Analisis Pengaruh Budaya Organisasi Dan Kompensasi Terhadap Kinerja Karyawan Dengan Motivasi Sebagai Variabel Intervening (Studi Kasus Pada PT. Lg Bagian Penjualan Indonesia Semarang)

The problems that occurred in the employee portion of sales LG Indonesia Semarang is adecline in performance is indicated by not achieving the target for 2015. The employeeperformance and motivation is also thought to be influenced by factors of organizationalculture and also compensation deemed not feasible by most employees. This study aimedto analyze the influence of organizational culture on the motivation and compensationand employee performance parts sales LG Indonesia Semarang. The population used inthis study were all employees of LG Indonesia Semarang. The sampling technique usedwas purposive sampling. Criteria samples taken were all employees of the salesdepartment LG Indonesia Semarang who have worked more than two years are 71nurses. The method of collecting the data in this study using questionnaires andinterviews. Methods of data analysis using path analysis. Based on the research,organizational culture and compensation have a positive effect on motivation andperformance, while motivation is also a positive effect on performance. Based on theresults Sobel Test to determine whether there is mediating the relationship between theindependent and dependent variables, it is known that motivation mediates the effect ofcompensation and organizational culture on performance

Kajian Pengelolaan Lahan Subdas Secang Kulonprogo YOGYAKARTA

Penelitian ini bertujuan untuk mengevaluasi kemampuan lahan, menyusun arahan penggunaanlahan dan mengkaji pengelolaan lahan SubDAS Secang. Metode yang digunakan dalam penelitianadalah sampel terpilih pada 48 satuan lahan. Penelitian menunjukkan bahwa kemampuan lahanSubDAS Secang terdiri atas kelas lahan I seluas 187 ha, kelas lahan II seluas 147 ha, kelas lahan IIIseluas 515,2 ha, kelas lahan IV seluas 1522,7 ha, kelas lahan V seluas 7,3 ha dan kelas lahan VI seluas1223,2 ha. Arahan penggunaan lahan SubDAS Secang berupa pertanian lahan basah seluas 326,85 ha,kawasan permukiman dan budidaya tanaman semusim seluas 200,55 ha, kawasan budidaya tanamanlahan kering seluas 525,81 ha, kawasan budidaya tanaman tahunan seluas 1981,31 ha, kawasanpenyangga seluas 716,54 ha. Pengelolaan lahan memberikan pedoman pemanfaatan lahan; daerah hilirsebagai daerah pemanfaatan untuk pertanian irigasi; daerah tengah diperuntukan permukiman danpembudidayaan tanaman lahan kering; serta daerah hulu sebagai daerah imbuhan diperuntukkanwanatani dan hutan penyangga

博士学位論文内容の要旨及び審査の結果の要旨 : 平成23年9月28日授与分

Full details on spike-in control oligos. (PDF 226 kb

Regulation of Lineage Specific DNA Hypomethylation in Mouse Trophectoderm

<div><p>Background</p><p>DNA methylation is reprogrammed during early embryogenesis by active and passive mechanisms in advance of the first differentiation event producing the embryonic and extraembryonic lineage cells which contribute to the future embryo proper and to the placenta respectively. Embryonic lineage cells re-acquire a highly methylated genome dependent on the DNA methyltransferases (DNMTs) Dnmt3a and Dnmt3b that are required for <i>de novo</i> methylation. By contrast, extraembryonic lineage cells remain globally hypomethylated but the mechanisms that underlie this hypomethylation remain unknown.</p> <p>Methodology/Principal Findings</p><p>We have employed an inducible system that supports differentiation between these two lineages and recapitulates the DNA methylation asymmetry generated <i>in vivo</i>. We find that <i>in vitro</i> down-regulation of <i>Oct3/4</i> in ES cells recapitulates the decline in global DNA methylation associated with trophoblast. The <i>de novo</i> DNMTs Dnmt3a2 and Dnmt3b are down-regulated during trophoblast differentiation. Dnmt1, which is responsible for maintenance methylation, is expressed comparably in embryonic and trophoblast lineages, however importantly in trophoblast giant cells Dnmt1fails to be attracted to replication foci, thus allowing loss of DNA methylation while implicating a passive demethylation mechanism. Interestingly, Dnmt1 localization was restored by exogenous Np95/Uhrf1, a Dnmt1 chaperone required for Dnmt1-targeting to replication foci, yet DNA methylation levels remained low. Over-expression of <i>de novo</i> DNMTs also failed to increase DNA methylation in target sequences.</p> <p>Conclusions/Significance</p><p>We propose that induced trophoblast cells may have a mechanism to resist genome-wide increases of DNA methylation, thus reinforcing the genome-wide epigenetic distinctions between the embryonic and extraembryonic lineages in the mouse. This resistance may be based on transcription factors or on global differences in chromatin structure.</p> </div

Differential expression of DNA methyltransferases in the first two lineages.

<p>mRNA expression of DNMTs and Np95 in E9.5 conceptus (Em: embryo proper, Tr: trophoblast cells), ZHBTc4 ES cells (ES), ZHBTc4-derived trophoblast cells (+Dx) and ZHBTc4-derived embryonic cells (-Lf). The value of ES cell is set to 1.0. Values are means ± SD of biological replicates (n=3-4). ***: <i>p</i><0.001, **: <i>p</i><0.01, *: <i>p</i><0.05, ns: not significant; Mann-Whitney and ANOVA followed by Tukey HSD post-hoc tests when appropriate. The data for +Dx and –Lf were collected at day 2 for <i>Dnmt3a2</i> and at day 4 for <i>Dnmt3a1</i>, <i>Dnmt3b</i>, <i>Dnmt1</i> and <i>Np95</i>. Data for marker gene expression and the time course experiments are shown in supporting information (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068846#pone.0068846.s003" target="_blank">Figure S3</a>).</p

Asymmetric DNA methylation in the first two lineages.

<p>(A) The percentage of CpG methylation analyzed by Sequenom which averages the methylation of CpG methylation across each region. Embryo proper (Em) and trophoblast cells (Tr) are from E9.5 conceptus. ZHBTc4 ES-derived trophoblast cells (+Dx) and embryonic cells (-Lf) are differentiated by addition of doxycycline or removal of LIF respectively. Genomic DNA was collected at day 4 after differentiation. Values are means ± standard deviation (SD) of biological replicates (n=3-4). ***: p<0.001, **: <i>p</i><0.01, *: <i>p</i><0.05, ns: not significant; t-test and ANOVA followed by Tukey HSD post-hoc tests when appropriate. (B) DNA methylation of minor satellite and MMLV analyzed by southern blotting. Genomic DNA was collected from undifferentiated ES cells (0), trophoblast cells day 4 (4) or day 9 (9) after doxycycline treatment (+Dx), and embryonic cells day 5 (5) after the removal of LIF (-Lf). Genomic DNA was digested with methylation sensitive restriction enzyme HpaII and analyzed with each probe. Digestion with methylation-insensitive restriction enzyme MspI (M) is a control.</p

Overexpression of the Np95 gene fails to restore somatic levels of DNA methylation in trophoblast differentiation.

<p>(A) Immunostaining analysis of ZHBTc4-derived trophoblast cells (+Dx), Np95-KO ES cells, ZHBTc4 ES cells overexpressing exogenous Np95 (+MycNp95 ES) and ZHBTc4-derived trophoblast cells overexpressing exogenous Np95 (+MycNp95+Dx) using antibodies against Dnmt1, Np95 or Myc. DNA and replication sites were visualized with DAPI and EdU respectively. Scale bar, 10 µm. (B) mRNA expression of <i>Np95</i>, <i>Oct3/4</i>, <i>Rhox6</i> and Plate <i>1</i> genes during ZHBTc4 trophoblast differentiation with (+Np95) or without (ZH) overexpression of exogenous Np95. A representative clone for Np95-overexpressing ZHBTc4 cells is shown in the figure. Values are means ± SD of technical replicates (n=3). Other independent clones also showed similar results for marker gene and Np95 expression. (C) Dnmt1 localization at replication site in Np95-overexpressing ZHBTc4-derived trophoblast cells at day 4 after differentiation. The control data in ZHBTc4-derived trophoblast cells (ZH+Dx) is identical to ZHBTc4+Dx of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068846#pone-0068846-g004" target="_blank">Figure 4C</a>. Values are means ± SD of biological replicates (n=3). ***: <i>p</i><0.001, **: <i>p</i><0.01, ns: not significant; ANOVA and Bonferroni’s multiple comparison test. (D) DNA methylation analysis by Sequenom in ZHBTc4 ES (Day0) and ZHBTc4-derived trophoblast cells (Day4+Dx) with (+Np95) or without (ZH) overexpression of exogenous Np95. The control data (+emp) is identical to the data of +emp in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068846#pone-0068846-g003" target="_blank">Figure 3B</a>. Values are means ± SD of biological replicates (n=3-5). ***: <i>p</i><0.001, **: <i>p</i><0.01; paired t-tests.</p

Overexpression of the Dnmt3 genes fail to restore somatic level of DNA methylation in trophoblast differentiation.

<p>(A) mRNA expression of <i>Dnmt3</i> genes, <i>Oct3/4</i>, <i>Rhox6</i> and Plate <i>1</i> during ZHBTc4 trophoblast differentiation with/without overexpression of exogenous Dnmt3a1 (3a1), Dnmt3a2 (3a2) or Dnmt3b (3b). A representative clone from each group of Dnmt3-expressing stable clone is shown in the figure. Values are means ± SD of technical replicates (n=3). Other clones in each group showed similar levels of expression of Dnmt3 genes and marker genes. (B) DNA methylation analysis by Sequenom in ZHBTc4 ES cells and trophoblast cells with/without overexpression of exogenous Dnmt3a1 (3a1), Dnmt3a2 (3a2) or Dnmt3b (3b). emp: empty vector. A gray column indicates ES cell data (Day 0). A black column indicates data from trophoblast cells induced by the addition of doxycycline (Day4+Dx). Values are means ± SD of biological replicates (n=3-5) except for the value of major satellite for empty vector day 4 whose value is shown as the mean of biological duplicate. So, there is no stats for the value of major satellite at day 4. ****: p<0.0001, ***: <i>p</i><0.001, **: <i>p</i><0.01, *: <i>p</i><0.05, ns: not significant; paired t-tests.</p

DSB requirement for AID nuclear localisation.

<p>A) Western blot showing γH2AX levels in NIH/3T3 cells treated for 2 hours with 200µM etoposide or 200µg/ml bleomycin, the concentration used in [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082110#B47" target="_blank">47</a>]. Histone H3 and α-tubulin are shown as loading controls, quantification of γH2AX is relative to tubulin. B) Western blot of γH2AX levels in NIH/3T3 cells after 2 hours treatment with given concentrations of etoposide and bleomycin. Controls as in A, quantification of γH2AX is relative to tubulin. C) Localisation of AID in a stable NIH/3T3 cell line expressing FLAG-AID, treated with bleomycin and etoposide at different doses for 2 hours and then allowed to recover for 5 hours. White arrows indicate cells with nuclear AID. Scale bars at bottom right of each image indicate 10µm. D) Quantification of cells shown in C with nuclear AID under different drug treatments. E) AID nuclear localisation with increasing etoposide concentration, cells treated as in C. Error bars represent ±1 s.e., * p<0.05 for Student’s <i>t</i>-test, n=3 samples per drug treatment except n=2 for 20µM etoposide and n=5 for 200µM etoposide, >100 cells were counted in each sample.</p

Etoposide Induces Nuclear Re-Localisation of AID

<div><p>During B cell activation, the DNA lesions that initiate somatic hypermutation and class switch recombination are introduced by activation-induced cytidine deaminase (AID). AID is a highly mutagenic protein that is maintained in the cytoplasm at steady state, however AID is shuttled across the nuclear membrane and the protein transiently present in the nucleus appears sufficient for targeted alteration of immunoglobulin loci. AID has been implicated in epigenetic reprogramming in primordial germ cells and cell fusions and in induced pluripotent stem cells (iPS cells), however AID expression in non-B cells is very low. We hypothesised that epigenetic reprogramming would require a pathway that instigates prolonged nuclear residence of AID. Here we show that AID is completely re-localised to the nucleus during drug withdrawal following etoposide treatment, in the period in which double strand breaks (DSBs) are repaired. Re-localisation occurs 2-6 hours after etoposide treatment, and AID remains in the nucleus for 10 or more hours, during which time cells remain live and motile. Re-localisation is cell-cycle dependent and is only observed in G2. Analysis of DSB dynamics shows that AID is re-localised in response to etoposide treatment, however re-localisation occurs substantially after DSB formation and the levels of re-localisation do not correlate with γH2AX levels. We conclude that DSB formation initiates a slow-acting pathway which allows stable long-term nuclear localisation of AID, and that such a pathway may enable AID-induced DNA demethylation during epigenetic reprogramming. </p> </div