• The Korean Journal of Physiology and Pharmacology
• /
• v.21 no.2
• /
• pp.241-249
• /
• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.

• The Korean Journal of Physiology and Pharmacology
• /
• v.23 no.5
• /
• pp.381-392
• /
• 2019

Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.

• Preventive Nutrition and Food Science
• /
• v.3 no.3
• /
• pp.216-220
• /
• 1998

Lipoxygenase inhibitory acitivity of the methanolic extract of 60 different kinds of plant seeds was determined by a spectrophotometric method using a soybean lipoxigenase(SLO) and linolenic acid. Among the extracts examined, the methanolic extract of nutmeg(Myristical fragrans)seed showed the most potent SLO inhibitory activity. To isolate SLO inhibitor, hence, the defatted methanol extract was further partitioned with ether, ehtylacetate , and n-butanol , stepwise. The ether souble fraction was successively chromatographed on silica gel, Sephadex LH-20 and preparative TLC. Three phenolic compounds were isolated , and one of them showing a strong SLO inhibition activity was identified as a 2,6-dihydroxy-9-(3', 4', -dihydroxyphenyl)nonylphenone (IC50a=0.39$\mu\textrm{g}$/ml) by 1H-& 13C0NMR, IR, and MS spectroscopy.

• Preventive Nutrition and Food Science
• /
• v.3 no.4
• /
• pp.315-319
• /
• 1998

Previously, the methanolic extract of Paeonia moutan seeds was found to potently inhibit soybean lipoxy-genase (SLO). Hence to isolate SLO inhibitor, the defattd methaniolic extract of the seeds was consecutively partitioned wiht ether, ethyl acetate,n-butanol ,adn water. The ether souble fraction showing strong inhibitory activity against SLO was further fractionated into a strongly acidic, a weakly acidic, and a neutral fractions. The strongly acidic components of the ether extract were successively subjected to chromatography on a silica gel, Sephadex LH-20, and preparative HPLC. Four phenolic compounds were isolated , and twio of them showing a strong SLO inhibition activity were identified as luteolin (IC50=2.32$\mu\textrm{g}$/ml) and 5,6,4'-trihydroxy-7,3'- dimethoxylflavone (IC50=0.31$\mu\textrm{g}$/ml) by UV, IR, 1H-& 13C-NMR, and MS spectroscopy. In addition, two flavonoids showed significantly antioxidative activity as strong as that of of $\alpha$-tocopherol (p<0.05) in the autoxidation system of linoleic acid. These results suggest that luteolin and 5,6,4'-trihydroxy-7,3'-dimethoxy-flavone may be used as a potential source of anti-inflammatory agents with antioxidative activity.

• Preventive Nutrition and Food Science
• /
• v.4 no.3
• /
• pp.163-166
• /
• 1999

Previously, the methanolic extract of Paeonia lactiflora seeds was shown to have strong ingibitory activity against soybean liposygenase (SLO). Four phenolic compounds were isolated from the seeds by solvent fractionation Sephadex LJ-20 column chromatography and preparative HPLC, and three of them showed strong SLO inhibitio and were characterized as trans-resveratrol, $\varepsilon$-viniferin and luteolin by UV, IR, 1H-NMR, 13C-NMR and MS spectrometry. trans-Resveratrol (IC50=1.02$\mu$M), $\varepsilon$-viniferin (IC50=0.81$\mu$M) and luteolin (IC50=10.01$\mu$M), first found in the above seeds, exhibited a potent SLO inhibitory activity although their activity was lower than that of a well-known lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA) (IC50=0.57$\mu$M). These results suggest that Paenia lactiflora seeds, now an unused plant seed, may be developed into useful sources of anti-inflammatory drugs.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.5 no.1
• /
• pp.37-43
• /
• 1969

A genetrc study was made on plant height of indica rices with a few segregating F2 populations involving three semi-dwarf varieties such as T(N)I, CP231-SLO17, and B569A12. These populations were grown in breeding nursery at the International Rice Research Institute (IRRI) during several seasons. 20 to 25 day old seedlings grown at upland seedbed were transplanted to the paddy in a single plant hill spacing 30 cm ${\times}$ 25cm. Measurements of plant height were made from the juncture between culm and roots to the tip of the longest panicle of a plant pulled out from the paddy when they are matured. The results are summarized as follows: 1. Tall indica varieties originated from the south-east Asian countries could be classified into two groups depending upon their allelism whether they showed monogenic segregating ratio of 3:1 or not when they were crossed to T(N)1. 2. Most of U.S. varieties, especially semi-dwarf breedirg materials such as CP231 ${\times}$ SIO17 and B569A12, did not show monogenic segregating mode of 3:1 ratio when they were crossed to T(N)1 or to other varieties bearing the same genetic allele of T(N)1 such as Peta and Sigadis.

• Korean Journal of Food Science and Technology
• /
• v.25 no.6
• /
• pp.637-642
• /
• 1993

Changes in the physicochemical characteristics and triglyceride molecular species of corn oil under the following condition of hydrogenation; temperature $180^{\circ}C,\;H_{2}$, pressure $2.0{\pm}0.3bar$, the amount of Ni catalyst 0.048%(Ni/oil by wt.) and agitation speed 300 rpm. The rate of hydrogenation, expressed as the reduction rate of the iodine value with respect to time, is first order and high (K>0.01). When the reduction rate of the iodine value was 39.9%, hydrogenation time was 30 min, 18:1 was highest(77.06%), thereafter that was decreased and 18:0 increased. In the triglyceride composition, OLL, LLL were reduced markedly in 10 min, thereafter reduced slightly. And PLO, PLL, OLO were eliminated in first 30 min. On the other hand, POO, PLS(CN52) and OOO, SLO(CN54) were increased sharply, and then that showed little change. The melting point(MP) of hydrogenated corn oil were $27.8^{\circ}C\;and\;44.1^{\circ}C$ after 20 min and 60 min, respectively. Trans isomer content increased to 46.8% during 40 mins of hydrogenation and then decreased insignificantly. The solid fat content were linearly increased with hydrogenation time. Accordingly, it is confirmed that this condition of hydrogenation was selective, preferential elimination of polyunsaturated fatty acid went stepwise and trans isomer was formed promotedly. These results suggest that fat modification techniques can be used for practical application.

• Korean Journal of Food Science and Technology
• /
• v.27 no.3
• /
• pp.271-280
• /
• 1995

These studies examined the production of oligosaccharides by ${\beta}-galactosidase$originated from Aspergillus oryzae and Kluyveromyces fragilis mixed. In addition, heat resistance and acid stability of transgalactosylated oligosaccharides were measured. When ${\beta}-galactosidase$ from A. oryzae, K. fragilis and mixed ${\beta}-galactosidase$ were added to 30%(w/v) lactose solution, maximum production of transgalactosylated oligosaccharides were 26.9%, 37.05 and 27.2%, respectively. The ratios of disacchride, trisacchride and tetrasaccharide in transgalactosylated oligosaccharides were 20.5 : 5.4 : 0.6, 20.4 : 10.5 : 4.2 and 21.0 : 4.1 : 1.9, respectively. Nine different oligosaccharides were recovered with 30% and 40% ethanol fractions. When the 30% ethanol fraction was treated at $150^{\circ}C$ for 10 min more than 90% of oligosaccharides remained stable. More than 90% of the oligosaccharides were stable at $130^{\circ}C$ for 3 min with pH 3.0, whereas there of Kluyveromyces was more than 90% with pH 3.5.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.7
• /
• pp.1025-1035
• /
• 2014

This study assessed the effect of immobilized lipase on weak base styrene resin using polyethyleneimine (PEI) with cross-linking. Two procedures were used in this study. The first one, "mono-layer" lipase immobilization, involves washing PEI after adsorption. The second procedure, "multi-layer" lipase immobilization, has no washing before the cross-linking step. Treverlite XS-100200 (weak base styrene resin) was immersed with PEI solution (2.2 mg/mL). Lipase AH (from Burkholderia cepacia) was adsorbed onto the support coated with PEI before cross-linking with glutaraldehyde. Structured lipid was synthesized by immobilized lipase-catalyzed interesterification using canola oil, palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). Total fatty acid contents of triacylglycerol (TAG) in structured lipids were analyzed to investigate activity, properties, and reusability of immobilized lipases. Activities of immobilized lipases on the multi-layer and mono-layer increased at a high concentration (8 mg/mL) of lipase solution used for immobilization. The results show that immobilized lipase with the mono-layer method at pH 8.0 on resin had the highest total saturated fatty acid content (26.17 area%). Activity of immobilized lipase with the multi-layer method at pH 7.5 on support was lower than that of the mono-layer, but total saturated fatty acid content was 16.79 area% higher than that of lipase AH (15.01 area%).