• Reproductive and Developmental Biology
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• 제39권4호
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• pp.117-125
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• 2015

Iron is required for cell viability but is toxic in excess. While the iron-mediated malfunction of testicular cells is well appreciated, the underlying mechanism(s) of this effect and its relationship with fertility are poorly understood. Ferritin is a ubiquitous intracellular protein that controls iron storage, ferroxidase activity, immune response, and stress response in cells. Ferritin light chain protein (FTL) is the light subunit of the Ferritin. Previously, we had identified the FTL in bovine spermatozoa following capacitation. In present study, to investigate the role of Ferritin in sperm function, mice spermatozoa were incubated with multiple doses (1, 10 and $100{\mu}M$) of sodium nitroprusside (SNP), an iron donor. SNP was increased Ferritin levels in a dose-dependent manner. The Ferritin was detected on the acrosome in spermatozoa by immunocytochemistry. Short-term exposure of spermatozoa to SNP increased tyrosine phosphorylation and the acrosome reaction (AR). Finally, SNP affected a significant decrease in the rate of fertilization as well as blastocyst formation during early embryonic development. On the basis of these results, we propose that the effects of Ferritin on the AR may reduce overall sperm function leads to poor fertility in males and compromised embryonic development.

• The Korean Journal of Physiology and Pharmacology
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• 제71권4호
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• pp.525-536
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• 2020

Transcellular calcium transport is an essential activity in mineralized tissue formation, including that in nervous systems. Dysregulation of Ca²⁺ homeostasis can induce excitotoxicity and neurodegeneration in the central nervous system. Nckx3, a potassium-dependent Na⁺/Ca²⁺ exchanger, is most abundant in the brain and has a critical role in the transport of intracellular calcium across the cell membrane. However, the roles of Nckx3 in neuron development and function remain unreported. Herein, we examined the behaviors of Nckx3-knock-out mice at the age of six weeks. Detailed behavioral analyses showed Nckx3^-/- mice exhibited an increase in moving distances in the open field test. Additionally, the rotarod test revealed motor learning defects in Nckx3^-/- mice. Both Nckx3^+/- and Nckx3^-/- mice also exhibited deficits in sociability and social novelty preference. Furthermore, Nckx3^-/- mice displayed increased depression-related behavior. However, there was no significant change in cognition function detected in Nckx3^-/- mice. This study demonstrates that NCKX3 is involved in behavior and neuronal function

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.101-109
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• 2010

Efficient L-lactic acid production from Jerusalem artichoke tubers, by Lactobacillus casei G-02, using simultaneous saccharification and fermentation (SSF) in a fed-batch culture, is demonstrated. A kinetic analysis of the SSF revealed that the inulinase activity was subjected to product inhibition, whereas the fermentation activity of G-02 was subjected to substrate inhibition. It was also found that the intracellular NADH oxidase (NOX) activity was enhanced by the citrate metabolism, which dramatically increased the carbon flux of the Embden-Meyerhof-Parnas (EMP) pathway, along with the production of ATP. As a result, when the SSF was carried out at $40^{\circ}C$ after an initial hydrolysis of 1 h and included a sodium citrate supplement of 10 g/l, an L-lactic acid concentration of 141.5 g/l was obtained after 30 h, with a volumetric productivity of 4.7 g/l/h. The conversion efficiency and product yield were 93.6% of the theoretical lactic acid yield and 52.4 g lactic acid/l00 g Jerusalem artichoke flour, respectively. Such a high concentration of lactic acid with a high productivity from Jerusalem artichokes has not been reported previously, making G-02 a potential candidate for the economic production of L-lactic acid from Jerusalem artichokes on a commercial scale.

• Biomolecules & Therapeutics
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• 제20권6호
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• pp.544-549
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• 2012

To examine the neuroprotective effects of ginsenoside R0, we investigated the effects of ginsenoside R0 in PC12 cells under an anoxic or oxidative environment with Edaravone as a control. PC12 neuroendocrine cells were used as a model target. Anoxic damage or oxidative damage in PC12 cells were induced by adding sodium dithionite or hydrogen peroxide respectively in cultured medium. Survival ratios of different groups were detected by an AlamarBlue assay. At the same time, the apoptosis of PC12 cells were determined with flow cytometry. The putative neuroprotective effects of ginsenoside R0 is thought to be exerted through enhancing the activity of antioxidant enzymes Superoxide dismutases (SOD). The activity of SOD and the level of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS), were measured to evaluate the protective and therapeutic effects of ginsenoside R0. Ginsenoside R0 treated cells had a higher SOD activity, lower MDA level and lower ROS, and their survival ratio was higher with a lower apoptosis rate. It is suggested that ginsenoside R0 has a protective effect in the cultured PC12 cells, and the protection efficiency is higher than Edaravone. The protective mechanisms of these two are different. The prevent ability of ginsenoside R0 is higher than its repair ability in neuroprotection in vitro.

• Journal of Chest Surgery
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• 제18권1호
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• pp.1-6
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• 1985

Isolated sinus node cells of the rabbit were used to assess the effects of extracellular Na, K, Ca and Mg concentrations on cardiac pacemaker activity. With intracellular glass micro-electrodes spontaneous action potentials of SA node were recorded and the effects of various ions and their blockers were analyzed in terms of the cycle length, the amplitude and the duration of action potentials, the results obtained were as follows. 1. Sodium reduction [up to 30%] decreased the amplitude of action potential and lengthened the cycle length. TTX, specific blocker of Na channel slightly lengthened the cycle length. 2. Increasing potassium ion concentration, the duration of action potential decreased and the frequency increased in 6mM, however, spontaneous action potential was stopped in 24 mM. Barium ion known to be decreasing K conductance increased the duration of action potential but no significant change in the cycle length was noticed. 3. Calcium ion has shortening effect on the duration and the cycle length of action potential but not with dose-dependent manner. Cadmium ion .[0.02mM] lengthened cycle length and the duration of action potential. 4. Increasing the concentration of magnesium ion the cycle length was lengthened, significantly.

• 약학회지
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• 제55권3호
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• pp.213-218
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• 2011

The aim of this study is to evaluate the sedative effects of ethanol extract and the three major constituents of Cimicifugae Rhizoma. They decreased locomotor activity significantly, and enhanced sleeping duration induced by thiopental sodium. The ethanol extract of Cimicifugae rhizoma and 24-epi-7,8-didehydrocimigenol-3-xyloside (24-epi.) increased the $Cl^-$ influx into the intracellular area of SH-SY5Y neuroblastoma cells significantly. The present results demonstrate that the sedative effects of Cimicifugae rhizoma are mediated via the GABA-gated $Cl^-$ channel, partly by 24-epi.

• The Korean Journal of Physiology
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• 제22권1호
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• pp.13-29
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• 1988

In order to elucidate the regulatory mechanism of intracellular calcium ion concentrations, contractions or contractures induced by $Na^{+}-removal$, calcium-application or ouabain-treatment as an index of $Na^+/Ca^{2+}$ exchange activity were studied in atrial muscle or vascular smooth muscle (aorta and renal artery) of the rabbit. The magnitude of low sodium contractures in atrial trabeculae increased with sigmoid shape when external sodium concentrations were reduced to sodium-free condition, whereas that of calcium contracture intensified in a parabolic pattern when external calcium concentrations were elevated to 8 mM. $Na^{+}-removal$ contractures were induced in a duration-dependent manner to $K^{+}-free$ exposure and same findings were observed with ouabain treatment. $Na^{+}-free$ contractures were not affected by verapamil treatment, but stimulated by $100{\mu}M\;Mn^{2+}$ and inhibited by high concentrations of $Mn^{2+}\;(2{\sim}8mM)$ in a dose-dependent manner. Ryanodine which is known to suppress the release of calcium from internal store abolished spontaneous twitch contractions induced by $K^{+}-free$ solution, but had no effect on the development $Na^{+}-free$ contractures. Na-free contractures were not always induced in vascular smooth muscle preparations. Contractures by $O\;mM\;Na^+$ were usually seen in aorta, but not often in renal artery.$50\;mM\;K^+$, noradrenaline (NA) and angiotensin II (AII) always evoked very large contraction in all preparations of vascular smooth muscle. Contractures developed by $O\;mM\;Na^+$ were not sensitive to verapamil treatment as in atrial trabeculae, but were abolished by $100{\mu}M\;Mn^{2+}$. In contrast to $Na^{+}-free$ contractures, $Mn^{2+}(100{\mu}M)$ had no effect on the contractures induced by NA or 50 mM$K^+$. Caffeine in the concentration of 10 mM evoked transient contracture in the distal renal artery. The rate of spontaneous relaxation in caffeine contracture was dependent upon the concentrations of external sodium, and had double component of relaxation when the rate of relaxation was plotted in the semilogarithmic scale of relative tension versus time. Especially late components of relaxation had more direct relation to $Na^+$ concentrations. It could be concluded that $Na^+/Ca^{2+}$ exchange mechanism in the heart has a large capacity, inhibited by $Mn^{2+}$ but not by verapamil and ryanodine, while $Na^+/Ca^{2+}$ exchange system in vascular smooth muscle has a very low capacity especially in small artery, inhibited by low concentration of $Mn^{2+}\;(100{\mu}M)$ but not affected by verapamil and ryanodine.

• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.249-255
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• 2015

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that $Ca^{2+}$ may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced $[Ca^{2+}]_i$ increase with nonspecific $Ca^{2+}$ channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive $[Ca^{2+}]_i$ elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a $Ca^{2+}$ chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that $Ca^{2+}$ may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream $Ca^{2+}$ regulation of the WNK-OSR1 pathway in intact cells.

• Applied Microscopy
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• 제22권2호
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• pp.118-126
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• 1992

The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.

• 한국식품위생안전성학회지
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• 제39권3호
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• pp.273-280
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• 2024

본 연구에서는 천연 항균제인 광귤 추출물의 항균 활성과 항균 메커니즘을 조사해 즉석섭취식품인 샐러드에서 Salmonella Typhimurium을 제어하기 위한 세척수로써 적용 가능성을 평가하였다. 액체배지희석법으로 S. Typhimurium에 대한 광귤 추출물의 최소 억제 농도(MIC)를 구했다. 그런 다음 다양한 농도(1/16 MIC-2 MIC)에 해당하는 광귤 추출물에 S. Typhimurium을 접종하고 성장곡선을 분석해 대조군과 성장값을 비교하여 항균 활성을 확인하였다. 광귤 추출물을 처리한 후, S. Typhimurium의 세포 내 활성산소종 수준과 막 전위 및 손상도의 변화, 핵산 누출량을 측정하여 광귤 추출물의 항균 메커니즘을 확인하였다. 최종적으로 S. Typhimurium을 인위적으로 접종한 샐러드에 다양한 농도의 광귤 추출물을 다양한 시간 동안 침지 방법으로 항균 처리하여 저감화 효과를 확인했다. S. Typhimurium에 대한 광귤 추출물의 MIC는 195.313 mg/L으로, 1 MIC와 2 MIC의 광귤 추출물은 S. Typhimurium의 성장을 완전히 억제하였다. 광귤 추출물의 처리농도가 높아질수록, 세포 내 ROS 수준과 막 전위, 막 손상도 그리고 핵산 방출량은 증가하였다. 마지막으로, 세척수인 광귤 추출물의 농도가 높고 처리 시간이 길수록 샐러드의 S. Typhimurium의 수가 감소하였다. 따라서 광귤 추출물은 S. Typhimurium를 효과적으로 제어할 수 있음을 입증했다. 광귤 추출물은 차아염소산나트륨과 비교하였을 때 효과적인 항균 활성을 보이며 안전한 샐러드 세척수로 사용될 수 있다. 이는 샐러드와 같은 식품에서 광귤 추출물이 식중독 발생을 미연에 방지하는데 기여할 수 있음을 시사한다.