1. Field of Invention
The present invention relates to color dividing optical device. More particularly, the present invention relates to optical device both of beam splitting and color dividing.
2. Description of Related Art
Digital image is formed from an array of pixels, so as to display an image. Further, for the color image, each pixel is, for example, composed by three sub-pixels of three primary colors in red (R), green (G), and blue (B), so as to express the color shading. Usually, in order to produce the sub-pixels of RGB. Each sub-pixel needs to be implemented with color filter with respect to the desired color. In this manner, the effective portion of the incident light source in white is reduced to ⅓ and the other portion of non-intended colors is absorbed.
For the application in flat displaying apparatus, the back light of the TFT-LCD needs to associate with the gray level modulation of liquid crystal and the color filter, so as to display the color shading of an image. The image sensor in a digital camera, such CCD or CMOS device, also needs the dye-type color filter to display the full-color image of an object. The other applications includes, for example, projector, color video, back-projection TV, . . . , and so on. No matter of single plate, three-plate (TFT-LCD, LCOS) or two-plate prism, all they need to uses the dye-type color filter to express the full color of image. When the system uses the conventional dye-type color filter, each pixel in the color filter can only display single color of the three primary colors R, G, and B. Basically, about ⅔ of the incident white light is absorbed and the light usage rate is reduced. In addition, the battery duration in use for the potable electronic apparatus is also reduced. Further, the fabrication process for the dye-type color filter is rather complicate and the material consumption. The cost for the product and the fabrication factor still remains high.
Some technologies in the art have been proposed for solving the issues, respectively. However, the advantages and disadvantages are both exiting. Therefore, further technologies are still under development.


INTRODUCTION {#s1}
============

Apoptosis is a complex process resulting in the regulated destruction of a cell, which plays a major role in normal development, cellular response to injury and carcinogenesis \[[@R1]\]. It has been suggested that apoptosis is one of the major mechanisms of cell death in response to cancer therapy \[[@R2]\]. The Bcl-2 family of proteins are among the most studied molecules in the apoptotic pathway. In this gene family, some are apoptosis inducers, including, bax, bak, bcl-Xs, bad, bid, bik and hrk, and others, such as bcl-2, bcl-XL, bcl-w, bfl-1, brag-1, Mcl-1 and A1 are apoptosis suppressors \[[@R3]\]. Bcl-2 or bcl-xL levels are elevated in a broad range of human cancers, indicating that these molecules may have a role in raising the apoptotic threshold in a broad spectrum of cancerous disorders. Expression of Bcl-xL in the NCI 60 cell lines strongly correlated with resistance to most chemotherapy agents \[[@R4]\]. It has been suggested that a decrease in Bcl-xL levels or the inhibition of Bcl-xL activity might provoke apoptosis or at least sensitise cells to apoptotic death. In the absence of a clearly defined biochemical mechanism of action or activity for this family of cell-death regulatory proteins (for which conventional inhibitors could therefore be developed), gene therapy and antisense approaches have become a reasonable alternative. For example, antisense oligonucleotides to bcl-xL have been shown to be active in down-regulation of the bcl-xL expression, leading to an increased chemosensitivity in a range of cancer cells \[[@R5]\].

An ideal oligonucleotide-based gene inactivation agent targeting RNA would combine the self-sufficient RNA digestion capability, such as "hammerhead" and "hairpin" ribozymes, with the relative biological resilience of the antisense oligodeoxynucleotides (ODN). Although DNA molecules with RNA cleavage activity have not been observed in nature, some have been derived as a result of an artificial evolutionary system known as *in vitro* selection \[[@R6]-[@R9]\]. In an *in vitro* selection system, DNA liberated from its complementary strand is free to explore a full range of structural possibilities, some of which have been found to be capable of catalytic activity, including site specific RNA cleavage and ligation \[[@R10], [@R11]\]. The 10-23 DNA enzyme or DNAzyme was named from its origin as the 23rd clone characterised from the 10th cycle of *in vitro* selection \[[@R10]\]. This enzyme has a number of features, which endow it with tremendous potential for applications both *in vitro* and *in vivo*. These include its ability to cleave almost any RNA sequence with high specificity provided it contains a purine-pyrimidine dinucleotide. This can be accomplished at very high kinetic efficiency with rates approaching and even exceeding those of other nucleic acid and protein endoribonucleases \[[@R10]\]. The ability of the 10-23 DNAzyme to specifically cleave RNA with high efficiency under simulated physiological conditions has fuelled expectation that this agent may have useful biological application in a gene inactivation strategy \[[@R12]-[@R16]\]. Recent studies showed promising clinical efficacy of the DNAzymes targeting c-jun and EBV-encoded oncogene for treating various cancers \[[@R17], [@R18]\], which demonstrated the feasibility to use DNAzymes as therapeutics.

Here, we identified an active DNAzyme against the bcl-xL gene by a multiplex selection. This DNAzyme was shown to effectively down-regulate the Bcl-xL expression in a range of cancer cells, thus overcome the anti-apoptotic block and sensitized the cancer cells to chemotherapy in xenograft animal models.

RESULTS {#s2}
=======

Identification of Cleavable sites in bcl-xL mRNA {#s2_1}
------------------------------------------------

DNAzymes were designed based on "10-23" model (Fig [1A](#F1){ref-type="fig"}). A bcl-xL cDNA clone (926 bp) was produced from the RNA sample of PC3 cells and used for screening the potential cleavable sites of DNAzymes. By bioinformatics analysis, 81 potential AU or GU sites were found and subjected to analyses on the thermodynamic stability of the enzyme-substrate heteroduplex as predicted by the hybridisation free energy \[[@R19]\]. DNAzymes with the greatest heteroduplex stability indicated by a low free energy of hybridisation (calculated using the nearest neighbour method), was often found to have the greatest kinetic activity. As a result, 26 DNAzymes with Δ G°\<- 16 kcal/mol were chosen for the further multiplex cleavage assay ([sTable 1](#SD1){ref-type="supplementary-material"}).

In order to select active DNAzymes, *in vitro* selection was performed using a multiplex method, which enables a pool of DNAzymes to be screened for their ability to access and cleave RNA substrate under simulated physiological conditions \[[@R20]\]. Based on the bioinformatics and physiochemical analyses as shown in sTable [1](#T1){ref-type="table"}, twenty-six DNAzymes (0nM, 5nM, 50nM and 500nM) and RNA substrate (400nM) were incubated for cleavage reaction and primer extension was then performed with Superscript II reverse transcriptase to define the cleavage sites within the bcl-xL transcript. The sequencing ladders were used as a guide to attribute cleavage bands to specific DNAzymes. The relative cleavage strength of each DNAzyme was determined by intensity of the cleavage products. A representative gel was shown in Fig [1B](#F1){ref-type="fig"}. DNAzymes were ranked according to their cleavage ability at lowest concentration (5nM). The multiplex selection resulted in 10 active DNAzymes that could efficiently cleave the bcl-xL mRNA (sTable [1](#T1){ref-type="table"}).

![Schematic description of "10-23" DNAzyme\
A, unmodified DNAzyme (PO-DNAzyme); B, phosphorothioate-modified DNAzyme (PS-DNAzyme) as indicated by \*. C, Multiplex *In vitro* selection of bcl-xL DNAzymes. Bcl-xl DNAzymes incubated with its RNA substrate for 60 minutes in the presence of 10 mM Mg^2+^ at 37^o^C. Primer extension was performed using the sequence-specific primers along the bcl-xl mRNA. The reactions were analysed alongside with DNA sequencing on a polyacrylamide gel. A representative gel is presented. V, molecular markers; C/A/T/G, sequencing ladders; 0/5/50/500 nM, a mixture of DNAzymes at the defined concentrations.](oncotarget-05-9039-g001){#F1}

Chemical modification of DNAzymes {#s2_2}
---------------------------------

To increase DNAzyme stability in cells, 1-5 phosphorothioate (PS) linkages were incorporated into each of the arms in DNAzyme (PS-Dz, DT882). The DNAzymes\' stability in human serum was significantly improved with the increase of number of PS linkages (Fig [2A](#F2){ref-type="fig"}). To examine if the modification had any impact on DNAzyme\'s catalytic activity, we further performed a single turnover kinetic analysis of the modified DNAzyme. As shown in Fig [2B](#F2){ref-type="fig"}, the PS modification had decreased the cleavage efficiency (*k*~obs~) of the modified DNAzymes, compared with the unmodified one. To strike a balance between the stability and catalytic activity, we selected the 3+3 PS modification scheme for the further cell and *in vivo* studies.

![Effect of phosphorothioate-modification on DNAzyme stability in human serum and cleavage kinetics\
A: the modified DNAzymes were incubated with the serum, extracted at the defined time points, and ^32^P-labelled. Intact DNAzyme (%) was presented as a ratio of the band intensity at different time points to the band intensity at the 0 time point. B: the kinetic efficiency was measured under a single turnover condition and *k*~obs~ was used as an indicator for cleavage activity.](oncotarget-05-9039-g002){#F2}

Effect of bcl-xL DNAzymes on bcl-xL expression {#s2_3}
----------------------------------------------

Using the modified version of 16 DNAzymes derived from the multiplex selection, we performed further testing for their ability to down regulate the bcl-xL expression in cells. The assays were performed in PC3 cells (a prostate cancer cell line). After screening all 16 DNAzymes, three DNAzymes (DT 882, DT883, and DT884) exhibited a very strong inhibitory effect on bcl-xL protein expression as shown in a representative gel (Figure [3A](#F3){ref-type="fig"}). An antisense control (ctggatccaaggctctaggt) and a negative control (scrambled arms of DT882, tggtgtgtgggctagctacaacgagttaataaa) were used in the screening. Based on the results of *in vitro* cell screening, we next selected one of the most active DNAzyme DT882 for further validation of its activity of down-regulation of bcl-xL expression in a panel of cancer cell lines (PC3, prostate cancer; T24, bladder cancer; A549, lung carcinoma; CNE-1, nasopharyngeal carcinoma; HCT116, colon cancer). Figures [3B](#F3){ref-type="fig"} showed that anti-bcl-xL DNAzyme DT882 reduced the level of the bcl-xL gene expression in the cell lines tested, which indicated that DT882 could be effectively transfected into the cells, found its target mRNA and inhibited bcl-xL expression in cells.

![Inhibition of bcl-xL protein expression by bcl-xL DNAzymes\
A. Screening of the selected DNAzymes (2 μM) using Western blotting was performed in PC3 cells and a representative blot was presented. Bcl-xL antisense or a scrambled control were used as controls. B. Validation of inhibitory activity of Bcl-xL DNAzyme (DT882) in different cancer cells was performed in PC3 (prostate cancer), T24 (bladder cancer), A549 (lung cancer), CNE-1 (nasopharyngeal carcinoma) and HCT116 (colon cancer) cells. TMP, transfection reagent; Control, scrambled DT882.](oncotarget-05-9039-g003){#F3}

Induction of apoptosis by Bcl-xL specific DNAzyme {#s2_4}
-------------------------------------------------

Having shown the DT882 inhibition of bcl-xL expression in cancer cells, its effect on apoptosis of PC3 cells was further examined. In the assay, same transfection procedure as in Western blotting assay was used, except those cells were subject to Annexin-V staining after the overnight incubation with the DNAzyme. Fig [4A](#F4){ref-type="fig"} clearly showed that there was a substantial increase in apoptosis population in the DNAzyme treated cells (23.17%), as compared with the control-treated (an inactivated DNAzyme with scrambled arm sequences of DT882), indicating that the cells treated with the bcl-xL DNAzyme was provoked to undergo apoptosis. To determine the effect of bcl-xL DNAzyme on the early events of apoptosis caused by down-regulation of bcl-xL, the measurement of Cytochrome C release from the cells was performed. As shown in Figure [4B](#F4){ref-type="fig"}, DNAzyme-mediated reduction of bcl-xL in PC3 cells led to an increased release of Cytochrome C. This result not only confirmed the data from FACS analysis, but also validated the specificity of the DNAzyme in induction of apoptosis via mitochondria pathway in PC3 cells.

Chemosensitization of Cancer Cells with anti-bcl-xL DNAzyme {#s2_5}
-----------------------------------------------------------

The Bcl-xL protein has been shown in a number of cell lines as a potent protector of cellular apoptosis induced by anti-neoplastic agents. To examine if DT882 could sensitise cancer cells to the effect of cytotoxic chemotherapy by down-regulating bcl-xL expression, cell survival was measured using MTS assays in a panel of cancer cells treated with either the DNAzyme alone or DNAzyme plus anti-cancer agent Taxol. The results showed that the anti-bcl-xL DNAzyme DT882 sensitised all the cancer cells to Taxol treatment (Table [1](#T1){ref-type="table"}). This sensitization led to about 150\~260% increase of cell death in a range of cancer cell lines treated with DT882 plus Taxol, compared with that treated with Taxol alone. To examine if DT882 could reverse the Taxol-resistance, a Taxol-resistant cell line CNE2R (nasopharyngeal carcinoma) were assayed for possible drug resistance reversal. As shown in Figure [4C](#F4){ref-type="fig"}, while the Taxol treatment in a range of concentrations did not cause cell death of the resistant cells or the control-treated resistant cells, DT882 treated resistant cells showed a significant reduction in cell survival, indicating that down-regulation of the bcl-xL in the Taxol-resistant cells could restore the cellular response to the chemotherapeutics.

###### Chemosensitization of cancer cells to Taxol by bcl-xL DNAzyme DT882

  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Cancer Cells   Cell Death (%)[\*](#tfn_001){ref-type="fn"}   Chemosensitization\
                                                               Index (%) [\#](#tfn_002){ref-type="fn"}
  -------------- --------------------------------------------- ----------------------------------------- ------------ ------------ ------------ ------------ ------------ -----
  PC3            1.75±0.27                                     16.33±3.10                                23.17±4.11   46.51±6.66   19.33±2.07   38.51±3.33   15.20±2.10   205

  T24            6.99±1.23                                     21.01±1.78                                31.33±3.95   56.71±6.12   22.53±2.03   41.78±4.01   18.78±2.78   202

  MDA-MB-231     3.10±0.45                                     12.09±2.11                                19.88±3.28   39.89±3.51   18.64±2.83   35.02±3.44   10.90±1.11   266

  CNE1           2.99±0.31                                     15.55±3.87                                25.76±5.12   42.06±5.42   21.54±4.01   39.55±3.81   16.66±2.65   152

  B9-58          1.44±0.64                                     13.67±2.45                                19.66±3.33   41.15±6.65   19.01±2.11   37.47±2.60   11.76±3.10   250

  HCT116         2.89±0.41                                     22.51±5.50                                24.83±2.21   57.34±7.25   24.69±4.23   41.28±3.55   21.89±2.89   162

  A549           4.67±0.87                                     19.23±2.99                                19.88±1.56   42.78±2.11   21.22±3.34   39.56±6.01   17.44±2.67   145
  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------

Cell death (%) was calculated as (untreated -- treated)/untreated.

Sensitization index was expressed as (DT882+Taxol -- Taxol) / Taxol x 100.

![Induction of apoptosis by bcl-xL DNAzyme\
PC3 cells were transfected with DT882 and control. The cells were subjected to either Annexin-V assay by FACS (A) or Western blot assay for cytochrome C (B). TMP, transfection reagent; Control, scrambled DT882 control. C, a Taxol-resistant cell line of nasopharyngeal carcinoma (CNE2R) was used for MTS assays in the presence of a series of Taxol concentrations.](oncotarget-05-9039-g004){#F4}

Chemosensitization in human tumour xenograft model by anti-bcl-xL DNAzyme DT882 {#s2_6}
-------------------------------------------------------------------------------

In order to demonstrate if DNAzyme-mediated down-regulation of the bcl-xL expression resulted in chemosensitization of tumour cells to anticancer drugs *in vivo*, a murine model with human PC3 prostate cancer xenograft was used to determine the sensitivity to the chemotherapeutic. In the experiments, seven groups of mice (8 mice per group) (Saline, DNAzyme, AS, Control, Taxol, Taxol + DNAzyme, and Taxol + AS) were employed. When tumours reached an average volume of 100-200 mm^3^, an Alzet osmotic pump, which was used as a delivery vehicle for DNAzyme oligonucleotide in tumour bearing mice, was surgically implanted in the peritoneum of the mouse via the abdominal route. As shown in Fig [5A](#F5){ref-type="fig"}, the combinational use of DT882 and Taxol markedly inhibited PC3 tumour growth compared with the groups of DNAzyme alone, Taxol alone or control plus Taxol.

To examine the *in vivo* stability of the DNAzyme and the pump delivery efficiency, the DNAzyme oligonucleotide was extracted from the tumor tissues, plasma and the remaining solution of the pumps in the mice treated with DT882 plus Taxol (day 14), and labelled with γ\--^32^P-ATP. Analysis showed that the DNAzyme was stable over the treatment period and the Alzet osmotic pump provided a consistent delivery of the DNAzyme *in vivo* (Fig [5B](#F5){ref-type="fig"}).

![Chemosensitization of PC3 human xenograph prostate cancer by anti-bcl-xL DNAzyme\
A: Nude mice bearing established, subcutaneously growing PC3 tumour xenograft either remained untreated (saline) or were treated with DT882, antisense control, Taxol or DNAzyme + Taxol and AS+ Taxol. DNAzyme DT882 or AS was delivered using an osmotic pump and Taxol was administrated via i.p. route weekly. Tumour size was measured at the time points indicated. AS, antisense control (25). B: DNAzyme (DT882) was extracted from tumor tissues, plasma and the remaining solution of the pumps, labelled and assayed on PAGE.](oncotarget-05-9039-g005){#F5}

DISCUSSION {#s3}
==========

DNAzymes have been shown to effectively suppress gene expression both *in vitro* and *in vivo* \[[@R21]\]. Limited therapeutic efficacy has been demonstrated in patients in studies targeting c-jun and EBV-LMP1 \[[@R17], [@R18]\]. In the present report, we present a comprehensive study in which an active DNAzyme targeting bcl-xL gene was identified and validated through *in vitro* selection to *in vivo* testing, showing clear experimental evidence that the Bcl-xL DNAzyme could be an effective chemo-adjuvant agent for potential clinical applications in cancer chemotherapy.

Targeting a DNAzyme to a particular mRNA is generally a multi-step process. In our previous study, a multiplex selection approach was devised to facilitate the selection of the accessible sites for DNAzymes within the target mRNA \[[@R20]\], but only at the *in vitro* level. Here, we successfully applied this methodology and further proved that there was a consistency between the *in vitro* cleavage activity and *in vivo* efficacy of the DNAzymes. Thus, our study provided a streamline approach to therapeutic uses of DNAzymes.

Despite the range of anti-cancer studies using nucleic acid-based agents, one common factor necessary for the success of therapeutic uses of those agents is the ability to delivery of the agents in a precise and dependable manner. The antitumor effects of the therapeutic agents are influenced by schedule of drug administration. In majority of the studies testing anti-cancer oligonucleotides, intra-tumoral injection has been the method of the choice to deliver oligonucleotides \[[@R22]\], which was somewhat invasive and less efficient. In this study, we surgically implanted the ALZET osmotic pump adjacent to the tumor, permitting continuous, controlled dosing, thus enabling us to achieve steady state conditions and accurate delivery of the DNAzyme over prolonged periods of time and to show a significant effect of chemosensitization by the DNAzyme to Taxol treatment.

To date, there have been substantial amount of preclinical and clinical studies in search for the inhibitors of Bcl-2 family, among which one Bcl-2 antisense and three small molecule inhibitors are being tested in clinical trials \[[@R23], [@R24]\]. However, it has been well recognized that the agents with high specificity tend to induce cancer drug resistance, while broader acting agents could cause unwanted toxicity. Considering that the anti-apoptotic Bcl-2 family is a large and redundant family of proteins, some of which have been shown to contribute to drug resistance in cancer cells \[[@R25], [@R26]\], inhibition of multiple Bcl-2 family members would be necessary in order to optimize the therapeutic effect. Importantly, previous studies showed that paclitaxel (Taxol) causes phosphorylation and inactivation of Bcl-2 \[[@R31]-[@R34]\]. Phosphorylation of Bcl-2 takes place only during mitotic arrest and the duration of the arrest is important for cell killing by paclitaxel/Taxol \[[@R35], [@R36]\]. However, Bcl-2 can be substituted by Bcl-xL \[[@R37]\], which underscores the importance to decrease Bcl-xL levels in Taxol-treated cells. This decrease actually increases sensitivity to Taxol as now shown by current work. Thus, our strategy further confirm that one anti-apoptotic protein can be substituted by the other, providing evidence that the Taxol resistance can be overcome by targeting Bcl-xL.

In conclusion, we present a novel agent that catalytically cleave the Bcl-xL mRNA, one of the important members of Bcl-2 family, thus add another candidate of inhibitors of anti-apoptotic Bcl-2 family with new mechanisms for potential multimodality treatment of cancers.

MATERIALS AND METHODS {#s4}
=====================

Cell cultures {#s4_1}
-------------

PC3 (prostate adenocarcinoma), T24 (bladder carcinoma), MDA-MB-231 (Breast cancer), A549 (lung carcinoma), B9-58 (lymphoma) and HCT116 (colon carcinoma) human cancer cell lines were obtained from the American Type Culture Collection. CNE1 (nasopharyngeal carcinoma) is the EB virus negative low differentiated nasopharyngeal squamous carcinoma cell line \[[@R27]\]. All cell lines were maintained according to vendors\' recommendations.

DNAzyme synthesis {#s4_2}
-----------------

All the oligonucleotides were made by TriLink BioTechnologies (San Diego, CA) and purified by gel electrophoresis for *in vitro* studies and by HPLC for cell-based assays and *in vivo* studies.

Design and thermodynamic analysis of DNAzymes {#s4_3}
---------------------------------------------

DNAzyme sequences for each target are assembled using the 10-23 catalytic motif \[ggctagctacaacga\] and hybridising arms specific for each site along the target RNA transcript (Fig. [1A](#F1){ref-type="fig"}). The length of each arm is usually fixed at 9 bases; however, these can be shortened or lengthened depending on their individual predicted hybridisation free energy. Each DNAzyme oligonucleotide is designed to target purine-uracil (RU). In most cases we ignore purine-cytosine sites as in our experience they are less reactive than RU sites, particularly AC junctions \[[@R28]\]. The potential DNAzymes were subjected to thermodynamic analyses (hybridization free energy and Tm) \[[@R19]\].

Multiplex selection of active DNAzymes *in vitro* {#s4_4}
-------------------------------------------------

DNAzyme target site selection was described previously \[[@R20]\]. Briefly, linear template DNA for the production of the bcl-xL transcript was prepared by PCR in a mixture containing 10 pM cDNA, 1 μM of the forward T7 phage promoter primer \[TAATACGACTCACTATAGGGAGA-AAGATTCTGAAGGGAGAG\] and 1 μM of the reverse primer \[CGGGTTTCTCCTGGTGGCAATG\]. The reaction was carried out over 25 temperature cycles at 95oC for 30 s, 60oC for 90 s, and 72oC for 60 s in a solution containing 16.6 mM (NH4)2SO4, 67 mM Tris-HCl (pH 8.8), 6.7 mM MgCl2, 0.87 units of Ampli*Taq* DNA polymerase (Perkin Elmer) and 300 μM each of dGTP, dATP, dTTP, dCTP. PCR products were electrophoresed in a 1% agarose gel alongside appropriate size/ mass standards and visualised by ethidium fluorescence. The illuminated amplicon bands were then excised from the agarose slab and extracted using Gene Clean (Bio 101). Template DNA was then redissolved in diethyl pyrocarbonate (DEPC) treated RNase free water and the concentration adjusted to approximately 100ng.μl^−1^.

For RNA transcription, reaction was performed with 100 ng of purified template DNA and T7 RNA polymerase at 37oC for 3 h using an RNA transcription kit (Epicentre). After the 3 hour incubation the DNA template was degraded by the addition of 1 MBU of RNase-free DNase I and further 15 min incubation at 37^o^C. This reaction was then terminated by adding an equal volume of phenol/ chloroform/ isoamyl alcohol. The RNA contained in the aqueous phase could be used directly in the multiplex cleavage reaction or precipitated in 0.3M sodium acetate and two volumes of absolute ethanol.

Multiple DNAzyme oligonucleotides (0.5 nM-0.5 μM) and synthetic RNA substrate (0.2 μM) were pre-equilibrated separately for 10 min at 37oC in equal volumes of 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 150 mM NaCl and 0.01 % SDS. The reaction was then initiated by mixing the DNAzymes and substrate together. After an hour the reactions were stopped by emersion in ice and precipitation in 0.3 M sodium acetate and two volumes of ethanol.

Primer extension analysis was then performed on DNAzyme cleaved RNA using SuperScriptII reverse transcriptase (Life Technologies). Two primers were designed for the extension reactions along the bcl-xL RNA \[PE1: GTTTCTCCTGGTGGCAATG; PE2: CCATCCCGGAAGAGTTCA\]. In each reaction 2 pmol of ^32^P-labelled primer was combined with 400 nM of RNA and denatured at 90oC for 5 min. For extension with Superscript reverse transcriptase, the primer was allowed to anneal slowly between 65oC and 45oC before adding the 1st strand buffer, dithiothreitol, deoxyribonucleotides and enzyme (according to the manufacturer\'s instructions). This mixture (final volume 20 μl) was then incubated for 1h at 45oC before being transferred to ice.

Sequencing fragments corresponding to each segment of the target were also generated by primer extension on the double stranded DNA template in the presence of chain terminating dideoxynucleotides (ddNTP). In these four reactions the dNTP concentration was reduced to 2.5 μM while being supplemented by either 10 μM ddGTP or 100 μM ddATP or 200 μM ddTTP or 100 μM ddCTP in 16.6 mM(NH4)2SO4, 67 mM Tris-HCl (pH 8.8), 6.7 mM MgCl2, 0.87 units of *Taq* DNA polymerase and 1.0 pmol of the 32P-labelled primer. This reaction was performed as a linear amplification over 25 temperature cycles at 95o C for 30 s, 60° C for 90 s, and 72° C for 60 s.

After primer extension, samples were combined with an equal volume of stop buffer (formamide/EDTA/loading dye) before electrophoresis on a 6% denaturing polyacrylamide gel. The corresponding image could then be revealed and the band intensity quantified at each position with Phosphorimager and ImageQuant software (Molecular Dynamics).

DNAzyme stability assay and kinetic analysis {#s4_5}
--------------------------------------------

The DNAzyme (DT882) was modified with 1, 3 or 5 phosphorothioate (PS) linkages respectively. Unmodified and PS-modified DNAzymes were incubated at a final concentration of 10 μM in 100% human AB serum (Sigma, St. Louis, MO) at 37°C. Aliquots (10 μl) of the incubation mixture were removed at 0, 2, 8, 24 and 48 hours and diluted in 290 μl of 10 mM Tris/1 mM EDTA, pH 8.0, buffer (TE buffer). The diluted DNAzyme was extracted by sequentially adding 150 μl phenol and 150 μl chloroform with vortexing. Samples were centrifuged at 11,300*g* for 10 minutes, and 100 μl supernatant was collected. Aliquots of the aqueous supernatant were labeled with ^32^P and electrophoresed on a 16% polyacrylamide gel. Labeled DNAzyme bands were quantified using Molecular Dynamics ImageQuant software (Sunnyvale, CA), and the recovered DNAzyme was expressed as a percentage of the DNAzyme diluted directly into TE buffer.

The *in vitro* activity of DNAzymes was determined by measuring the rate of RNA cleavage under single turnover condition. The ^32^P-labelled RNA transcripts to be used in the cleavage reaction were produced utilising the RNA transcription kit (Epicentre). The transcripts were electrophoresed on a 16% denaturing PAGE and bands were excised and gel purified. The observed constant (k~obs~) is the rate of cleavage and it was calculated by regression analysis of the progress curve using the following equation:

\[P\] = \[P\]∞(1 - e^−1kobst^)

where \[P\] is the cleavage yield, \[P∞\] is the final cleavage yield, and t is the reaction time \[[@R29]\]. Cleavage reactions were performed over multiple time points.

DNAzyme transfections {#s4_6}
---------------------

To facilitate delivery of DNAzyme oligonucleotides into cells, a cationic porphyrin, tetra meso (4-methylpyridyl) porphyrin (TMP), was used as a transfection reagent for intracellular delivery \[[@R30]\]. Briefly, 0.8 × 10^5^ cells were seeded in a 60-mm culture dish and incubated at 37^o^C, 5% CO~2~ overnight. DNAzyme and control molecules were transfected into cells with TMP at a charge ratio of 3. The final concentration of DNAzyme in the transfection mix was 2 μM in 2 ml of medium without serum. After 4 hours, the cells were replaced with fresh culture medium. Twenty-four hours after transfection, the cells were harvested for Western blot and cell biology analyses.

Western analysis {#s4_7}
----------------

Protein was extracted from the transfected cells in RIPA buffer (50 mM Tris-HCl, pH 8, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% sodium deoxycholate, 0.57 mM PMSF and 1 μg/ml aprotinin). Protein samples (40 μg) were run on 12% SDS-PAGE gels (BioRad), transferred to nylon membranes and immunoblotted with the following antibodies: anti Bcl-xL mouse monoclonal antibody (H-5, Santa Cruz), anti-Cytochrome C rabbit polyclonal antibody (H-104, Santa Cruz) and anti-β-actin monoclonal antibody (AC-74, Sigma). Binding of primary antibodies was detected with goat anti-rabbit or anti-mouse horse radish peroxidase conjugated secondary antibodies (Santa Cruz). This binding was visualized with the ECL western blotting detection reagents (Amersham Pharmacia Biotech, England) on exposure to chemiluminescent film.

Apoptosis and cell survival assays {#s4_8}
----------------------------------

To determine apoptosis induced by DNAzymes, Annexin-V-FUOS Kit was used to detect apoptotic cells by FACS, according to the manufacturer\'s instruction (Roche). Apoptotic cells were quantified using a FACScan flow cytometer (Becton--Dickinson).

Chemosensitization effect of DNAzymes on cancer cells was examined using MTS assay. Cancer cells were treated with DNAzyme/TMP complex for 4 hours. The medium was then replaced with fresh DMEM containing 10% FBS and 100 nM Taxol, and further incubated for 72 hours. MTS assays were performed for cell viability of all the samples.

Detection of cytosolic Cytochrome C expression in PC3 cells {#s4_9}
-----------------------------------------------------------

Transfected cells were harvested by centrifugation at 200 *g* for 5 minutes at 4° C. The cell pellets were washed once with ice-cold phosphate-buffered saline (PBS) and resuspended with 5 volume of buffer containing 20 mM HEPES-KOH, pH 7.5; 10 mM KCl; 1.5 mM MgCl2; 1 mM sodium EDTA; 0.1% SDS; 1 mM dithiothreitol; and 0.1 mM phenylmethyl sulfonyl fluoride containing 250 mM sucrose. The cells were homogenized with a 22-gauge needle, and the homogenates were centrifuged at 1000*g* for 10 minutes at 4°C to remove nuclei, unbroken cells, and large membrane fragments. The supernatant was centrifuged once more at 10,000*g* at 4 °C for 30 min. The supernatant cytosolic fraction was collected, quantitated and used in Western analysis.

*In vivo* mouse model and DNAzyme treatment {#s4_10}
-------------------------------------------

Four-week-old female Balb/C nude mice were obtained from the Animal Resources Center (Perth, Australia) and were housed and maintained in compliance with the National Health and Medical Research Council Statement on Animal Experimentation and the requirements of the New South Wales State legislation. Four groups of mice (8 mice per group) (Saline, DNAzyme, Taxol, Taxol + DNAzyme) were employed. At day 1: acclimatised nude male Balb/C athymic mice were injected with 1x 10^6^ PC3 cells suspended in 0.1 ml Matrigel in the right hind leg under methoxyfluorane anesthesia. Tumour growth was measured twice weekly using digital callipers and tumour volume is calculated using the (l \* w \* h\* π/6) formula. When tumours reached an average volume of 100-200 mm^3^, an Alzet osmotic pump (BioScientific), which was used as a delivery vehicle for DNAzyme oligonucleotide in tumour bearing mice, was surgically implanted in the peritoneum of the mouse via the abdominal route. The Alzet model1002 pump is a capsule shaped pump (1.5 × 0.6 cm) and delivered a total volume of 0.5 ml at a rate of 0.25 μl/hr over a period of 14 days. The pump was filled with a saline solution containing DNAzyme oligonucleotide, which resulted in a dose rate of 12.5 mg/kg/d. Some mice received sub-optimal doses of 25 mg/kg Taxol by intraperitoneal route in a 200μl injection once weekly post-surgery for the duration of the study. At the end of 14d, mice were euthanized by cervical dislocation and tumours harvested. Tumours were weighed, snap-frozen and stored at --80^o^C till analysis. To analyse the concentration of the DNAzyme, the DNAzyme oligonucleotide was extracted from tumor tissue, plasma and remaining solution in the pumps with phenol/chloroform and precipitated with ethanol. The extracted DNAzyme was labelled with γ-^32^P-ATP by T4 polynucleotide kinase and assayed on PAGE.

SUPPLEMENTARY TABLE
-------------------

This work was partially supported by the National Science Foundation China (81172188) and Specialized Research Fund for the Doctoral Program of Higher Education, Ministry of Education, China (20110162110010). Part of work was performed at formerly Johnson and Johnson Research Sydney.


If you’re looking for new and innovative ways to gain prominence at events and exhibitions, then look no further. Locus Displays’ Pop Out Outdoor Signs offer maximum publicity at a very affordable cost.
Our patent horizontal outdoor signs make your message prominent, whereas vertical outdoor signs add professionalism, and circle and triangle outdoor signs give your display an artistic touch of perfection. These unique signs are tailor-made to stand the test of time.
A high quality tension fabric embossed over a sturdy aluminum frame prevents the structure from collapsing and keeps it safe from wear and tear. Through dye sublimation, you can customize text, graphics and other designs to your liking. These double-sided print outdoor signs also offer dual sides for sublimation, allowing you to promote your brand and experiment with creativity simultaneously.

When it comes to making an impact on event attendees, you can use our tower outdoor signs. In addition to their innovative structure and artistic appeal, this popular and trendy outdoor signage is lightweight, easy to install, durable and can easily be dismantled, packed away and transported.
Velcro-fitting enables the tension fabric to adhere to the base structure. Even in strong wind, rain, snow or any other weather condition, your fabric stays in its place and does not tear easily.
For added support and stability, all our event outdoor signs are customizable with sand or water-filled weight bases. These allow your tension fabric to pop out when required and fold back easily into the base, while remaining intact and in position throughout the show.

Our outdoor signs are also compatible with flags, flagpoles and literature dispensers. This allows you to setup a multi-purpose outdoor exhibition display that combines the best of aesthetic appeal with lightweight options and improved durability.
Whether they are used for a tradeshow display, sales exhibition, job fare, educational event, recruitment center, exposition or even a simple presentation, our outdoor displays take advertising to a whole new level – with a series of budget-friendly solutions.
We offer a complete collection of customized and stock signs that sway to and fro beautifully in the breeze and advertise your brand to every passerby and potential customer. In addition, we offer a hard plastic case for storing and transporting the structure. This is offered with a premium trolley bag in which you can fold and store your banner with ease. As the summers approach, your outdoor displays will become impossible to ignore with Locus.


2014 RC

2014 RC is a sub-kilometer asteroid, classified as near-Earth object of the Apollo group. On September 7, 2014, it passed within  (0.1 LD) of the Earth. The asteroid is approximately the diameter of the Chelyabinsk meteor, and passed almost as close to Earth as 367943 Duende (2012 DA14) did in 2013.

With an absolute magnitude of 26.8, the asteroid is about  in diameter depending on the albedo. Observations by the NASA Infrared Telescope Facility conclude the asteroid is a fairly bright Sq-class asteroid which have an average albedo of around 0.24, and would give the asteroid a spherical equivalent diameter of . Measurements by multiple telescopes indicate that the asteroid rotates in 15.8 seconds making it one of the fastest rotating asteroids so far discovered. Using the 15.8 second rotation period, more accurate radar observations by Goldstone shows the asteroid has a largest axis of at least . Due to the asteroid's fast rotation, it is a monolith and not a rubble pile.

On 8 September 2115 the asteroid will pass about  from the moon. On 5 September 1973, the asteroid passed between  and  from Earth. 2014 RC was removed from the JPL Sentry Risk Table on 5 September 2014 and there are no known possible impact dates in the next 100 years.

2014 approach
It made a close approach to Earth of  (0.1 lunar distances) around 18:02 UTC on 7 September 2014. The asteroid briefly brightened to about apparent magnitude 11.5, but it was still not visible to the naked eye or common binoculars. At the peak brightness the asteroid had a declination of –47, and was most easily visible over New Zealand. During 2014, asteroids 2014 AA and  have come closer to Earth.

The Managua explosion on 6 September 2014 may or may not have been created by a bolide that was missed by millions of people, but either way it was not caused by the close approach of 2014 RC.

Orbital shift

During the 2014 Earth close approach the orbital period of 2014 RC was reduced from 600 days to 549 days. The orbital eccentricity decreased while the orbital inclination increased.

Close-approach table

See also
List of fast rotators (minor planets)

References

External links
 ASTEROID 2014 RC Tracking
 Close Approach of Asteroid 2014 RC (Remanzacco Observatory)
 Small Asteroid Will Pass Earth Closely but Safely on Sunday (Phil Plait)

#
Category:Minor planet object articles (unnumbered)
#
#
#
20140907
20140907
20140901


Muller, who won the 2010 Golden Boot in South Africa, has incredibly scored more goals at World Cups than Diego Maradona with 8 goals in 7 World Cup games. Perhaps more startling is that fact that Muller’s 8 goals have come from just nine recorded shots on target.

Looking at Muller’s stats in another way, at 24[years-young the Bayern attacking midfielder has more World cup goals than Lionel Messi, Cristiano Ronaldo, Wayne Rooney and Zlatan Ibrahimovic combined, while his goals per game rate in the World Cup of 1.14 is superior to that of Gerd Muller (1.08).

Both striker also cashed in with braces in their matchday one of the competition, and with qualification for both nations looking likely, the first-named centre-forwards for the Oranje and Les Bleus will also fancy their chances of adding more goals in Brazil.


Article content continued

Since the aircraft belonged to Malaysian Airlines and the incident is presumed to have started in Malaysian airspace, the lead nation in the investigation is Malaysia. In my opinion, this is the Achilles heel of the inquiry. The majority owner of Malaysian Airlines Systems (MAS) is PMB, a Malaysian government holding company. MAS has clocked up net losses of $1.3 billion in the past three years. This is a clear conflict of interest, which has resulted, intentionally or otherwise, in a bungled investigation. If the bungling is intentional, then might it have something to do with the cargo that MH370 was carrying? (More of which later.) Until this matter is resolved, the disappearance will continue to be surrounded by conspiracy theories. For me, the answer is clear: the one party benefiting from the continuing state of confusion surrounding MH370 is Malaysia.

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The disappearance of this twin-engine wide-body airliner is without parallel in modern aviation, a mystery replete with questions. But what is certain is that something fast and furious occurred on that aircraft as it flew over the South China Sea.

There is always the possibility of design flaw in anything mechanical, and there is an established procedure by which aircraft manufacturers and regulators handle these issues. Service Bulletins (SBs) issued by manufacturers, and Airworthiness Directives (ADs) issued by regulators keep the industry informed. The Boeing 777 has had its share of such notices, and there are two in particular that are relevant to MH370 – one involving a short-circuit in the hose feeding emergency oxygen to the crew, and one warning of possible rupturing of the aircraft pressure vessel due to the mounting of a satellite communications antenna.


Flat White Vs Latte: What Is the Difference?

So you love coffee. You like it white and creamy. You’ve tried cappuccinos, but you’re not keen on the foam mustache. You, my friend, are a dedicated latte lover.

But then along comes this other coffee. The flat white. You’ve heard people rave about it, but you’re not sure what it is.

Should you abandon your latte and give it a try?

Before you decide on flat white vs latte, take a look at our explanation of the differences between the two. Covering everything from strength to flavor, we’ll give you the information you need to make your choice.

The origin stories

As you might expect, the latte has a longer history than the flat white.

It’s believed to have been invented in the US in the 1950s. The Caffe Mediterraneum in Berkeley, California, lays claim to its creation. The story goes that one of the café owners, Lino Meiorin, adapted the latte from a traditional Italian cappuccino.

Lino, who was himself Italian, saw that many customers found the cappuccino too strong. He’d therefore instruct the barista to add more “latte” – milk, in Italian – to the cup. After a while, his variation became so popular that he added it to the menu as a new drink. The caffe latte was born (1).

At first, the latte was served in a bowl. That was soon replaced with a pint glass, and later with the wide rimmed cup we’re familiar with today. It grew in popularity in the 1980s, thanks to Seattle’s coffee houses, and today it’s one of America’s favorite coffees (2).

It wasn’t until around 30 years later that the flat white came onto the scene. The question of who invented it is the subject of heated debate. There are three contenders for the honor: Australian Alan Preston and two New Zealanders, Derek Townsend and Fraser McInnes (3).

Preston claims that white coffees served “flat” were popular in Queensland in the 1960s and ’70s. When he moved to Sydney, he says, he opened his own coffee house and served the flat white there from 1985.

Townsend claims to have served the first flat white in his café in Auckland a year later. He admits to having taken the name – though not the recipe – from a café in Melbourne, Australia.

Finally, McInnes says that he invented the coffee by accident in 1989 in Wellington. His story was that the milk in the cappuccino he was making was low-fat and hadn’t created much foam. When he presented it to the customer, he apologized for it being a “flat white” (4).

When Starbucks added the flat white to its American coffee menu in 2015, the accompanying note said it had originated in Australia. You can imagine the furore that followed!

Flat white vs latte: Size matters

If you look at a flat white and a latte next to each other there’s one difference you’ll notice straight away – the size.

Traditionally a flat white is served in a 5 to 6-ounce cup. That makes it smaller than a latte (although much bigger than an espresso). In some larger coffee chains, the size is increased to 10 ounces – but beware. The difference is almost always made up with milk, which dilutes the flavor.

A latte, on the other hand, is typically prepared in an 8-ounce cup.

What about the coffee content?

For once, the amount of an espresso in the drink doesn’t make any difference. You can have either a single or double espresso shot in a latte or a flat white. Double shots are more commonly used in both, however, to create a good balance between the coffee and the creaminess of the added milk.

That’s not to say that individual coffee houses won’t use different methods. Some baristas use a ristretto for their flat whites, i.e. an espresso prepared with half the usual amount of water. Using an espresso in their lattes distinguishes more strongly between the two drinks (5).

Starbucks serve their version of a flat white with a double shot of ristretto to balance out the milk in its supersized 12-ounce drink. Those in the know, however, can go off-menu and order a “short flat white” as an 8-ounce serving (6).

The difference is in the milk

The main thing that makes the flat white experience different from a latte is the way the milk is added.

When you heat up milk, whether on the stove or with a steam wand, you’ll get three different elements. These are the liquid milk, micro-foam and froth.

Flat white

Latte

At the bottom is the hot milk in its liquid form. As it heats up it remains smooth and silky, and becomes slightly thicker than it was when it was cold.

On top of the liquid milk is what’s known as micro-foam. These are small bubbles which still have a fairly liquid consistency. The uppermost layer is the froth – big bubbles that are stiffer than micro-foam.

Creating the foam is an important part of the barista’s art. The process of heating and frothing the milk is known as “stretching”. A good barista will spend a lot of time learning how to get the technique just right.

The milk for a flat white is frothed, folded and swirled to blend together the bubbles and the liquid milk. This creates a microfoam mix with a velvety consistency throughout the whole drink.

In a latte, on the other hand, the foam remains at the top of the drink with a smaller layer of micro-foam and the liquid milk beneath it. That’s why lattes are often served in glasses rather than cups, allowing you to see the different layers.

In this YouTube video, Todd from wholelattelove.com demonstrates the art of frothing and pouring milk for a range of espresso-based drinks.

Pouring for perfection

It isn’t just heating and stretching the milk that is important to get the right results. If the barista pours the milk incorrectly into the cup or glass, he or she can undo all that good work by damaging the foam.

To prepare for pouring a flat white, the barista will swirl the milk smoothly inside the pitcher, mixing the milk with the micro-foam. The ideal texture should be similar to gloss paint (7).

He or she may also give the pitcher a sharp tap on the counter to burst any large bubbles remaining on the surface of the milk. However, this shouldn’t be necessary if the milk has been swirled well and smoothly.

The most experienced baristas will be able to get the perfect foam for any drink simply by adjusting the speed and angle of the pour. This is a technique known as “free pouring”.

For the rest of us, the flat white is the easiest option to pour. It should be poured as soon as possible after stretching is complete to avoid the milk separating. As long as the micro-foam and milk have been properly mixed, it’s just a case of pouring the liquid from the pitcher into the cup.

For a latte, the simplest option is to pour the milk from the pitcher to the cup, using a spoon to hold back the froth. Then add a thin layer of foam on top of the cup.

These finishing touches create the distinctive appearances of the two drinks. The flat white – as its name suggests – has a smooth, glossy surface. The latte, on the other hand, has a thin layer of froth on top.

Pouring the milk slowly is key to getting the crema to rise to the top of the cup. Which brings us to the next important distinction between a flat white and a latte…

Crema

The crema is the golden-brown foam that sits on top of a shot of espresso. It has a sweeter taste than the liquid coffee and a silky texture.

Because a latte is topped in a thin layer of foam, the crema is usually lost underneath it. In a well-made flat white, on the other hand, the crema will merge with the textured milk. The result will be a smooth golden surface to the drink.

Skilled baristas can make the top of either a latte or a flat white look like a work of art. This YouTube video shows some great examples.

Flavor and strength

We’ve talked about texture and appearance – but what about the difference in the taste?

The different approaches to the preparation and pouring of a flat white and latte create two different drinking experiences. As a flat white also has a higher coffee to milk ratio than a latte, it has a stronger taste.

The results? A latte essentially tastes like a milky coffee, while a flat white is more like a creamy espresso.

The price of fashion

A flat white is often a slightly more expensive option in coffee shops than a latte. In the UK, the price difference has caused quite a stir, with the television programme Supershoppers claiming that customers were being ripped off.

Both drinks contain the same amount of coffee, and a flat white has less milk. Surely that means that, if anything, it’s the latte that should be more expensive?

Part of the reason for the flat white’s higher price tag is its reign as the king of coffee cool. Simply put, customers are prepared to pay more for the new kid on the coffee block.

But others contend that there’s a good reason to pay a premium. The art of producing the smooth micro-foam that characterizes the flat white takes skill. And if you want a pretty flower on top of your coffee, that’s another trick for the barista to learn.

Both those mean that for the perfect flat white, baristas will need training and time to master new techniques. It’s only reasonable, the coffee shops argue, that this is reflected in the price (8).

In reality, despite the controversy, the price difference isn’t huge. In the UK, a flat white in Starbucks costs 20 pence – about 28 cents – more than a tall latte.

Calorie: flat white vs latte

For those watching their waistlines, the calorie content of the drinks will be of interest. Is choosing a latte or a flat white the more diet-friendly option?

The bad news is that since both drinks are made with milk they contain fat. Limiting the number of cups you drink is a good idea if you’re watching your weight. Swapping from full- or half-fat to skimmed milk will also make a big difference.

Lattes and flat whites are almost the same in their proportions of coffee and milk. That means that for the same size serving, there’s virtually no difference in their calorie and fat contents.

A 7.75-ounce latte made with full-fat milk contains 123 calories and 4.4 grams of saturated fat. The same sized flat white made the same way also has 123 calories but 4.6 grams of saturated fat.

The small difference is the result of the foam on top of the latte. That includes more air, so there’s slightly less fat than in the flat white. Bear in mind too, that flat whites are usually served in smaller sizes (9).

As ever, though, beware of differences to the norm. In Starbucks, ordering a tall latte will get you a 12-ounce drink with 150 calories.

Order a grande flat white in the same chain and you’ll receive a whopping 16-ounce drink with 220 calories.

Of course, if you want to get a lower calorie, you can try to invest a Nespresso Machine, and make the low-calorie latte or flat white at home.

Conclusion

So there you have it. From the history to the ingredients and flavor, to the impact on your pocket and your waistline – you now have all the information you need to choose between a flat white and a latte.

Of course, the best way to make a decision is to go down to your local coffee shop and sample both drinks! You might even want to take a blind taste test to make sure your preference is based on flavor alone. But if you want to brew at home, A home latte machine could help you speed up.


Q:

Using Integer.parseInt crashes my program

I have looked all over the internet and cannot find my out why this is happening to my program.
Basically I am trying to make a TextField that that makes a string of numbers, and that string is turned into an integer and the label is changed to the number +1
Whenever I attempt to use Integer.parseInt It crashed
Here is my Code
public class dfadsfa {

    private JFrame frame;
    private JTextField textField;

    /**
     * Launch the application.
     */
    public static void main(String[] args) {
        EventQueue.invokeLater(new Runnable() {
            public void run() {
                try {
                    dfadsfa window = new dfadsfa();
                    window.frame.setVisible(true);
                } catch (Exception e) {
                    e.printStackTrace();
                }
            }
        });
    }

    /**
     * Create the application.
     */
    public dfadsfa() {
        initialize();
    }

    /**
     * Initialize the contents of the frame.
     */

    String Text;
    int Number = Integer.parseInt(Text); //This line Screws it up

    private void initialize() {
        frame = new JFrame();
        frame.setBounds(100, 100, 450, 300);
        frame.setDefaultCloseOperation(JFrame.EXIT_ON_CLOSE);
        frame.getContentPane().setLayout(null);

        JLabel lblNumber = new JLabel("");
        lblNumber.setBounds(198, 181, 61, 16);
        frame.getContentPane().add(lblNumber);

        textField = new JTextField();
        textField.addActionListener(new ActionListener() {
            public void actionPerformed(ActionEvent e) {
                Text = textField.getText();
                lblNumber.setText(Number+1+"");
            }
        });

        textField.setBounds(163, 140, 130, 26);
        frame.getContentPane().add(textField);
        textField.setColumns(10);

    }

}

This is the message that shows up-
java.lang.NumberFormatException: For input string: ""
at java.lang.NumberFormatException.forInputString(NumberFormatException.java:65)
at java.lang.Integer.parseInt(Integer.java:592)
at java.lang.Integer.parseInt(Integer.java:615)
at dfadsfa.<init>(dfadsfa.java:42)
at dfadsfa$1.run(dfadsfa.java:21)
at java.awt.event.InvocationEvent.dispatch(InvocationEvent.java:311)
at java.awt.EventQueue.dispatchEventImpl(EventQueue.java:756)
at java.awt.EventQueue.access$500(EventQueue.java:97)
at java.awt.EventQueue$3.run(EventQueue.java:709)
at java.awt.EventQueue$3.run(EventQueue.java:703)
at java.security.AccessController.doPrivileged(Native Method)
at java.security.ProtectionDomain$JavaSecurityAccessImpl.doIntersectionPrivilege(ProtectionDomain.java:76)
at java.awt.EventQueue.dispatchEvent(EventQueue.java:726)
at java.awt.EventDispatchThread.pumpOneEventForFilters(EventDispatchThread.java:201)
at java.awt.EventDispatchThread.pumpEventsForFilter(EventDispatchThread.java:116)
at java.awt.EventDispatchThread.pumpEventsForHierarchy(EventDispatchThread.java:105)
at java.awt.EventDispatchThread.pumpEvents(EventDispatchThread.java:101)
at java.awt.EventDispatchThread.pumpEvents(EventDispatchThread.java:93)
at java.awt.EventDispatchThread.run(EventDispatchThread.java:82)

A:

You have an unintialized String text. This is guaranteed to fail when trying to parse an int.
String Text;
int Number = Integer.parseInt(Text);

You should initialize this text before trying to parse the int.

A:

Text is not initialized , So it gives a NumberFormatException.
Here Integer.parseInt() method will give the NumberFormatException if the passed string does not contain a parsable integer.
Use
String Text = "0" to solve the problem


They comin’. The road to Iran is through Syria…but they damn sure comin’ for ’em. Gwenyth Todd, former political adviser to the US Navy’s 5th Fleet and former US National Security Council staffer in the Clinton and first Bush administration, knows this. Those cock suckers at the FBI tried to muscle her the first time...


Dibenzalacetone h nmr. ChemSpider SyntheticPages 2019-02-02

Dibenzalacetone h nmr Rating:
9,2/10
1757
reviews

DIBENZYLIDENEACETONE(538

Die Analyse gab keine verständlichen Zahlen, daher ist die Substanz wahrscheinlich ein Gemenge. You need to or later to see this applet. We thanks for providing the predicting engine. The last peaks are a doublets of two protons, which is the most downfield proton; the α hydrogen is indeed the most deshielded proton, as it is next to a carbonyl group — a withdrawing group. The Claisen-Schmidt is an aldol condensation reaction, which takes place between an aromatic aldehyde and a ketone to result in an enone.

Dibenzylideneacetone

Benzaldehyde is used in a slight excess 2. The reaction uses the simple starting materials, acetone and benzaldehyde, to form the 15-carbon product, dibenzalacetone, in diastereoselective fashion. The rate determining step is addition of the enolate to the aldehyde. As for the low yield 40. The oxygen atom then attacks a proton on the newly formed water to create a hy droxide group on the carbon of the enolate. It is a pale-yellow solid insoluble in water, but soluble in ethanol.

Notes on NMR Solvents

A modification of a classic experiment is described that incorporates a discovery approach to organic synthesis. It will calculate the same spectrum but taking into account the second order effects. If you are really in hurry, just click to get a predicted spectrum of ethylvinylether. This explanation should be relevant to consider the multiplets are divided to 5H each, and to consider that these two benzene rings gives different signals as they conjugate, so the readings shifted a bit from each other. The reaction works best when there is no possibility of the aldehyde forming an enolate, e.

trans,trans

Therefore, signals will be observed for the solvent and this must be accounted for in solving spectral problems. Journal of Magnetic Resonance 2011. The mechanism eq 3-5 involves deprotonation of an α-hydrogen on the ketone to form an enolate eq 3 , which then adds nucleophilically to the aldehyde carbonyl to form a β-hydroxy ketone eq 4. Recall that deuterium has a spin quantum number I of 1, so n deuterium atoms will split a proton signal into 2In+1 lines. Therefore, the aldehyde has to react with itself to yield one product. .

Predict 1H proton NMR spectra

Dibenzylideneacetone is a component of the catalyst. What is the purpose of the water washes in step 2 of the work up procedure? It was first prepared in 1881 by the German chemist 1851—1930 and the Swiss chemist Charles-Claude-Alexandre Claparède 14 April 1858 — 1 November 1913. Sorry, but copying text is forbidden on this website! Draw the molecule with the correct steriochemistry and using numbers, indicate which carbons are in equivalent environments. Zuerst bildet sich aber ein öliger, unzersetzt flüchtiger Körper, der einen an Cumarin erinnernden Geruch besitzt, und der höchst wahrscheinlich das Methylaceton der Zimmtsäure ist: …. The doublets peak mirrors a vicinal carbon with one hydrogen; the hydrogen mention earlier in the analysis. Dibenzylideneacetone is used as a component in and as a in.

trans,trans

The difference of the first doublets is 0. It might be caused by the solvent used during recrystallization was a little bit more than advised in the lab manual ~15 mL instead of ~12 mL of ethanol. The condensation occurs extraordinarily quickly, after just a few minutes, and after some time the whole mass becomes resinous. Is this the normal absorption for this functional group? The next peaks about to be discussed are the multiplets between 7. It is a labile ligand that is easily displaced by , hence it serves a useful entry point into 0 chemistry. Using excess aldehyde ameliorates this problem because the aldehyde never gets all used up and the concentration always stays well above zero. Castillo, Luc Patiny and Julien Wist.

ChemSpider SyntheticPages

It is a doublets because there is a vicinal carbon with a proton, so it makes sense, according to the n + 1 rule. Acetone is used as the enolate forming compound, adding to the benzaldehyde followed by the dehydration to form a benzal group. Berichte der Deutschen Chemischen Gesellschaft in German. Acetone has α-hydrogens on both sides and, thus, can react in aldol fashion with two equivalents of benzaldehyde to give the compound known as dibenzalacetone eq 1. This would yield a different product. Preparation of Dibenzalacetone by a Double Aldol Condensation Relevant textbook reading - Smith, Chapter 24. There are two doublets consisting of two hydrogen at 7.

Notes on NMR Solvents

The crystal was also wasted in the Erlenmeyer flask and some on the filter paper used to dry it. This allows the reaction to reach completion in a reasonable amount of time. An enone is a conjugated ketone product such as dibenzalacetone, which can be used as a good way to make C-C bonds 1 due to the positive charge that can be produced at the unsaturated Dibenzalacetone is an organic compound , which in this case was yellow solid after recrystallization. Bibliography Organic Chemistry 7th EdFrancis A. This is because the product of this reaction contains both an aldehyde and alcohol. The chemical shift of the alkene protons is downfield from the expected resonance. The carbon-carbon bond-forming reaction is referred to as aldol addition.


Ted Nelson wrote:
> m, M => extends a cursorlike cell in d.mark.
> By default, it will contain the characters "m.1"
> for the first marked sequence.
> Each time this is done, such a cell will be extended
> in d.mark from the marked cell.
> (Presumably these are clones.)
> Meanwhile, all these cells containing "m.1"
> will be put on a list in d.marklist.
Clones of what?
And Xanni, how on EARTH would I keep the d.marklist lists working?
Gossamer
--
: Gossamer - gossamer@xxxxxxxxxxxxxx - http://www.tertius.net.au/~gossamer/
: *** Link of the week: http://www.gnuhoo.com/ ***
: Be glad of life. It gives you the chance to love and to work and to
: play and to look up at the stars. -- H Van Dyke


# Copyright (c) Facebook, Inc. and its affiliates.
#
# This source code is licensed under the MIT license found in the
# LICENSE file in the root directory of this source tree.

# Test: Add pyre-strict (change mode)
#!/usr/bin/env python3

from typing import Any

def foo(x: Any) -> str:
    return x


Q:

ReactJs 0.14 - TreeView

Upgrading this example to React.js 0.14 .....
I am receiving an error when toggling:

Uncaught TypeError: Cannot read property 'setState' of null

import React, { Component } from 'react';
import ReactDOM from 'react-dom';
import classNames from 'classnames';
import { Link } from 'react-router';

export default class TreeNode extends Component {

  constructor(props) {
    super(props);
    this.state = {
      visible: true
    };
  }

  render() {
    let childNodes;
    let classObj;

    if (this.props.node.childNodes !== undefined) {
      childNodes = this.props.node.childNodes.map((node, index) => {
        return (

<TreeNode node={ node } />

        );
      });
    }

    classObj = classNames({
      'togglable': true,
      'togglable-down': this.state.visible,
      'togglable-up': !this.state.visible
    });

    let style;
    if (!this.state.visible) {
      style = { display: 'none' };
    }

    return (

          { this.props.node.title }

          { childNodes }

    );
  }

  toggle() {
    this.setState({ visible: !this.state.visible });
  }
}

TreeNode.propTypes = {
  node: React.PropTypes.object
};

ReactDOM.render(<TreeNode node={ treeData } />, document.querySelector('#reactRoot'));

Anyone see my error?
Many thanks in advance if you can point it out.

A:

Nevermind, dumb, dumb, dumb .....


Pages

Monday, May 20, 2013

what type of karma do you choose?

Verse 3.9: Work done as a sacrifice for Viṣṇu has to be performed, otherwise work causes bondage in this material world. Therefore, O son of Kuntī, perform your prescribed duties for His satisfaction, and in that way you will always remain free from bondage.

If you had a choice, what would you choose - good or bad karma?

Whether we realize it or not, that's the question we face on a daily basis. Sometimes, however, the best choice may not be the one that has been presented, which is exactly the case here. That's why the Gita is offering us the other option. The one that many of us would not have thought of.

Good karma is definitely the way to go if the options were only limited to good and bad. But even then, there's something important to realize. The thing with good and bad karma, is that they both bind one to a temporary material body. That's because we don't always receive our karma in the body we are inhabiting in presently. Simply put, if we do something good, we may not reap that benefit now.

The goal of bhakti yoga is to free ourselves from the captivity of impermanence which is what we face here in the material world. Why is that? Because we are eternal souls and therefore all the temporality we experience doesn't sit right with any of us and that's why we are always longing for ever-lasting pleasure.

So whether it be good or bad karma, it still serves to trap us in an endless cycle of impermanence. The key to escaping this prison is by performing akarmic activities, those that result in no karma. The amazing aspect of performing akarmic work is the fact that it doesn't require one to give up their occupation or their talents. All it requires is a shift in consciousness.

This topic was briefly discussed in Chapter 2 when we discussed the secret behind working without attachment to the result. Instead of acting in a consciousness of "it's all for me", instead we work in a spirit of gratitude. Now we get a better understanding of why this consciousness shift actually benefits us. As Krsna says, it's because we will always remain free from bondage by working in this way. Or in other words, it won't tie us to having to take birth in another material body.

So....the choice is yours. The next time someone asks you whether you would choose good or bad karma, go ahead and choose akarma! That's the path by which we can experience our eternally blissful true selves.

About Me

I am an adventurer at heart. I feed off of travelling, discovering and uncovering lost treasures. I invite you join me as I set out to explore and uncover the gems of each verse of the Bhagavad gita-as it is.


Q:

SQL Database scripting table changes

I'm working on a project where we are updating client databases from older version to newer versions of the database and the related schema.
I'm looking for a way do the following for all of the tables in the database that our program uses.
If the table doesn't exist
  create table
else
  for each column in the table
    bring the column up to date or create it

I can't seem to find a way to do this through management studio and I haven't turned anything up through Google.
Something that would either give me this option in SQL Management Studio or a program that would generate the scripts for me would be good.
Thanks.

A:

You can use tools like RedGates SQL Compare or Apex SQLDiff to compare and sync your schemas. It will compare the old and new database and generate the script for you.


Clean intermittent catheterization and prevention of renal disease in spinal cord injury patients.
Upper urinary tract complications have been reported in about 20-30% of spinal cord injury patients. Their pathogenesis is linked to the presence of high-pressure uninhibited detrusor contractions, high leak point pressure and low bladder compliance. The aim of this study was to evaluate the incidence of upper urinary tract complications in two homogeneous groups of spinal cord injury patients with different bladder emptying modalities (intermittent catheterization vs. tapping, abdominal straining, Crede's manoeuvre) and the relationship with clinic and urodynamic features. A total of 17 patients (22%) had upper urinary tract complications. The incidence of both urinary tract dilatation and vesicoureteral reflux was significantly lower in patients having intermittent catheterization (P = 0.03 and 0.04 respectively). Intermittent catheterization thus seems to be effective in preventing upper urinary tract disease in spinal cord injury patients requiring mechanical bladder emptying modalities. The finding of upper urinary tract complications also in patients having intermittent catheterization showing high intravesical pressures stresses the need of adding anticholinergic medications to the rehabilitation regimen of these patients.


package com.raw.utils;

public class Camera {

	public static float DEFAULT_VIEW_ANGLE = (float) Math.toRadians(45);
	public int width, height;

	float viewAngle;
	float dist;

	public Camera(int width, int height) {
		this(width, height, true);
	}

	public Camera(int width, int height, boolean init) {
		this.width = width;
		this.height = height;

	}

	public void setViewAngle(float viewAngle) {
		this.viewAngle = viewAngle;
		this.dist = (this.width / 2)
				/ (float) Math.tan(viewAngle / 2);
	}

	public void setViewAngle(int width, int height, float viewAngle) {
		this.viewAngle = viewAngle;
		this.dist = (width / 2) / (float) Math.tan(viewAngle / 2);
	}

	@Override
	public String toString() {
		return " ";
	}
}


Sunday, November 21, 2010

Not only are these gold nanoparticles gorgeous to look at – they may one day act as microscopic powerhouses for molecular machines. Researchers at the Nano/Bio Interface Center at the University of Pennsylvania recently discovered a novel to way to generate solar power by shining light onto gold nanoparticles. The discovery has far-reaching implications in the realm of nanotechnology, and may open the door for everything from self-powering molecular circuits to super-efficient data storage.

To generate current the researchers first packed a bunch of light-sensitive gold nanoparticles together on a glass substrate and then exposed them to optical radiation (light). This knocks conductive electrons free from the gold particles, which run along the surface to create surface plasmons, which in turn induce an electrical current across the molecules.

The amount of electricity generated is minute, but the researchers believe that by optimizing size, shape, and orientation of the nanparticles they could create a current strong enough to power nano-sized circuits. Professor Bonnell, who participated in the experiment, said “If the efficiency of the system could be scaled up without any additional, unforeseen limitations, we could conceivably manufacture a 1A, 1V sample the diameter of a human hair and an inch long“.

Turning sunlight into electrical power is all but a new problem, but recent advancements made by researchers at the University of Pennsylvania have given a new twist to the subject. While not currently aimed at solar panel technology, their research has uncovered a way to turn optical radiation into electrical current that could lead to self-powering molecular circuits and efficient data storage.

Professor of materials science Dawn Bonnell and colleagues placed light-sensitive gold nanoparticles on a glass substrate, minimizing the distance between the nanoparticles. The team then stimulated conductive electrons with optical radiation to ride the surface of the gold nanoparticles, creating so-called "surface plasmons" that induce electrical current across molecules.

Under these conditions, surface plasmons were found to increase the efficiency of current production by a factor of four to 20. The size, shape and separation of the array of golden nanoparticles can be customized independently of the optical characteristics of the molecule, and optimization of these parameters could, the researchers say, produce enhancement factors of thousands, and the resulting electrical current could be easily transported to the outside world.

"If the efficiency of the system could be scaled up without any additional, unforeseen limitations, we could conceivably manufacture a 1A, 1V sample the diameter of a human hair and an inch long," Prof Bonnell explained.

The results may lead to better nano-sized circuits that can power themselves, potentially through sunlight. Another interesting application suggested by the researchers could be for data storage, where a photovoltaic circuit could encode bits using wavelengths of light rather than electrical charge.

The study, published in the current issue of the journal ACS Nano, was supported by the Nano/Bio Interface Center, National Science Foundation, the John and Maureen Hendricks Energy Fellowship and the US Department of Energy.


To confirm the presence of the EPEL repository, run the following rpm command.

# rpm -qa | grep epel

Confirm EPEL Installation

Next, run the command below to add the Remi repository.

# dnf install https://rpms.remirepo.net/enterprise/remi-release-8.rpm

Install Remi Repo on CentOS 8

Once again, to verify the existence of the Remi repository, run the command.

# rpm -qa | grep remi

Confirm Remi Repo Installation

Step 2: Install PHP 7.4 on CentOS 8

Upon successful addition of EPEL and Remi repositories, execute the command below to get a list of available PHP module streams.

# dnf module list php

List PHP Modules

From the output above, remi-7.4 is the latest PHP stream, and therefore we are going to enable the module stream as shown below.

# dnf module enable php:remi-7.4

Enable PHP Remi Stream

Once the PHP remi-7.4 module has been enabled, you can then proceed and install PHP using the command below. This will also install a host of other packages such as Apache and Nginx modules.

# dnf install php php-cli php-common

Install PHP on CentOS 8

To check the version of PHP installed, run the command.

# php -v

Verify PHP Installed Version

From the output, we can clearly see that we have installed PHP version 7.4! You can now proceed to test and deploy your applications using PHP. And that’s just about it for now. We do hope you found this guide helpful.

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Valeria Lukyanova: ‘Human Barbie’ Says She Doesn’t Need Food Or Water

Well this is an interesting life choice. The Ukrainian model, Barbie doll look-alike, revealed in a shocking interview that she’s well on her way to being able to subsist solely on air and light.

Ukrainian model Valeria Lukyanovahad some shocking things to say in a new interview, admitting that she hasn’t eaten in weeks and is almost able to subsist solely on air and light. Would you expect anything less from a life-sized version of a Barbie doll?

Valeria Lukyanova: Model No Longer Eating

Many people have accused Valeria of Photoshopping her pictures to get those doll-like proportions, but her scary, miniscule waistline may be from all that eating she’s NOT doing.

“In recent weeks I have not been hungry at all,” the human Barbie told the International Business Times. “I’m hoping it’s the final stage before I can subsist on air and light alone.”

Valeria’s no water, no food diet is part of her philosophy as a member of Breatharianism, a cult-like sect that she’s allegedly part of. The sect advocates living without food and water, and subsisting solely on what they call “cosmic micro-food.”

The Human Barbie’s Odd Confessions

If you can believe it, this isn’t the first time the human Barbie has made grandly weird confessions about her life.

Valeria has also declared that she is a spiritual teacher who can speak with aliens through light (clearly, light is an important part of her life). She also believes she’s from another planet and that she has the ability to time travel have out-of-body experiences.

We… we don’t really even know what else to say, except that we hope Valeria is sneaking some cubes of cheese or something in between her doses of light and air.


Sunday, March 13, 2016

Southern Comfort ***

Kate Davis’s 2001 documentary of the same title is an extremely moving film about Robert Eads, a female-to-male transgender person dying, ironically, of ovarian cancer in rural Georgia, surrounded by his mostly transgender friends. Robert Dusold and Thomas Caruso have conceived the work for the stage, with music by Julianne Wick Davis and book and lyrics by Dan Collins. During his final year of life Eads (Annette O’Toole) finds true love with Lola Cola, (Jeff McCarthy) a tall transexual who has not yet committed to hormones or surgery. Jackson (Jeff Kuhn) is a younger F-to-M who has been friends with Robert for 10 years and regards him as his spiritual father. Sam (Donnie Cianciotto), another F-to-M transexual, and Melanie (Robin Skye), a biological woman, are a devoted couple who are also friends of Robert’s. Carly (Aneesh Sheth) is a sexy M-to-F transexual who is currently seeing Jackson. Four of the five musicians (David M. Lutken, Lizzie Hagstedt, Joel Waggoner and Elizabeth Ward Land) also sing and step into the action to play secondary characters such as Robert’s parents and Jackson’s father. (David Morse, the pianist, does not.) Southern Comfort is the name of the annual transgender event in Atlanta that Robert and his friends regularly attend. While the book is mostlly faithful to the essentials of the film, it makes things a bit more schematic. As I am not a fan of country/bluegrass music, I did not really enjoy the score. To my ears, much of the music seemed monotonous and repetitive. The present Public Theater production features the same cast as the 2011 CAP 21 version except for Cianciotto and Sheth who are transgendered. The song list is almost identical too. The rustic set by James J. Fenton is dominated by a large tree filled with boxes of tchotchkes a la Joseph Cornell. Patricia Doherty’s costumes are spot-on. Thomas Caruso’s direction is seamless. The entire cast, especially O’Toole and McCarthy, are excellent. Although, for me, the music did not really enhance the story, it is still a moving and timely tale that I am glad will be seen by new audiences. I highly recommend renting the film too. Running time: 2 hours, 20 minutes including intermission.


Q:

How to use drools in grails3 without any plugin?

Is it possible to use drools rule engine in GRAILS3 without any plugin installation? I ask that because I know drools is implemented in java and  actual official plugin by Ken Siprell for GRAILS is (apparently) no more working.

A:

After much investigation and many attempts, I got a small GRAILS3 API service through which it is possible to process data using DROOLS engine without any plugin. All that is possible because DROOLS is based on java and because of perfect compatibility between GRAILS and Java.
All you need is the following:

include minimal DROOLS dependency in build.gradle
choose a folder to store files.drl
have some entities to use as facts to process (not mandatory to persist them, so I skip hibernate configuration)
implement a service to build rule base knowledge and to get a session
implement some methods into API controller to process data through DROOLS (having got its session from service)

Below there is a simple example of all that:
DROOLS dependencies (in build.gradle):
runtime "org.drools:drools-compiler:6.5.0.Final"
compile "org.drools:drools-core:6.5.0.Final"
compile "org.drools:knowledge-api:6.5.0.Final"

DRL storage in src/rules (reference to this path will be in the service, see below): myrules.drl
import my.entities.Book;
import java.util.List;

rule "Find author"
   salience 10
   when
    $book: Book( author=="Shakespeare" )
   then
    System.out.println("Book found, date:"+$book.getDate0());
end

Some entity, for example Book:
package my.entities
import java.util.Date

class Book {
    String title, author
    Date date0
}

Service to build DROOLS knowledge and get session (I prepared a stateless engine, lighter than the stateful one):
package my.services
import grails.converters.*

import org.kie.api.runtime.*;
import org.kie.internal.io.ResourceFactory;
import org.kie.api.*;
import org.kie.api.io.*;
import org.kie.api.builder.*;

class DroolsService  {

def getSession() {
        def path    = "src/rules"
        def lru = ["myrules.drl"]
        def rules   = []
        lru.each{
            rules.add("${path}${it}")
        }
        StatelessKieSession ksess   = buildSession(rules)
        return ksess
    }
}

private buildSession(def lfile) {
    println "Building DROOLS session..."
    try {
        def lres    = []
        lfile.each{
            Resource resource   = ResourceFactory.newFileResource(new File(it));
            lres.add(resource)
        }

        KieContainer kieContainer = buildKieContainer(lres)
        StatelessKieSession kieSession = kieContainer.newStatelessKieSession()
        return kieSession
    } catch(Exception e) {
        e.printStackTrace()
        return null
    }

protected KieContainer buildKieContainer(def lres) {
    KieServices kieServices = KieServices.Factory.get()
    KieFileSystem kieFileSystem = kieServices.newKieFileSystem()
    lres.each{
        kieFileSystem.write("src/main/resources/rule.drl", it)
    }

    KieBuilder kieBuilder = kieServices.newKieBuilder(kieFileSystem).buildAll()
    Results results = kieBuilder.results
    if (results.hasMessages(Message.Level.ERROR)) {
        throw new IllegalStateException(this.class.name + ": " + results.messages.toString())
    }
    KieContainer kieContainer = kieServices.newKieContainer(kieServices.repository.defaultReleaseId)
    kieContainer
}

}

And use of service in API controller:
class ApiController  {

def droolsService

def proc = {
    def sess    = droolsService.getSession()

    def mess    = "ok DROOLS proc from JSON"
    Book book   = null

    if (params.contbook) {
        book = new Book(JSON.parse(params.contbook))
        sess.execute book
    }

    response.status  = 200
    render mess
}

In the controller I take json data from parameter and populate an entity by them, in order to execute it with the rule engine initialised by the DROOLS service. Of course this is a very simple solution, but it is working.
Some notes:

the RHS part of each drl rule (after "then") must be java, so, as  in the example, you cannot access directly private property of the entity but you have to use a getter or setter (implicitly created by GRAILS)
getSession create a new build of the knowledge and a new session and that is not optimal, you may redesign it in order to store a DROOLS session and then reuse it through a user session
you can have several files.drl in the same folder and then you have to list them all in the service in order to pack them in the knowledge engine

Hope all that is useful.


MUSKEGON, MI - A video which appears to have been taken by a Muskegon resident showing two Muskegon Police Department officers in uniform playing football with neighborhood children is going viral on Facebook and other social media sites.

Muskegon Director of Public Safety Jeffrey Lewis said Friday, April 17, that he is proud of the two officers and hopes the Muskegon community realizes that it's a reflection of the department as a whole.

Lewis said he had a chance to see the video this morning. The officers in the video, taken around 7:30 p.m. on Thursday, April 16, are Muskegon Police Sgt. Tim Bahorski and Officer Kevin Kattoula, Lewis said.

"We are pretty impressed," Lewis said. "It shows that there are really a lot of good officers out there."

The city of Muskegon Facebook page also posted the video.

Lewis said he is looking into more details about the events of that evening. He said the officers in the video work the night shift and may have simply wanted to interact with the neighborhood children there.

The video was posted by Jessica Lindsay who appears to be filming the video from her home. Under the video she captioned: "Come outside and this is wht I seen...two Muskegon police officer was plyn football wit the kids."

MLive Muskegon Chronicle was unable to reach Lindsay early Friday. It wasn't clear where in the city the interaction was recorded.

Lewis said he was going to find out later this afternoon.

Heather Lynn Peters covers police and fire, and writes a statewide food column, The Spunky Kitchen, for MLive/Muskegon Chronicle. Email her at hpeters@mlive.com and follow her on Twitter @HLPNEWS.


1. Field of the Invention
The present invention relates to a steering control system for use in a track-laying vehicle and, more particularly, to a steering control system for use in a track-laying vehicle such as a bulldozer, power shovel or crane which is most suitably applied to a steering control system in which a steering motor driven by the rotation of a power source drives crawler belts provided at both sides of the vehicle such that a relative travel difference is caused between the crawler belts, thereby turning the vehicle rightwards or leftwards.
2. Description of the Prior Art
In a prior art steering control system for the above-type track-laying vehicle, the revolution speed of the steering motor is unitarily determined and varied according to the displacement distance of a steering lever alone, regardless of the revolution speed of the power source. Such a steering lever is one form of a steering instructing means which can be displaced from a neutral position by operation and which instructs, according to the direction and distance of its displacement, a turning direction and a turning radius for the vehicle, the turning radius decreasing as the displacement distance increases.


Q:

ConcurrentSkipList? That is, not a ConcurrentSkipListSet

I need a very fast (insert, remove, contains) highly concurrent list that can be sorted using a comparator/comparable.
The existing ConcurrentSkipListSet would be ideal, if it was a list and not a set.  I need to insert multiple items which are equal into the data structure.
I'm currently thinking of using a LinkedDeque if I can't find anything better, but that structure is considerably slower than a skiplist at high contention.
Any suggestions?
EDIT: What I actually need, bare minimum, is something that is sorted using compareTo, can insert concurrently and can remove/get items using object identity. All other concurrent requirements mentioned in comments still apply.

A:

The existing ConcurrentSkipListSet would be ideal, if it was a list and not a set.

So the SkipList data-structure at it's core is a linked list.  If you are worried about order and the ability to traverse it easily and in order, the SkipList will work very well for that as well.  It is also a probabilistic  alternative to a balanced tree which is why it can also be a Set or a Map.  The data structure in memory looks something like the following:

To quote from the Javadocs:

This class implements a concurrent variant of SkipLists providing expected average log(n) time cost for the containsKey, get, put and remove operations and their variants. Insertion, removal, update, and access operations safely execute concurrently by multiple threads. Iterators are weakly consistent, returning elements reflecting the state of the map at some point at or since the creation of the iterator. They do not throw ConcurrentModificationException, and may proceed concurrently with other operations. Ascending key ordered views and their iterators are faster than descending ones.

If you explain more about what features you want from List, I can answer better whether ConcurrentSkipListSet will be able to work.

Edit:
Ah, I see.  After some back and forth in the comment, it seems like you need to be able to stick two objects that are equivalent into the Set which isn't possible.  What we worked out is to never have compareTo(...) return 0.  It's a bit of a hack but using AtomicLong to generate a unique number for each object, you can then compare those numbers whenever the real comparison field (in this case a numerical timeout value) is equal.  This will allow objects with the same field to be inserted into the Set and kept in the proper order based on the field.


Q:

Homotopy dimension of homotopy domination

The homotopy dimension of a space $X$ is the smallest covering dimension of any space homotopy equivalent to $X$.
Assume that for topological spaces $X$ and $Y$, there exist maps $f:X\to Y$ and $g:Y\to X$ so that $g\circ f\simeq 1_X$. If $X\not \simeq Y$, then is it true  that homotopy dimension of $X$ is  strictly  smaller than homotopy dimension of $Y$?

A:

No. Take $X=S^n$ and $Y=S^n\vee S^n$, with $f$ the inclusion into one of the wedge summands. Then it is clear that $X\not\simeq Y$, and that the homotopy dimensions of $X$ and $Y$ are equal.


    <?= $this->tabs ?>

    <?= $this->form; ?>


Cooking Directions:

Preheat oven to 375°F (190°C). Set up three bowls with the following: 1) flour mixed with a pinch of salt and pepper; 2) an egg-wash; made by beating the eggs and milk together; 3) cornflakes mixed with cheese.

Coat a 13 x 9-inch baking pan with non-stick cooking spray.

Dip chicken pieces in flour, then into the egg-wash, then roll in the cheesy-flakes mixture, coating the entire piece of chicken and place on the baking pan. Discard any unused cornflake mixture after coating chicken.