• Food Science of Animal Resources
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• v.42 no.6
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• pp.1061-1073
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• 2022

Receptor activator of NF-kB ligand (RANKL) is known to play a major role in bone metabolism and the immune system, and its recombinant form has been expressed in bacterial systems for research since the last two decades. However, most of these recombinant forms are used after purification or directly using living cells. Here, there were cell extracts of recombinant Lactococcus lactis expressing mouse RANKL (mRANKL) used to evaluate its biological activity in mice. Mice were divided into three groups that were fed phosphate-buffered saline (PBS), wild-type L. lactis IL1403 (WT_CE), and recombinant L. lactis expressing mRANKL (mRANKL_CE). The small intestinal transcriptome and fecal microbiome were then profiled. The biological activity of mRANKL_CE was confirmed by studying RANK-RANKL signaling in vitro and in vivo. For small intestinal transcriptome, differentially expressed genes (DEGs) were identified in the mRANKL_CE group, and no DEGs were found in the WT_CE group. In the PBS vs. mRANKL_CE gene enrichment analysis, upregulated genes were enriched for heat shock protein binding, regulation of bone resorption, and calcium ion binding. In the gut microbiome analysis, there were no critical changes among the three groups. However, Lactobacillus and Sphingomonas were more abundant in the mRANKL_CE group than in the other two groups. Our results indicate that cell extracts of mRANKL_CE can play an effective role without a significant impact on the intestine. This strategy may be useful for the development of protein drugs.

• Restorative Dentistry and Endodontics
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• v.49 no.1
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• pp.6.1-6.16
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• 2024

Objectives: This study aimed to investigate the elemental analysis and microhardness of a bioactive material (Activa) and marginal tooth structure after storage in different media. Materials and Methods: Fifteen teeth received cervical restorations with occlusal enamel and gingival dentin margins using the tested material bonded with a universal adhesive, 5 of them on the 4 axial surfaces and the other 10 on only the 2 proximal surfaces. The first 5 teeth were sectioned into 4 restorations each, then stored in 4 different media; deionized water, Dulbecco's phosphate buffered saline (DPBS), Tris buffer, and saliva. The storage period for deionized water was 24 hours while it was 3 months for the other media. Each part was analyzed by scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) analysis for different substrates/distances and the wt% of calcium, phosphorus, silica, and fluoride were calculated. The other 10 teeth were sectioned across the restoration, stored in either Tris buffer or saliva for 24 hours or 3 months, and were evaluated for microhardness of different substrates/areas. Data were analyzed using analysis of variance and Tukey's post hoc test. Results: Enamel and dentin interfaces in the DPBS group exhibited a significant increase in calcium and phosphorus wt%. Both silica and fluoride significantly increased in tooth structure up to a distance of 75 ㎛ in the 3-month-media groups than the immediate group. Storage media did not affect the microhardness values. Conclusions: SEM-EDS analysis suggests an ion movement between Activa and tooth structure through a universal adhesive while stored in DPBS.

• Analytical Science and Technology
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• v.23 no.6
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• pp.531-536
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• 2010

A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.6
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• pp.538-544
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• 1985

This study was carried cut to evaluate the water quality of spring waters in Pusan area(see Fig. 1). In this experiment, twenty-five water samples were collected from 5 stations from December 1983 to August 1984. Range and mean values of constituents of the samples are as follows: pH $5.80{\sim}7.25$, 6.60; water temperature $6.0{\sim}23.0^{\circ}C,\;12.9^{\circ}C$; total residue $33.0{\sim}325mg/l$, 121.2mg/l; alkalinity $4.75{\sim}51.6mg/l$, 24.1mg/l; hardness $9.47{\sim}85.0mg/l$, 30.3mg/l; electrical conductivity $0.495{\sim}2.750{\times}^2{mu}{\mho}/cm,\;1.239{\times}10^2{\mu}{\mho}/cm$;turbidity $0.54{\sim}7.80$NTU, 2.04NTU; $KMnO_4$ consumed $0.51{\sim}8.47mg/l$, 1.96mg/l; chloride ion $4.91{\sim}36.0mg/l$, 12.55mg/l; fluoride ion ND-0.30ppm, 0.08ppm; nitrate-nitrogen ND-8.94mg/l, 1.94m:g/l; nitrite-nirogen ND-0.10mg/l, 0.03mg/l; ammonia-nitrogen ND-0.16mg/l, 0.03mg/l: phosphate-phosphorus ND-0.09mg/l, 0.03mg/l; silicate-silicious $0.42{\sim}22.7ng/l$, 7.96mg/l; copper ND-10.5ppb, 2.46ppb; lead ND-22.7ppb, 3.54ppb; zinc ND-103ppb, 21.33ppb; iron $20.3{\sim}2,800ppb$, 801.72ppb, respectively. Arsenic, cyan, cadmium, manganese, mercury, chrome and phenol were not detected. Total residue, electrical conductivity, turbidity and chloride ion of station 1 (Milrakdong) were higher than others as 178.1mg/l, $2.127{\times}10^2{\mu}{\mho}/cm$, 3.16NTU and 16.32mg/l. The concentration of silicious had a great influence on precipitation. The concentration of fluoride ion of spring waters was lower as 0.08ppm than the criterion for drinking water as 1ppm, while iron was exceed 2.7 times as 801.72ppb.

• Journal of dental hygiene science
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• v.18 no.4
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• pp.218-226
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• 2018

The aim of this study was to compare fluoride release and surface changes according to different orthodontic bracket adhesives the application of fluoride products. We used non-fluoridated composite resin Transbond fluoridated composite resins Blugloo and LightBond, resin-modified glass ionomer Rely $X^{TM}$ Luting 2, and conventional glass ionomer Fuji $I^{(R)}$. Fluoride release of five orthodontic bracket adhesives and fluoride release ability after application of three fluoride products (1.23% acidulated phosphate fluoride gel, Tooth Mousse $Plus^{(R)}$, Fluor Protector, and a toothbrush with sodium fluoride-containing toothpaste) were measured using a fluoride electrode that was connected to an ion analyzer. After 4 weeks of fluoride application, the surface roughness and surface morphology were examined using a surface roughness tester and field emission scanning electron microscopy. The amounts of fluoride release were observed not only on application of Tooth Mousse $Plus^{(R)}$ and Fluor Protector on resin-modified glass ionomer Rely $X^{TM}$ Luting 2 and Fuji $I^{(R)}$, but also during tooth brushing using fluoride-containing toothpaste. After application of Tooth Mousse $Plus^{(R)}$, except Transbond XT, the surface roughness increased, and all orthodontic adhesives showed a partial drop of micro-particle filler. On application of 1.23% acidulated phosphate fluoride gel on all orthodontic bracket adhesives, their surface roughness increased. To bond the orthodontic bracket, resin-modified glass ionomer Rely $X^{TM}$ Luting 2 and Fuji $I^{(R)}$ adhesives are highly recommended if the amount of fluoride release is considered to confer a preventative effect on dental caries, and among the fluoride products, Tooth Mousse $Plus^{(R)}$ and Fluor Protector are better than 1.23% acidulated phosphate fluoride gel, and these are expected to prevent dental caries even during tooth brushing with fluoride-containing toothpaste.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.4
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• pp.347-355
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• 1986

The Sonakdong river is very important water source not only for agricultural water of Kimhae field but also for fishery water. Recently the middle and lower areas of the river have the tendency to be heavily contaminated by domestic sewage and agricultural chemicals. Fifty six water samples were collected from 8 stations from July to December in 1985 for the experiment (Fig. 1). To evaluate the water quality, pH, water temperature, electrical conductivity, chloride ion, nutrients ($NO^{-}_{2}-N,\;NO^{-}_{3}-N,\;NH^{+}_{4}-N,\;PO^{3-}_{4}-P,\;SiO_2-Si$), total coliform, fecal coliform, and fecal streptococcus were determined. Range and mean value of the samples were as follows; pH $6.3{\sim}9.4$, 7.91; water temperature $6.1{\sim}34.8^{\circ}C,\;23.88^{\circ}C$; electrical conductivity (from St. A to G) $1.575{\times}10^2{\sim}30.50{\times}10^2{\mu}{\mho}/cm,\;6.57{\times}10^2{\mu}{\mho}/cm$; chloride ion $23.5{\sim}14,300mg/l$, 770.0mg/l; nitrite-nitrogen $0.007{\sim}0.110mg/l$, 0.053mg/l; nitrate-nitrogen $0.001{\sim}1.638mg/l$, 0.649 mg/l; ammonia-nitrogen $0.017{\sim}4.200mg/l$, 0.497mg/l; phosphate-phoshorus $0.011{\sim}0.281mg/l$, 0.086mg/l; and silicate-silicious $2.4{\sim}6.5mg/l$, 4.43mg/l. Electrical conductivity and chloride ion of the station F(Chomanpo) were $2.676{\times}10^2{\mu}{\Omega}cm$ and 123.99mg/l which were lower than those of others. Among the analyzed nutrients, silicate-silicious concentration was the highest through all the samples. The bacterial density of the samples ranged $36{\sim}110,000/100ml$ for total coliform, $15{\sim}46,000/100ml$ for fecal coliform and $3.6{\sim}15,000/100ml$ for fecal streptococcus. The range and the mean of the TC/FC ratio were $3.0{\sim}9.6$, 5.51 and those of the FC/FS ratio were $1.1{\sim}9.2$, 6.19, respectively. On the other hand, fecal coliform was not detected in about $78\%$ of the water samples examined. Composition of coliform was $52\%$ Escherichia coli group, $3\%$ Citrobacter freundii group, $13\%$ Enterobacter aerogenes group and $31\%$ others.

• Journal of Life Science
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• v.21 no.7
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• pp.1032-1038
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• 2011

Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.583-591
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• 1997

The functional derangement of skeletal muscles which may be attributed to chronic hypoxia has been accepted as a possible mechanism of exercise impairment in patients with chronic obstructive pulmonary disease (COPD). The metabolic changes in skeletal muscle in patients with COPD are characterized by impaired oxidative phosphorylation, early activation of anaerobic glycolysis and excessive lactate and hydrogen ion production with exercise. But the cause of exercise limitation in patients with chronic lung disease without hypoxia has not been known. In order to evaluate the change in the skeletal muscle metabolism as a possible cause of the exercise limitation in chronic lung disease patients without hypoxia, we compared the muscular metabolic data of seven male patients which had been derived from noninvasive $^{31}P$ magnetic resonance spectroscopy(MRS) with those of five age-matched normal male control persons. $^{31}P$ MRS was studied during the sustained isometric contraction of the dominant forearm flexor muscles up to the exhaustion state and the recovery period. Maximal voluntary contraction(MVC) force of the muscle was measured before the isometric exercise, and the 30% of MVC force was constantly loaded to each patient during the isometric exercise. There were no differences of intracellular pH (pHi) and inorganic phosphate/phosphocreatine(Pi/PCr) at baseline, exhaustion state and recovery period between two groups. But pHi during the exercise was lower in patients group than the control group (p < 0.05). Pi/PCr during the exercise did not show significant difference between two groups. These results suggest that the exercise limitation in chronic lung disease patients without hypoxia also could be attributed to the abnormalities in the skeletal muscle metabolism.

• Journal of Korean Society of Forest Science
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• v.74 no.1
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• pp.37-46
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• 1986

To study the changes of soil and vegetational composition of Pinus densiflora community by campsite impact of its users, located in Kumo Mountain Provincial Park, the South part of Korea, three sites were selected according to the density of users. The heavy-use sites in this study has changed in the following ways. (1) The concentration of total nitrogen, available phosphate and Ca ion have decreased greatly. (2) From 68 to 73 percent of the undergrowth vegetation has been removed from the $2.000m^2$ camp area, leaving a sparse vegetation cover quite dissimilar in composition to nonuse campsite. (3) The soil hardness was $38.9kg/cm^2$ in heavyuse site and $1.76kg/cm^2$ in nonuse site. (4) The tolerant trees to campsite impact is Acer palmatum, Stephanandra incisa, Rhus trichocarpa, Rubus crataegifolitus and Parthenocissus tricuspidata. (5) Species diversity decreased to a degree at the heavy-use site. Similarity index was 25.38% between the heavy-use and nonuse site.

• Journal of agricultural medicine and community health
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• v.22 no.1
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• pp.61-74
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• 1997

In case of tissue invading nematode, proteolytic enzyme was required at their parasitic life. Proteinases obtained from these parasites(Toxocara canis, Ansakis spp. and Trichinella spiralis) were extracted, isolated and further purified. And then the analysis for activity and inhibitory effect of proteinases were performed by appropriate substrate. Determination of protein as a circulating antigen was done in use of infected animal serum with above parasites, respectively. For above experimental objects, following procedures were performed. First, enzymatic activity was measured in use of azocasein and inhibitory effect of porteinase were studied by various inhibitors. Second, partially purified proteins containing enzymatic activity were obtained by ion exchange chromatography, ultrafiltration and electrophoretic elution. Third, role of the partially purified protein as a circulating antigen. The results obtained were as follows : 1. Enzymatic activity of each nematode proteinase was varied according to pH. Optimal pH of Toxocara canis, Ansakis spp. and Trichinella spiralis were pH 6.0, pH 5.5 and pH 6.5, respectively. The optimal molarity of buffer was 0.1M phosphate buffer. Although little difference between these proteinases was observed, temperature stability was at least maintained at $4^{\circ}C$ until 5 days. 2. In case of Ansakis spp. and Toxocara canis, enzymatic activity of these proteinases was considerably inhibited by Leupeptin and EDTA. For maximum enzymatic activity of above proteinases, it was required that cysteine residue of enzyme should be protected. And it was suggested that metallo type was contained in enzyme active site. Proteinase of Trichinella spiralis contained metallo type also. 3. Although partial purification was performed in Ansakis spp. and Toxocara canis, proteins maintaining enzymatic activity were identified as a circulating antigen. From SDS-PAGE and immunoblot, 25 kDa was presented in Ansakis spp.. Specific antigen of Toxocara cains was 110 kDa protein fraction. 55 and 42 kDa proteins were reacted with normal serum. Trichinella spiralis 60 kDa protein fraction was successfully purified from excretory materials in culture. As a result of immune-reaction with Trichinella spiralis infected serum, highly purified 60 kDa protein was maintained antigenicity until final purification step.