• Animal cells and systems
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• v.13 no.3
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• pp.283-288
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• 2009

The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.3
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• pp.269-275
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• 2018

$Na^+/H^+$ exchangers (NHEs) have been shown to be involved in regulating cell volume and maintaining fluid and electrolyte homeostasis. Pooled evidences have suggested that loss of $Na^+/H^+$ exchanger isoform 8 (NHE8) impairs intestinal mucosa. Whether NHE8 participates in the pathology of infectious colitis is still unknown. Our previous study demonstrated that somatostatin (SST) could stimulate the expression of intestinal NHE8 so as to facilitate $Na^+$ absorption under normal condition. This study further explored whether NHE8 participates in the pathological processes of infectious colitis and the effects of SST on intestinal NHE8 expression in the setting of infectious colitis. Our data showed that NHE8 expression was reduced in Citrobacter rodentium (CR) infected mice. Up-regulation of NHE8 improved diarrhea symptom and mucosal damage induced by CR. In vitro, a similar observation was also seen in Enteropathogenic E. coli (EPEC) infected Caco-2 cells. Seglitide, a SST receptor (SSTR) 2 agonist, partly reversed the inhibiting action of EPEC on NHE8 expression, but SSTR5 agonist (L-817,818) had no effect on the expression of NHE8. Moreover, SST blocked the phosphorylation of p38 in EPEC-infected Caco-2 cells. Taken together, these results suggest that enhancement of intestinal NHE8 expression by SST could ameliorate the symptoms of mice with infectious colitis.

• Molecules and Cells
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• v.28 no.2
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• pp.81-85
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• 2009

$Na^+-H^+$ exchanger (NHE) is the main acid extruder in cardiac myocytes. We review the experimental findings of ion-dependency of NHE activity, and the mathematical modeling developed so far. In spite of extensive investigation, many unsolved questions still remain. We consider that the precise description of NHE activity with mathematical models elucidates the roles of NHE in maintaining ionic homeostasis, especially under pathophysiological conditions.

• Biomolecules & Therapeutics
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• v.25 no.6
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• pp.593-598
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• 2017

The $Na^+/H^+$ exchanger-1 (NHE-1) is a ubiquitously expressed pH-regulatory membrane protein that functions in the brain, heart, and other organs. It is increased by intracellular acidosis through the interaction of intracellular $H^+$ with an allosteric modifier site in the transport domain. In the previous study, we reported that glutamate-induced NHE-1 phosphorylation mediated by activation of protein kinase C-${\beta}$ (PKC-${\beta}$) in cultured neuron cells via extracellular signal-regulated kinases (ERK)/p90 ribosomal s6 kinases (p90RSK) pathway results in NHE-1 activation. However, whether glutamate stimulates NHE-1 activity solely by the allosteric mechanism remains elusive. Cultured primary cortical neuronal cells were subjected to intracellular acidosis by exposure to $100{\mu}M$ glutamate or 20 mM $NH_4Cl$. After the desired duration of intracellular acidosis, the phosphorylation and activation of PKC-${\beta}$, ERK1/2 and p90RSK were determined by Western blotting. We investigated whether the duration of intracellular acidosis is controlled by glutamate exposure time. The NHE-1 activation increased while intracellular acidosis sustained for >3 min. To determine if sustained intracellular acidosis induced NHE-1 phosphorylation, we examined phosphorylation of NHE-1 induced by intracellular acidosis by transient exposure to $NH_4Cl$. Sustained intracellular acidosis led to activation and phosphorylation of NHE-1. In addition, sustained intracellular acidosis also activated the PKC-${\beta}$, ERK1/2, and p90RSK in neuronal cells. We conclude that glutamate stimulates NHE-1 activity through sustained intracellular acidosis, which mediates NHE-1 phosphorylation regulated by PKC-${\beta}$/ERK1/2/p90RSK pathway in neuronal cells.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.242-248
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• 2015

This study was performed to investigate the microbiological contamination on toothbrushes, toothbrush caps, and tooth cleaning cups in the child care centers and to evaluate the toxin genes, toxin production ability and antibiotic resistance of food-borne pathogens. The average number of total aerobic bacteria and fungi were 5.3 log CFU and 3.2 log CFU. Coliform bacteria were detected in 41 (54.7%) of 75 toothbrushes, 13 (44.8%) of 29 toothbrush caps, and 29 (44.6%) of 65 tooth cleaning cups. Salmonella spp. was not detected in all of samples but Bacillus cereus was isolated from 1 (1.3%) of 75 toothbrushes and 2 (3.1%) of 65 tooth cleaning cups. Staphylococcus aureus was detected in 1 (1.5%) of 65 tooth cleaning cups. The nheA, nheB, nheC, hblC, hblD, hblA and entFM toxin genes were possessed in B. cereus isolated from toothbrush which also produce NHE and HBL enterotoxins. S. aureus was resistant to ampicillin and penicillin, while B. cereus was resistant to ${\beta}-lactam$ antibiotics. These results indicated that the sanitary conditions of toothbrushes and tooth cleaning cups in the child care centers should be improved promptly. The UV sterilization after drying and then storage in dried condition is required to improve the sanitary condition of toothbrushes and tooth cleaning cups in the child care center.

• Health Policy and Management
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• v.12 no.3
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• pp.99-111
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• 2002

This paper draws the determinants of National Health Expenditures(min) and collectivizes OECD countries which are positioned by same conditions using the decision tree analysis. Major findings are summarized as follows. We find that the power of influence of income level on NHE has been 58.35% in 1985, 65.37% in 1990, 66.90% in 1995, and 66.47% in 1997. The power of influence of public share in NHE has been on the increase during that period: 19.50% in 1985, 19.91% in 1990, 22.81% in 1995 and 26.88% in 1997. The two factors(income level, public share) tells for the most part of NHE: 77.85% in 1985, 85.28% in 1990, 89.71% in 1995, 93.35% in 1997. Our results support the hypothesis that NHE could be explained mostly by the income level and show that public share is negatively correlated with the growth of NHE.

• Bulletin of the Korean Chemical Society
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• v.30 no.11
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• pp.2621-2625
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• 2009

To identify a non-acylguanidine NHE-1 inhibitor, an acylguanidne group was replaced with an imidazole group in the potent NHE-1 inhibitors with furan or benzothiphene core template, found from our previous studies. We synthesized and biologically evaluated 4-heteroaryl-2-amino-5-methylimidazole derivatives. All those imidazole compounds (16-18) represented the potent NHE-1 inhibitory activities, similar to the corresponding acylguanidine compounds.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.3
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• pp.187-191
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• 2003

$Na^+/H^+$ exchanger (NHE) has a critical role in regulation of intracellular pH (pHi) in the renal proximal tubular cells. It has recently been shown that dopamine inhibits NHE in the renal proximal tubules. Nevertheless, there is a dearth of information on the effects of long-term (chronic) dopamine treatment on NHE activities. This study was performed to elucidate the pHi regulatory mechanisms during the chronic dopamine treatments in renal proximal tubular OK cells. The resting pHi was greatly decreased by chronic dopamine treatments. The initial rate and the amplitude of intracellular acidification by isosmotical $Na^+$ removal from the bath medium in chronically dopamine-treated cells were much smaller than those in control. Although it seemed to be attenuated in $Na^+$-dependent pH regulation system, $Na^+$-dependent pHi recovery by NHE after intracelluar acid loading in the dopamine-treated groups was not significantly different from the control. The result is interpreted to be due to the balance between the stimulation effects of lower pHi on the NHE activity and counterbalance by dopamine. Our data strongly suggested that chronic dopamine treatment increased intrinsic intracellular buffer capacity, since higher buffer capacity was induced by lower resting pHi and this effect could attenuate pHi changes under extracellular $Na^+$-free conditions in chronically dopamine-treated cells. Our study also demonstrated that intracellular acidification induced by chronic dopamine treatments was not mediated by changes in NHE activity.

• Journal of Food Hygiene and Safety
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• v.36 no.6
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• pp.520-527
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• 2021

In this study, toxin gene and antibiotic resistance of food poisoning strains isolated from indoor air in the cafeteria were analyzed to prevent food poisoning. Staphylococcus aureus (16 strains) and Bacillus cereus (37 strains) isolated from indoor air in child care center were tested. The toxin genes of S. aureus and B. cereus were detected by PCR assay. The antimicrobial susceptibility test followed the disc diffusion method described by the Clinical and Laboratory Standard Institute. The seg and sei toxin genes were detected in 11 of 16 S. aureus strains (68.6%). The nheA and nheB toxin genes were detected in 37 B. cereus strains. In this study, a total of 12 toxin gene patterns of B. cereus were found, among which the nheA-nheB-nheC toxin gene was found to be the most frequent pattern. The result of the antimicrobial susceptibility test of S. aureus revealed 93.8% and 87.5% resistance to ampicillin and penicillin antibiotics, but methicillin resistance S. aureus and vancomycin resistance S. aureus were not detected. All 37 B. cereus tested in this study were resistant to ampicillin and penicillin antibiotics. Based on the result of this study, it was judged that regular ventilation and air quality management were necessary to prevent food poisoning caused by S. aureus and B. cereus contaminated in the indoor air of child care centers.

• Parasites, Hosts and Diseases
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• v.54 no.1
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• pp.21-29
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• 2016

The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of $Na^+$ and $H^+$ ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of $Ca^{2+}$. In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.