Visible-light Induced Reduction of Graphene Oxide Using Plasmonic Nanoparticle

Present work demonstrates the simple, chemical free, fast, and energy efficient method to produce reduced graphene oxide (r-GO) solution at RT using ...

Korea University

Low-density lipoprotein (LDL) induces cell proliferation in human aortic smooth muscle cells (hAoSMCs), which may be involved in atherogenesis and intimal hyperplasia. Recent studies have demonstrated that Cl- channels are related to vessel cell proliferation induced by a variety of stimuli. In this study, we investigated a potential role of Cl-channels in the signaling pathway of LDL effects on hAoSMC proliferation with a focus on the activation of Erk1/2-PI3K/Akt and the subsequent upregulation of Egr-1. Cl- channel blockers, DIDS, but neither NPPB nor Furosemide, completely abolished the LDL-induced DNA synthesis and cell proliferation. Moreover, DIDS, but not NPPB, significantly decreased LDL-stimulated Cl- concentration, as judged by flow cytometry analysis using MQAE as a Cl- -detection dye. DIDS pretreatment completely abolished the activation of Erk1/2 and PI3K/Akt in a dose-dependent manner that is the hallmark of LDL activation, as judged by Western blot and proliferation assays. Moreover, pretreatment with DIDS (Cl- channel blockers) but not LY294002 (PI3K inhibitors) completely abolished the LDL-induced upregulation of Egr-1 to the same extent as PD98059 (MEK inhibitors to inhibit Erk), as judged by Western blot and luciferase reporter assays. This is the first report, to our knowledge, that DIDS-sensitive Cl- channels play a key role in the LDL-induced cell proliferation of hAoSMCs via the activation of Erk1/2 and PI3K/Akt and the upregulation of Egr-1.

Cl- -channel is essential for LDL-induced cell proliferation via the activation of Erk1/2 and PI3k/Akt and the upregulation of Egr-1 in human aortic smooth muscle cells.

By screening C. elegans mutants for severe defects in germline proliferation, we isolated a new loss-of-function allele of cdc-25.1, bn115. bn115 and another previously identified loss-of-function allele nr2036 do not exhibit noticeable cell division defects in the somatic tissues but have reduced numbers of germ cells and are sterile, indicating that cdc-25.1 functions predominantly in the germ line during postembryonic development, and that cdc-25.1 activity is probably not required in somatic lineages during larval development. We analyzed cell division of germ cells and somatic tissues in bn115 homozygotes with germline-specific anti-PGL-1 immunofluorescence and GFP transgenes that express in intestinal cells, in distal tip cells, and in gonadal sheath cells, respectively. We also analyzed the expression pattern of cdc-25.1 with conventional and quantitative RT-PCR. In the presence of three other family members of cdc-25 in C. elegans defects are observed only in the germ line but not in the somatic tissues in cdc-25.1 single mutants, and cdc-25.1 is expressed predominantly, if not exclusively, in the germ line during postembryonic stages. Our findings indicate that the function of cdc-25.1 is unique in the germ line but likely redundant with other members in the soma.

A mutation of cdc-25.1 causes defects in germ cells but not in somatic tissues in C. elegans.

We report the construction and analysis of 4,836 heterozygous diploid deletion mutants covering 98.4% of the fission yeast genome providing a tool for studying eukaryotic biology. Comprehensive gene dispensability comparisons with budding yeast--the only other eukaryote for which a comprehensive knockout library exists--revealed that 83% of single-copy orthologs in the two yeasts had conserved dispensability. Gene dispensability differed for certain pathways between the two yeasts, including mitochondrial translation and cell cycle checkpoint control. We show that fission yeast has more essential genes than budding yeast and that essential genes are more likely than nonessential genes to be present in a single copy, to be broadly conserved and to contain introns. Growth fitness analyses determined sets of haploinsufficient and haploproficient genes for fission yeast, and comparisons with budding yeast identified specific ribosomal proteins and RNA polymerase subunits, which may act more generally to regulate eukaryotic cell growth.

Analysis of a genome-wide set of gene deletions in the fission yeast Schizosaccharomyces pombe.

Four new alleles, bn116, bn117, bn118, and bn119, on LG I were isolated in C. elegans with defects in germline stem cell proliferation. Using genetic mapping and snip-SNP mapping, bn116, bn117, bn118, and bn119 were located 5.0 cM, 1.3 cM, 2.3 cM, and 5.0 cM, respectively, to the right of dpy-5 on LG I. Further, bn116 and bn119 were grouped into the same complementation group by a complementation test. They are loss-of-function recessive alleles that produce homozygous sterile worms whose germ cells do not proliferate during larval development. However, the worms contained normal somatic gonadal structures including distal tip cells and gonadal sheath cells, suggesting that the defect in germline proliferation was not caused by the absence of somatic signaling. Although DAF-16 was localized to the nucleus in all four mutants, the life span was extended only in the three mutants except bn116. These results suggest that the defect in germline stem cell proliferation, the presence of normal somatic gonadal tissues, and DAF-16 nuclear translocation were sufficient for extending the lifespan of the bn117, bn118, and bn119 mutants, but not the bn116 mutant. Intriguingly, bn116 and bn119 were identified as two different mutations on the same gene, pab-1, which encodes a poly(A)-binding protein. Therefore, although the bn116 and bn119 mutations cause similar defects in germ cell proliferation, their effects on life span are different.

Two mutations in pab-1 encoding poly(A)-binding protein show similar defects in germline stem cell proliferation but different longevity in C. elegans.

Pseudoprogression is a major diagnostic dilemma in current treatment protocols for malignant gliomas that involve concurrent chemoradiotherapy. We hypothesized that methylation-specific multiplex ligation probe amplification (MS-MLPA), an assay that permits semiquantitative evaluation of promoter methylation, may be used to diagnose pseudoprogression based on the quantification of the methylation status of the O(6)-methylguanine DNA methyltransferase (MGMT) promoter. We examined the methylation ratio of the MGMT promoter with MS-MLPA in 48 samples from glioblastoma patients. The results were compared with those from methylation-specific polymerase chain reaction (MSP), and protein levels were confirmed by immunohistochemical staining. We then evaluated the correlation between those molecular signatures and clinical outcomes. With regard to radiological progression after chemoradiotherapy, the diagnostic accuracy of the MS-MLPA method was 80% (using a cut-off value of 0.2). These results are better than those obtained with MSP (diagnostic accuracy of 68%). Combining the MS-MLPA and MSP methods resulted in a diagnostic accuracy of 93% for the identification of pseudoprogression among patients to whom these results were coherent. These results demonstrate that MS-MLPA is a useful method to predict radiological progression vs pseudoprogression in glioblastoma patients and that the interpretation of these results in combination with MSP results will provide good practical guidelines for clinical decision making in glioblastoma treatment.

Usefulness of MS-MLPA for detection of MGMT promoter methylation in the evaluation of pseudoprogression in glioblastoma patients.

&#x5C3D;&#x7BA1;&#x4F17;&#x6240;&#x5468;&#x77E5;&#x7684;&#x56DB;&#x500D;&#x4F53;&#x80DA;&#x80CE;&#x53EA;&#x6709;&#x52A9;&#x4E8E;&#x80CE;&#x76D8;&#xFF0C;&#x4E3A;&#x4EC0;&#x4E48;&#x56DB;&#x500D;&#x4F53;&#x80DA;&#x80CE;&#x5206;&#x5316;&#x4E3A;&#x80CE;&#x76D8;&#x7684;&#x95EE;&#x9898;&#x4ECD;&#x7136;&#x4E0D;&#x6E05;&#x695A;&#x3002;&#x4EE5;&#x7814;&#x7A76;&#x5BF9;&#x5C0F;&#x9F20;&#x56DB;&#x500D;&#x4F53;&#x5375;&#x6BCD;&#x7EC6;&#x80DE;&#x53D1;&#x5C55;&#x7535;&#x878D;&#x5408;&#x7684;&#x5F71;&#x54CD;&#x3001; &#x5C0F;&#x9F20; 2-&#x7EC6;&#x80DE;&#x80DA;&#x80CE;&#x88AB;&#x878D;&#x5408;&#x548C; Chatot-Ziomek-Bavister &#x4ECB;&#x8D28;&#x4E2D;&#x4F53;&#x5916;&#x57F9;&#x517B;&#x7EC6;&#x80DE;&#x3002;&#x7535;&#x878D;&#x5408;&#x540E;, &#x4E24;&#x4E2A;&#x67D3;&#x8272;&#x4F53;&#x7EC4;&#x4ECE;&#x56DB;&#x500D;&#x4F53;&#x5375;&#x88C2;&#x7403;&#x88AB;&#x5355;&#x72EC;&#x91CD;&#x590D;&#x524D;&#x6838;&#x805A;&#x53D8;&#x3002;&#x5728;&#x7535;&#x878D;&#x5408;&#x540E; 8-10 &#x5C0F;&#x65F6;&#xFF0C;&#x6BCF;&#x4E2A;&#x67D3;&#x8272;&#x4F53;&#x7EC4;&#x51DD;&#x6C7D;&#x5F0F;&#x548C;&#x6838;&#x819C;&#x88AB;&#x6253;&#x7834;&#x4E86;&#x3002;&#x7EA6; 12-14 &#x5C0F;&#x65F6;&#x540E;&#x7535;&#x878D;&#x5408;&#xFF0C;&#x4E24;&#x4E2A;&#x67D3;&#x8272;&#x4F53;&#x96C6;&#x4E86;&#x7EC4;&#x5408;&#x5728;&#x4E00;&#x8D77;&#x5E76;&#x8FBE;&#x6210;&#x4E86;&#x7B2C;&#x4E8C;&#x6B21;&#x6709;&#x4E1D;&#x5206;&#x88C2;&#x4E2D;&#x671F;&#xFF0C;&#x6B64;&#x65F6;&#x4E0E; 8n &#x8BBE;&#x7F6E;&#x7684;&#x67D3;&#x8272;&#x4F53;&#x3002;&#x6709;&#x8DA3;&#x7684;&#x662F;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x8868;&#x8FBE;&#x7684; OCT4&#x3001; &#x5185;&#x7EC6;&#x80DE;&#x5927;&#x89C4;&#x6A21;&#x7EC6;&#x80DE;&#x751F;&#x7269;&#x6807;&#x5FD7;&#x7269;&#xFF0C;&#x635F;&#x5931;&#x7684;&#x56DB;&#x500D;&#x4F53;&#x6269;&#x5927;&#x56CA;&#x80DA;&#xFF0C;&#x4F46;&#x8FD9; CDX2&#xFF0C;trophectoderm &#x7EC6;&#x80DE;&#x751F;&#x7269;&#x6807;&#x8BB0;&#x7269;&#xFF0C;&#x5728;&#x8FD9;&#x4E00;&#x9636;&#x6BB5;&#x8868;&#x793A;&#x5F3A;&#x70C8;&#x3002;&#x8FD9;&#x4E00;&#x89C2;&#x5BDF;&#x63D0;&#x4F9B;&#x4E86;&#x8BC1;&#x636E;&#x6F84;&#x6E05;&#x4E3A;&#x4EC0;&#x4E48;&#x53EA;&#x5BF9; trophectoderm &#x7684;&#x56DB;&#x500D;&#x4F53;&#x80DA;&#x80CE;&#x4F5C;&#x51FA;&#x8D21;&#x732E;&#x3002;

&#x67D3;&#x8272;&#x4F53;&#x91CD;&#x6784;&#x548C;&#x5206;&#x5316;&#x7684;&#x56DB;&#x500D;&#x4F53;&#x80DA;&#x80CE;&#x690D;&#x5165;&#x524D;&#x7684;&#x53D1;&#x80B2;&#x8FC7;&#x7A0B;&#x3002;

Although it is known that the tetraploid embryo contributes only to the placenta, the question of why tetraploid embryos differentiate into placenta remains unclear. To study the effect of electrofusion on the development of mouse tetraploid oocytes, mouse two-cell embryos were fused and cultured in vitro in Chatot-Ziomek-Bavister medium. After electrofusion, two chromosome sets from the tetraploid blastomere were individually duplicated before nuclear fusion. At 8-10 hr after electrofusion, each chromosome set was condensing and the nuclear membrane was breaking down. Around 12-14 hr after electrofusion, the two chromosome sets had combined together and had reached the second mitotic metaphase, at this point with 8n sets of chromosomes. Interestingly, we discovered that expression of OCT4, an inner cell mass cells biomarker, is lost by the tetraploid expanded blastocysts, but that CDX2, a trophectoderm cells biomarker, is strongly expressed at this stage. This observation provides evidence clarifying why tetraploid embryos contribute only to trophectoderm.

Chromosome remodeling and differentiation of tetraploid embryos during preimplantation development.

Mais on sait que l&#39;embryon t&eacute;traplo&iuml;de contribue seulement au placenta, la question de pourquoi les embryons t&eacute;traplo&iuml;des se diff&eacute;rencient en placenta demeure incertaine. Pour &eacute;tudier l&#39;effet de l&#39;&eacute;lectrofusion sur le d&eacute;veloppement des ovocytes t&eacute;traplo&iuml;de de souris, les embryons de deux cellules de souris ont &eacute;t&eacute; fondues et cultiv&eacute;es in vitro dans un milieu Chatot-Ziomek-belmir. Apr&egrave;s electrofusion, deux ensembles de chromosomes du blastom&egrave;re t&eacute;traplo&iuml;de ont &eacute;t&eacute; individuellement dupliqu&eacute;s avant la fusion nucl&eacute;aire. 8-10 H apr&egrave;s &eacute;lectrosoudables, chaque ensemble de chromosomes a condensation et la membrane nucl&eacute;aire &eacute;tait briser. Environ 12-14 hr apr&egrave;s electrofusion, le chromosome deux ensembles &eacute;taient regroup&eacute;s et avaient atteint la deuxi&egrave;me m&eacute;taphase mitotique, &agrave; ce stade avec 8n d&eacute;finit des chromosomes. Fait int&eacute;ressant, nous avons d&eacute;couvert qu&#39;expression de OCT4, un biomarqueur de cellules masse cellulaire interne, est perdue par les t&eacute;traplo&iuml;des blastocystes &eacute;largis, mais cette CDX2, un biomarqueur de cellules trophectoderme, est fortement exprim&eacute; &agrave; ce stade. Cette observation d&eacute;montre de clarifier la raison pour laquelle les embryons t&eacute;traplo&iuml;des contribuent seulement &agrave; trophectoderme.

Remodelage des chromosomes et la diff&eacute;renciation des embryons t&eacute;traplo&iuml;des au cours du d&eacute;veloppement pr&eacute;implantatoire.

Es ist zwar bekannt, dass die tetraploide Embryonen nur zu der Plazenta beitr&auml;gt, bleibt die Frage, warum tetraploide Embryonen in Plazenta differenzieren unklar. Um die Wirkung der Elektroschweiss auf die Entwicklung der Maus tetraploide Eizellen zu studieren, waren Maus zwei Zellen Embryonen verschmolzen und in-vitro Chatot-Ziomek-Bavister Medium kultiviert. Nach dem Heizwendelschwei&szlig;en wurden zwei Chromosomens&auml;tze von der tetraploide Blastomer einzeln vor die Kernfusion dupliziert. Mit 8-10 h nach Heizwendelschwei&szlig;en jedes Chromosom-Set wurde kondensiert und die Kernmembrane Abbau war. Ca. 12-14 hr nach Heizwendelschwei&szlig;en, das zwei Chromosom Sets hatten miteinander kombiniert und erreichte die zweite mitotische Metaphase, an diesem Punkt mit 8n Chromosomen setzt. Interessant ist, entdeckten wir, dass Ausdruck des OCT4, eine innere Zelle Masse Zellen Biomarker, ist durch die tetraploide erweiterten Blastozysten verloren, aber diese CDX2, Trophectoderm Zellen Biomarker, wird in diesem Stadium stark ausgedr&uuml;ckt. Diese Beobachtung liefert Beweise Kl&auml;rung Warum tetraploide Embryonen nur zur Trophectoderm beitragen.

Chromosom Umbau und Differenzierung der tetraploide Embryonen w&auml;hrend der Pr&auml;implantationsdiagnostik Entwicklung.

Anche se &egrave; noto che l&#39;embrione tetraploide contribuisce solo alla placenta, la questione del perch&eacute; gli embrioni tetraploidi differenziano in placenta rimane poco chiara. Per studiare l&#39;effetto di elettrofusione sullo sviluppo degli ovociti tetraploidi mouse, embrioni del mouse due celle erano fuse e coltivati in vitro in Chatot-Ziomek-Bavister medio. Dopo elettrofusione, due set di cromosomi dal blastomero tetraploide individualmente sono stati duplicati prima fusione nucleare. A 8-10 ore dopo elettrofusione, ciascun set di cromosomi era condensa e membrana nucleare era abbattere. Intorno al 12-14 ore dopo elettrofusione, il cromosoma due set aveva combinato insieme e aveva raggiunto la seconda metafase mitotica, a questo punto con 8n set di cromosomi. &Egrave; interessante notare che, abbiamo scoperto che espressione di OCT4, un biomarcatore di cellule massa cellulare interna, &egrave; perso per le blastocisti espanse tetraploidi, ma quel CDX2, un biomarcatore di cellule trophectoderm, &egrave; fortemente espresso in questa fase. Questa osservazione fornisce prove chiarire perch&eacute; gli embrioni tetraploidi contribuiscono solo a trophectoderm.

Cromosoma rimodellamento e la differenziazione degli embrioni tetraploidi durante lo sviluppo preimpianto.

&#x305D;&#x308C;&#x306F; 4 &#x500D;&#x4F53;&#x80DA;&#x306F;&#x80CE;&#x76E4;&#x306B;&#x306E;&#x307F;&#x5BC4;&#x4E0E;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x306A;&#x305C; 4 &#x500D;&#x4F53;&#x80DA;&#x306F;&#x80CE;&#x76E4;&#x306B;&#x533A;&#x5225;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x8CEA;&#x554F;&#x306F;&#x4E0D;&#x660E;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x30DE;&#x30A6;&#x30B9; 4 &#x500D;&#x4F53;&#x5375;&#x6BCD;&#x7D30;&#x80DE;&#x306E;&#x767A;&#x9054;&#x306B;&#x53CA;&#x307C;&#x3059;&#x96FB;&#x6C17;&#x7684;&#x878D;&#x5408;&#x3092;&#x52C9;&#x5F37;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x30DE;&#x30A6;&#x30B9; 2 &#x7D30;&#x80DE;&#x671F;&#x80DA;&#x306F;&#x878D;&#x5408;&#x3057;&#x3001;Chatot Ziomek Bavister &#x57F9;&#x5730;&#x3067;&#x57F9;&#x990A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x96FB;&#x6C17;&#x878D;&#x5408;&#x5F8C;&#x56DB;&#x500D;&#x4F53;&#x306E;&#x5272;&#x7403;&#x304B;&#x3089;&#x306E; 2 &#x3064;&#x306E;&#x67D3;&#x8272;&#x4F53;&#x30BB;&#x30C3;&#x30C8;&#x306F;&#x500B;&#x5225;&#x306B;&#x6838;&#x878D;&#x5408;&#x3059;&#x308B;&#x524D;&#x306B;&#x91CD;&#x8907;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;8-10 &#x6642;&#x9593;&#x96FB;&#x6C17;&#x878D;&#x5408;&#x5F8C;&#x5404;&#x67D3;&#x8272;&#x4F53;&#x30BB;&#x30C3;&#x30C8;&#x3092;&#x51DD;&#x7E2E;&#x3068;&#x6838;&#x819C;&#x6253;&#x7834;&#x3067;&#x3042;&#x3063;&#x305F;&#x3002;&#x7D04; 12-14 &#x6642;&#x9593;&#x5F8C;&#x96FB;&#x6C17;&#x7684;&#x878D;&#x5408;&#x30BB;&#x30C3;&#x30C8;&#x4E00;&#x7DD2;&#x306B;&#x7D44;&#x307F;&#x5408;&#x308F;&#x305B;&#x3066;&#x3044;&#x305F;&#x3057;&#x3001;2 &#x756A;&#x76EE;&#x306E;&#x7D30;&#x80DE;&#x5206;&#x88C2;&#x4E2D;&#x671F;&#x306B;&#x9054;&#x3057;&#x3066;&#x3044;&#x305F; 2 &#x3064;&#x306E;&#x67D3;&#x8272;&#x4F53;&#x306F;&#x3053;&#x306E;&#x6642;&#x70B9;&#x3067;&#x306E;&#x67D3;&#x8272;&#x4F53; 8n &#x3092;&#x8A2D;&#x5B9A;&#x3057;&#x307E;&#x3059;&#x3002;&#x8208;&#x5473;&#x6DF1;&#x3044;&#x3053;&#x3068;&#x306B;&#x3001;OCT4 &#x306F;&#x5185;&#x90E8;&#x7D30;&#x80DE;&#x8CEA;&#x91CF;&#x7D30;&#x80DE;&#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x306E;&#x767A;&#x73FE;&#x306F;&#x56DB;&#x500D;&#x4F53;&#x306E;&#x62E1;&#x5F35;&#x80DA;&#x76E4;&#x80DE;&#x3067;&#x3001;&#x5931;&#x308F;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x305D;&#x306E; CDX2&#x3001;&#x6804;&#x990A;&#x5916;&#x80DA;&#x8449;&#x7D30;&#x80DE;&#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x306F;&#x5F37;&#x304F;&#x3053;&#x306E;&#x6BB5;&#x968E;&#x3067;&#x8868;&#x73FE;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x306E;&#x89B3;&#x6E2C;&#x306F; 4 &#x500D;&#x4F53;&#x80DA;&#x306E;&#x6804;&#x990A;&#x5916;&#x80DA;&#x8449;&#x306B;&#x3060;&#x3051;&#x306F;&#x306A;&#x305C;&#x304B;&#x660E;&#x767D;&#x306B;&#x306A;&#x308B;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;

&#x5206;&#x5316; 4 &#x500D;&#x4F53;&#x80DA;&#x306E;&#x7740;&#x5E8A;&#x524D;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x306B;&#x67D3;&#x8272;&#x4F53;&#x3092;&#x6539;&#x9020;&#x3057;&#x3001;&#x3002;

Tetraploid &#xD0DC;&#xC544; &#xD0DC; &#xBC18;&#xC5D0;&#xB9CC; &#xAE30;&#xC5EC; &#xD558;&#xB294; &#xAC83;&#xC774; &#xC54C;&#xB824;&#xC838;, &#xBE44;&#xB85D; tetraploid &#xD0DC;&#xC544; &#xD0DC; &#xBC18;&#xC73C;&#xB85C; &#xAD6C;&#xBD84; &#xD558;&#xB294; &#xC774;&#xC720;&#xC758; &#xC9C8;&#xBB38; &#xBD88;&#xBD84;&#xBA85; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB9C8;&#xC6B0;&#xC2A4; tetraploid oocytes &#xAC1C;&#xBC1C;&#xC5D0; electrofusion&#xC758; &#xD6A8;&#xACFC; &#xC5F0;&#xAD6C; &#xD558;&#xB824;&#xBA74; &#xB9C8;&#xC6B0;&#xC2A4; &#xB0C9;&#xB3D9; &#xBC30;&#xC544; &#xC735;&#xD569; &#xB418;&#xC5C8;&#xACE0; Chatot-Ziomek-Bavister &#xB9E4;&#xCCB4;&#xC758; &#xCCB4; &#xC678; &#xBC30;&#xC591;. Electrofusion, &#xD6C4; &#xB450; &#xC5FC;&#xC0C9;&#xCCB4; &#xC138;&#xD2B8; tetraploid blastomere&#xC5D0;&#xC11C; &#xD588;&#xB2E4; &#xD575; &#xC735;&#xD569; &#xD558;&#xAE30; &#xC804;&#xC5D0; &#xAC1C;&#xBCC4;&#xC801;&#xC73C;&#xB85C; &#xBCF5;&#xC81C; &#xB429;&#xB2C8;&#xB2E4;. Electrofusion &#xD6C4; 8-10 &#xC2DC;&#xAC04;&#xC5D0; &#xAC01; &#xC5FC;&#xC0C9;&#xCCB4; &#xC138;&#xD2B8;&#xB294; &#xC751;&#xCD95;&#xACFC; &#xD575; &#xB9C9; &#xB728;&#xB294;. &#xC57D; 12-14 &#xC2DC;&#xAC04; &#xD6C4;&#xC5D0; electrofusion, &#xB450; &#xAC1C;&#xC758; &#xC5FC;&#xC0C9;&#xCCB4; &#xC9D1;&#xD569;&#xACFC; &#xD568;&#xAED8; &#xACB0;&#xD569; &#xD588;&#xB2E4; &#xB450; &#xBC88;&#xC9F8; mitotic &#xBD84;&#xC5F4;&#xC5D0; &#xB3C4;&#xB2EC; &#xD588;&#xB2E4; &#xADF8;&#xB7EC;&#xBA74; 8n &#xC138;&#xD2B8;&#xC758; &#xC5FC;&#xC0C9;&#xCCB4;. &#xD765;&#xBBF8;&#xB86D;&#xAC8C;&#xB3C4;, &#xC6B0;&#xB9AC;&#xB294; OCT4, &#xB0B4;&#xBD80; &#xC140; &#xB300;&#xB7C9; &#xC140; biomarker &#xD45C;&#xD604;&#xC758; tetraploid &#xD655;&#xC7A5;&#xB41C; blastocysts&#xC5D0; &#xC758;&#xD574; &#xC190;&#xC2E4; &#xB429;&#xB2C8;&#xB2E4; &#xD558;&#xC9C0;&#xB9CC; &#xADF8; CDX2, trophectoderm &#xC140; biomarker &#xAC15;&#xB825; &#xD558; &#xAC8C;&#xC774; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; &#xD45C;&#xD604; &#xB41C;&#xB97C; &#xBC1C;&#xACAC; &#xD588;&#xB2E4;. &#xC774; &#xAD00;&#xCC30; tetraploid &#xBC30;&#xC544; trophectoderm&#xC5D0;&#xB9CC; &#xAE30;&#xC5EC; &#xD558;&#xB294; &#xC774;&#xC720;&#xB97C; &#xBA85;&#xD655;&#xD788; &#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;.

&#xC5FC;&#xC0C9;&#xCCB4; &#xAC1C;&#xC870;&#xC640; preimplantation &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; tetraploid &#xBC30;&#xC544;&#xC758; &#xCC28;&#xBCC4;&#xD654;.

Aunque se sabe que el embri&oacute;n tetraploide contribuye s&oacute;lo a la placenta, la pregunta de por qu&eacute; embriones tetraploides se diferencian en la placenta sigue siendo confusa. Para estudiar el efecto de electrofusi&oacute;n en el desarrollo de ovocitos de rat&oacute;n tetraploide, embriones de dos c&eacute;lulas de rat&oacute;n se fundieron y cultivados in vitro en medio Chatot-Ziomek-Bavister. Despu&eacute;s de electrofusi&oacute;n, dos conjuntos de cromosomas de la blast&oacute;mera tetraploide individualmente fueron duplicados antes de la fusi&oacute;n nuclear. En 8-10 horas despu&eacute;s de electrofusi&oacute;n, cada juego de cromosomas se condense y era romper la membrana nuclear. Alrededor de 12-14 horas despu&eacute;s de electrofusi&oacute;n, el cromosoma dos conjuntos hab&iacute;an combinado juntos y hab&iacute;an alcanzado la segunda metafase mit&oacute;tica, en este momento con 8n juegos de cromosomas. Curiosamente, descubrimos que la expresi&oacute;n de OCT4, un biomarcador de las c&eacute;lulas de la masa celular interna, se pierde por los blastocistos expandidos tetraploides, pero eso CDX2, un biomarcador de las c&eacute;lulas del trofectodermo, se expresa fuertemente en esta etapa. Esta observaci&oacute;n proporciona evidencia clarificar por qu&eacute; embriones tetraploides contribuyen s&oacute;lo a trofectodermo.

Remodelaci&oacute;n del cromosoma y diferenciaci&oacute;n de embriones tetraploides durante el desarrollo preimplantacional.

This study was designed to investigate quality of life (QOL) differences between younger (<50 years) and older (≥50 years) breast cancer survivors and to determine the unique effect of age on QOL after adjusting age-correlated variables.

Quality of Life Differences between Younger and Older Breast Cancer Patients.

Postovulatory mammalian oocyte developmental potential decreases with aging in vivo and in vitro. Aging oocytes typically show cellular fragmentation and chromosome scattering with an abnormally shaped spindle over time. Previously, it was shown that histone acetylation in the mouse oocyte increased during aging and that treatment with trichostatin A (TSA), an inhibitor for class I and II histone deacetylases (HDACs), enhanced the acetylation, that is, aging. In this study, we examined the effect of nicotinamide (NAM), an inhibitor for class III HDACs, on in vitro aging of mouse oocytes as well as TSA. We found that treatment with NAM significantly inhibited cellular fragmentation, spindle elongation and astral microtubules up to 48 h of culture. Although presence of TSA partially inhibited cellular fragmentation and spindle elongation up to 36 h of culture, treatment with TSA induced chromosome scattering at 24 h of culture and more severe cellular fragmentation at 48 h of culture. Further, we found that α-tubulin, a nonhistone protein, increased acetylation during aging, suggesting that not only histone but nonhistone protein acetylation may also increase with oocyte aging. Thus, these data indicate that protein acetylation is abnormally regulated in aging oocytes, which are associated with a variety of aging phenotypes, and that class I/II and class III HDACs may play distinct roles in aging oocytes.

Nicotinamide: a class III HDACi delays in vitro aging of mouse oocytes.

Previous studies show that treatment of zygotes with trichostatin A (TSA), a histone deacetylase inhibitor (HDACi), impacts the subsequent development to a blastocyst as well as full-term development. To reveal the dynamics of protein acetylation, with and without TSA treatment during one-cell stage, we examined oocytes and zygotes by immunofluorescence and Western Blot analyses using anti-acetylated lysine and acetylated α-tubulin antibodies. In unfertilized oocytes, lysine acetylation level was extremely low over all but faintly detected in the spindle. Once oocyte activation occurs, a dramatic increase of lysine acetylation signal was observed mostly in the pronuclei and a fiber-like structure, the so called midbody, suggesting activation coupled up-regulation of lysine acetylation presumably in histones and α-tubulin. TSA treatment resulted in significantly more hyperacetylation not only in the midbody structure and pronuclei but also in the whole cytoplasm. Consistently, Western Blot analysis revealed that acetylation of proteins about 53 kDa and 11 kDa in size, corresponding to α-tubulin and histone H4 sizes respectively, were increased mainly after oocyte activation and exclusively enhanced by TSA treatment in zygotes. To confirm this behavior of acetylated nonhistone proteins, acetylated α-tubulin was examined and found to be faintly detected in the spindle of MII oocytes but later in whole in the cell of zygotes including the midbody, which was enhanced by TSA treatment. To elucidate the mechanism underlying up-regulation of lysine acetylation following oocyte activation, we assayed the HDAC activity, and found significant reduction of HDAC activity from MII to zygotic stages. Taken together, our data indicate that HDACs play an important role in maintaining low acetylated status in a MII oocyte. However, once an oocyte has been activated, histone and nonhistone proteins including α-tubulin are hyperacetylated partly due to a reduction of HDAC activity. TSA treatment of zygotes enhances their acetylation, which could affect subsequent embryonic development.

Dynamics and regulation of lysine-acetylation during one-cell stage mouse embryos.

In mammals, a diploid genome following fertilization of haploid cells, an egg, and a spermatozoon is unique and irreproducible. This implies that the generated unique diploid genome is doomed with the individual's inevitable demise. Since it was first reported in 1997 that Dolly the sheep had been cloned, many mammalian species have been cloned successfully using somatic-cell nuclear transfer (SCNT). The success of SCNT in mammals enables us not only to reproduce offspring without germ cells, that is, to "passage" a unique diploid genome, but also to address valuable biological questions on development, nuclear reprogramming, and epigenetic memory. Successful cloning can also support epigenetic reprogramming where the aging clock is reset or reversed. Recent work using iPS cell technology has explored the practicality and led to the recapitulation of premature aging with iPSCs from progeroid laminopathies. As a result, reprogramming tools are also expected to contribute to studying biological age. However, the efficiency of animal cloning is still low in most cases and the mechanism of reprogramming in cloned embryos is still largely unclear. Here, based on recent advances, we describe an improved, more efficient mouse cloning protocol using histone deacetylase inhibitors (HDACis) and latrunculin A, which increases the success rates of producing cloned mice or establishing ES cells fivefold. This improved method of cloning will provide a strong tool to address many issues including biological aging more easily and with lower cost.

Using somatic-cell nuclear transfer to study aging.

Marine red macroalgae have emerged to be renewable biomass for the production of chemicals and biofuels, because carbohydrates that form the major component of red macroalgae can be hydrolyzed into fermentable sugars. The main carbohydrate in red algae is agarose, and it is composed of D-galactose and 3,6-anhydro-L-galactose (AHG), which are alternately bonded by β1-4 and α1-3 linkages. In this study, a novel β-galactosidase that can act on agarooligosaccharides (AOSs) to release galactose was discovered in a marine bacterium (Vibrio sp. strain EJY3); the enzyme is annotated as Vibrio sp. EJY3 agarolytic β-galactosidase (VejABG). Unlike the lacZ-encoded β-galactosidase from Escherichia coli, VejABG does not hydrolyze common substrates like lactose and can act only on the galactose moiety at the nonreducing end of AOS. The optimum pH and temperature of VejABG on an agarotriose substrate were 7 and 35°C, respectively. Its catalytic efficiency with agarotriose was also similar to that with agaropentaose or agaroheptaose. Since agarotriose lingers as the unreacted residual oligomer in the currently available saccharification system using β-agarases and acid prehydrolysis, the agarotriose-hydrolyzing capability of this novel β-galactosidase offers an enormous advantage in the saccharification of agarose or agar in red macroalgae for its use as a biomass feedstock for fermentable sugar production.

A novel agarolytic β-galactosidase acts on agarooligosaccharides for complete hydrolysis of agarose into monomers.

Genome-wide targeted gene deletion, a systematic method to study gene function by replacing target genes with deletion cassettes, using serial-PCR or block-PCR requires elaborate skill. We developed a novel gene-synthesis method to systematically prepare deletion cassettes on a 96-well basis in fission yeast. We designed the 2129-bp deletion cassette as three modules: a central 1397-bp KanMX4 selection marker module and two flanking 366-bp gene-specific artificial linker modules. The central KanMX4 module can be used in multiple deletion cassettes in combination with different sets of flanking modules. The deletion cassettes consisted of 147 oligonucleotides (93 for the central module+25 for each of the flanking modules+4 for the joints) and the oligonucleotides were designed as ~29mers using an in-house program. Oligonucleotides were synthesized on a 96-well basis and ligated into deletion cassettes without gaps by ligase chain reaction, which was followed by two rounds of nested PCR to amplify trace amounts of the ligated cassettes. After the artificial linkers were removed from the deletion cassettes, the cassettes were transformed into wild-type diploid fission yeast strain SP286. We validated the transformed colonies via check PCR and subjected them to tetrad analysis to confirm functional integrity. Using this method, we systematically deleted 563 genes in the fission yeast Schizosaccharomyces pombe with a >90% success rate and a point-mutation rate of ~0.4 mutations per kb. Our method can be used to create systematic gene deletions in a variety of yeasts especially when it included a bar-code system for parallel analyses.

Ah-Reum Lee

KU-KIST Graduate School of Converging Science and Technology

Korea University

Visible-light Induced Reduction of Graphene Oxide Using Plasmonic Nanoparticle

Ah-Reum Lee

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}KU-KIST Graduate School of Converging Science and Technology

Korea University

Visible-light Induced Reduction of Graphene Oxide Using Plasmonic Nanoparticle

KU-KIST Graduate School of Converging Science and Technology