sampled from 3 sites in Moon-Chon reservoir were analyzed by in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotide probes from August 2000 until July 2001. In addition, physico-chemical parameters such as temperature, pH, chi-a and DOC were measured to determine correlations between those factors and the size of nitrifying bacterial populations. Total bacterial numbers varied in the range of $0.8{\sim}1.5{\times}10^6\;cells/ml$ independent of sites and had the maximal values in March at all 3 stations. The ratio of eubacteria to total bacteria ranged from 44.9% to 79.5%, and the ratio of each nitrifying bacteria to eubacterial numbers reached only $1.0{\sim}7.4%$. The variations of ammonia-oxidizing bacteria ranged from $1.1{\times}10^4$ to $3.0{\times}10^4\;cells/ml$ without noticeable peak values whereas those of nitrite-oxidizing bacteria varied in $1.3{\sim}5.7{\times}10^4\;cells/ml$ with the increasing tendency in winter regardless of the sites. Moreover it was observed that the numbers of nitrite-oxidizing bacteria were higher than those of ammonia-oxidizing bacteria. Total bacterial numbers correlated with water temperature (r = 0.355, p<0.05) and DOC (r = 0.58G, p<0.01) positively whereas nitrite-oxidizing bacteria correlated with temperature (r = -0.416, p<0.05) and pH (r = -0.568, p = 0.001) negatively. In addition, DOC represented good correlations with eubacterial numbers (r = 0.448, p<0.01). These results indicate that temperature, DOC and pH might be one of the main factors affecting variations of bacterial populations in the aquatic ecosystem. It was also suggested that FISH method is a useful tool for detection of slow growing nitrifying bacteria.
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