Purpose: $^{13}N$-ammonia is a well known radiopharmaceutical for the measurement of a myocardial blood flow (MBF) non-invasively using PET-CT. In this study, we investigated a correlation between MBF obtained from dynamic imaging and myocardial perfusion score (MPS) obtained from static imaging for usefulness of cardiac PET study. Methods: Twelve patients (11 males, 1 female, $57.9{\pm}8.6$ years old) with suspicious coronary artery disease underwent PET-CT scan. Dynamic scans (6 min: $5\;sec\;{\times}\;12,\;10\;sec\;{\times}\;6,\;20\;sec\;{\times}\;3,\;and\;30\;sec\;{\times}\;6$) were initiated simultaneously with bolus injection of 11 MBq/kg $^{13}N-ammonia$ to acquire rest and stress image. Gating image was acquired during 13 minutes continuously. Nine-segment model (4 basal walls, 4 mid walls, and apex) was used for a measurement of MBF. Time activity curve of input function and myocardium was extracted from ROI methods in 9 regions for quantification. The MPS were evaluated using quantitative analysis software. To compare between 20-segment model and 9-segment model, 6 basal segments were excluded and averaged segmental scores were used. Results: There are weak correlation between MBF (rest, 0.18-2.38 ml/min/g; stress, 0.40-4.95 ml/min/g) and MPS (rest 22-91%, stress, 14-90%), however the correlation coefficient between corrected MBF and MPS in rest state was higher than stress state (rest r=0.59; stress r=0.80). As a thickening increased, correlation between MBF and MPS also showed good correlation at each segments. Conclusions: Corrected and translated MPS as its characteristics using $^{13}N$-ammonia showed good correlation with absolute MBF measured by dynamic image in this study. Therefore, we showed MPS is one of good indices which reflect MBF. We anticipate PET-CT could be used as useful tool for evaluation of myocardial function in nuclear cardiac study.
Purpose: Multidrug resistance (MDR) of the cancer cells related to mdr1 gene expression can be effectively treated by selective short hairpin RNA for mdr1 gene (shMDR). Sodium/iodide symporter (NIS) gene is well known to have both reporter and therapeutic gene characteristics. We have co-transfected both shMDR and NIS gene into colon cancer cells (HCT15 cell) expressing MDR and Tc-99m sestamibi and I-125 uptake were measured. In addition, cytotoxic effects of doxorubicin and I-131 therapy were also assessed after transfection. Material and Methods: At first, shMDR was transfected with liposome reagent into human embryonic kidney cells (HEK293) and HCT cells. shMDR transfection was confirmed by RT-PCR and western blot analysis. Adenovirus expressing NIS (Ad-NIS) gene and shMDR (Ad-shMDR) were co-transfected with Ad-NIS into HCT15 cells. Forty-eight hours after infection, inhibition of P-gycoprotein (Pgp) function by shMDR was analyzed by a change of Tc-99m sestamibi uptake and doxorubicin cytotoxicity, and functional activity of induced NIS gene expression was assessed with I-125 uptake assay. Results: In HEK293 cells transfected with shMDR, mdr1 mRNA and Pgp protein expressions were down regulated. HCT15 cells infected with 20 MOI of Ad-NIS was higher NIS protein expression than control cells. After transfection of 300 MOI of Ad-shMDR either with or without 10 MOI of Ad-NIS, uptake of Tc-99m sestamibi increased up to 1.5-fold than control cells. HCT15 cells infected with 10 MOI of Ad-NIS showed approximately 25-fold higher I-125 uptake than control cells. Cotransfection of Ad-shMDR and Ad-NIS resulted in enhanced cytotoxic by doxorubicin in HCT15 cells. I-131 treatment on HCT15 cells infected with 20 MOI of Ad-NIS revealed increased cytotoxic effect. Conclusion: Suppression of mdr1 gene expression, retention of Tc-99m sestamibi, enhanced doxorubicin cytotoxicity and increases in I-125 uptake were achieved in MDR expressing cancer cell by co-transfection of shMDR and NIS gene. Dual therapy with doxorubicin and radioiodine after cotransfection shMDR and NIS gene can be used to overcome MDR.
Purpose: Our purpose was to evaluate spinal bony metastasis which could be missed on an F-18 FDG PET/CT (FDG PET/CT) alone, and to characterize discordant metastatic lesions between FDG PET/CT and bone scan. Material and Methods: FDG PET/CT and bone scans of 43 patients with spinal bony metastasis were analyzed retrospectively. A McNemar test was performed comparing the FDG PET/CT alone to the FDG PET/CT plus bone scan in the spinal bony metastases. A one-way chi-square test was performed to characterize the metastases that were missed on the FDG PET/CT alone. To evaluate discordant lesions between FDG PET/CT and bone scan, we performed logistic regression analyses. The independent variables were sites (cervical, thoracic, and lumbar), size (large and small), and maximum SUVs, and the dependant variable was bone scan uptake (positive and negative MDP uptake). Results: A significant difference was found between the FDG PET/CT alone and the FDG PET/CT combined with the bone scan (p < 0.01). Using the FDG PET/CT only, diffuse osteoblastic metastasis was missed with a significantly higher frequency (p = 0.04). In the univariate analysis, cervical vertebra and small size were related to negative MDP uptake, and thoracic vertebra and large size were related to positive MDP uptake. However, in the multivariate analysis, only the large size was related to positive MDP uptake. Conclusion: A bone scan in addition to the FDG PET/CT increased the ability to evaluate spinal bony metastases, especially for diffuse osteoblastic metastasis. Large metastasis was related to positive bone scan uptake in spinal bony metastasis.
A strain GP32 which produces a highly viscous extracellular polysaccharide was conducted with soil samples and identified as Pseudomonas species. The culture flask conditions for the production of extracellular polysaccharide by Pseudomonas sp. GP32 were investigated. The most suitable carbon and nitrogen source for extracellular polysaccharide production were galactose and (NH4)2SO4. The optimum carbon/nitrogen ratio for the production of extracellular polysaccharide was around 50. The optimum pH and temperature for extracellular polysaccharide production was 7.5 and 32℃, respectively. In batch fermentation using a jar fermentor, the highest extracellular polysaccharide content (15.7 g/l) was obtained after 70 hr of cultivation. The extracellular polysaccharide produced by Pseudomonas sp. GP32 (designated Biopol32) was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography. Biopol32, which has an estimated molecular weight of over 3×107 datons, is a novel polysaccharide derived from sugar components consisting of galactose, glucose, gulcouronic acid and galactouronic acid in an approximate molar ratio of 1.85 : 3.24 : 1.00 : 1.42. The solution of Biopol32 showed non-Newtonian characteristics. The viscosity of Biopol32 exhibited appeared to be higher at all concentration compared to that of zooglan from Zoogloea ramigera. An analysis of the flocculating efficiency of Biopol32 in industry wastewater (food, textile, and paper wastewater) revealed chemical oxygen demand (COD) reduction rates 58.4-67.3% and suspended solid (SS) removal rates 82.6-91.3%. Based on these results, Biopol32 is a possible candidate for industrial applications such as wastewater treatment.
Equine chorionic gonadotropin (eCG) consists of highly glycosylated ${\alpha}-$ and ${\beta}-subunits$ and is a unique member of the gonadotropin family, because it elicits the response characteristics of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. To directly assess the biological function of $rec-eCG{\beta}/{\alpha}$, we constructed mammalian expressing vectors of equine luteinizing hormone/chorionic gonadotropin receptors (eLH/CGR). The activity of $rec-eCG{\beta}/{\alpha}$ in vitro assayed in transient transfected CHO-K1 cells and in stably transfected PathHunter Parental cells with eLH/CGR was investigated. $rec-eCG{\beta}/{\alpha}$ was efficiently secreted in the CHO-K1 suspension cell media, and the quantity detected was about 200 mIU/ml from 1 to 7 days after transfection. In the western blot analysis, the $rec-eCG{\beta}/{\alpha}$ protein was broadly identified to be about 40~45 kDa molecular weight. The cAMP stimulation in CHO-K1 cells expressing eLH/CGR was determined to evaluate the activity of $rec-eCG{\beta}/{\alpha}$. The cAMP concentration increased in direct proportion to the concentration of the $rec-eCG{\beta}/{\alpha}$. The $EC_{50}$ value in the transient transfected CHO-K1 cells was $8.1{\pm}6.5ng$. The stable cell lines of eLH/CGR were established in the PathHunter Parental cells expressing ${\beta}-arrestin$. We found that $rec-eCG{\beta}/{\alpha}$ had full LH activity in the PathHunter Parental cells expressing eLH/CGR. The $EC_{50}$ value in transient and stable cells was $5.0{\pm}4.7ng/ml$ and $4.5{\pm}5.2ng/ml$, respectively. These results suggest that $rec-eCG{\beta}/{\alpha}$ has a biological activity in a cell expressing eLH/CGR. These stable cells expressed in PathHunter Parental cells could be useful for elucidating the functional mechanisms of deglycosylated $rec-eCG{\beta}/{\alpha}$ mutants.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.
Characteristics related to grain quality and physiochemical components such as mineral, total amino acid, free amino acid, and free sugar composition were investigated in Protox inhibitor resistanttransgenic rice (MX, PX, and AP37) and its nontransgenic counterpart (WT). Head rice, palatability, protein, and whiteness (except for MX and AP37) of milled transgenic rice were high or similar to those of the non-transgenic counterpart. Immature rice, unfilled grain, and cracked kernels (PX and AP37) of milled transgenic rice were lower than those of its non-transgenic counterpart. However, there were no significant differences in damaged grain between the transgenic rice lines and its counterpart. Potassium content in PX and copper contents in PX and AP37 were only low compared with their non-transgenic counterparts, but other mineral contents in transgenic rice lines were high or showed no significant differences compared with non-transgenic counterparts. Contents of most total amino acid composition in transgenic rice lines were high or similar to those in non-transgenic counterparts, but the content of isoleucine in AP37 was only low compared with its non-transgenic counterpart. On the other hand, free amino acid, leucine and tyrosine in PX and AP37, and total free amino acid in PX were low compared with their non-transgenic counterparts. However, the content of free amino acid in other kinds in transgenic rice lines were similar to those in their non-transgenic counterparts. Contents of sucrose in MX and PX were low compared with non-transgenic counterpars, but contents of fructose, glucose, and maltose in transgenic rice lines were high or similar compared with their non-transgenic counterparts. This results indicated that Protox genes had no negative affect on the nutritional composition of rice.
Journal of Korea Technical Association of The Pulp and Paper Industry
/
v.48
no.2
/
pp.34-45
/
2016
Global warming and climate change have been caused by combustion of fossil fuels. The greenhouse gases contributed to the rise of temperature between $0.6^{\circ}C$ and $0.9^{\circ}C$ over the past century. Presently, fossil fuels account for about 88% of the commercial energy sources used. In developing countries, fossil fuels are a very attractive energy source because they are available and relatively inexpensive. The environmental problems with fossil fuels have been aggravating stress from already existing factors including acid deposition, urban air pollution, and climate change. In order to control greenhouse gas emissions, particularly CO2, fossil fuels must be replaced by eco-friendly fuels such as biomass. The use of renewable energy sources is becoming increasingly necessary. The biomass resources are the most common form of renewable energy. The conversion of biomass into energy can be achieved in a number of ways. The most common form of converted biomass is pellet fuels as biofuels made from compressed organic matter or biomass. Pellets from lignocellulosic biomass has compared to conventional fuels with a relatively low bulk and energy density and a low degree of homogeneity. Thermal pretreatment technology like torrefaction is applied to improve fuel efficiency of lignocellulosic biomass, i.e., less moisture and oxygen in the product, preferrable grinding properties, storage properties, etc.. During torrefacton, lignocelluosic biomass such as palm kernell shell (PKS) and empty fruit bunch (EFB) was roasted under an oxygen-depleted enviroment at temperature between 200 and $300^{\circ}C$. Low degree of thermal treatment led to the removal of moisture and low molecular volatile matters with low O/C and H/C elemental ratios. The mechanical characteristics of torrefied biomass have also been altered to a brittle and partly hydrophobic materials. Unfortunately, it was much harder to form pellets from torrefied PKS and EFB due to thermal degradation of lignin as a natural binder during torrefaction compared to non-torrefied ones. For easy pelletization of biomass with torrefaction, pellets from PKS and EFB were manufactured before torrefaction, and thereafter they were torrefied at different temperature. Even after torrefaction of pellets from PKS and EFB, their appearance was well preserved with better fuel efficiency than non-torrefied ones. The physical properties of the torrefied pellets largely depended on the torrefaction condition such as reaction time and reaction temperature. Temperature over $250^{\circ}C$ during torrefaction gave a significant impact on the fuel properties of the pellets. In particular, torrefied EFB pellets displayed much faster development of the fuel properties than did torrefied PKS pellets. During torrefaction, extensive carbonization with the increase of fixed carbons, the behavior of thermal degradation of torrefied biomass became significantly different according to the increase of torrefaction temperature. In conclusion, pelletization of PKS and EFB before torrefaction made it much easier to proceed with torrefaction of pellets from PKS and EFB, leading to excellent eco-friendly fuels.
The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).
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