Original Article

99 cases were operated while we could not use antibiotics. The author traced X-ray photos on paper and measured areas of the peeled cavities with a planimeter. Results were as follows. 1) 66 cases had increasing stage and the rates were more than 30 %. 2) Cases with good developments showed larger original areas (50〜100cm^2) and smaller increasing rates (less than 30 %). 3) Also their X-ray photos showed coinciding or almost coinciding lines of the apices of lungs and the bases of cavities, but we had to take precautions against suppuration when they showed a horizontal line several days after operation. 4) Most of too high degree of adhesion or thickning of pleura did not show good results. When we found a cord which we must manage with some procedures by pneumolysis we must attend to suppuration too. 5)We ought to resect 4th or 5th rib more than 20 cm and 5th or 4th several cm supplementary. 6) As a method of constriction we commend the INVAGI.NATION method. 7) The author noticed in a considerable number of cases that the areas of cavities increased again after they kept long balanced stages

Atherosclerosis V, Proceeding of the Fifth International Symposium, A.M. Gotto, L.C. Smith, B. Allen, Spring Verlag, 1979(BOOK REVIEW)

Antiviral effect of micafungin on three strains of human rhinoviruses. H1HeLa cells were infected with human rhinovirus type 14 (A), 21 (B), or 71 (C) (100 CCID50) and immediately treated with indicated concentrations of micafungin. Three days after compound treatment antiviral activity was determined by the reduction of cytopathic effect using MTT assay. Cell viability of DMSO-treated cells was set to 0 % and that of uninfected cells was set to 100 %. (TIF 100 kb

miR-6734 induces cell cycle arrest in HCT-116 cells.

<p>HCT-116 cells were transfected with mock, dsCon or the indicated concentrations of miR-6734 for 48 h. (A) Cell cycle distribution was examined by flow cytometry. (B) The percentage of cells in G0/G1, S, and G2/M phases were calculated using the ModFit program. Data are presented as mean ± S.D. of triplicate experiments. Statistical significance was analyzed by one-way ANOVA and Dunnett’s t-test (** p < 0.01; *** p < 0.001 versus mock). (C) Protein levels of p21, cyclin-A, p-Rb and GAPDH in the total cell lysates were determined by Western Immunoblot analysis.</p

miR-6734 induces apoptosis in HCT-116 cells.

<p>HCT-116 cells were treated with mock, dsCon or the indicated concentrations of miR-6734 for 72 h. (A) Transfected cells were stained with propidium iodide and annexin V-FITC and apoptotic cells were measured by flow cytometry. (B) The percentage of early and late apoptotic cells were calculated using the WinMDI program. (C) Culture supernatants were collected, and the activity of caspase-3/7 was determined via caspase-Glo 3/7 assay kit. Data are presented as mean ± S.D. of triplicate experiments. Statistical significance was analyzed by one-way ANOVA and Dunnett’s t-test (** p < 0.01; *** p < 0.001 versus mock).</p

miR-6734 targets p21 promoter and induces p21 gene expression.

<p>(A) Sequence of dsP21-322 and miR-6734 target site located at nucleotide -322 relative to the transcriptional start site in p21 promoter. (B) Five human cancer cells were transfected with miR-6734 at 10nmol/L for 72h. The mRNA expression of p21 was analyzed by qPCR. (C) HCT-116 cells were transfected with mock and miR-6734-5P inhibitor at 30 nmol/L for 72 h. The mRNA expression of p21 was analyzed by qPCR. Data are presented as mean ± S.D. of quadruplicate experiments. Statistical significance was analyzed by one-way ANOVA and Dunnett’s t-test (* p < 0.05; ** p < 0.01 versus mock). (D) HCT-116 cells were transfected with biotin-labeled miR-6734 and chromatin immunoprecipitation (ChIP) assays were performed by using antibodies against biotin to pull down associated DNA. The precipitated DNA was amplified by PCR using primer sets specific to miR-6734 target sites (-360/-260) of p21 promoter. Input DNA was amplified as a loading control. DNA pulled down by the anti-IgG antibody was served to identify background amplification. Duplicate samples were analyzed for each treatment.</p

Inactivation of human DGAT2 by oxidative stress on cysteine residues

<div><p>Diacylglycerol acyltransferases (DGATs) have a crucial role in the biosynthesis of triacylglycerol (TG), the major storage form of metabolic energy in eukaryotic organisms. Even though DGAT2, one of two distinct DGATs, has a vital role in TG biosynthesis, little is known about the regulation of DGAT2 activity. In this study, we examined the role of cysteine and its oxidation in the enzymatic activity of human DGAT2 <i>in vitro</i>. Human DGAT2 activity was considerably inhibited not only by thiol-modifying reagents (NEM and IA) but also by ROS-related chemicals (H₂O₂ and β-lapachone), while human DGAT1 and GPAT1 were little affected. Particularly, ROS-related chemicals concomitantly induced intermolecular disulfide crosslinking of human DGAT2. Both the oxidative inactivation and disulfide crosslinking were almost completely reversed by the treatment with DTT, a disulfide-reducing agent. These results clearly demonstrated the significant role of ROS-induced intermolecular crosslinking in the inactivation of human DGAT2 and also suggested DGAT2 as a redox-sensitive regulator in TG biosynthesis.</p></div

Multimeric complex of human DGAT2 formed by H₂O₂-induced disulfide crosslinking in human cells.

<p>Huh-7 cells were transfected with plasmid overexpressing human DGAT2 for 47 hours and further incubated with indicated concentrations of H₂O₂ for 1 hour. Cell extracts were harvested in a way described in Materials and Methods section and subjected to Western blot analysis using anti-DGAT2 antibody (A). The amount of monomeric human DGAT2 proteins presented as redDGAT2 in (A) was quantified and the amount of relative redDGAT2 protein was calculated by setting the values from samples treated with PBS to 100% (B). The mean values and standard deviations were determined from three independent experiments. Asterisks indicate non-specific bands.</p

Susceptibility of human DGAT2 activity to cysteine-specific modifying reagents.

<p>(A) Membrane extracts from human DGAT2-overexpressing Sf9 insect cells were treated with indicated concentrations of NEM or IA. Human DGAT2 activity was measured by using the conventional extraction-based <i>in vitro</i> DGAT assay. The relative DGAT2 activity in percentage was calculated by setting the value from DMSO-treated sample to 100%. (B) Selective inhibitory effect of NEM on human DGAT2 activity compared to that on human DGAT1 and GPAT1. Membrane extracts from human DGAT2-, DGAT1-, or GPAT1-overexpressing Sf9 insect cells were treated with indicated concentrations of NEM or DMSO. Human DGAT1, DGAT2, and GPAT1 activity was measured by using the conventional extraction-based <i>in vitro</i> assays which are described in detail in the Materials and Methods section. The relative enzyme activity in percentage was calculated by setting the value from DMSO-treated sample to 100%. The mean values and standard deviations were determined from four independent assays.</p

Inhibitory effect of ROS and ROS generator on human DGAT2 catalytic activity.

<p>Membrane extracts from human DGAT2-overexpressing Sf9 insect cells were treated with indicated concentrations of H₂O₂ (A) or β-lapachone (B) in the presence or absence of 20 mM DTT. Human DGAT2 activity was measured by using the conventional extraction-based <i>in vitro</i> assays which are described in detail in the Materials and Methods section. The activities of membrane extracts treated with PBS (instead of H₂O₂) or DMSO (instead of β-lapachone) in the absence of DTT were defined as 100%. The mean values and standard deviations were determined from four independent experiments.</p

Multimeric complex of human DGAT2 formed by ROS-induced intermolecular disulfide crosslinking <i>in vitro</i>.

<p>Membrane extracts from human DGAT2-overexpressing Sf9 insect cells were treated with H₂O₂ (A) or β-lapachone (B) in the presence or absence of 20 mM DTT and subjected to Western blot analysis using anti-DGAT2 antibody. The amount of monomeric human DGAT2 proteins presented as redDGAT2 in (A) and (B) was quantified and the amount of relative redDGAT2 protein was calculated by setting the values from samples treated with PBS (C) or DMSO (D) to 100%. Asterisk indicates a non-specific band.</p