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Transcriptomic and metabolomic shifts in rice roots in response to Cr (VI) stress
Widespread use of chromium (Cr) contaminated fields due to careless and inappropriate management practices of effluent discharge, mostly from industries related to metallurgy, electroplating, production of paints and pigments, tanning, and wood preservation elevates its concentration in surface soil and eventually into rice plants and grains. In spite of many previous studies having been conducted on the effects of chromium stress, the precise molecular mechanisms related to both the effects of chromium phytotoxicity, the defense reactions of plants against chromium exposure as well as translocation and accumulation in rice remain poorly understood.
Detailed analysis of genome-wide transcriptome profiling in rice root is reported here, following Cr-plant interaction. Such studies are important for the identification of genes responsible for tolerance, accumulation and defense response in plants with respect to Cr stress. Rice root metabolome analysis was also carried out to relate differential transcriptome data to biological processes affected by Cr (VI) stress in rice. To check whether the Cr-specific motifs were indeed significantly over represented in the promoter regions of Cr-responsive genes, occurrence of these motifs in whole genome sequence was carried out. In the background of whole genome, the lift value for these 14 and 13 motifs was significantly high in the test dataset. Though no functional role has been assigned to any of the motifs, but all of these are present as promoter motifs in the Database of orthologus promoters.
These findings clearly suggest that a complex network of regulatory pathways modulates Cr-response of rice. The integrated matrix of both transcriptome and metabolome data after suitable normalization and initial calculations provided us a visual picture of the correlations between components. Predominance of different motifs in the subsets of genes suggests the involvement of motif-specific transcription modulating proteins in Cr stress response of rice.
Chromium (Cr) is the seventh most abundant metal in the earth's crust and an important environmental contaminant released mainly by leather, paint and fertilizer industries
Several studies have been carried out on the uptake and translocation of Cr from soil by different plants
In plants, a predominant model for detoxification of heavy metals is by the formation of complexes of heavy metals with phytochelatins (PCs). As a detoxification mechanism, such complexes are compartmentalized in vacuoles. However, unlike other heavy metals such as Cu, Pb and Cd, Cr is unable to induce PCs, and thus detoxification mechanism for this metal is poorly understood
Transcriptional regulation, also known as transcriptome reprogramming, is essential for plant adaptation to biotic and abiotic stresses. Use of "omic" studies have provided more inside about the physiological and molecular effects of environmental stress which led to the identification of genes involved and their expression patterns during the course of stress perception and response
High-throughput genomic technologies have made it possible to analyze expression of thousands of genes at a time. We have earlier reported comparative genome-wide transcriptome analysis of rice seedlings treated with As(III) and As(V) and identified molecular processes and networks associated with response to these metals
Results and discussion
Physiological responses of rice seedling to chromium stress
Seedlings grown on media containing different concentrations of Cr (VI) showed retardation in the growth of the seedlings with increasing concentration of Cr (VI). The shoot and root growth in terms of root length, shoot length, root and shoot weight was significantly inhibited at 100 μm Cr (VI) with respect to control. Significant decline (40-50%) in the length and weight of roots (Table
Table 1
Effect of Cr (VI) on root and shoot length and biomass of rice plant (IR-64)
0 μM
25 μM
50 μM
100 μM
250 μM
Shoot Length (mm)
23.8 ± 0.273
22.8 ± 0.447
22.2 ± 0.570
17.4 ± 0.547**
17.2 ± 0.447**
Root Length (mm)
7.3 ± 0.836
6.9 ± 0.741
6.6 ± 0.547*
6.5 ± 0.655*
5.8 ± 0.447**
Shoot FW (mg)
160.95 ± 0.636
134 ± 0.282*
90.8 ± 0.282**
58.5 ± 0.282***
47.4 ± 0.565***
Shoot DW (mg)
27.45 ± 0.63
26.55 ± 0.424
24.2 ± 0.848*
15.15 ± 0.494***
14 ± 0***
Root FW (mg)
52.25 ± 0.353
48.5 ± 0.707
41.6 ± 0.565*
32 ± 0.707***
26.2 ± 0.282***
Root DW (mg)
3.91 ± 0.296
3.35 ± 0.353
2.43 ± 0.240**
2.345 ± 0.360**
1.95 ± 0.494***
Shoot (Conc. Cr accumulation mg kg-1 dw)
46.84 ± 4.05*
211.95 ± 50.91**
679.76 ± 133.70**
1298.80 ± 146.37***
Root (Conc. Cr accumulation mg kg-1 dw)
994.31 ± 21.99***
1963.03 ± 465.94***
2560.77 ± 150.80***
4106.58 ± 29.58***
The values are means of triplicates ± SD. *, ** and *** = significantly different at P ≤ 0.05, P ≤ 0.01 and P ≤ 0.001, respectively, according to Student's unpaired t-test.
Chromium accumulation in roots and shoots increased with increasing concentration of Cr (VI). The chromium accumulation was higher in roots as compared to shoot at all the concentrations (Table
Figure 1
Effect of 100 mM Cr (VI) on the level of Cr accumulation (A), MDA (B), Proline (C), Non-protein thiol (D) in rice root after 24 h and 7 days treatment
Effect of 100 mM Cr (VI) on the level of Cr accumulation (A), MDA (B), Proline (C), Non-protein thiol (D) in rice root after 24 h and 7 days treatment. The rice variety IR-64 was germinated and allowed to grow for 5 d at 37°C and then transferred to Hewitt solution for growth. After 10 d of growth, seedlings of uniform size and growth were treated with Cr (VI) (100 μM) under standard physiological conditions of 16 h light (115 μmol m-2 s-1) and 8 h dark photoperiod at 25 ± 2°C. The values are means of triplicates ± SD. ** and *** = significantly different at P ≤ 0.01 and P ≤ 0.001, respectively, according to Student's unpaired t-test.
Significant increase in lipid peroxidation was observed after 7 d treatment (Figure
Free proline levels in roots were higher in 24 h treatment in comparison to 7 d post treatment (Figure
The level of non-protein thiols was statistically higher in roots after 24 h and 7 d treatment with 100 μM Cr (Figure
General features of Cr (VI)-stress expression profiles
Roots are the main organ for heavy metal penetration. We exposed roots of 7 d old seedlings for 24 h in 100 μM Cr (VI) to analyze the change in transcriptome. The higher metal exposures yielded a linear increase in root tissue accumulation, leading to higher degree of toxicity. However, our results evidenced the importance of the concentrations used in works of this nature, since in exposures to environmentally relevant concentrations, which are themselves representative of high contaminations; plants were able to cope with stress without displaying any noticeable phytotoxic effects, despite some accumulation of the metal accumulation. However, short-term exposure to higher Cr concentration indicated the vulnerability of this species to high Cr. Genome wide gene expression analysis was conducted using RNA from rice roots exposed to Cr (VI) (100 μm) for 24 h in hydroponic solution. An average of 47% probe sets hybridize to the transcripts, as analyzed by GCOS. Utilizing high-density Affymetrix gene-chip based microarray representing 51,279 genes, we observed that the expression of 1138 genes was up-regulated, and that of 1610 genes was down-regulated by Cr (VI). Functional analysis of these genes was done to relate the genome response to the shift in biological activities. The differentially expressed genes belong to various gene families related to transporters, cellular stress response, regulatory proteins, growth and development and secondary metabolism. A list of the up- and down-regulated probe sets under Cr stressed rice seedling roots with their annotations are provided in Additional files
Additional File 1
Table S1. List of genes up-regulated during Cr (VI) stress in rice.
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Additional File 2
Table S2. List of genes down-regulated during Cr (VI) stress in rice.
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Oligonucleotide primers for the 22 genes related to transport, stress, and defense functions were used for validation of the microarray data through RealTime PCR (Table
Table 2
Comparison of microarray and RT-PCR analysis of selected genes, differentially regulated during Cr (VI) stress in rice
Locus ID
PDR-like ABC transporter, putative, expressed
33.59 ± 0.25
ABC transporter, ATP-binding protein, putative, expressed
12.84 ± 3.31
ABC transporter, ATP-binding protein, putative, expressed
93.05 ± 0.009
Multidrug resistance-associated protein 4, putative, expressed
20.11 ± 0.06
Oligopeptide transporter 9, putative, expressed
3.83 ± 0.08
Peptide transporter PTR2, putative, expressed
3.83 ± 0.24
Sulfate transporter 3.5, putative, expressed
16.45 ± 0.04
Cytochrome P450 72A1, putative, expressed
139.10 ± 0.08
Cytochrome P450 76C2, putative, expressed
448.82 ± 0.08
Cytochrome P450 72A1, putative, expressed
80.45 ± 0.15
Cytochrome P450 74A2, putative, expressed
120.26 ± 0.002
Cytochrome P450 72A1, putative, expressed
182.28 ± 0.2
Cytochrome P450 81E1, putative, expressed
87.43 ± 0.18
Cytochrome P450 CYP99A1, putative, expressed
171.25 ± 0.27
Glutathione S-transferase GSTU6, putative, expressed
347.29 ± 0.56
Glutathione S-transferase GSTU6, putative, expressed
4.96 ± 0.03
Glutathione S-transferase GSTU6, putative, expressed
74.02 ± 0.19
Glutathione S-transferase GSTU6, putative, expressed
15.24 ± 0.03
Glutathione S-transferase, putative, expressed
30.06 ± 0.13
Heat shock cognate 70 kDa protein (DnaK family protein), putative, expressed
2683.69 ± 12.4
Heat shock cognate 70 kDa protein (DnaK family protein), putative, expressed
5.43 ± 0.81
17.4 kDa class I heat shock protein 3, putative, expressed
7.88 ± 0.09
Gene annotation and detail gene response to Cr (VI)-stress
Functional distribution of the submitted probe sets (Up-regulated and down-regulated) in the 2nd level of the GeneBins ontology is provided in Figure
Figure 2
Functional groups of differentially expressed genes following Cr (VI) stress
Functional groups of differentially expressed genes following Cr (VI) stress. The rice variety IR-64 was germinated and allowed to grow for 5 d at 37°C and then transferred to Hewitt solution for growth. After 10 d of growth, seedlings of uniform size and growth were treated with different concentrations of Cr (0 and 100 μM) under standard physiological conditions. Total RNA was extracted from the treated rice roots using and microarray was performed using one-cycle target labeling and control reagents. The hybridization data were analyzed using dCHIP software. For the genes that were differentially regulated, the gene ontology classification provided in
Among the up-regulated genes, the most affected genes are biosynthesis of secondary metabolites (122), specially flavonoid biosynthesis (58), lipid metabolism (95), amino acid metabolism (111), carbohydrate metabolism (170), biodegradation of xenobiotics (127), ascorbate and aldarate metabolism (42), membrane transport (53) specially ABC-2.A ABC-2 type transport system ATP-binding protein (11) and ABC transporters (17), glutathione metabolism (18), MAPK signaling pathway (75) and a large number of glutathione S-transferase (18) etc. On percentage basis, the most effected genes that are down-regulated during Cr (IV) stress are those for energy metabolism, cell growth and death, development. The most up-regulated genes are related to amino acid metabolism, biosynthesis of secondary metabolites and xenobiotics, membrane transport and signal transduction.
The genes coding for proteins such as glutathione S- transferase (GSTs; eighteen genes; Os01g37750, Os09g20220, Os10g38350, Os10g38495, Os01g72150, Os10g38610, Os01g49710, Os01g49710, Os10g38600, Os10g38600, 001071656, Os10g38140, Os01g72160, Os10g38740, Os10g38501, Os10g38150, AF402802, Os10g38489) which are involved in xenobiotic metabolism, are found to be unregulated in Cr stress. GSTs are known to be induced by a number of intracellular and environmental factors that include heavy-metal stress
Additional File 3
Figure S1. Expression profiles of differentially expressed GST during 100 μM Cr (VI) in rice roots. The microarray data from different rice tissues/organs and developmental stages including seedling root (R), mature leaf (ML), leaf (YL; leaf subtending the shoot apical meristem), shoot apical meristem (SAM), and various stages of panicle (P1-P6) and seed (S1-S5) development. The data were taken from (GSE6893), GSE7951 (expression profiling of stigma), GSE6901 (expression data for stress treatment), GSE7256 (expression data for virulent infection by
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Figure 3
Modulation in expression and activity of glutathione-S- transferase during Cr-stress
Modulation in expression and activity of glutathione-S- transferase during Cr-stress. Expression profiles of differentially expressed GSTs after Cr (VI), As (V), Cd and Pb stress in rice root (A). The rice variety IR-64 was germinated and allowed to grow for 5 d at 37°C and then transferred to Hewitt solution for growth. After 10 d of growth, seedlings of uniform size and growth were treated with 100 μM of Cr (VI), As (V), Cd, and Pb under standard physiological conditions. Total RNA was extracted from the treated rice roots and microarray was performed using one-cycle target labeling and control reagents. The hybridization data were analyzed using dCHIP software. The color scale (representing log signal values) is shown at the bottom. Effect of 100 μM Cr (VI) on the level of GST-activity (B) in rice root after 24 h and 7 days treatment.
A large number of cytochrome P450-related genes (35 nos.) were up-regulated in Cr stress. Cytochrome P450 s constitutes a large superfamily of enzymes, with 351 members in rice that are involved in the metabolism of plant biosynthetic pathways, including those for UV protectants (flavonoids, coumarins, sinapoylesters), pigments (anthocyanins), defense compounds (isoflavonoids, phytoalexins, hydroxamic acids), fatty acids, hormones (gibberellins, brassinosteroids), signaling molecules (salicylic acid, jasmonic acid, and so on), accessory pigments (carotenoids) and structural polymers (lignins)
Numerous genes related to the peroxidase family were down-regulated in Cr stress. It suggests that the thylakoid membranes are susceptible to lipid peroxidation caused by chromium stress. It is presumed that the down regulation of peroxidases may be due to chromium stress-induced damage to thylakoid membranes caused by lipoxygenases leading to increase in lipid peroxidation as evidenced by higher MDA content in root tissue (Figure
Three heat shock proteins (HSPs: Os03g16920, Os05g38530, Os03g16030) are up-regulated in Cr stress. Out of the 3 heat shock proteins, two are DnaK family proteins and one is Hsp20/alpha crystalline family protein. DnaK is the prokaryotic analogue of eukaryotic Hsp70. Out of these, two (Os03g16920, Os03g16030) have been shown to be up-regulated during As stress in rice
Two cysteine synthase (Os06g05690, Os01g59920) and one glutathione synthetase (Os12g16200) were down-regulated in rice roots following Cr stress in our study. Glutathione synthetase is the second enzyme in the glutathione biosynthesis pathway. It catalyses the condensation of gamma-glutamylcysteine and glycine to form glutathione
A large number of transporters were differentially expressed after challenge with Cr. Four members of sulfate transpoters were differentially expressed in our study. Out of these, two (Os01g52130, Os03g06520) were up and two (Os03g09980, Os03g09940) were down-regulated respectively during Cr stress. There are many different sulfate transporters in plants that differ in their intracellular locations, expression patterns, and kinetic properties. Sulfate can enter plants through a high-affinity sulfate transporter present in the plasma membrane
Five MATE-efflux family proteins (Os03g37470, Os06g29844, Os10g20350, Os03g37490, Os10g20350) were up-regulated on challenge with Cr. Plant MATE are localized in both vacuoles and the plasma membrane. They transport secondary metabolites and xenobiotics probably through H+-exchange. The rice genome possesses 59 MATE orthologs
Ten ABC transporter family proteins (Os01g50100, Os01g50100, Os01g50100, Os08g30770, Os08g30740, Os09g39910, Os09g39910, Os06g38950, Os01g07870, and Os04g13220) were up-regulated in Cr stress. Polypeptides encoded by ABC transporter gene family transport a wide variety of substrates across extra- and intracellular membranes, including metabolic products, lipids and sterols, and drugs. The results suggest that ABC transporters might play an important role in transport of chromium into the cell. One transporter of a major facilitator family (Os08g06010), two metal cation transporter (Os07g12890, Os03g46470), one potassium transporter (Os01g70490), two multidrug resistance-associated proteins (Os04g49890, Os04g13210) were up-regulated specifically in Cr stress. One peptide transporter PTR2 (Os04g50940), One oligopeptide transporter (Os06g03560), and one POT family protein (Os01g65110) were also up-regulated. Moreover, six aquaporin proteins (Os06g12310, Os04g44060, Os01g74450, Os10g36924, Os02g41860, and Os06g22960) and six heavy metal-associated domain containing proteins (Os08g31340, Os01g20830, Os01g48710, Os01g48710, Os01g48710, and Os01g48710) were down-regulated in this study.
Thirty six transcription factors were differentially expressed following chromium stress. Among these, eight
In our microarray experiments, we compared datasets for shift in expression at different developmental stages of rice and observed that 30 and 247 probe sets up- and down-regulated respectively were root specific during Cr-stress (Additional files
Additional File 4
Table S3. List of root-specific genes up-regulated during Cr (VI) stress in rice.
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Additional File 5
Table S4. List of root-specific genes down-regulated during Cr (VI) stress in rice.
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Figure 4
Expression profile of one of the root-specific putative ABC transporter (Os08g30770) gene up-regulated during Cr-stress
Expression profile of one of the root-specific putative ABC transporter (Os08g30770) gene up-regulated during Cr-stress. Semi-quantitative RT-PCR analysis during different heavy metal stresses in root (A). The rice variety IR-64 was germinated and allowed to grow for 5 d at 37°C and then transferred to Hewitt solution for growth. After 10 d of growth, seedlings of uniform size and growth were treated with 100 μM of Cr (VI), As (V), Cd, and Pb (100 μM) under standard physiological conditions. The expression profiles of ABC transporter (B) in seedling root (R), mature leaf (ML), shoot apical meristem (SAM), various stages of panicle development (P1-P6) and seed development (S1-S5). The microarray data was collected for different rice tissues/organs and developmental stages from the Gene Expression Omnibus database (GSE6893) at the National Center for Biotechnology Information and analyzed using dChip software. The color scale (representing log signal values) is shown at the bottom.
Large numbers of genes related to MAPK signaling pathway are down-regulated during chromium stress. Mitogen-activated protein (MAP) kinases are serine/threonine-specific protein kinases that respond to extracellular stimuli (mitogens) and regulate various cellular activities, such as gene expression, mitosis, differentiation, proliferation, cell survival/apoptosis and are involved in cell response to growth factors
Cross talk between the Cr (VI)-stress responses and abiotic stress has also been explored which revealed a high degree of similarity. Specifically, cold, draught and salt stress was found to induce many of the same genes as did chromium stress. Plant response to chromium and abiotic stresses is very similar, with both disrupting the ion and osmotic homeostasis of the plant, thus the signaling pathways would be expected to be similar. Expression of several NAC domain-containing proteins, WRKY transcription factors, cytochrome P450 and basic region/Leu Zipper (bZIP) also got induced similar to what was observed during abiotic stress
Metabolic shift following Cr (VI) stress
Metabolites in rice roots exposed to 100 μM Cr (VI) for 24 h analyzed mainly by GC-MS and NMR. Assignment of the compounds was done comparing the GC-MS profile and 1H spectra of reference compounds together with Biological Magnetic Resonance Data Bank
Additional File 6
Figure S2. The COSY spectrum of aqueous fraction of rice roots.
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Table 3
Metabolic profiling of rice root during Cr (VI) stress
MS/NMR signals
Control mg/g db
Cr mg/g db
aMyristic acid ME
0.50 ± 0.07
0.10 ± 0.02
aPentadecanoic acid ME
0.85 ± 0.12
0.10 ± 0.03
aPalmitic acid ME
4.58 ± 0.52
2.33 ± 0.07
aLinoleic acid ME
1.74 ± 0.22
4.89 ± 0.12
aErgo Sterol
0.27 ± 0.05
0.12 ± 0.02
aStigma Sterol
0.45 ± 0.03
0.18 ± 0.02
0.30 ± 0.07
0.16 ± 0.08
a2-Hydroxy Propanoic acid (TMS)2
219(M+-CH3),147, 117,73 (Me3Si.)
0.93 ± 0.09
1.21 ± 0.04
0.71 ± 0.10
0.30 ± 0.02
aBenzoic acid-TMS
0.14 ± 0.12
0.16 ± 0.03
aSuccinic acid-(TMS)2
0.69 ± 0.08
0.41 ± 0.02
aMalic acid- (TMS)3
335(M+-CH3), 233,147,73(Me3Si.)
0.10 ± 0.03
0.21 ± 0.01
aβ-Hydroxy β- Methayl Glutaric acid- (TMS)3
0.18 ± 0.02
0.44 ± 0.05
a4-Hydroxy Benzoic acid- (TMS)2
282(M+),267(M+-CH3),223,193 (M+-Me3SiO), 73(Me3Si.)
0.21 ± 0.07
0.33 ± 0.02
a Suberic acid- (TMS)2
0.21 ± 0.03
2.10 ± 0.12
aAzelaic acid-(TMS)2
3.31 ± 0.13
2.28 ± 0.15
a4-Hydroxy Cinnamic acid-(TMS)2
308 (M+),293 (M+-CH3),249,219,179,73(Me3Si.)
1.61 ± 0.12
2.10 ± 0.08
δ1.48, δ 3.77
0.37 ± 0.03
1.19 ± 0.05
δ 3.19, δ 3.52
0.25 ± 0.03
0.91 ± 0.06
aD-Fructose- (5 TMS, MeOXI)
364, 307, 217,147,103,73(Me3Si.)
0.81 ± 0.07
1.61 ± 0.08
δ 1.91, δ 2.29, δ 3.01
0.66 ± 0.05
1.66 ± 0.09
a D-galactose
554(M+-CH3), 319, 217, 205, 147, 73(Me3Si.)
2.10 ± 0.10
4.87 ± 0.22
aD-gluconic acid
613(M+-CH3), 333, 292, 217, 147, 73(Me3Si.)
0.63 ± 0.06
1.89 ± 0.09
δ 1.33, δ 4.11
0.16 ± 0.02
1.09 ± 0.08
612(M+), 507, 318, 305, 217, 147, 73(Me3Si.)
0.22 ± 0.02
0.55 ± 0.05
δ 1.6-1.8
1.04 ± 0.05
3.33 ± 0.42
273(M+), 258(M+-CH3), 230, 156, 147, 73(Me3Si.)
0.35 ± 0.02
1.12 ± 0.03
bPhenyl alanine
δ 7.3-7.45
0.45 ± 0.03
0.88 ± 0.05
δ 5.95, δ 7.75
0.45 ± 0.04
1.39 ± 0.60
δ 0.98, δ 1.04, δ 2.25
0.54 ± 0.05
1.44 ± 0.08
aIndentified and quantified by GC-MS, b Identified and quantified by NMR.
Higher accumulation of various fatty acids was observed in Cr exposed rice roots. Cellular fatty acid composition is the result of a sum of complex phenomena, required for maintaining optimal viability of against external challenges. The biosynthesis of a reactive target, such as linoleic acid and its subsequent peroxidation by the radicals generated by cells after stress exposure prevents major damage to cellular DNA
Simultaneous analysis of microarray and metabolite content using PlantMetGenMAP software
Figure 5
Altered sucrose degradation pathway using Plant MetGenMAP software
Altered sucrose degradation pathway using Plant MetGenMAP software
Metabolic networks derived from transcriptome expression profiles
The derivatisation of potential metabolic networks from microarray data is a very useful method for maximizing information
Figure 6
Functional network predicted from the genes expressed differentially during Cr (VI) stress in rice (For detail pathway please visit
Functional network predicted from the genes expressed differentially during Cr (VI) stress in rice (For detail pathway please visit
Identification of over-represented motifs in promoters of differentially regulated genes during chromium stress
As described in materials and methods, the promoters of the up-regulated, down-regulated and the total rice genes were analysed for significant abundance of the 6, 7 and 8nt motifs. The lift, support and confidence criteria used were described by Doi et al.,
Table 4
Putative motifs predicted in promoters of genes up-regulated during chromium stress
Motif Logo
Table 5
Putative motifs predicted in promoters of genes down-regulated during chromium stress
Motif Logo
Additional File 7
Figure S3 a-e. K-means clustering and GO annotations of genes up-regulated during chromium stress. The clustering was done according to the presence of the motifs identified in this study. a(i) cluster1 a(ii) GO annotation of cluster1. b(i) cluster2 b(ii) GO annotation of cluster2 c(i) cluster3 c(ii) GO annotation of cluster3 d(i) cluster4 d(ii) GO annotation of cluster4 e(i) cluster5 e(ii) GO annotation of cluster5
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Additional File 8
Figure S4 a-e. K-means clustering and GO annotations of genes down-regulated during chromium stress. The clustering was done according to the presence of the motifs identified in this study. a(i) cluster1 a(ii) GO annotation of cluster1. b(i) cluster2 b(ii) GO annotation of cluster2 c(i) cluster3 c(ii) GO annotation of cluster3 d(i) cluster4 d(ii) GO annotation of cluster4 e(i) cluster5 e(ii) GO annotation of cluster5
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Clusters of up-regulated datasets
The promoters containing the Cr specific motifs were used for functional clustering. The promoters were clustered according to the presence of various motifs. Five clusters for the up and down-regulated genes were generated by k-means clustering and functional annotation was carried out using GO annotation
Cluster 1 of the up-regulated promoters had an over-representation of genes related to transcription factor activity, DNA binding, catalytic activity and response to stress (Figure S3a). Four motifs (AGCTAGC, CTAGCTA, TAGCTAG and AGCTAGCT) were highly represented in this set. Two other motifs (GCTAGCT and TCGATCG) were also present in these genes, though at a lower frequency. The second cluster had motifs TCGTCGA and CGATCGA (Additional file
Clusters of down-regulated datasets
Cluster 1 has an over-representation of the motifs TTATCC, CTTATC, CTTATGC and CCTTATC (Additional file
A significant effect on root growth being observed at 24 h at 100 μM Cr (VI) this treatment is further used for transcriptomics and metabolomics analyses. Cr (VI) treatment was associated with lipid peroxidation and an increased in proline synthesis. Transcriptomics analysis revealed that the expression of 1138 genes was up-regulated, and that of 1610 genes was down-regulated in roots by Cr (VI). Most of the genes differentially expressed under both Cr (VI) stress were related to glutathione metabolism, transport, and signal-transduction pathways. However, somewhat unexpectedly, up-regulation of phytochelatin synthase was not detected by microarray analysis suggesting that PCs are not involved in Cr (VI) detoxification. This might be due to their nonresponsive behavior to Cr (VI) stress, which is in agreement with previous report (3, 10). On the contrary, in our study it is clear that glutathione plays an important role for detoxification of Cr-stress. Simultaneous analysis of microarray and metabolite content suggested that sucrose degradation pathway was modulated in Cr stress response involving three main fermentation pathways operating as a rescue mechanism when respiration is arrested. We also analyzed presence of cis-acting elements in differentially regulated genes during Cr (VI) stress. To check whether the Cr-specific motifs were indeed significantly over represented in the promoter regions of Cr-responsive genes, occurrence of these motifs in whole genome sequence was carried out which suggests significant co-relation between differentially expressed genes and identified motifs.
Materials and methods
Plant Material and growth parameters
The rice variety IR-64 was germinated and allowed to grow for 5 d at 37°C and then transferred to Hewitt solution for growth. After 10 d of growth, seedlings of uniform size and growth were treated with different concentrations of Cr (0, 25, 50, 100, 250 μM) under standard physiological conditions of 16 h light (115 μmol m-2 s-1) and 8 h dark photoperiod at 25 ± 2°C. Different concentrations of Cr were prepared using K2Cr2O7 (Cr VI - Merck). Length of the main root and shoot of seedlings was measured at different time intervals following Cr exposure. Roots treated with 100 μM Cr after 24 h were taken to evaluate early expressed genes subjected to Cr stress. After 10 d of growth followed by Cr treatment, shoot length, root length, fresh root weight and shoot weight were measured. All the samples were ground in liquid N2 and stored at -80°C.
Metal accumulation
Harvested plants were washed thoroughly with distilled water for quantification of total metal accumulation. The roots and shoots of the plants were separated manually, dried in an oven at 80°C for one week and kept for metal analysis. For the estimation of Cr in roots and shoots of rice, 0.5g oven dried (at 70°C) grinded plants tissue were taken and digested in 3ml of HNO3 at 120°C for 2 h and 140°C for 4 h then filtered in 10 ml of Milli Q water and stored at 4°C till the estimation. Cr was quantified with the help of Inductively Coupled Plasma Mass Spectrometer (ICP-MS, Agilent 7500 ce) at SGS India Pvt. Ltd, Gurgaon, Haryana. The standard reference materials of metal (E-Merck, Germany) were used for the calibration and quality assurance for each analytical batch. Recovery of Cr from the plant tissue was found to be more than 95.5% as determined by spiking samples with a known amount of Cr. The detection limit for Cr was 5 μg l-1.
Measurement of lipid peroxidation
Lipid peroxidation was estimated as malondialdehyde (MDA) produced using thio barbituric acid (TBA) method as described by Heath and Packer
Measurement of NP-TH (non protein thiol)
NP-TH content was assayed according to Patra et al.
For non- protein -SH group determination, the proteins were removed by TCA (12% w/v) precipitation, and after centrifugation at 12,000 × g for 10 min, thiol groups were assayed by reaction with DTNB, as mentioned above. Standard plots of L-cystein (5-40 μM ml-1) were used for determination of non- protein -SH group.
Measurement of proline
Proline was determined following Bates et al.
GST activity
Activity of GST in the tissue samples were measured by adopting the protocol of Habig et al.
Total RNA extraction and transcriptome analysis
Total RNA was extracted from the treated rice roots using the QIAGEN RNeasy Plant Maxi Kit (QIAGEN, MD). The yield and RNA purity were determined spectrophotometrically (NanoDrop, Wilmington, DE) and by formaldehyde-agarose gel electrophoresis. The microarray was performed using one-cycle target labeling and control reagents (Affymetrix, USA) using 5
The microarray data was collected for different rice tissues/organs and developmental stages, including germinating seedling (GS), seedling root (R), mature leaf (ML), leaf (YL; leaf subtending the shoot apical meristem), shoot apical meristem (SAM), and various stages of panicle (P1-P6) and seed (S1-S5) development (Jain et al., 2007; GSE6893). GSE7951 (expression profiling of stigma), GSE6901 (expression data for stress treatment), GSE7256 (expression data for virulent infection by
Expression analysis by Real-Time PCR
Total RNA was extracted as per procedure given in previous section. First strand cDNA was synthesised using 5 μg purified total RNA and RevertAid First Strand cDNA synthesis Kit (Fermantas, Life Sciences, USA). Real Time PCR was performed in 25 μl for set of selected genes using Power SYBR Green PCR Master Mix (ABI, USA). List of selected genes and oligonucleotide primers (MWG, India) used for each gene are listed in the Additional file
Additional File 9
Table S5. List of primers used for RT-PCR analysis of Cr (VI) stress responsive genes in rice roots.
Click here for file
Pathway analysis
The differentially expressed transcripts ranked by the magnitude of the moderated t-statistic determined from the statistical analysis were analyzed for cellular pathway and transcriptome interactions using the program Pathway Studio (Ariadne Genomics, USA). The pathway diagram was further filtered to show the proteins involved in cellular processes associated with the Cr response network.
Samples preparation for metabolom analysis
Fresh roots of rice were collected after Cr (VI) treatment and washed with MiliQ water. Adherent water was removed using blotting papers and immediately frozen in liquid nitrogen followed by crashing with mortar and pastel. Crashed tissue were lyophilized to remove moisture from the sample and kept at -20°C until use for analysis. For metabolomics investigation dried plant material was extracted with ten times of its weight of hexane. The solvent portion was collected by filtration and this procedure was repeated five more times until the hexane layer became almost colourless. Collected hexane layer was concentrated under reduced pressure and resulting sticky mass was stored at -20°C until analyzed by GC-MS. The remaining solid plant material was further extracted thrice with five fold excess (w/w) of 90% and then with 70% warm methanol-water. Volume of the extract was reduced to 1/3 rd using rotavapour and defatted with equal volume of hexane. Defatted water-methanol layer was partitioned (liquid-liquid) with equal volume of EtOAc (five times). EtOAc layers was separated and dried over sodium sulphate, concentrated to semisolid mass and stored at -20°C till GC-MS analysis. Remaining methanolic water layer was lyophilized to dryness and the resulting solid was again saved similarly for NMR and GC-MS analysis.
All the solvents used for the extraction of phytochemicals from rice root tissues were purchased from Qualigen (ExcealR). All detureted solvents for NMR were purchased from Sigma Chemical Company (USA).
GC-MS Method
GC-MS analysis was performed using Thermo Trace GC Ultra coupled with Thermo fisher DSQ II mass spectrometers (Thermo Scientific, USA) with electron impact ionisation at 70 eV to generate mass spectra. Thermo TR50 column 30 m × 0.25 mm (polysiloxane column coated with 50% methyl and 50% phenyl groups) was used for chromatographic separation of metabolites. To prepare the sample for GC-MS analysis of non polar hexane extract, 10 mg portion was heated at 60°C for 6 h with 5 ml of methanolic sulphuric acid (5%, v/v). After cooling, the reaction mixture was diluted and vigorously shaken with 25 ml hexane and 20 ml water. Separated hexane layer was washed with 20 ml water containing 5% (w/v) sodium bicarbonate followed by equal volume of 5% (w/v) sodium chloride solution. Hexane layer was collected and concentrated using rota vapour after drying over anhydrous sodium sulphate. The resulting oily mass was dissolved in 1 ml of GC-grade
NMR analysis
1H NMR spectra of the hexane and aqueous methanolic extracts were obtained on Bruker Biospin Avance 400 MHz NMR spectrometer (Bruker Biospin, USA) using a 5 mm broad band inverse probe head, equipped with shielded z-gradient accessories. One-dimensional 1H-NMR spectral analyses of hexane extracts were carried out using one-pulse sequence by dissolving samples in 500 ml deuterated chloroform taken in 5-mm NMR tubes. A reusable sealed capillary tube containing 30 ml of 0.375% trimethylsilyl phosphate (TSP) in deuterium oxide was inserted into the NMR tube before recording the spectra. TSP served as chemical shift reference as well as internal standard for quantitative estimation. Typical parameters for both the extractions were: spectral width: 6000 Hz; time domain data points: 32 K. For quantification purpose the effective flip angle of 45° was used, optimized and standardized instead of 90° using total relaxation delay of 7.73 s for complete recovery of the magnetization by taking consideration of our earlier studies on amino acids
Each of the characteristic peaks of specific metabolite was integrated with respect to signals of known amount of TSP and metabolites were quantified by following equation
Identification of over-represented motifs in promoters of differentially regulated genes during Cr (VI) stress
Microarray data sets of cold, drought and salinity stress (Gene Expression Omnibus database at the National Center for Biotechnology Information under the series accession numbers GSE6901) were compared with data set following chromium stress. Chromium-specific rice up-regulated genes (599) and down-regulated genes (388) were selected. One kb upstream regions from these genes were extracted from rice genome database (TIGR) and used as the test dataset. As control dataset one kilobase upstream regions from all rice promoters (66710) were used To find novel cis-regulatory elements specific to chromium stress, a database was constructed which consist of all possible combinations of 6, 7 & 8 base lengths. Perl Script was written to count the number of genes in which a given motif was present in the dataset of up-regulated, down-regulated and all rice promoters (66710). The above result was filtered for motifs with minimum of 10 and 20 counts for down and up-regulated genes respectively in the test dataset. Lift, Support and Confidence values were calculated for the filtered dataset
The up and down-regulated genes were clustered as per to the presence of the different motifs. For each of the up and down regulated sets, 5 K-means cluster group were created using the MeV software
Authors' contributions
SD, PM, SC and SD performed the experiments; DC, SB, SM and MHA performed the data analysis; DC, PKT, RDT and RT wrote the manuscript; all of the authors contributed to the research design, discussed the results and commented on the manuscript.
We thank other members of our research group for technical assistance and discussions. This work was supported by grants from the CSIR network project. We thank Profs. Raja Ray and C.L. Khetrapal (Centre of Biomedical Magnetic Resonance, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India) for providing the NMR facility to carry out our experiments.
Chromium in natural and human environments
The biological and environmental chemistry of chromium
Chromium interactions in plants: current status and future strategies
Heavy metal content of rice and Shellfish in Taiwan
Chromium toxicity in plants
Chromium uptake by rice and accumulation in soil amended with municipal solid waste compost
Study of chromium concentration in tarrom rice cultivated in the Qaemshahr region and its daily intake
Chromium, environment health criteria
Heavy metals induce lipid peroxidation and affects antioxidants in wheat leaves
Chromium stress in plants
Chromium-mediated oxidative stress and ultrastructural changes in root cells of developing rice seedlings
The uptake and translocation of tin and hexavalent chromium and effects on growth of oat in flowering nutrient solution and in soil
Phytoremediation of chromium using Salix species: Cloning ESTs and candidate genes involved in the Cr response
Abiotic Stress in Rice. An "Omic" Approach
Crosstalk between abiotic and biotic stress responses: a current view from the points of convergence in the stress signaling networks
Comparative transcriptome analysis of arsenate and arsenite stresses in rice seedlings
Free radical scavenging and senescence in Iris tepals
Thiol metabolism and antioxidant systems complement each other during arsenate detoxification in
Effect of arsenic on growth, oxidative stress and antioxidant system in rice seedlings
Effect of a free radical scavenger, 3,4,5-trichlorophenol, on ethylene production and on changes in lipids and membrane integrity during senescence of petals of cut carnations (
Lead bioaccumulation potential of an aquatic macrophyte Najas indica are related 3 to antioxidant system
Nitrogen containing compounds and adaptation of plants to salinity stress
Protection of plasma membrane of onion epidermal cells by glycine betaine and proline against NaCl stress
Chromium-mediated oxidative stress and ultrastructural changes in root cells of developing rice seedlings
Is there a role for reactive oxygen species in the mechanism of chromium(VI) carcinogenesis?
Genome-wide identiWcation and characterization of putative cytochrome P450 genes in the model legume
Transcriptional profiling of Arabidopsis seedlings in response to heavy metal lead (Pb)
Transgenic Rice Plants Expressing Human P450 Genes Involved in Xenobiotic Metabolism for Phytoremediation
Differential accumulation of high and low molecular weight heat shock proteins in Basmati rice (
Expression of stress-related genes in tomato plants exposed to arsenic and chromium in nutrient solution
Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins
Differential accumulation of dehydrin-like proteins by abiotic stresses in Deschampsia antarctica Desv
Cellular mechanisms for heavy metal detoxification and tolerance
Ascorbate is the principal reductant of chromium(VI) in rat liver and kidney ultrafiltrates
Heavy metal stress and sulfate uptake in maize roots
Constitutive expression of a high-affinity sulfate transporter in Indian mustard affects metal tolerance and accumulation
EDS5, an essential component of salicylic acid-dependent signaling for disease resistance in Arabidopsis, is a member of the MATE transporter family
Interactions of chromium with micro-organisms and plants
MAPK cascades in plant defense signaling
Prolyl-4-hydroxylase (
Effect of proline on lactate dehydrogenase activity: testing the generality and scope of the compatibility paradigm
Biochemical responses to drought stress in mulberry (
Long-term effect of NaCl on the activity of uridine and uracil salvage for nucleotide synthesis in cultured mangrove (
Alteration in cellular fatty acid composition as a response to salt, acid, oxidative and thermal stresses in Lactobacillus helveticus
Changes in plasma membrane lipids, aquaporins and proton pump of broccoli roots, as an adaptation mechanism to salinity
The effect of low temperature on metabolism of membrane lipids in plants and associated gene expression
Development of a novel data mining tool to find cis-elements in rice gene promoter regions
Identification of cis-Elements That Regulate Gene Expression during Initiation of Axillary Bud Outgrowth in Arabidopsis
Sugar response sequence in the promoter of a rice -amylase gene serves as a transcriptional enhancer
Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation
Tolerance and co-tolerance of the grass Chloris barbata Sw. to mercury, cadmium and zinc
Rapid determination of free proline for water-stress studies
Glutathione S-transferases: The first enzymatic step in mercapturic acid formation
Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection
GeneBins: a database for classifying gene expression data: Application to plant genome arrays
Improved Quantification from 1H-NMR Spectra Using Reduced Repetition Times
Metabolic profile of lettuce leaves by high-field NMR spectra
Quantitative NMR spectroscopy in the quality evaluation of active pharmaceutical in gredients and excipients


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|2 Fast 2 Furious Trailer
|Former cop, Brian O'Conner is finally arrested after letting his leader escape the law. To avoid the consequences, he must now work with an old college friend and help the police arrest a local drug exporter.
|Related Titles:
|2 Fast 2 Furious
|Related Names:
|James Remar, John Singleton, Thom Barry, Mark Boone Junior, Mo Gallini, Cole Hauser, Ludacris, Eva Mendes, Tyrese Gibson, Paul Walker, Amaury Nolasco, Michael Ealy, Devon Aoki, Jin Au-Yeung, Roberto 'Sanz' Sanchez, Edward Finlay
|Content Rating:
|Not Rated
|Video URL:
|Courtesy of:
|Screenplay, Inc.


text:
BigEasyBaby11's Mom Answers Activity
BigEasyBaby11 is a star in these topics:
Pregnancy > Emotional Health During Pregnancy (42 helpful answers)
Does stress affect your pregnancy? Watch this
Most recent answer (06/09/2015):
Stress makes me cry all the time in my pregnancy. I am an emotional fool girl don't know why everyone thinks that i am full of negati...posted by a BabyCenter Member


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Built in 2008, this charming design hotel comprises a total of 11 rooms including 3 junior suites. Facilities on offer to guests include a bar and conference facilities. Guests can also take advantage of the room service.
This hotel is located in the beautiful district of Cannaregio in the north west of Venice, just 100 m from the Ca D'Oro Palace. Guests will find restaurants, bars and shops in the immediate vicinity of the hotel and there are links to the public transport network 200 m away. Sights of interest to tourists in the surrounding area include the Rialto Bridge (400 m) and San Marco (800 m). Cannaregio is easily accessible via waterbus routes which run down the Grand Canal, along the Cannaregio canal, and along the northern shore of the island. With its central location and easy accessibility, it is a great choice for business and pleasure travellers alike.
All rooms come with a private bathroom with a shower/bathtub and a hairdryer and offer a king-size or double bed. They are equipped with a direct dial telephone, satellite TV, Internet access, a safe and a minibar/fridge. Furthermore individually regulated air conditioning and heating are provided in all accommodation as standard and all rooms feature either a balcony or a terrace.
Guests can select their breakfast from a buffet.
American Express, Diners Club, MasterCard and VISA are accepted at this hotel.
Room Facilities:
Direct dial telephone
Satellite cable TV
Internet access
Mini fridge
Double bed
King size bed
Individual airconditioned
Individual heating
Wheelchair accessible


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Windows 7 x64
WestTech replied to sux010's topic in Windows VistaIssues with Windows 7 Media Center - does not recognize the Xbox 360, (this is with the Kinect dashboard update – November 2010) - go to "Network and Sharing Center" and click on "Choose homegroup and sharing options" and then click "Change advanced sharing options" - Ensure media sharing is enabled, and click "Choose media streaming options" - This will display a list of devices allowed or not allowed to communicate with your PC's media. - If you Xbox 360 is appearing as a device allowed, and still not communicating with your PC, double click on it, and confirm the IP address. If it is different than your Xbox 360's actual IP address, you will need to 'remove' it from the list of known devices. Don't worry, it is going to reappear. - Open command prompt with elevated rights and run an "ipconfig /flushdns" on your PC. - restart your Xbox 360, go to Settings – Computer – and remove any PC connections that may be listed. Then select 'setup' - Open Windows Media Center, go to settings and click 'Extender' add new extender, and your Xbox 360 will appear.


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Some lessons for wiki publishing from Gutenberg's time
Take a look at this page:
It is from a rare Blackletter Bible (1497) printed in Strasbourg by J.R. Grueninger. The coloured chapter initials were hand written after the page was printed. Other famous incunabula include the Gutenberg Bible of 1455, the Peregrinatio in terram sanctam of 1486, printed and illustrated by Erhard Reuwich.
If you look closely at the image you can see the several letters have been painted in by hand. This is because the publishers of the time were trying to recreate the effects of the handwritten manuscripts. In other words, apart from the extra expense and technological challenges of dealing with a new medium of publishing – the printing press – they were also reintroducing the problems and labor intensiveness of handwriting.
Think about this the next time you are trying to use a wiki page as a Word document. This is a new publishing medium and its advantages are in authoring that is fast and collaborative. The presentation is supposed to be minimal. Do not spend your time trying to recreate a Word document's formatting.
Incidentally, there are a couple of other lessons that can be drawn from Gutenberg's time. First, Europe's scientific community benefited from synchronicity. Prices of books did drop, eventually, but the real early gain was that one scientist's book would be simultaneously available for purchase by other European scientists. For example, in the 13th century it took decades for Fibonnaci's Liber Abaci to be copied and reach European countries outside of Italy. This was a shame because only with its arrival in a country did the local learned community get to learn the multiplications tables that you and I took for granted during primary school. By contrast the printing press meant that Principia Mathematica could spread very quickly.
Do not underestimate the utility of the daily e-mail feature in enterprise wiki software like Socialtext. In my company's research team we use the daily e-mail to learn what each of us has been up to in their research and to build on each other's work.
Finally, it is worth remembering that Gutenberg had to declare bankruptcy and that he did not make money from his bible publishing activities. On the other hand, perhaps his most lucrative activity was printing indulgences for the Catholic church. New technology requires new business models and perhaps the one thing that won't make you money is reproducing the products of the old, established technologies.


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3x Aluminium Limited Edition Olympic Games Coca Cola Bottles New 250ml Boxed
- Sold for: Start Free Trial or Sign In to see what it's worth.
- Item Category: Sports
- Source: eBay UK
- Sold Date: Sep 12, 2012
- Channel: Auction House Sell similar item on eBay
3 x Aluminium Limited Edition Coca Cola Bottles. The set consists of three olympic designs gymnastics, swimming and athletics, one of each design is included within this auction. Each bottle is 250ml, unopened and boxed. Any questions then please ask :-) Will post overseas, please email for overseas postage costs
Posted with eBay Mobile


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Posted by anonymous
When all else failed, he was one I could afford. I'll never forget him. In 1986, he listed his services in the Detroit Free Press. At the time he was handling Wayne County Divorces only. For $215.00 total, I was set free of a loser. Thanks Mr R.K. Mayer!!!


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Here's an interview that NBC Miami did with local DJ, Jessica Who?. When asked about her current top 3 songs that she's spinning, here's what she said:
"…Jay-Z's "On to the Next One" is one of the hottest tracks out right now. It has a great beat, and everyone loves Jay-Z even if they don't know it. A local producer, Sonic C, has a track called "Stickin'" that is just insane. It's definitely what most DJs call "car alarm" music, but it just gets people crazy even if they have no idea what it is. And another one like that is A-trak's remix of the Yeah Yeah Yeah's "Heads Will Roll." It has a great big room vibe and gets a great reaction even from people who have never heard it…"
Read the rest of the interview by clicking here.


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The Beavers open up the 2007 slate in Reser Stadium against Utah in an evening game set for 7 p.m. The following week OSU travels to Cincinnati where they will face the Big East's Bearcats in a 4:30 p.m. showdown on ESPN.
Record on TV under Riley
2006: 8-3 2005: 4-5 2004: 3-5 2003: 4-5 1998: 1-3 1997: 0-2 ----- Total: 20-23
The following weekend brings the Men in Black back to Corvallis for their final non-conference match up against Idaho State in a 3:30 p.m. game televised by FoxSports NW.
Conference tilts at USC on Nov. 3 and at Oregon on Dec. 1 will be televised by ESPN or ABC with the Nov. 10 home game against Washington being shown live by FoxSports Net.
Oregon State appeared on TV in 11 of their 14 games last year. OSU posted a 4-1 record on FoxSports NW, a 1-1 record on ESPN, a 2-1 record on FoxSports Net and a 1-0 record on CBS.
Last year was the first time the Beavers posted a winning record (8-3) in televised games under coach Mike Riley.
See the full 2007 schedule here or download desktop wallpaper with the 2007 schedule.


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|18 Dec 2010
Ibm t41 win 7 sound and aero
I have a standard ibm T41 ultraportable and base. (Not tablet)
I tried win 7 but no sound and no aero peek and desktop rsolution not very clear.
Fronm all ive seen from google and going to ibm driver downlaod site this machine cannot run win 7.... but in utube have seen guys who say its possible.
Has anyone succesfully run IBM t41 on win 7 ultimate ???
If so could you point me in direction of working drivers please.
(Some boards I saw previously said the initial beta copies of windows had drivers which worked, and these drivers later dissapeared)
|My System Specs
|19 Dec 2010
Systems 1 and 2: Windows 7 Enterprise x64, Win 8 Developer
This is the page for drivers:
Lenovo Support - Right navigation for documents
Lists Vista, but not Win 7.
Try using Vista drivers.
If that doesn't work, try using compatibility mode.
Finally, try to identify video and audio hardware by using Device Manager. Then you can try to find updates outside of Lenovo.
|My System Specs
|19 Dec 2010
Win 7 Ultimate x64
GWM121, welcome to the Seven Forums.
This your laptop,
Lenovo Support - Overview - ThinkPad T41, T41p
if so there is not now and never will be a Windows 7 driver for the video, doesn't matter whether it has the 7500 or 9000 in it as both are just too old. That and there hasn't been a driver update for either of them since late 2006, and there is no Vista driver either (sorry Lemur, the Vista driver on that download page is listed as N/A). Even if there was a driver available for it Aero wouldn't work as it requires a minimum of DirectX 9 hardware support, and at best the 9000 supports up to DirectX 8.1 and the 7500 is even older and only supports DirectX 7. IBM/Lenovo won't have a driver as ATI never made one.
You can try the XP driver from Lenovo, but I would be more inclined to go with the last one from ATI,
and use this,
Mobility Modder - ATI Version, Install the Latest Drivers on your ATI laptop
on it so that it will (or at least should) work with the video card in the laptop. In either case you will probably have to use compatibility mode,
Compatibility Mode
to get the driver to install.
The audio is of equal vintage as the audio, and from looking at the driver from Lenovo is based on the Soundmax audio chip. No idea which version of Soundmax is in it, so can't recommend a driver to try for it. You can try the XP driver from Lenovo.
|My System Specs
|20 May 2012
win xp
how to install drivers for ibm t41 in win 7
Hi there,Good news for those who have been trying to use windows 7 in ibm t41.the problem countered was with sound and videos.rest assured the problem has a easy solution.The drivers which u have for ur xp version ,open it in win 7 by right clicking it and checking on open in compatible mode for [previous version.Wwill surely help.feel free to ask any doubts
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Ok so I built myself a new PC and everything was running fine. Then I decided to change the graphics card from an ASUS Radeon HD 6770 to an ASUS Radeon HD6850 1GB. Upon restart and every restart since, the Aero effects do not work. At first I just figured I hadnt got the right drivers. Games...
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Aero Theme/Aero system files Crashed.. basic theme only.. plzz help..
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Hi all. This is my first post since upgrading to Windows 7. I am currently looking a way to enable Aero in my machine. (as note, currently I am missing the Aero Glass, Aero Peek, and Aero Flip.) My specs: Axioo NVS M720S Laptop Genuine Intel Pentium Dual Core T2130 @ 1.86GHz 1024MB DDR2...
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After killing the process of Aero (wich had a memory leak, it was reaching 1 GB by itself and using all of one core). I restarted the Aero process only to be greeted with something like "your performance analisis was reset" (all my windows7 performance scores were reset) also , aero glass...
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|Message view
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|« Date » · « Thread »
|Regina Henschel <rb.hensc...@t-online.de>
|Re: [Calc] Feature request: An easier way of laying out form elements
|Sat, 23 Jun 2012 15:06:14 GMT
Hi Dwayne, Dwayne Henderson schrieb: > What do you guys think? > > http://i.imgur.com/iSrzf.png vs http://i.imgur.com/esY1o.png > > https://issues.apache.org/ooo/show_bug.cgi?id=119671 > I personally like this approach: 1. I like to get something like "Anchor as character" is in Writer for Calc too. Such anchored object would always be completely inside its parent cell (which may be merged cells too). The position would be relative to the anchor, the same way as it is now for "anchor to frame" in Writer. Snapping to grid while creating or modifying is not persistent. You loose alignments, when the column widths or row heights are changed. Nevertheless the currently available snapping can be improved. 2. I like to get formatting styles for form controls for all modules, not only in Calc. Such formatting style would contain all the settings, which are now already possible for graphic objects in Draw/Impress. 3. I like to get formatting styles for graphic objects in Writer and Calc, so that graphic objects and form controls are handled equally in all modules. Kind regards Regina --------------------------------------------------------------------- To unsubscribe, e-mail: firstname.lastname@example.com For additional commands, e-mail: email@example.com
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Case is damaged
I bought this for my son so I have no idea if it's a good game or not, but I am sure it is with they way he has gone on and on about for months now. But I want to comment on the fact that when we got it in the mail the case was damaged. Smashed. Since they are sold out he has decided to go ahead and keep it and try it to see if the cds are fine. One was not even on it's little holder it was just sliding around loose in the case. :( We will be returning it if they dont work. They shipped it in a bag not a box. No protection. It could be a manufacturer defect because my other son bought one as well and said his was the same way. Not sure where he bought his from though. Not happy about this at all.
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Owner Financing Homes in 46772, IN
|3 Listings Found
This property will be available for a lease purchase with owner financing. No bank qualifying
Bluffton, INDetails »
in Walton Addition.We offer seller financing. Perfect Credit NOT Needed.Banks Are Not Lending But You Can
Fort Wayne, IN, 46806Details »
5,250 sq-ft residential lot for sale with owner financing. Ready to build a home- Need to lock down
Fort Wayne, IN, 46807Details »
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Luna Traders
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Ida's Maryland Crabby Dip
Recipe by Lindas Busy Kitchen
My friend Ida shared this recipe with me... These are her comments: I served this dip in a rosemary and olive oil bread bowl. I toasted the bits of bread from the center, and served them along side. This was SO GOOD! Ida
Top Review by Caroline Cooks
Wonderful appie! I made the recipe into 2 casseroles--one for immediate consumption. LOL and one for the freezer and Holidays. This dip is great hot; but had a couple of bites left and it was great cold, too. Made for Christmas Spelling Bee Event.
See Discussion:1 Review
Ingredients Nutrition
- 11 ounces cream cheese, softened
- 1 small onion, finely chopped
- 5 tablespoons mayonnaise
- 2 (6 ounce) cans crabmeat, drained, and flaked
- 1⁄8 teaspoon garlic powder
- 1 lemon, juice of
- 2 teaspoons Old Bay Seasoning
- 1 tablespoon Worcestershire sauce
- salt and pepper, to taste
- 1 bread, bowl and bread, chunks for dipping or 1 cracker
Directions See How It's Made
- Preheat oven to 350.
- In a med. bowl, combine cream cheese, onion, lemon, Worcestershire sauce, Old Bay Seasoning, mayonnaise, crabmeat, garlic powder, salt and pepper.
- If cream cheese is still too hard, place all ingredients into a bowl, and use a mixer, on low speed, to combine.
- Spread mixture into a 1 quart baking dish.
- Bake for 20 minutes in a preheated oven.
- While the dip is baking, cut a circle in the top of the bread, and scoop out the inside, to create a bread bowl.
- Tear the removed bread into pieces for dipping.
- Remove baked crab dip from the oven, and stir well.
- Spoon the mixture into the hollowed out loaf.
- Place bread bowl and chunks of bread on a med. baking sheet, and bake for an additional 10 minutes.
- Serve hot.


text:
Exeter, UK (SPX) Feb 25, 2009
A team of scientists, led by the University of Exeter, has used game theory to explain the bizarre behaviour of a group of ravens. Juvenile birds from a roost in North Wales have been observed adopting the unusual strategy of foraging for food in 'gangs'.
New research explains how this curious behaviour can be predicted by adapting models more commonly used by economists to analyse financial trends.
This is the first time game theory has been used to successfully predict novel animal behaviour in the real world. The researchers believe this analysis could also shed light on the variation in feeding strategies in different populations in other species.
Ravens feed on the carcasses of large animals. Most populations live in temperate forests, where individuals search for carcasses and finds are then defended by a pair of territorial adults. Unpaired younger birds, on the other hand, gather at communal roosts from which they search individually for carcasses on adult territories and recruit each other to overwhelm adult protectionism.
However, at one raven roost on Anglesey, things work differently: juveniles forage in gangs. This level of coordination had not been seen before in a raven population.
The researchers built a mathematical model to understand how this behaviour evolved and why it might occur in some roosts and not others. The model designed for this study was based on techniques used in other game theory models, which identify the most profitable behaviours of individuals in different situations to predict what would be favoured by evolution.
The study revealed two strategies as being most profitable for ravens to find food. One is for birds to search independently for food and recruit each other. The other is for the birds to forage in gangs.
The findings showed that gang foraging should occur when searching for food individually is no more efficient than foraging in groups. This is likely to be the case if the roost covers landscape that can be thoroughly explored by a gang over the course of a day. The deforested Welsh countryside offers just such conditions.
The study also identifies the availability of food as a key factor. The roost in Anglesey is situated in an agricultural area, which means that the carcasses of farm animals are often available so food is more plentiful than in wild locations.
When food is abundant, the opportunity for social advancement becomes more important. These ravens seem to be using foraging behaviour, not only to find food, but also to gain social status, which could help in other aspects of their lives, including finding a mate.
Lead author Dr Sasha Dall of the University of Exeter said: "This is a rare example of how game theory has been used to predict behaviour in animals in the real world. Our study shows the potential for game theory to help biologists understand how different social structures and behaviours evolve in different environments and in response to human activities."
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Urban elephants ply Bangkok streets in search of tourist dollars
Bangkok (AFP) Feb 24, 2009
A drunken tourist staggers about as he repeatedly drops his bags of elephant feed on Bangkok's Soi Cowboy boulevard.
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text:
I just wanted to know if I can combine class abilities/actions without having to switch between the classes. In other words, I am a gladitor and I want to be able to heal myself using conjurer abilities without having to switch to a conjurer class during a fight.
My stats:
Gladiator: 10
Miner: 10
Goldsmith: 8
Blksmith: 8
Armor: 5


text:
Royal Icing Transfer
Decorating By doreenre Updated 3 Dec 2008 , 2:03am by Daisys_Cakes
I've seen great things w/ the buttercream transfers that people have done on cakes and was wondering if it was possible to do something similar using royal icing. Anyone ever try a royal icing transfer to fondant?
i have no idea, but im as curious as you are to find out!
...........waiting patiently................
I don't see how it could work, but wouldn't that be the same as a RI plaque, or color flow?
You use RI to do the outline for color flow and then fill in the spaces with color flow or diluted RI. So, I guess the answer is yes.
I guess I"m not familiar w/ color flow then. Can someone elaborate?
Would you do the same as you would a buttercream transfer...decorating onto wax paper then transferring?
See my posts on this thread:
Daisy -- thank you so much for the link...it's exactly what I was looking for!
You can also use candy melts or colored white chocolate to do a transfer. The good thing about this is that it sets almost instantly in the fridge so it's much faster. And it tastes good.
....... do the same as you would a buttercream transfer...decorating onto wax paper then transferring?.......
It is best done on *plastic wrap* especially if the piece is rather large.
As was mentioned make sure you 'pillow' the flow into the outline. It settles down as it dries & if you haven't pillowed it enough it actially will dry w/a depression
Yes, it can be done right on a cake but there is no room for error here
Chocolate (CT) is a great alternative.
Daisy -- thank you so much for the link...it's exactly what I was looking for!
No problem!
Quote by @%username% on %date%


text:
Look inside for:
- Free information and tips about mounting 3D photographs yourself
- Links to sources of stereo mounting equipment, supplies, and services
- Instructions for sending your film to us for our all-new mounting service!
Be sure not to miss our unbelievable
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Are we CRAZY??!! How can we make this offer?


text:
Remake / Hoffman and Kim
Wilhelm Heinrich Otto Dix's "Portrait of Sylvia Von Harden" remade by Stephan Hoffman & SoYeon Kim. See all the Remake submissions, here.
Instructions for submitting your own, here.
Wilhelm Heinrich Otto Dix's "Portrait of Sylvia Von Harden" remade by Stephan Hoffman & SoYeon Kim. See all the Remake submissions, here.
Instructions for submitting your own, here.
My friends at Drawn and Quarterly have sent over two amazing books for me to giveaway! "The Death Ray" by Daniel Clowes, and "The Great Northern Brotherhood of Canadian Cartoonists" by Seth.
So who wants to win these two books?
Already getting amazing submissions to our Remake project with Adobe!
"Behind the Gare Saint-Lazare, 1932" by Henri Cartier-Bresson.
"Behind the Gare Saint-Lazare, 1932" remake by Ollie Jarman.
"Narcissus" by Caravaggio.
"Narcissus" remake by Max Zerrahn.
Make one of your own! Full details HERE.
Adobe recently put forth a challenge to students in the UK with a cash prize of £10,000. The deadline is soon so if you want in on it you better work quick! Since so many of our readers live outside the UK I thought it might be fun to work with Adobe on our own project. They thought so too, so here it is!
The Booooooom + Adobe Remake photo project.
I have one of those cool Gizmon Half-D cameras to give away courtesy of the gang at Four Corner Store. It's a mini sized digital camera in the spirit of the original Olympus Pen, take a look at the second image to see just how small it is!
We are officially on Tumblr! http://booooooom.tumblr.com/
Nikon recently launched a contest in search of the coolest concert photos and I thought we could have our own – just for Booooooom readers! With so many amazing photographers frequenting this site and so many awesome concerts going on I thought it would be a fun way to wrap up the summer!
So here is the Booooooom + Nikon "Shoot The Show" Contest.
I just got a copy of Mike Mills' new book that accompanies his latest film "Beginners". I'm gonna give away another copy of the book to you, as well as a digital download of the film soundtrack!


text:
1 hour wellness consultation
I will empower you to incorporate productive habits in basic nutrition, sleep management, time management and positive thinking, one at a time, so you can reach for the greatest version of yourself. This service is a great fit for individuals that want to invest in their personal development and have a long term vision for the person they can become. Do you feel that you never have enough time? Do you feel anxious thinking about work, relationships or food? Do you wish you could change just one thing in your life you know would make all the difference in the world? Do you know there is more to life than where you're at, but don'know where to start?
I used to be an abusive drinking, pill poping insomniac surrounded by toxic relationships and a job that no longer served my true self. I've made a big change, so I know you can do it to. And I'm here to help 😊
Training & Qualifications
I share what I know. I have successfully embraced, applied and kept every habit and life changing shifts I educate my clients on.
I grew up in France with a confident relationship with food and the pleasures we can embrace in life. My relentless focus on improvements and wellness have given me a lifestyle where I can productively navigate work and personal life, cultural influence and my authentic self, being involved yet staying aware of self care.
That balancing act took me years to design. The amazing results are the reason why I want to help individuals enjoy the lifestyle they dream of. I know you can do it!
Availability & Preferences
I'm usually free on Tuesday afternoon and evenings, Thursdays during the day and evening, Sunday late afternoon and evening.
Reviews of Paola
Joy provided unit of Virtual Assistance for PaolaJul 17, 2017
Stephen received hour of 1 hour wellness consultation from PaolaJun 30, 2017
Joy provided unit of Virtual Assistance for PaolaJun 11, 2017
Joy provided unit of Virtual Assistance for PaolaMay 01, 2017
Lauren received hour of 1 hour wellness consultation from PaolaMar 09, 2017
Joy provided unit of Virtual Assistance for PaolaMar 08, 2017
Alii received Help with Zipper Repair from PaolaFeb 24, 2017
Lauren received hour of 1 hour wellness consultation from PaolaFeb 23, 2017
Lauren received hour of 1 hour wellness consultation from PaolaFeb 11, 2017
Joy provided unit of Virtual Assistance for PaolaFeb 09, 2017
About Paola Aguillon
Paola has had a diverse professional life spanning two continents, France and United States, and several industries, including hospitality, fashion, retail, wellness and dance. The common denominat...or was always a drive for excellence supported by a solid foundation of integrity, consistency and honesty, with the absolute certitude that hard work and earned trust is the path to success and making a positive difference in people's life. The goal is to share that experience to empower others and help them find harmony and balance in their life. show moreOpen to other proposals
Paola Aguillon is looking for:
- Language Lessons
- Business
- Photography
- Music & Audio
- Art & Design


text:
| Clubhouse: Bear Watchers
belongs to Wikiality.com's
Bear Watchers Club
This is the future home of the BEAR WATCHERS CLUBHOUSE. grrrr. It will be manly, and there will probably be taxidermied dead things hanging on the mantle. If you want, you can extend the "Bear Watchers Club" to include the "Watching" of all Godless Killing Machines on the Wikiality.com Tubes, or you can make the other GKM-Watchers get their own club. Entirely up to you...
Anyway, watch this space for more Bear Watching updates.
And if you're interested in being the Bear Watchers Club Captain, leave us a note here. Required for Club Captainship: excellent Bear Watching skills, Godliness, Stepheneration, and Americanness; Bear Huntin' skills a plus. Apply today.
Some pages of potential interest for the Wikiality.com Bear Watchers Club (grrr):


text:
Senior diplomats from North Korea and China held strategic talks in Beijing yesterday as they work to repair strained relations, but offered little specific indication they will lead to a resumption of nuclear disarmament talks any time soon.
North Korean First Vice Foreign Minister Kim Kye Gwan told Deputy Foreign Minister Zhang Yesui that Pyongyang was willing to develop friendly and co-operative relations with Beijing, Xinhua reported, adding that the country was willing to engage in talks on nuclear issues.
"DPRK is willing to resolve the dispute over its nuclear programme peacefully through talks, including returning to the six-party talks process as well as any other format of dialogues," Xinhua quoted Kim as saying.
Zhang said China supported dialogue among all concerned parties and wished the stalled six-party talks could resume as soon as possible. He also said China would work with North Korea to forge stable bilateral relations.
The meeting followed Pyongyang's surprise offer on Sunday to hold direct talks with the US after months of threats that raised the possibility of fresh fighting on the Korean peninsula.
The North's nuclear programme was also believed to be on the agenda for the meeting between United Nations Secretary General Ban Ki-moon and President Xi Jinping in Beijing yesterday. Ban met Xi after visiting a centre that trains China's UN peacekeepers. Xi said: "The UN carries the expectations of people of all countries."
In its offer to Washington, North Korea's National Defence Commission, the powerful governing body led by leader Kim Jong-un, insisted that there be no preconditions to talks and no demands that Pyongyang give up its nuclear assets unless Washington was willing to do the same.
The Obama administration responded that it was open to dialogue, but wanted "credible negotiations" that involved North Korean compliance with UN resolutions and would lead to a nuclear-free North.
The proposal is expected to be discussed in meetings this week in Washington involving US, Japanese and South Korean officials.
Cui Zhiying , director of the Korean Peninsula Research Centre at Tongji University in Shanghai, said Beijing wanted to push for dialogue.
"China will call on Pyongyang to go back to the negotiating table," Cui said. "Beijing is expected to ask both Pyongyang and Washington to offer sincere gestures in having dialogue."
Cui said Beijing knew that denuclearisation on the Korean peninsula could not be achieved in a short period of time.
Cai Jian , deputy director of the Centre for Korean Studies at Fudan University in Shanghai, said it was unlikely that South Korea and the US would immediately engage in talks with Pyongyang.
Associated Press, Reuters


text:
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Free Shipping in US over $40 (exceptions apply)
SKU# deep-v-neck-gown
Regular Price:
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My favorite dress. Very comfortable and flattering. Getting another one in a different color. On a side note, seems that Yummie has improved their shipping speed.
I look forward to wearing this gown/jammie every night. It is so comfortable and cosy. It keeps you cool while you sleep and I even feel a little sexy at times when it curves my body!
Love, love LOVE this long sleeve V Neck Nightgown. Tempted to order another 1 in gray
I thought this was going to be a comfy nightgown I could throw in my suitcase and chill in. Well it's too cute for that! I plan on wearing it with a long sweater and booties this fall. Maybe I'll buy a Medium and see if it's looser to wear to bed LOL!
elegant and comfortable. the look of this gown is casual and can also be dressed up. I love that it hugs my body yet doesnt feel "clingy".
Wonderful feel to the fabric. Strange tight fit in hips. Struggled with the decision, but had to return it. Had it been longer in length, would have been seduced to keep it.
Sexy & fabulous. I love it.
I was disappointed with this product which is hard for me to say because I have so many yummie products and absolutely LOVE them all. But this garment is thin and stretches out quickly for the amount of money it was extremely disappointing!
This gown is beyond expectations! It is cozy & comfy. Well worth the price. You will be amazed. Love it so much went back and purchased the matching robe!!! Wonderful treat for myself. No guilt here !! Love it especially with the socks I received free! Perfect for winter nights...
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I haven't had a chance to do price checks on these items, but we have some huge stacks with Checkout51 cashback and some high-value coupons. You may want to print these coupons now and price check the items next time you shop.
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text:
The Vikings placed a first-round tender on Edwards, but that that might not hold up when a new collective bargaining agreement is reached and he could be headed for unrestricted free agency, where he would get his wish to look elsewhere. The Vikings may be willing to let Edwards get away when free agency begins, but they aren't going to let the DE position become a weakness. Brian Robison, whose role in the defense has been increasing every year, now looks to be the opening-day starter after signing a three-year contract worth $14.1 million, including a $6.5 million signing bonus.
The Vikings had placed a second-round tender on Robison Wednesday, but it was clear that he was in the future plans. Whether it is a portent of things to come with Edwards or not, the Vikings have invested in Robison and it would seem that he is the man in and Edwards is the man out.
"I think I've done everything I possibly can to show that I can be a starter. I feel like I've taken advantage of the opportunities I've been given," Robison said after the 2010 season. "I feel like I've done very well showing them that I can play the run as well as the pass. I think I've done everything I could possibly do to show them that I deserve to be a starter and every-down player in this league."
John Holler has been writing about the Vikings for more than a decade for Viking Update. Follow Viking Update on Twitter and discuss this topic on our message boards. To become a subscriber to the Viking Update web site or magazine, click here.


text:
what is meant by collegium system
The collegium system of appointment of judges to the country's constitutional courts was evolved in order to ensure judicial independence so that no other branch of the state, including the legislature and the executive, would have any say in the appointment of judges. It is a system under which appointments and transfers of judges is decided by a forum comprising the Chief Justice of India and the four seniormost judges of the Supreme Court. There is no mention of the collegium either in the original Constitution of India or in successive amendments.
Article 124 of the Constitution, dealing with the appointment of Supreme Court judges, says the appointment should be made by the President after consultation with such judges of the high courts and the Supreme Court as the President may deem necessary. The CJI is to be consulted in all appointments, except his or her own. Article 217 that deals with the appointment of high court judges says a judge should be appointed by the President after consultation with the CJI and the governor of the state. The Chief Justice of the high court concerned too should be consulted.
The collegium system is based on three judgments of the Supreme Court that are collectively called the 'Three Judges Cases'. The SP Gupta vs Union of India case of 1981 is called the 'First Judges Case'. It declared that the "primacy" of the CJI's recommendation to the President can be refused for "cogent reasons". This brought a paradigm shift in favour of the executive having primacy over the judiciary in judicial appointments for the next 12 years. The collegium system has been in use since the judgment in the 'Second Judges Case' was issued in 1993. On October 6, 1993, a nine-judge bench decision in the Supreme Court Advocates-on Record Association vs Union of India case (also called the 'Second Judges Case') said "justiciability" and "primacy" required that the CJI be given the "primal" role in such appointments. It overturned the SP Gupta judgment, saying "the role of the CJI is primal in nature because this being a topic within the judicial family, the executive cannot have an equal say in the matter.
Here, the word 'consultation' would shrink in a mini form. Should the executive have an equal role and be in divergence of many a proposal, germs of indiscipline would grow in the judiciary." For the next five years, there was confusion on the roles of the CJI and the two judges in judicial appointments and transfers. In many cases, CJIs took unilateral decisions without consulting two colleagues. Besides, the President became only an approver.
The third judgment was pronounced in the year 1998 when President K.R. Narayanan issued a presidential reference to the Supreme Court as to what the term "consultation" really means in Articles 124, 217 and 222 (transfer of HC judges) of the Constitution.
Legal experts point to systemic errors in the collegium system of appointment of judges. Some of them point out that there is immense administrative burden of appointing and transferring judges without a separate secretariat or intelligence-gathering mechanism dedicated to collection of and checking personal and professional backgrounds of prospective appointees. The system is also criticised for being a closed-door affair without a formal and transparent mechanism.
A National Judicial Commission (NJC) is a suggested alternative to the collegium system. The Constitution (98th Amendment) Bill was introduced in the Lok Sabha by the NDA government in 2003. It provided for the constitution of an NJC to be chaired by the CJI and with two of the seniormost judges of the Supreme Court as its members. The Union law minister would be a member along with an eminent citizen to be nominated by the President in consultation with the Prime Minister. The Commission would decide the appointment and transfer of judges and probe cases of misconduct by judges, including those from the highest judiciary.


text: In 2010, Matchbox introduced a special model in 2010's regular package with a '08 Ford E-350 Ambulance as a promotion model. They are found at meijer stores, and the car is in white with a white box and Green / Gold Tampos and Life EMS. It came with a Zamac Base, as usual, and came with disc hubs. This is the only vehicle released for the Meijer Promotion.


text:
Latest Jobs
Data entry clerkFreelance, Any location
We are looking fo someone who capable of entry data We are looking for a person who is qualified for data entry clerk, using a computer. The candidate will be able to find datas, reproduce them and range them ... Read More
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We are looking for a PHP developer who is passionate, like to learn, knows Wordpress environment. The candidate will have to work on our new project. The candidate should speak at least two of the following languages: english, french, spanish. Read More
Michael Jordan's girl is proud to be lesbian
NBA icon Michael Jordan's daughter, Jasmine Jordan announces her homosexuality. She said she's so proud to be lesbian... Read More
Serena Williams and Patrick Mouratoglou spend their holidays in Europe
After winning the 2012 US Open, Serena Williams is enjoying some holidays with her man and coach Patrick Mouratoglou. She was photographed while shopping with her friends in luxury boutiques like Gucci and Dolce & Gabbana Read More
Sirai Kong, a vixen
Sirai Kong is a vixen, a video girl known for her performances hip hop videos, scene, exotic photos for magazines. She has a different type of beauty because of her asian Read More
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Adrienne Bailon fabulous with her red bikini; she was enjoying the sun with kids and exposing her beautiful body at a Puerto Rican beach Read More
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Beyonce's young sister, Solange Knowles married Alan Ferguson. It was a All-White theme ceremony, a small one with freinds and relatives only. Beyonce, Jay-Z, Tina Knowles Read More
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Karrueche Tran is a young woman who can be called a wannabe. She has dated Chris Brown while he was withe singer Rihanna. She is an aspiring model who has a cover for the Rolling Stone Magazine revealing her feminine jewels ... Read More
Angela Simmons was glowing in Nigeria
Fashionista Angela Simmons was in Nigeria. She was wearing an African dress when she attended an event. She made box braids for Thanksgiving ... Read More
Kevin Hart, Laura Govan, Draya Michele and more at the Wedding Ring premiere
Kevin Hart and his fiancee, Laura Govan and her sister, Draya Michele and more attended the Wedding Ring movie premiere. They were all well dressed ont the red carpet. Everyone was laughing at the cameras when Laura Govan's ... Read More
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Rapper Nicki Minaj was for the party she was hosting in las Vegas. She was celebrating the Memorial Day. Nicki Minaj is preparing her future album the The Pink Print ... Read More


text:
Decision makers
- The 112th Congress of The United States of America
Ban of all automatic & semiautomatic guns, rifles and assault weapons.
There is thought to be an approximate 250 million guns in the US civilian population right now and as the violence in this country increases, many people think that if they buy more guns, they will somehow be safer. This is a fallacy. A society where every Tom, Dick and Harry is wielding an assault weapon for self protection is not a safe, nor a very sane one.... While people will always find a way to acquire guns, that is not a reason to throw up our hands and say, "why bother with government regulation and control?" Sometimes a nation needs to re-interpret its laws and constitution in accordance with a societies greater good. In our democracy, it's all about 'individual choice'. But when a nation is suffering from a gun problem so pervasive that it has become a national security and public health problem, individual choice no longer takes precedence. Banning all guns? ... No, that's not what this petition is about. Specifically, ... I'm talking about banning the purchase of automatic and semiautomatic guns, rifles and assault weapons within the civilian population, the likes of which can easily be attained at gun trade shows and your local handgun wielding hardware store. Increasingly, it's these very weapons that violently kill innocent law abiding men, women and children. They float around, are traded and sold .... stolen, trafficked and traded again with no record of ownership and or registration. Automatic and semiautomatic handguns, rifles and assault weapons that fire many rounds in a minute do not do much more to enhance self-defense than an ordinary pistol or rifle. Such weapons are also not much use for hunting and gaming. They are however very good for killing lots of people very fast.... Let's ban the purchase of automatic and semiautomatic handguns, rifles and assault weapons within our civilian population now! Let's do this for those who have suffered such violent deaths in the presence of these weapons.
- The 112th Congress of The United States of America
Ban of all automatic & semiautomatic guns, rifles and assault weapons now.
JoAnn Yurchesyn started this petition with a single signature, and now has 58 supporters. Start a petition today to change something you care about.
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text:
HTC has revealed a new mobile phone based on the forthcoming data transfer protocol called WiMAX. Called the HTC MAX 4G, the new handset currently has a distinctly Russian flavour because the company has otimised the new phone with Scartel for the Russian market. In fact, the mobile will be released into that market on 26th November.
A launch into the UK is surely also on the table, however. When that might occur is uncertain at this time. However, you can access a video of the new mobile phone for yourself via the link at the bottom of this story. The video spans a generous four and a half minutes in length and details a comparison between the new HTC MAX 4G and the popular Apple iPhone.
Be warned, however, the the video is in Russian so this fact will reduce the effectiveness of the comparison just a tad. However, you will still be able to extract a flavour of the new HTC handset at least.
Click here to see the video


text:
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Lee Women's Black Ivy Tee - Black
$24.99 for this stylish wardrobe essential - get it now!
Available in women's sizing - see size chart for correct fit!
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- Rolled sleeves
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- All over ivy print
- Cropped length
- Materials: 100% cotton
- Machine washable
- Our model wears a size 8 and is 188cm tall
- Country of origin: China


text:
I made a plaster mold of a piece and cast it in a low fire white pouring slip. The pieces were handpainted using Amaco Velvets in the greenware stage. Fired to cone 05, applied a clear glaze and fired to cone 05 for the glaze firing. I made four pieces. Three turned out without any issues. The one had some parts of the paint flake off when I picked it up. I have never had this happened to me and have worked with molds and casting. Any suggestions as to what happened and why only the one had some areas that flaked off? Thanks in advance for any advise! 101.JPG 2.85MB 27 downloads
PanamaxMember Since 17 Apr 2010
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Ceramics and all art forms.
Contact Information
- Website URL http://www.marie-elena.com


text: BAG Collection Agency, offer a full range of collections services to meet the specific needs of any client, we collect all types of debt both Consumers and Commercials accounts. We handle all of our accounts with respect and integrity. We are also a full-service notory provider and offer legal support.