Significance of the Extracapsular Spread of Metastatic Lymph Nodes in Papillary Thyroid Carcinoma.

OBJECTIVES: The extracapsular spread (ECS) of metastatic lymph nodes is associated with aggressive tumor behavior, and is regarded as a major risk factor for local recurrence in patients with head and neck squamous cell carcinoma. However, the significance of ECS of metastatic lymph nodes has not been well established in well-differentiated thyroid carcinoma. The purpose of this study was to examine this question. METHODS: A retrospective review was performed of 335 patients with papillary thyroid carcinoma who underwent total thyroidectomy with lymph node dissection from April 2001 to December 2009. We analyzed various clinical characteristics, pathologic factors, and the size, number, and ECS of foci in metastatic lymph nodes. RESULTS: On pathologic review, 201 of the patients (56.6%) had lymph node metastasis. This was significantly related to age and tumor size. ECS was noted in 64 of these 201 patients (31.8%), and was significantly related to male gender, tumor size, presence of extrathyroidal extension, metastatic lymph node size, and focus size. Recurrence occurred in 13 patients (3.9%), and the presence of ECS was significantly related to recurrence. CONCLUSION: ECS of metastatic lymph nodes is an important prognostic factor for loco-regional recurrence in papillary thyroid carcinoma.

Site-Directed Mutagenesis-Based Functional Analysis and Characterization of Endolytic Lyase Activity of N- and C-Terminal Domains of a Novel Oligoalginate Lyase from Sphingomonas sp. MJ-3 Possessing Exolytic Lyase Activity in the Intact Enzyme.

A novel oligoalginate lyase from a marine bacterium, Sphingomonas sp. strain MJ-3, exhibited a unique alginate degradation activity that completely depolymerizes alginate to monomers through the formation of oligomers. In order to reveal the reason why MJ-3 oligoalginate can exhibit both endolytic and exolytic alginate lyase activities, ten mutants were developed and characterized on the basis of homology modeling. When the recombinant cell lysates containing the mutated proteins of MJ-3 oligoalginate lyase were allowed to react with alginate, the Asn177Ala, His178Ala, Tyr234Phe, His389Ala, and Tyr426Phe mutants showed reduced oligoalginate lyase activity, whereas the Arg236Ala mutant exhibited endolytic activity. Interestingly, the overexpressed Arg236Ala protein (79.6 kDa) was proteolytically cleaved into two fragments, i.e., the N-terminal 32.0-kDa and the C-terminal 47.6-kDa fragments. Both the purified N-terminal and C-terminal fragments showed endolytic lyase activity. They preferentially degraded a heteropolymeric (polyMG) block than poly-ß-D-mannuronate (polyM) or poly-α-L-guluronate (polyG) blocks. These results suggest that the oligoalginate lyase activity of MJ-3 enzyme is derived from the cooperative interaction between the N- and C-terminal endolytic alginate lyase domains in the intact enzyme.

Batch conversion of methane to methanol using Methylosinus trichosporium OB3b as biocatalyst.

Recently, methane has attracted much attention as an alternative carbon feedstock since it is the major component of abundant shale and natural gas. In this work, we produced methanol from methane using whole cells of Methylosinus trichosporium OB3b as the biocatalyst. M. trichosporium OB3b was cultured on NMS medium with a supply of 7:3 air/methane ratio at 30°C. The optimal concentrations of various methanol dehydrogenase inhibitors such as potassium phosphate and EDTA were determined to be 100 and 0.5 mM, respectively, for an efficient production of methanol. Sodium formate (40 mM) as a reducing power source was added to enhance the conversion efficiency. A productivity of 49.0 mg/l·h, titer of 0.393 g methanol/l, and conversion of 73.8% (mol methanol/mol methane) were obtained under the optimized batch condition.

Biosynthesis of glycerol carbonate from glycerol by lipase in dimethyl carbonate as the solvent.

Glycerol carbonate was synthesized from renewable glycerol and dimethyl carbonate using lipase in solvent-free reaction system in which excess dimethyl carbonate played as the reaction medium. A variety of lipases have been tested for their abilities to catalyze transesterification reaction, and Candida antartica lipase B and Novozyme 435 exhibited higher catalytic activities. The silica-coated glycerol with a 1:1 ratio was supplied to prevent two-phase formation between hydrophobic dimethyl carbonate and hydrophilic glycerol. Glycerol carbonate was successfully synthesized with more than 90% conversion from dimethyl carbonate and glycerol with a molar ratio of 10 using Novozyme 435-catalyzed transesterification at 70 °C. The Novozyme 435 [5% (w/w) and 20% (w/w)] and silica gel were more than four times recycled with good stability in a repeated batch operation for the solvent-free synthesis of glycerol carbonate.

Growth-inhibitory effects of a Bulnesia sarmienti aqueous extract on A549 cells in vitro and S180 cells in vivo.

The effects of Bulnesia sarmienti (BS) aqueous extract on the cell growth of A549 cell lines were investigated. BS has strong cytotoxic activity on the A549 cell lines (IC(50); less than 100 microg/mL) in MTT assay. HPLC confirmed that BS contains catechins as major compound. Cell cycle analysis by flow cytometry indicated that BS arrested the cell cycle in the sub-G(1) phase. BS induced DNA fragmentation, and increased the expression of the p53 protein in immunoblot analysis. These results indicated that the anticancer effect of BS was mediated via the process of apoptosis and growth-inhibition.

Anticancer activity and apoptotic effects of Bulnesia sarmienti against human lung cancer H460 cells.

Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. The effects of BS aqueous extract (100, 200, and 400 microg/ml) on H460 cell lines were investigated. High-performance liquid chromatography (HPLC) confirmed that BS contains catechins as major compound. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell cycle analysis, DNA fragmentation, apoptosis, and immunoblot analysis on cells were carried out. BS has strong cytotoxic activity on the H460 cell lines (IC50; less than 100 microg/ml) in MTT assay. Flow cytometry indicated that BS arrested the cell cycle in the sub-G1 phase. When BS was treated on H460 cells, DNA fragmentation was increased, and early apoptotic cells were shown to be positive by annexin V staining. Also, the expressions of the p53 and Bax were increased and Bcl-2 protein was downregulated with BS treatment. These results indicated that the BS has anticancer activity on H460 cells and BS may be useful in future therapeutic applications for developing anticancer agents.

Functional expression of recombinant human ribonuclease/angiogenin inhibitor in stably transformed Drosophila melanogaster S2 cells.

A recombinant plasmid harboring heterologous genes coding human ribonuclease/angiogenin inhibitor (RAI) was expressed in stably transformed Drosophila melanogaster Schneider 2 (S2) cells. Stably transformed polyclonal cell populations expressing RAI were isolated after 4 weeks of selection with hygromycin B. Recombinant RAI with a molecular weight of 50 kDa was detected in the intracellular (cell) and extracellular (medium) fractions of S2 cells. Recombinant RAI was purified from the extracellular fraction using a two-step purification scheme comprised of Ni-NTA and ion-exchange chromatography. Purified RAI migrated on SDS-PAGE as a single band in the elution fraction containing 300 mM NaCl. The ribonuclease inhibitor activity of purified RAI was measured using yeast tRNA and RNase A. Purified RAI exhibited an activity of approximately 8 U mug(-1) for the inhibition of RNA degradation by RNase A. Cultivation of stably transformed S2 cells using HyQ((R))SFX-insect MP medium increased cell growth by 79% and approximately doubled the production of recombinant RAI.

Characteristics of styrene degradation by Rhodococcus pyridinovorans isolated from a biofilter.

A novel strain (PYJ-1) of Rhodococcus pyridinovorans that was isolated from a biofilter was able to degrade styrene at a maximum rate of 0.16 mg (mg protein)(-1) h(-1) in batch culture at 97 mg l(-1) of initial styrene gas concentration. The optimum pH and temperature for styrene degradation were 7 and 32 degrees C, respectively. The degradation kinetic constants were obtained using substrate inhibition kinetics. In a perlite-packed biofilter the maximum styrene removal rate by the strain was 279 gm(-3)h(-1). Styrene removal in the biofilter was more sensitive to the temperature than in the batch culture.

Characteristics of Rhodococcus pyridinovorans PYJ-1 for the biodegradation of benzene, toluene, m-xylene (BTX), and their mixtures.

The degradation rates of benzene, toluene, and m-xylene (BTX) by Rhodococcus pyridinovorans PYJ-1, were highest at 30, 50, and 25 mg/l, respectively. The degradation rate was highest for toluene (0.106 mg/mg-protein x h) followed by benzene and m-xylene at the optimum pH and temperature of 7 and 32 degrees C, respectively. For BTX mixtures, toluene was the preferred substrate, but degradation of each BTX was competitively inhibited by other BTX compounds. The degradation rate of each component of in the BTX mixture decreased by 57-89% depending on the concentration (1-5 mg/l) of the component compared with that of the component as a single substrate.

Concentrating rotavirus and recombinant-enhanced green fluorescent protein from E. coli using a pH-sensitive hydrogel.

A rotavirus and a recombinant-enhanced green fluorescent protein from E. coli were concentrated 1.7 times and 1.5 times, respectively, by ultrafiltration at 37 degrees C and pH 7 using a pH-sensitive hydrogel, poly(N-isopropylacrylamide-co-N,N'-dimethylaminopropyl methacrylamide). Recoveries were 77% and 69%, respectively, and separation efficiencies were 58% and 44%, respectively. The concentration increase of the protein was confirmed by SDS-PAGE.

13-Hydroxy-9Z,11E,15E-octadecatrienoic acid from the leaves of Cucurbita moschata.

A new unsaturated hydroxy fatty acid was isolated from the leaves of Cucurbita moschata through repeated silica gel column chromatography and chemical methods. The structure of the new fatty acid was determined as 13-hydroxy-9,11,15-octadecatrienoic acid on the basis of several spectral data including 2D-NMR. The stererostructures of double bonds were determined to be 9Z, 11E and 15E by coupling patterns of related proton signals in the 1H-NMR and NOESY experiments.

Effects of gas flow rate, inlet concentration and temperature on biofiltration of volatile organic compounds in a peat-packed biofilter.

The effects of incoming gas concentration, empty bed residence time (EBRT), and column temperature on the removal efficiency of volatile organic compounds (isoprene, dimethyl sulfide, chloroform, benzene, trichloroethylene, toluene, m-xylene, o-xylene and styrene) were studied for 101 d in a biofilter comprising two glass columns (I.D. 5.0 cm x height 62 cm) packed with peat. At an EBRT of 3 min the removal efficiency increased up to 90% 34 d after start up at both 25 degrees C and 45 degrees C when the incoming gas concentration was raised stepwise to 65 g.m(-3). When the incoming gas concentration increased to 83 g.m(-3), the removal efficiency was 93% at 25 degrees C, but dropped to 74% at 45 degrees C. At an incoming gas concentration of 92 g.m(-3) and an EBRT of 1.5 min, the removal efficiencies were 91% and 94% at 25 degrees C and 32 degrees C, respectively. However, at 1 min of EBRT, the removal efficiencies decreased to 68% and 81% at 25 degrees C and 32 degrees C, respectively. The removal rate per unit time and per unit volume of the biofilter was proportional to the incoming gas rate up to 3483 g VOC.m(-3).h(-1). Further increase of the incoming gas rate lowered the removal rate as compared to that predicted by the proportionality. The maximum removal rate was 3977 g.m(-3).h(-1) at 32 degrees C. At an EBRT of 1.5 min, the removal efficiency was highest for isoprene (93%), and lowest for chloroform (84%). Aromatic compounds (benzene, toluene, and xylene) were removed by 93-94%. The cell concentration increased 100-fold from the initial value, and reached 1.12 x 10(8) cells.(g of dry peat)(-1). At 32 degrees C, 67% of the incoming VOC was removed in the first quarter of the column.