• Journal of Microbiology and Biotechnology
• /
• v.12 no.5
• /
• pp.819-825
• /
• 2002

Methanol dehydrogenase (MDH) is a key enzyme in the process of methanol oxidation in methylotrophic bacteria. However, information on MDH genes from genus Methylobacillus is limited. In this study, a 6.5-kb HindIII DNA fragment of Methylobacillus sp. SK-5 chromosomal DNA was isolated from the genomic library of the strain by using a degenerate oligonucleotide probe that was designed based on JV-terminal amino acid sequence of the MDH $\alpha$ subunit purified from the strain. Molecular analysis of the fragment revealed four tightly clustered genes (mxaFJGI) involved in the methanol oxidation. The first and fourth genes were very similar to mxaF (77% identity for nucleotides an 78% identity for amino acids) and mxaF (67% Identity for nucleotides and 68% Identity for amino acids) genes, respectively, from Methylovorus sp. SSI. Genes mxaF and mxaI encode $\alpha$ and $\beta$ subunits of MDH, respectively. The two subunits were identified from purified MDH from Methylobacillus sp. SK-5. A dendrogram constructed by comparison of amino acid sequences of MDH u subunits suggests that MxaF from Methylobacillus sp. SK-5 belongs to a subfamily cluster of MDH u subunits from $\beta$-subgroup Proteobacteria. The subfamily cluster is separated from the other subfamily that consists of $\beta$- and $\gamma$-subgroup Proteobacteria. This study provided information on mn genes from a methylotrophic bacterium in $\beta$-subgroup Proteobacteria, which would aid to better develop a gene probe to detect one-carbon metabolizing bacteria.

• Korean Journal of Microbiology
• /
• v.38 no.3
• /
• pp.192-197
• /
• 2002

Fluorescent in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes was used to compare the community structures of free-living and aggregated bacteria at thawing period in Lake Baikal. Targeted groups were Eubacteria, $\alpha$-, $\beta$-, $\gamma$- proteobacteria groups, Cytophaga-Flavobacterium group and Planctomycetales. Total bacterial numbers of free-living bacteria were ranged from $0.2{\times}10^6\cells{\cdot}ml^-1$ to $3.2{\times}10^6\cells{\cdot}ml^-1$, which were decreasing with depth, while the aggregated bacterial numbers were dramatically increasing from $0.4{\times}10^4 to 3.3{\times}10^4 \cells{\cdot}ml^-1$ with depth. The ratios of EUB probe binding cells to DAPI counts were ranged from 52.3 to 74.1% in free-living bacteria, and from 39.6 to 66.7% in the aggregated bacteria, respectively. Community structures of the aggregated bacteria were very different from each free-living bacteria at every depth. At 25 m depth, where the chlorophyll a concentration was highest, both structures were quite different from those of surface layers, rendering the fact that the community structures might be affected by phytoplankton. The vertical profile of community structure of aggregated bacteria is particular. The proportion of $\beta$-proteobacteria group was increasing with depth and it was 51.8% at 100 m, but the dominant group was $\gamma$-pro-teobacteria group at 250 m. Taken together, the biodiversity and succession of aggregated bacteria are quite different from free-living bacteria.

• Journal of Microbiology
• /
• v.42 no.4
• /
• pp.278-284
• /
• 2004

Changes in the soil bacterial community of a coniferous forest were analyzed to assess microbial responses to wildfire. Soil samples were collected from three different depths in lightly and severely burned areas, as well as a nearby unburned control area. Direct bacterial counts ranged from $3.3­22.6\times10^8\;cells/(g{\cdot}soil).$ In surface soil, direct bacterial counts of unburned soil exhibited a great degree of fluctuation. Those in lightly burned soil changed less, but no significant variation was observed in the severely burned soil. The fluctuations of direct bacterial count were less in the middle and deep soil lay­ers. The structure of the bacterial community was analyzed via the fluorescent in situ hybridization method. The number of bacteria detected with the eubacteria-targeted probe out of the direct bacterial count varied from $30.3\;to\;84.7\%,$ and these ratios were generally higher in the burned soils than in the unburned control soils. In the surface unburned soil, the ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ Cytoph­aga-Flavobacterium group, and other eubacteria groups to total eubacteria were 9.9, 10.6, 15.5, 9.0, and $55.0\%,$ respectively, and these ratios were relatively stable. The ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ and Cytophaga-Flavobacterium group to total eubacteria increased immediately after the wildfire, and the other eubacterial proportions decreased in the surface and middle layer soils. By way of contrast, the composition of the 5 groups of eubacteria in the subsurface soil exhibited no significant fluctuations dur­ing the entire period. The total bacterial population and bacterial community structure disturbed by wildfire soon began to recover, and original levels seemed to be restored 3 months after the wildfire.

• Journal of Biomedical Engineering Research
• /
• v.5 no.2
• /
• pp.167-172
• /
• 1984

In the present study, we developed a nuclear stethoscope interfaced with a personal computer (APPLE- II) and evaluated it's performances as compared with the gamma camera. This single-probe, ECG-gated system has been used for displaying the left ventricular time-activity curves, the peak ejection time and its rate, the peak filling time and its rate, and the R-R interval distribution. The radioactivity was measured at every 10msec around the ventricular region, where the activity curves shows the maximal peak to peak variations. The background activity was measured around the lung area showing its counts approximately 50% of the end-diastolic count with minimal variations. The average time-activity curves of 100 beats were used for analysis in the equilibrium study after intravenous injection of 15-20mCi of Tc. The ejection fractions measured by the nuclear stethoscope(Y) were compared with those measured by gamma camera(X) in 47 patients with various heart diseases. The correlation coefficient between two measurements was 0.766 with a relation of Y=1.04 x-8.48. Also, the high reproducibility was obtained for the same patient. Also, the high reproducibility was obtained for the same patient. From this study, we conclude that this device is useful for continuous monitoring in the intensive care unit, as it is portable, compact, and inexpensive.

• Biomolecules & Therapeutics
• /
• v.22 no.1
• /
• pp.27-34
• /
• 2014

Curcumin is naturally occurring polyphenolic compound found in turmeric and has many pharmacological activities. The present study was undertaken to evaluate anti-allergic inflammatory activity of curcumin, and to investigate its inhibitory mechanisms in immunoglobulin E (IgE)/Ag-induced mouse bone marrow-derived mast cells (BMMCs) and in a mouse model of IgE/Ag-mediated passive systemic anaphylaxis (PSA). Curcumin inhibited cyclooxygenase-2 (COX-2) dependent prostaglandin $D_2$ ($PGD_2$) and 5-lipoxygenase (5-LO) dependent leukotriene $C_4$ ($LTC_4$) generation dose-dependently in BMMCs. To probe the mechanism involved, we assessed the effects of curcumin on the phosphorylation of Syk and its downstream signal molecules. Curcumin inhibited intracellular $Ca^{2+}$ influx via phospholipase $C{\gamma}1$ ($PLC{\gamma}1$) activation and the phosphorylation of mitogen-activated protein kinases (MAPKs) and the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) pathway. Furthermore, the oral administration of curcumin significantly attenuated IgE/Ag-induced PSA, as determined by serum $LTC_4$, $PGD_2$, and histamine levels. Taken together, this study shows that curcumin offers a basis for drug development for the treatment of allergic inflammatory diseases.

• Biomolecules & Therapeutics
• /
• v.16 no.4
• /
• pp.328-335
• /
• 2008

This study was undertaken to investigate whether honokiol could enhance the pentobarbitalinduced sleeping behaviors through $\gamma$-aminobutyric acid (GABA) receptor $Cl^-$ channel activation. Thirty minutes after the oral administration of honokiol, mice were received sodium pentobarbital (42 mg/kg, i.p.). The time elapsed from pentobarbital injection to the loss of the righting reflex was taken as sleeping latency. The time elapsed between the loss and voluntary recovery of the righting reflex was considered as the total sleeping time. Western blot technique and $Cl^-$ sensitive fluorescence probe were used to detect the expression of $GABA_A$ receptor subunits and $Cl^-$ influx in the primary cultured cerebellar granule cells. Honokiol (0.1 and 0.2 mg/kg) prolonged the sleeping time induced by pentobarbital (42 mg/kg) in a dosage-dependent manner. Honokiol (20 and 50 ${\mu}M$) increased $Cl^-$ influx in primary cultured cerebellar granule cells, and selectively increased the $GABA_A$ receptor $\alpha$-subunit expression, but had no effect on the abundance of $\beta$ or $\gamma$-subunits. Chronic treatment with 20 ${\mu}M$ honokiol in primary cultured cerebellar neurons did not affect the abundance of GAD65/67. The results suggested that honokiol could potentiate pentobarbital-induced sleeping through $GABA_A$ receptor $Cl^-$ channel activation.

• Journal of Adhesion and Interface
• /
• v.14 no.1
• /
• pp.13-20
• /
• 2013

Palladium plays a pivotal role in the production of dental and medicinal devices, medicinal substances, jewellery, automobile and high-performance adhesives. Despite the frequent and fruitful use of such reactions, one major setback is the high level of palladium in the resultant compounds which can harm the human body. Among the palladium species, $PdCl_2$ is the most toxic. As a consequence it is desirable to detect the $Pd^{2+}$ cations by fluorescence spectra because it can provide an operationally simple and cost-effective detection method together with high sensitivity and selectivity. Herein, an ${\alpha}$-carbonyl substituted pyrene derivative, ${\gamma}$-oxo-1-pyrenebutyric acid (OPBA), was demonstrated to be a highly sensitive and selective fluorescent probe for $Pd^{2+}$ among the metal cations examined in aqueous solutions.

• Clinical and Experimental Reproductive Medicine
• /
• v.38 no.2
• /
• pp.68-74
• /
• 2011

Objective: Previously, we found that oocyte specific homeobox (Obox) 4 plays significant role in completion of meiosis specifically at meiosis I-meiosis II (MI-MII) transition. The purpose of this study was to determine the mechanism of action of $Obox4$ in oocyte maturation by evaluating downstream signal networking. Methods: The $Obox4$ dsRNA was prepared by $in$ $vitro$ transcription and microinjected into the cytoplasm of germinal vesicle oocytes followed by $in$ $vitro$ maturation in the presence or absence of 0.2 mM 3-isobutyl-1-metyl-xanthine. Total RNA was extracted from 200 oocytes of each group using a PicoPure RNA isolation kit then amplified two-rounds. The probe hybridization and data analysis were used by Affymetrix Gene-Chip$^{(R)}$ Mouse Genome 430 2.0 array and GenPlex 3.0 (ISTECH, Korea) software, respectively. Results: Total 424 genes were up (n=80) and down (n=344) regulated after $Obox4$ RNA interference (RNAi). Genes mainly related to metabolic pathways and mitogen-activated protein kinase (MAPK) signaling pathway was changed. Among the protein kinase C (PKC) isoforms, PKC-alpha, beta, gamma were down-regulated and especially the MAPK signaling pathway PKC-gamma was dramatically decreased by $Obox4$ RNAi. In the cell cycle pathway, we evaluated the expression of genes involved in regulation of chromosome separation, and found that these genes were down-regulated. It may cause the aberrant chromosome segregation during MI-MII transition. Conclusion: From the results of this study, it is concluded that $Obox4$ is important upstream regulator of the PKC and anaphase-promoting complex action for maintaining intact germinal vesicle.

• Journal of Radiation Industry
• /
• v.5 no.1
• /
• pp.25-33
• /
• 2011

Skin contact allergy (SCA) is not life-threatening, but a large number of people have been suffered from the reactions caused by various kinds of chemicals and products. Thus, in this study, radiation technology was employed to improve the traditional herb addition method on the SCA reduction of Rhus verniciflua sap (RVS). Rhus verniciflua has traditionally been used as an herbal medicine plant, but its urushiol derivatives are known as a major allergen for the SCA. The present study was commenced to assess the allergenicity of both gamma-irradiated and non-irradiated RVS by using guinea-pig maximization test (GPMT) in order to probe the mechanism of an SCA. In the acute dermal irritation assays, non-irradiated RVS caused erythema, but the irradiated RVS did not provoke any erythema on the abdominal skin of the guinea pigs. From the result of the GPMT, urushiols, the main chemical components of RVS, were identified as an extreme skin sensitizer, and the removal of urushiols by irradiation extremely reduced the erythema. These results suggest that radiation technology is a novel method to reduce SCA through the removal of urushiols of RVS.

• Korean Journal of Clinical Laboratory Science
• /
• v.37 no.3
• /
• pp.178-184
• /
• 2005

Carry over contamination has been reduced in some systems by flushing the internal and external surfaces of the sample probe with copious amount of diluent. It between specimens should be kept as small as possible. A built-in, continuous-flow wash reservoir, which allows the simultaneous washing of the interior and exterior of the syringe needles, addresses this issue. In addition, residual contamination can further be prevented through the use of efficient needle rinsing procedures. In discrete systems with disposable reaction vessels and measuring cuvets, any carry over is entirely caused by the pipetting system. In analyzers with reuseable cuvets or flow cells, carry over may arise at every point through which high samples pass sequentially. Therefore, disposable sample probe tips can eliminate both the contamination of one sample by another inside the probe and the carry over of in specimen into the specimen in the cup. The results of the applicative carry over experiment studied on 21 items for total protein (TP), albumin (ALB), total bilirubin (TB), alkaline phosphatase (ALP), aspratate aminotranferase (AST), alanine aminotranferase (ALT), gamma glutamyl transferase (GGT), creatinine kinase (CK), lactic dehydrogenase (LD), creatnine (CRE), blood urea nitrogen (BUN), uric acid (UA), total cholesterol (TC), triglyceride (TG), glucose (GLU), amylase (AMY), calcium (CA), inorganic phosphorus (IP), sodium (Na), potassium (K), chloride (CL) tests in chematology were as follows. Evaluation of process performance less than 1% in all tests was very good, but a percentage of ALB, TP, TB, ALP, CRE, UA, TC, GLU, AMY, IP, K, Na, and CL was 0%, implying no carry over. Other tests were ALT(-0.08%), GGT(-0.09%), CK(0.08%), LD(0.06%), BUN(0.12%), TG (-0.06%), and CA(0.89%).