• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.62-70
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• 2020

Zygosaccharomyces rouxii is an important yeast that is required in the food fermentation process due to its high salt tolerance. In this study, the responses and resistance strategies of Z. rouxii against salt stress were investigated by performing physiological analysis at membrane level. The results showed that under salt stress, cell integrity was destroyed, and the cell wall was ruptured, which was accompanied by intracellular substance spillover. With an increase of salt concentrations, intracellular Na⁺ content increased slightly, whereas intracellular K⁺ content decreased significantly, which caused the increase of the intracellular Na⁺/K⁺ ratio. In addition, in response to salt stress, the activity of Na⁺/K⁺-ATPase increased from 0.54 to 2.14 μmol/mg protein, and the ergosterol content increased to 2.42-fold to maintain membrane stability. Analysis of cell membrane fluidity and fatty acid composition showed that cell membrane fluidity decreased and unsaturated fatty acid proportions increased, leading to a 101.21% rise in the unsaturated/saturated fatty acid ratio. The results presented in this study offer guidance in understanding the salt tolerance mechanism of Z. rouxii, and in developing new strategies to increase the industrial utilization of this species under salt stress.

• The Korean Journal of Physiology
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• v.20 no.2
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• pp.225-235
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• 1986

Effects of anions on p-Aminohippurate (PAH) transport across the basolateral membrane (BLM) were studied. Basolateral membrane vesicles were isolated from rabbit renal cortex by using a self-orienting Percoll-gradient centrifugation and $Mg^{2+}-precipitation$. The characteristics of the membrane vesicles was examined by marker enzyme activity, membrane orientation and transport studies. The Na-K-ATPase activity in the fraction containing BLM vesicles was enriched 9·fold, and the alkaline phosphatase activity in the fraction containing BBM vesicles was increased 9-fold, compared with those of the homogenate. The transport properties of the two membrane preparations were studied by a rapid filtration technique. The uptake of PAH by BLM was sensitive to changes in medium osmolarity and inhibited by probenecid. When the uptake of $50{\mu}M$ PAH in voltage-clamped BLM vesicles was determined in the presence of various anions in the incubation medium, cis inhibitions by $SO_4\;and\;SSO_3$ were observed in the presence of sodium gradient (out>in). Sodium-dependent PAH uptake was inhibited competitively by external $SO_4$ PAH uptake in BLM vesicles loaded with 20 mM acetate and $SO_4\;or\;200\;{\mu}M$ PAH was significantly stimulated as compared with unloaded vesicles. The extent of trans-stmulation of PAH uptake by $SO_4$. was increased with the inside concentration of $SO_4$. This trans-stimulatory effect by $SO_4$, was observed to be additive in the presence of Na gradient and completely inhibited by 2 mM probenecid and 1 mM SITS. These results demonstrate that PAH/anion exchange is present in BLM of renal cortex and in this exchange mechanism inorganic as well as organic anions are involved as substrates.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.12
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• pp.1897-1906
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• 2019

Objective: An experiment was conducted to investigate the effects of dietary non-phytate phosphorus (nPP) deficiency on intestinal pH value, digestive enzyme activity, morphology, nutrient utilization, and gene expression of NaPi-IIb in meat ducks from 1 to 21 d of age. Methods: A total of 525 one-d-old Cherry Valley ducklings were fed diets (with 7 pens of 15 ducklings, or 105 total ducklings, on each diet) with five levels of nPP (0.22%, 0.34%, 0.40%, 0.46%, or 0.58%) for 21 d in a completely randomized design. Five experimental diets contained a constant calcium (Ca) content of approximately 0.9%. Body weight (BW), body weight gain (BWG), feed intake (FI), and feed to gain ratio (F:G) were measured at 14 and 21 d of age. Ducks were sampled for duodenum and jejunum digestion and absorption function on 14 and 21 d. Nutrient utilization was assessed using 25- to 27-d-old ducks. Results: The results showed ducks fed 0.22% nPP had lower (p<0.05) growth performance and nutrient utilization and higher (p<0.05) serum Ca content and alkaline phosphatase (ALP) activity. When dietary nPP levels were increased, BW (d 14 and 21), BWG and FI (all intervals), and the serum phosphorus (P) content linearly and quadratically increased (p<0.05); and the jejunal pH value (d 14), duodenal muscle layer thickness (d 14), excreta dry matter, crude protein, energy, Ca and total P utilization linearly increased (p<0.05); however, the serum ALP activity, jejunal $Na^+-K^+$-ATPase activity, and duodenal NaPi-IIb mRNA level (d 21) linearly decreased (p<0.05). Conclusion: The results indicated that ducks aged from 1 to 21 d fed diets with 0.22% nPP had poor growth performance related to poor intestinal digestion and absorption ability; but when fed diets with 0.40%, 0.46%, and 0.58% nPP, ducks presented a better growth performance, intestinal digestion and absorption function.

• The Korean Journal of Physiology
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• v.20 no.2
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• pp.175-183
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• 1986

Since it was proposed that vanadate may be an ‘ideal endogenous regulator of the $Na^+,\;K^+-ATPase$ activity (Cantley et at, 1979), vanadate has been a subject of intensive research and a variety of its physiological effects have been described (Nechay, 1984). In isolated guinea pig heart muscle vanadate shows a positive inotropic effect on ventricular muscle, while it induces a negative inotropic effect on atrial muscle. But its underlying mechanism has not been elucidated so far. Therefore, in this study the flux rates of calcium ion into and from guinea pig heart muscle were measured to throw some light on the underlying mechanism, because those rates have been known to be closely related to the cardiac contractility and the results are summarized as follows: 1) Calcium efflux rates from the intracellular $Ca^{++}$ pool (compartment 4) of both guinea pig left atrium and right ventricle were significantly reduced by vanadate and their pool sizes were significantly increased by vanadate. 2) The magnitude of calcium influx into left atrium was reduced by vanadate, While the magnitude of calcium influx into right ventricle was not affected by vanadate. From these results, it may be concluded that the positive inotropic effect of vanadate on the ventricular muscle was due to a reduced efflux rate of calcium ion and its negative inotropic effect on atrial muscle was resulted from a reduced influx of calcium ion.

• Journal of Life Science
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• v.26 no.2
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• pp.164-173
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• 2016

Isotocin (IT), a nonapeptide homolog of oxytocin in mammals, has been suggested to be involved in physiological processes including social behaviors, stress responses, and osmoregulation in teleost fish. To study its structure and function, the gene encoding the IT precursor was cloned from the genomic DNA and brain cDNA of the cinnamon clownfish, Amphiprion melanopus. The IT precursor gene consists of three exons separated by two introns, and encodes an open reading frame of 156 amino acid (aa) residues, comprising a putative signal peptide of 19 aa, a mature IT protein of 9 aa, a proteolytic processing site of 3 aa, and 125 aa of neurophysin. Tissue-specific analysis of the IT precursor transcript indicated its expression in the brain and gonads of A. melanopus. To examine its osmoregulatory effects, the salinity of the seawater (34 ppt) used for rearing A. melanopus was lowered to 15 ppt. Histological analysis of the gills indicated the apparent disappearance of an apical crypt on the surface of the gill lamella of A. melanopus, as pavement cells covered the surface upon acclimation to the lower salinity. The level of Na⁺/K^+-ATPase activity in the gills was increased during the initial stage of acclimation, followed by a decrease to its normal level, suggesting its involvement in osmoregulation and homeostasis. The only slight increase in the level of IT precursor transcript in the A. melanopus brain upon low-salinity acclimation suggested that IT played a minor role, if any, in the process of osmoregulation.

• Clinical and Experimental Pediatrics
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• v.49 no.3
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• pp.317-325
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• 2006

Purpose : This study was carried out to elucidate the effects of nitric oxide synthase(NOS) inhibitor, NG-monomethyl-L-arginine(L-NMMA) and nitric oxide precursor, L-arginine(L-Arg) on cerebral hemodynamics and energy metabolism during reoxygenation-reperfusion(RR) after hypoxia-ischemia(HI) in newborn piglets. Methods : Twenty-eight newborn piglets were divided into 4 groups; Sham normal control(NC), experimental control(EC), L-NMMA(HI & RR with L-NMMA), and L-Arg(HI & RR with L-Arg) groups. HI was induced by occlusion of bilateral common carotid arteries and simultaneously breathing with 8 percent oxygen for 30 mins, and followed RR by release of carotid occlusion and normoxic ventilation for one hour. All groups were monitored with cerebral hemodynamics and cytochrome $aa_3$ (Cyt $aa_3$) using near infrared spectroscopy(NIRS). $Na^+$, $K^+$-ATPase activity, lipid peroxidation products, and tissue high energy phosphate levels were determined biochemically in the cerebral cortex. Results : In experimental groups, mean arterial blood pressure, $PaO_2$, and pH decreased, and base excess and blood lactate level increased after HI compared to NC group(P<0.05). These variables subsequently returned to baseline after RR except pH. There were no differences among the experimental groups. In NIRS, oxidized hemoglobin($HbO_2$) decreased and hemoglobin(Hb) increased during HI(P<0.05) but returned to base line immediately after RR; 40 min after RR, the $HbO_2$ had decreased significantly compared to NC group(P<0.05). Changes of Cyt $aa_3$ decreased significantly compared to NC after HI and recovered at the end of the experiment. Significantly reduced cerebral cortical cell membrane $Na^+$, $K^+$-ATPase activity and increased lipid peroxidation products(P<0.05) were not improved with L-NMMA or L-Arg. Conclusion : These findings suggest that NO is not involved in the mechanism of HI and RR brain damage during the early acute phase of RR.

• Toxicological Research
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• v.17
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• pp.117-125
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• 2001

Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an important complication in patients bitten by Russells viper. Experimental work in animals and in vitro has elucidated pathophysiological mechanisms that contribute to life threatening complications and have suggested possibilities for therapeutic intervention. The evidence in experimental animals regarding mechanisms of venom action in relation to changes in either extrarenal or intrarenal factors is presented. The cardiovascular system and renal hemodynamics are affected by venom. Reductions of renal function including renal hemodynamics are associated directly with changes in general circulation during envenomation. Possible endogenous mechanisms for releasing the hormone inducing renal vasoconstriction after envenomation are evident. Hormonal factor such as the catecholamine, prostaglandin and renin angiotensin systems induce these changes. Direct nephrotoxicity of venom action is studied in the isolated per-fused kidney. Characteristic polarization of the cell membrane, changes of mitochondrial activity and Na-K ATPase in renal tubular cells are observed. Changes in renal function and the cardiovascular system are observed of ter envenomation and are reversed by the administration of Russells viper antivenom (purified equine immunoglobulin, $Fab_2$ fragment). The neutralizing effects are more efficient when the intravenous injection of antivenom is given within 30 min after the envenomation.

• Journal of Acupuncture Research
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• v.18 no.2
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• pp.111-122
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• 2001

Objectives ; This study was undertaken to determine if Salviae Radix herb-acupuncture (SRA) exerts protective effect against alterations in membrane transport function in rabbits with mercury chloride (Hg)-induced acute renal failure. Methods and Results ; The administration of Hg at a subcutaneous single dose of 10mg/kg caused a reduction in GFR to 9.4% of the basal value and an increase in fractional Na+ excretion to 10-fold, indicating generation of acute renal failure. When animals were acupunctured with $0.5m{\ell}$ of SRA extract (0.1%) in both sides of Shinsu(BL23) for 7 days prod to Hg administration, such changes were significantly attenuated. The fractional excretion of glucose and phosphate was increased to approximately 132-fold and 7-fold, respectively, in rabbits treated with Hg alone, but the fractional excretion of glucose was increased to 26-fold and that of phosphate was not different from the basal value in SRA-pretreated rabbits. Uptakes of glucose and phosphate in purified isolated brush-border membrane and $Na^+-K^+$-ATPase activity in microsomal fraction were inhibited in rabbits treated with Hg alone, suggesting that impairment in proximal reabsorption of glucose and phosphate is resulted from a direct damage of membrane transport carriers and disruption of the normal $Na^+$ gradient. Conclusions ; Such changes were prevented by SRA. Uptakes of organic ions, PAH and TEA, in renal cortical slices were inhibited by the administration of Hg, which was prevented by SRA. Pretreatment of an antioxidant DPPD attenuated the increase in the fractional excretion of glucose and phosphate induced by the administration of Hg.

• Journal of Acupuncture Research
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• v.19 no.5
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• pp.199-208
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• 2002

Objective : This study was undertaken to determine if Carthami Semen Aquacupunc- ture(CSA) exerts protective effect against alterations in membrane transport function rabbits with mercury chloride(HG)-induced acute renal failure. Methods : The administration of Hg at a subcutaneous single dose of 10 mg/kg caused a reduction in GFR and an increase in fractional Na excretion, indicating generation of acute renal failure. When CSA were given for 7 days prior to Hg administration, such changes were significantly attenuated. The fractional excretion of glucose and phosphate was increased in rabbits treated with Hg alone. Results : The increase in rabbits treated with Hg following CSA are significantly lower than that in animals treated with Hg alone. Uptakes of glucose and phosphate in purified isolated brush-border membrane and Na-K-ATPase activity in microsomal fraction were inhibited in rabbits treated with Hg alone. Such changes were prevented by CSA. Uptakes of organic ions, PAH and TEA, in renal cortical slices were inhibited by the administration of Hg, which was prevented by CSA. Exposure of renal cortical slices to Hg in vitro caused an increased LDH release and lipid peroxidation, which was significantly prevented by CSA extract. Conclusions : These results indicate that the administration of Hg causes impairment in reabsorption of solutes in the proximal tubule via the generation of reactive oxygen species. CSA provides the protection against the impairment in proximal reabsorption, and its effect may be resulted from its antioxidant effect.

• Bulletin of the Korean Chemical Society
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• v.32 no.6
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• pp.1925-1930
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• 2011

Psychrophilic bacteria have acquired cold-resistance in order to protect themselves against freezing temperatures, which would otherwise be lethal. DnaK/DnaJ/GrpE systems are molecular chaperones which facilitate proper folding of newly synthesized proteins. Efficient folding processes are of great importance especially in a cold environment, such as the Arctic. In order to understand the protection mechanisms of psychrophilic bacteria against cold temperatures, we have explored a genome of KOPRI22215, tentatively identified as Psychromonas arctica, whose genome sequence has not yet been discovered. With an aim of searching for a coding gene of DnaK from KOPRI22215, we have applied a series of polymerase chain reactions (PCR) with homologous primers designed from other Psychromonas species and LA PCR in vitro cloning. 1917 bp complete coding sequence of dnaK from KOPRI22215 was identified including upstream promoter sites. Recombinant plasmids to overexpress PaDnaK along with EcDnaK (DnaK of E. coli) were then constructed in pAED4 vector and the pET-based system to induce PaDnaK expression by IPTG. Characterization assays of expressed PaDnaK were carried out by measuring survival rates upon 4 day incubation at 4 $^{\circ}C$: a refolding assay as molecular chaperone, and ATPase assay for functional activity. Taking account of all the data together, we conclude that PaDnaK was identified, successfully expressed, and found to be more efficient in providing cold-resistance for bacterial cells.