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Zhang-Zhi Hu1*, Hongzhan Huang1, Amrita Cheema2, Mira Jung3, Anatoly Dritschilo3, Cathy H. Wu1
1Department of Biochemistry and Molecular & Cellular Biology
2Proteomics and Metabolomics Shared Resource
3Department of Radiation Medicine, Lombardi Comprehensive Cancer Center Georgetown University Medical Center, Washington, DC 20007, USA
Received Date: May 24, 2008; Accepted Date: May 24, 2008; Published Date: May 24, 2008
Citation: Zhag ZH, Hongzhan H, Amrita C, Mira J, Anatoly D, et al. (2008) Integrated Bioinformatics for Radiation-Induced Pathway Analysis from Proteomics and Microarray Data. J Proteomics Bioinform 1: 047-060. doi: 10.4172/jpb.1000009
Copyright: © 2008 Zhang ZH, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Functional analysis and interpretation of large-scale proteomics and gene expression data require effective use of bioinformatics tools and public knowledge resources coupled with expert-guided examination. An integrated bioinformatics approach was used to analyze cellular pathways in response to ionizing radiation. ATM, or mutated in ataxia-telangiectasia, a serine-threonine protein kinase, plays critical roles in radiation responses, including cell cycle arrest and DNA repair. We analyzed radiation responsive pathways based on 2D-gel/MS proteomics and microarray gene expression data from fibroblasts expressing wild type or mutant ATM gene. The analysis showed that metabolism was significantly affected by radiation in an ATM dependent manner. In particular, purine metabolic pathways were differentially changed in the two cell lines. The expression of ribonucleoside- diphosphate reductase subunit M2 (RRM2) was increased in ATM-wild type cells at both mRNA and protein levels, but no changes were detected in ATM-mutated cells. Increased expression of p53 was observed 30min after irradiation of the ATM-wild type cells. These results suggest that RRM2 is a downstream target of the ATM-p53 pathway that mediates radiation-induced DNA repair. We demonstrated that the integrated bioinformatics approach facilitated pathway analysis, hypothesis generation and target gene/protein identification.
Bioinformatics; Proteomics; Radiation; Purine metabolism; DNA repair; Pathway and network
ATM: Ataxia Teleangiectasis Mutated; RRM2: Ribonucleotide Reductase subunit M2, or Ribonucleoside-diphosphate Reductase subunit M2; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; iProXpress: integrated Protein eXpression system; UniProt: Unified Protein Resource; 2D-gel/ MS: Two-dimensional gel/Mass Spectrometry; PIR: Protein Information Resource.
The last decade has seen a rapid expansion of genomics, transcriptomics, proteomics, and other omics studies applied to all areas of biomedical research. High-throughput technologies such as DNA microarray and mass spectrometry (MS)-based proteomics allow generation of large amounts of data from a single experiment. However, high-throughput data are generally of high variation, low reproducibility, noisy (von Mering and Bork, 2002), thus analysis and interpretation of the omics data remain challenging and require effective bioinformatics approaches. Biological interpretation of high-throughput data for forming hypotheses and for guiding experimental validation is typically a downstream process of the omics workflow after the high-throughput raw data are processed for functional analysis. At the core of functional interpretation of omics data is the knowledge (such as annotations and literature data) provided to the biological objects, being genes, mRNAs, or proteins from various molecular databases. Meanwhile bioinformatics tools have been developed for analyzing and interpreting the large lists of genes or proteins, such as DAVID (Huang et al., 2007), BABELOMICS (Al-Shahrour et al., 2005), Ingenuity (http://www.ingenuity.com/) and GeneGO (Ekins et al., 2007) for function and pathway analysis of large-scale data.
While bioinformatics tools have greatly assisted data analysis, a careful review of the major steps and flow of data in a typical high-throughput analysis reveals gaps that need to be addressed. One issue is the lack of standardization when dealing with a large list of proteins or genes annotated in different sources. For example, different protein IDs/names may be used for the same protein in different sources, even different versions of the same database may result in different IDs if the database identifier is not stable. The lack of standards presents a continuing challenge for integrating annotations from heterogeneous databases. Consequently, expression analysis is often carried out in an ad hoc manner, with a fragmented and inefficient use of rich annotations available in various resources. In addition, the effectiveness of the bioinformatics analysis system often relies on the amount and the type of knowledge available for genes and proteins annotated in the databases. To provide effective protein or gene ID mapping and comprehensive annotations for the largescale data analysis, we integrated two databases, UniProt (UniProt Consortium, 2008) and iProClass (Wu et al., 2004), into an integrated bioinformatics analysis system, iProXpress recently developed at the Protein Information Resource (PIR) (Huang et al., 2007). UniProt is a central international repository of protein sequences and functional information and provides the most comprehensive annotations for all proteins. iProClass database is a protein knowledge base providing value-added annotations integrated from over 90 molecular biology databases. iProClass coupled with UniProtKB became a data powerhouse of the iProXpress system, serving as a basic infrastructure for the omics data mapping and as a knowledge source for data analysis and interpretation.
In this paper, we describe an integrated bioinformatics approach for the gene expression and proteomics studies of human fibroblasts derived from patients with ataxia teleangiectasis (AT) who are sensitive to ionizing radiation-induced DNA damage. Radiation induces a myriad of cellular responses, including genotoxic stress signaling, cell cycle arrest, activation of a complex DNA repair machinery, and metabolic changes (Valerie, et al., 2007; Jeggo and Löbrich 2006; Spitz et al., 2004). ATM, or ataxia-telangiectasia mutated was first identified in AT patients in 1995 (Savitsky et al., 1995). ATM plays critical roles in radiation- induced responses (Kastan et al., 2001; Kurz and Lees- Miller, 2004), and has been identified as a potential target for novel radiosensitizers (Sarkaria and Eshleman, 2001; Ahmed and Li, 2007). For example, small molecule inhibitors of ATM or downstream signaling molecules (Kim et al., 1999; Jung and Dritschilo, 2001) may offer a strategy to sensitize tumors to the lethal effects of ionizing radiation while sparing normal tissues.
To identify ATM-mediated pathways underlying cellular responses to ionizing radiation that lead to radiation resistance or sensitivity in cells, the AT-patient derived fibroblasts expressing mutated ATM genes or wild-type ATM were used as models. The two cell lines were subjected to proteomics and microarray experiments, analyzed by global expression profiling and pathway/ network analysis. We showed radiation-induced and ATMmediated major biological pathways and proposed proteins for further validation.
Experimental Data Source
The proteomics and gene expression data were obtained from radiation-treated AT5BIVA and ATCL8 cell lines. AT5BIVA was derived from human fibroblasts of ataxia teleangiectasis (AT) patient with mutated ATM (AT mutated) gene (Jung et al., 1995), while ATCL8 was derived by reintroducing the wild-type ATM gene into the AT5BIVA cells. The two cell lines were exposed to 10 Gy of ionizing radiation and analyzed at time intervals from 30 minutes to 24 hours. The proteomics data were obtained from two-dimensional gel electrophoresis (2D-gel) followed by MALDI-MS of the excised gel spots. The gene expression data were obtained using Affymetrix DNA microarray (U133A probe set of 14500 human genes) chip assays. The experimental procedures for cell culture and radiation treatment, 2D-gel, MALDIMS proteomics and microarray have been described elsewhere (Lee et al., 2001; Mewani et al., 2006). Protein identification from MALDI-MS was based on MASCOT search engine using UniProtKB/Swiss-Prot database. Lists of proteins were identified (with UniProtKB accession #) from differentially changed 2D-gel spots based on >=2-fold changes (p-value <=0.05), increased (including newly appeared spots after irradiation) or decreased (including spots only in control but disappeared after irradiation) for each time point and cell type. Lists of genes were identified (with Entrez Gene #) from differentially expressed mRNAs (increased or decreased) in microarray based on >= 1.5- fold changes (p-value <= 0.05).
Data Integration and Bioinformatics Analysis
We applied an integrated bioinformatics approach for the proteomics and gene expression data analysis. The iProXpress integrated protein expression analysis system (http:// pir.georgetown.edu/iproxpress/) was primarily used as a platform for the functional data analysis, coupled with the Ingenuity Pathway Analysis (IPA) tool for pathway and network analysis. A prototype of the iProXpress system has been applied to several previous high-throughput studies (Li et al., 2004; Chi et al., 2006; Hu et al., 2007). Below we briefly describe the bioinformatics analysis procedures.
Protein mapping: Gene or protein lists were mapped to UniProtKB protein entries primarily based on gene/protein identifiers. Genes with common identifiers such as GenBank, UniGene or Entrez Gene are mapped based on the PIR ID mapping service (http://pir.georgetown.edu/pirwww/search/ idmapping.shtml). For genes with no ID match, the mapping is based on sequence comparison, or name mapping if the sequence is not available. The protein and gene lists from AT5BIVA and ATCL8 cells were integrated into the iProXpress system after protein mapping.
Protein annotation: After protein mapping, rich annotations are described in a protein information matrix that captures salient features of proteins, such as functions and pathways, for given experimental data sets. These rich annotations are derived from comprehensive protein information that have been integrated into the UniProt and iProClass databases and from sequence analysis for homology-based inference.
Functional profiling: The gene and protein lists were divided into experimental groups based on cell types and time course for functional profiling using various functional attributes (i.e. annotation fields of the protein information matrix). Primarily used for functional profiling were GO slims (a subset of GO with high level terms at GO hierarchy) (http://www.geneontology.org/ GO.slims) and pathway information (e.g. from KEGG database).
Pathway and network analysis: Pathway visualization was based on pathway diagrams provided in source pathway databases such as KEGG and the IPA tool. An ATM protein interaction pathway map was also used, which was curated by scientists who initially discovered the ATM gene (Savitsky et al., 1995) and reflects the current state of knowledge for ATM-mediated pathways (available at http://www.cs.tau.ac.il/~spike/images/ 1.png). Network analysis was done using the IPA tool, which dynamically generates functional association networks based on curated literature information of protein-protein interaction, coexpression, and genetic regulation.
Figure 1 depicts genotypes the overview of an integrated bioinformatics approach to analyze and interpret the proteomics and gene expression data from irradiated cells with mutant or wild type ATM genotypes.
Figure 2 shows the iProXpress web interface for searching, browsing, and profiling the experimental groups of different cell types, time courses, and protein or mRNA level changes. The interactive graphical user interface provided several functionalities for data analysis, such as selecting data groups, browsing the proteins and associated annotations, and expression profiling using GO slims and pathways.
Figure 2: iProXpress web interface for browsing and profiling proteomics and microarray data.
1) Selected experimental groups can be chosen from the pull down menu. 2) Boolean operations can be used to query the data, such as "get all proteins identified from proteomics (A_8_30m*) OR microarray (B_8_30m*) in ATCL8 cell at 30 minutes." 3) The "protein information matrix" displays protein list from the selected groups, and provides annotations integrated from over 90 sources. 4) The "Display Option" allows selection of desired fields for display. 5) For the given list of proteins, the interface provides functional profiling "buttons" to show profiles in GO slim (molecular function, cellular component, and biological process) or KEGG pathways. 6) The interface also provides protein sequence analysis tools listed such as BLAST, FASTA and sequence alignment. 7) An example of GO biological process profile. 8) Comparative profiling across selected data groups based on given GO categories.
Radiation-Induced Changes in Expression Profiles
The 2D-gel/MS proteomics and DNA microarray data generated from radiation-treated AT5BIVA and ATCL8 cells are summarized in Table 1, which shows total numbers of UniProt protein entries mapped from proteomics and gene expression data. Most up-regulated proteins were observed at 3hr post-irradiation in both AT5BIVA and ATCL8 cells, and with many more up-regulated in ATCL8 than in AT5BIVA cells. In contrast, most downregulated proteins were seen at 30min in ATCL8 and at 24hr in AT5BIVA cells. At gene expression level, prominent responses to radiation at early time in ATCL8 cells were observed, for example, three times as many mRNAs were up-regulated at 30min in ATCL8 (33) as in AT5BIVA (11) cells, while up-regulation of most genes was only seen 1hr after radiation in AT5BIVA cells. These differences showed that ATCL8 was more radiation-responsive at both protein and mRNA levels at earlier time than the ATM-mutated AT5BIVA cells. Compared to AT5BIVA, ATCL8 cells were shown to quickly respond to irradiation at 30min by increasing more gene expressions and by decreasing the amounts and/or activities (presumably modification states) of more proteins, followed by increasing more at 3hr.
The profiling of the differentially changed proteins or genes from irradiated cells based on GO slims and the KEGG pathways provided global views of functional changes in these cells. Table 2 shows the major GO biological process categories of radiation induced protein changes. The total changed proteins (combined up- and down-) in the two cell lines generally showed similar profiles among top categories of GO biological processes. However, profiles based on up- or down-regulated proteins showed clear differences between the two cell lines. For example, in AT5BIVA cells, a higher percentage of proteins involved in cell cycle was down-regulated (8.3%) than up-regulated (4.8%), and more were up-regulated than down-regulated in RNA metabolism, transcription, and protein biosynthesis. In ATCL8 cells, a higher percentage of proteins were up-regulated in signal transduction and protein modification, while more were down-regulated in protein biosynthesis.
When profiling is performed using KEGG pathways for the total changed proteins, differences were observed in the percentages of proteins involved in purine metabolism, glycolysis/gluconeo-genesis, pyrimidine metabolism, and glutamate metabolism in the two cell lines. Pathway profiling based on the up- or downregulated proteins resulted in more differences between the AT5BIVA and ATCL8 cells. For example, higher percentages of down-regulated proteins in purine metabolism and of up-regulated proteins in starch and sucrose metabolism and folate biosynthesis were observed in AT5BIVA cells. Also consistent with GO process profiles, more cell cycle proteins were seen downregulated in AT5BIVA while more were up-regulated in ATCL8 cells. Overall, metabolic pathways were clearly affected, and purine metabolism was the most affected pathway in irradiated AT5BIVA and ATCL8 cells based on the expression profiling using iProXpress as well as from the Ingenuity pathway profiles (not shown).
Biological Pathways and Signaling Proteins in Response to Radiation
Although the general profiles in Table 2 provided global views of major functional changes in the two cell lines without regard to specific time points, profiles based on more specific or focused data groups, such as at certain time points, offered more biological insights. We selected a proteomics data set at 3hr from both AT5BIVA and ATCL8 cells and a microarray data set at 30min from ATCL8 only for further analysis, when most differentially changed protein or gene expressions were observed or most upregulation of proteins or genes occurred (Table 1). The comparative pathways profiling of four data groups representing the upand down-regulated proteins from AT5BIVA and ATCL8 cells at 3hr post-irradiation showed that purine metabolism is the most predominant pathway with 10 differentially expressed proteins, and major differences exist between the four data groups (Figure 3).
Table 3 lists proteins of purine metabolism from all time points (30min to 24hr) in AT5BIVA and ATCL8 cells. Most enzyme changes in this pathway occurred at 3hr in both cell lines, and those changed at other time points were mostly down-regulated in both cells. Strikingly, while most changed enzymes were downregulated at 3hr in AT5BIVA cells, all changed enzymes were up-regulated at 3hr in ATCL8 cells. Two enzymes with opposite changes were identifiet from the two cell lines, adenylate kinase 2 (up in ATCl8 and down in AT5Cl8 at 30min), and IMP dehdrogenase 2 (up in ATCl8 at 3hr and down in AT5BIVA at 24hr).
Figure 4 shows a diagram of the purine metabolism pathway with differentially expressed enzymes listed in Table 3 superimposed onto the pathway map. Interestingly, most of these enzymes are located at the biochemical steps surrounding the ADP/ATP or GDP/GTP synthesis. For enzymes involved in these steps, most were down-regulated in AT5BIVA cells, while most were up-regulated in ATCL8. This strongly suggests that the ATCL8 cells were able to respond to irradiation by increasing the amount or activities of nucleotide synthesis enzymes to prepare for increased DNA synthesis and repair.
Figure 4: Differentially expressed enzymes in purine metabolism identified from irradiated AT5BIVA and ATCL8 cells. Enzyme Commission numbers (EC#, e.g. 18.104.22.168) are used to represent enzymes in metabolism. Highlighted in green background are known human enzymes annotated in the KEGG database. Differentially expressed enzymes in purine metabolism (Table 3) are superimposed onto this pathway diagram: blue-boxed are enzymes changed in AT5BIVA cells, red-boxed those in ATCL8 cells, and pinkboxed those from both cells. Areas circled with broken lines highlight closely related biochemical steps surrounding ADP/ATP (left) or GDP/GTP (right) metabolisms, which include most of these differentially expressed enzymes from either cell type.
Because of the relatively low numbers of differentially expressed genes from the microarray experiment, expressing profiling using GO or KEGG pathways was usually not revealing for most of the experimental groups (Table 1). Instead we focused on the differentially expressed genes from ATCL8 cells at 30min postirradiation, when more genes were differentially expressed in ATCL8 than in AT5BIVA cells, and most up-regulated genes in ATCL8 occurred. Table 4 lists gene products from the top 3 GO biological process categories, signal transduction, protein modification, and transcription, from the microarray experiment. Among them, p53, BRCA1 and HDAC1 were all up-regulated at 30min in ATCL8 cells and are also well-known to be involved in DNA repair and cell cycle control.
Furthermore, despite the low numbers of differentially expressed genes from microarray experiment, it was interesting to correlate these genes with differentially changed proteins from proteomics data. A total of 103 proteins (UniProt entries) from AT5BIVA and 131 from ATCL8 cells were mapped from from the microarray data of both AT5BIVA and ATCL8 cells (Table 1). Table 5 shows the common protein set of 13 proteins, namely the overlapping genes/proteins between the proteomics and microarray data, 10 were from ATCL8 and 3 from AT5BIVA cells.
Interestingly, from above proteomics and microarray data RRM2 was shown to be increased at both mRNA and protein levels in ATCL8 cells, with mRNA level increased at 30min, and protein level increased at 1hr and 3hr (Tables 3 and 5, and Figure 4).
RRM2-Associated Functional Networks and Pathways
A critical rate-limiting enzyme in DNA synthesis, RRM2 expression increased in ATCL8 cells at 1hr and 3hr after irradiation, while no changes were detected in AT5BIVA cells. Since RRM2 is involved in DNA repair, we wanted to examine the functional association networks involving RRM2 in the context of current proteomic and gene expression data.
Figure 5 shows the network in which RRM2 is connected with several major DNA repair and cell cycle proteins, including HDAC1, p53, BRCA1, and CDKN2A, directly or indirectly. Except for CDKN2A, a negative regulator of cell cycle progression, the other three proteins were all differentially regulated in ATCL8 cells, suggesting that RRM2 play an important role in radiation-induced and ATM-mediated DNA repair processes and cell cycle control.
Since AT5BIVA and ATCL8 cell lines were specifically designed as models for examining ATM-mediated pathways, we used an ATM protein interaction pathway map to examine changed proteins or genes from proteomics and gene expression data. This pathway map (Figure 6, left) shows that two proteins directly interacted with and activated by ATM are p53 and BRCA1, which were up-regulated in ATCL8 cells. Based on the expression data and the network analysis, we hypothesize that RRM2 is involved in radiation induced ATM-p53-mediated DNA repair pathway in the ATCL8 cells (Figure 6, right). RRM2 directly binds p53 and upon irradiation dimerizes with RRM2 to form the ribonucleotide reductase (RR) holoenzyme complex. Increased RR activity will result in an increase in the pool of deoxyribonucleotide precursors for DNA synthesis which is required for DNA repair in response to radiation damage.
In this study we used an integrated bioinformatics approach (Figure 1) to analyze and interpret the proteomics and gene expression data from radiation treated cells with mutant or wild type ATM genotypes. The iProXpress system provides a protein-centric data integration for functional analysis and allows direct comparison of different molecules (mRNA vs. protein) from same samples under study. As functional understanding of the omics data is underpinned by the current knowledge annotated in databases for given lists of genes or proteins from high throughput experiments, it is crucial to maximize the use of known knowledge from heterogeneous databases and resources. The iProXpress system uses both iProClass and UniProtKB databases for data mapping, data analysis and interpretation, and also takes advantage of the extensive informatics infrastructure at PIR, e.g. the Text Search engine for data browsing and searching. One of the most useful features of the iProXpress system is to allow comparison of functional profiles across multiple data sets or groups obtained from different issue/cell types and time points, or from different omics experiments. In particular, while differential profiles with GO slim or pathway terms may not be evident when generated from combined data groups, profiles from more specific groups may reveal clearer differences. For example, purine metabolism became evident when examining individual time points from both AT5BIVA and ATCL8 cells.
While existing annotations from databases are critical for the omics analysis, the knowledge base is still limited. GO has become a common standard for annotation and functional analysis, but currently only about half of all human genes/proteins are annotated with GO terms, and even less with experimentally validated and manually annotated GO functions. Compared to GO profiling, pathway and network mapping provide more biological insight, however, an estimated <10% of human genes/ proteins have been annotated with biological pathways in databases. Therefore, as part of the integrated bioinformatics approach, expert-guided analysis should be coupled with review of scientific literature for functional interpretation of the large scale omics data and for formulation of scientific hypothesis.
The expression profiling and pathway/network analyses have shown that enzymes of purine metabolisms, especially surrounding steps of the ADP/ATP and GDP/GTP synthesis, were differentially affected in irradiated AT5BIVA and ATCL8 cells. RRM2 is a small subunit of the RR complex that is well known for its role in DNA synthesis. RR is the only enzyme responsible for the reduction of ribonucleotides to their corresponding deoxyribonucleotides, providing a balanced supply of precursors for DNA synthesis and repair. It has been shown that an increase in RRM2 protein levels and RR activity in human nasopharyngeal cancer cells results in ionizing radiation resistance, which appears mediated by enhanced ionizing radiation damage repair during G2 phase of the cell cycle. However, overexpression of the large subunit, RRM1, of RR in these cells did not affect RR activity or ionizing radiation response (Kuo et al., 2003). RRM2 over expression is also associated with gemcitabine chemoresistance in pancreatic adenocarcinoma cells, and that suppression of RRM2 expression using RNA interference enhances gemcitabine-induced cytotoxicity in vitro (Duxbury et al., 2004). Human RRM2 has been shown to be a target of p53 through direct protein-protein interaction that leads to the nuclear accumulation of RR subunits after UV exposure (Xue et al., 2003), and inhibition of RRM2 by hydroxyurea results in increased sensitivity to UV irradiation in prostate cancer (PC3) cells (Zhou et al., 2003). Our results suggest that RRM2 is involved in the ATM- and p53-mediated signaling pathway leading to DNA repair in response to radiation in ATCL8 cells, while the ATM-mutated AT5BIVA cells became more sensitive to radiation possibly due to the impaired activation of RRM2 expression.
Most of proteins in this study were derived from the 2D-gel/MS experiment, and not all identified proteins from given 2D-gel spots were responsible for the observed changes. We used this integrated bioinformatics to help rational selection of candidate proteins for validation. Based on common pathways (e.g. purine metabolism) and their differential expression patterns, we can preferentially select those proteins commonly associated with a pathway over those not associated with the pathway for validation. Indeed, the enzyme RRM2, identified from a spot with 40 identified proteins at 3hr and a spot with 12 identifications at 1hr in ATCl8 cells (not shown), was actually one that was most likely to have changed, also consistent with the finding that RRM2 mRNA was up-regulated at 1hr in the same cells.
It was noted that the intersection between changed proteins and genes from the proteomics and gene expression data in this study was small. The lack of direct correlation between changes in proteins and genes from gene expression and proteomics experiments has been previously observed (Jansen et al., 2002; Hewick et al., 2003). This is due in part to the experimental artifacts and in part to differential post-transcriptional or post-translational regulation of genes or proteins. For example, an increased or new 2D-gel spot may result from increased protein phosphorylation without corresponding mRNA changes.Constructing gene regulatory networks may potentially help identify correlations between proteomics and gene expression data when direct correlation between the two is not apparent (Perco et al., 2005).
Besides identifying RRM2 as a potential downstream target of the ATM-p53-mediated pathway for DNA repair in response to radiation, other enzymes in purine metabolism and several other metabolic pathways, such as AK2, IMPDH, and NDK, were differentially expressed as well in the two cell lines. Interestingly, three forms of NDKs (nucleoside diphosphate kinase) were observed to be down-regulated in AT5BIVA cells. NDKs have recently been found to have DNA binding and exonuclease activities (Yoon et al., 2005). It is not clear however whether this is related to the reduced DNA damage repair in ATM mutated cells. Their roles and significance of these metabolic enzymes in the ATM-mediated pathways and in radiation responses remain to be further examined. Currently we are extending this study by applying metabolomics measurement to the two cell lines after irradiation, aiming to identify changes in metabolites in response to irradiation and the anticipated differential patterns in wildtype ATM vs. mutant ATM-expressing cells. Our current proteomics and gene expression data will provide a valuable reference for future analysis and interpretation of radiation damage- induced metabolites. We envision that integration and correlation of proteomics, functional genomics and metabolomics data generated from the same experimental system will provide new biological insight.
In conclusion, we have demonstrated an integrated bioinformatics approach that includes expert-guided examination of data to define radiation-induced and ATM-mediated pathways in cell models with wild-type or mutant ATM genotype. We have shown that purine metabolic pathways were differentially affected in response to radiation, and that RRM 2 was up-regulated only in ATM-wild type but not in ATM-mutated cells. We hypothesize that in this cell model, ionizing radiation activates ATM-p53- mediated pathway that directly targets RRM2 and leads to DNA damage repair, thus increasing radiation resistance in the ATCL8 cells
This work is supported in part by NIH/NCI grant (P01CA074175). The bioinformatics infrastructure for this study was supported in part by NIH grant U01-HG02712.


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Here is the next chapter of this…series.
That is really all I have to say, enjoy!
And here is my little tid bit for recruitment: we are recruiting and we need more staff! We are in need of translators to help out on new projects we want to successfully pick up. Typesetters, cleaners/redrawers, and quality checkers to help out with more releases!
Usotsuki Paradox - Volume 08 Chapter 59: Moment Between Crises

subset: commoncrawl


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Even though Microsoft walked away from its bid to buy Internet search giant Yahoo!, the company is apparently still interested in buying a part of Yahoo!'s action. Microsoft CEO Steve Ballmer said during a technology and media summit on Thursday that he is still interested in striking a deal, according to CNBC.
Mr. Ballmer claimed that he has spoken with Yahoo!'s new CEO, Carol Bartz, and told her that he would like to talk about a potential deal between the two companies.
The two companies found themselves in a long battle in 2008 after Microsoft presented Yahoo! with an unsolicited take over bid. Yahoo! rejected the offer and said that Microsoft was undervaluing the company. The two companies began publicly feuding over Microsoft's buyout attempt that led to a threat of a hostile take over before the Redmond-based company changed its mind and walked away from the negotiating table.
If Microsoft does land a deal with Yahoo!, it would give the Windows and Office maker a substantial leg up against Google in the online search advertising market. So far, it appears that Microsoft and Yahoo! have not officially entered into buyout negotiations.


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This North Face Men's Windstopper Jacket Black Light Gray quilted lines on interior down coat keep the fill shifting, to help eliminate cold spots. North Face Windstopper Jacket hooded exterior shell features ample pockets and attached, adjustable hood for protection against the elements.
* Large, intuitive harness and pack-friendly chest pockets
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text: Potato, Carrot and Courgette Rosti Baby Weaning & baby recipes From 9 months 0 Prep: Under 20 minutes Cook: Under 10 minutes Age Suitability: 9 months Meal Type: LunchTea Portion: Freezable: Continues below ad These quick and easy fried rostis can be served as a whole meal or side dish, suitable from 9 months Ingredients 1 whole carrot 1 whole courgette 1 whole potato 30g butter (or 4 tbsp olive oil) Method Grate the carrot, courgette and potato and mix well in a bowl. Place the grated mixture into a clean tea towel and squeeze out all the moisture. Heat the butter or oil in a frying pan then add the mixed grated veg. You can make one single rosti or several - the mixture will hold together if shaped. Make sure that the mixture is flat and fry gently for 3-4 minutes on both sides or until the rosti's are crispy and a light golden brown colour. Transfer to a plate, mash roughly and serve. Hints & Tips Add some grated cheese to a cooked rosti whilst still warm to make a delicious all-round meal. Alternatively, the rosti can be a great side dish for an older toddler alongside some grilled chicken or fish. After you have squeezed out the moisture from the grated veg, put the tea towel aside for a couple of minutes, then go back to it and wring out the last few drops of water. Like this? Try these: Oven-roasted veggie sticks Roasted veggie kebabs Country vegetable casserole Baby weekly meal planner - 9 months Download our free, award-winning Weaning App Baby Weaning & baby recipes From 9 months Last updated: 15/08/2011 12:19:43 By 0 Comments Share Like this article? Showing of Uh-oh! You need to be logged in to do this... Sign In Sign Up Load more posts Write a comment Related Articles Homemade haddock fish fingers Country vegetable casserole Mediterranean vegetable pasta bake Mini fish pies with cheesy potato topping Oven-roasted veggie sticks Latest on Made For Mums Community Getting Pregnant Pregnancy Baby Toddler & Child Product Reviews & Shopping Competitions News & Gossip


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As one has the habit of doing.
Sometimes there are phrases in one of the three languages that I use on a fairly regular basis that fit better than any translation that I could undertake in another: comme d'habitude is one of these. It can be either slightly pejorative as in "she didn't get dressed until 10 – as was her habit", "he showed up again late for work, as was his want", "they went to the movies yet again last Saturday night – never doing anything out of the mold", etc.
It can also though denote stability or, in my current case, a wonderful state of affairs where one returns to one's habits!
As I can finally dress myself on the days where I don't need a shower or shampoo I am finally free to follow my normal (another way of expressing "comme d'habitude") schedule. And today all the criteria were met for me to finally walk up to the village as was my want before the accident: I could dress myself in a timely manner (without waiting until 10 or later); it was warm enough for the cape that I have to envisage walking outside; it wasn't raining although overcast and, most importantly, the phrase "comme d'habitude" ran through my head like a leitmotif. Habits can be wonderful.
It took me longer than it used to: what is it about a seemingly minor accident that affects one's whole system? But oh the joy of walking into my local bakery on my own two feet, sit down to my usual table, see my coffee buddies and pick up my mail – as usual.
OK that was enough for the morning, if not perhaps for the day, and I didn't refuse the offer of a lift home. Still comforting to know that soon "comme d'habitude" will be simply that – as usual or according to my normal pattern.


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Dearest One,
Mashallah, you are NINE months old! Where does the time go??!!
We had a busy month. You are trying to get SIX teeth out all at once. You visited with your great aunt, khalti Lina, my grandparents, your tete and your jiddo (first time meeting face-to-face) and your second cousins.
You are eating three solid meals. You get a bottle at night still. If we are home, you get breastfed. If baba is home with us, you get a bottle in the morning and another in the afternoon. You enjoy cuddling with baba and spending time with him, alhamdellah. You have lost some weight though, as your appetite seems to have been affected by your teeth coming on. But, you are still growing like a weed, mashallah. Most of your clothes are too small now. Even stuff that are sized for 12 month Olds can be too small on you now, mashallah.
You don't seem interested in crawling at all. The minute your belly is touching the ground, you flip yourself over onto your back with ease mashallah. But, you like walking. All anyone has to do is to grab a hold of your arms and you are putting one foot in front of the other mashallah. While we were visiting the family in New Jersey, we picked up a walker. You weren't sure at first, and it looked like you might not be interested. But it didn't take long for you to get the hang of it and run after us all!
Mashallah, mashallah, mashallah! You are an amazing traveler! The plane rides over to NJ and back, did not seem to bother you. You made friends easily with fellow travelers and flight attendants. The only issue was always keeping you entertained. But, all in all, for a first time travelor at almost nine months old, is an accomplishment. Not to mention that your great grandparents, my tete and jiddo, are huge fans of yours and love you to bits and pieces. The whole family was very happy to have you visiting with them and were enjoying your antics and words. You are more vocal now and try to copy as many sounds as you can hear. One thing that you have done since day one and is the cutest thing ever, is that you hold your right hand's three middle fingers straught and flick them up and down on things. It is charming and cute. Your jiddo, my baba, thinks it's the most beautiful thing ever and was the only one to notice it outside of myself and your baba.
I am attaching a picture of our electronic ticket to NJ, as a momento for this month, since it was your first time traveling on an airplane, the first time you met your jiddo face-to-face, the first time I was alone with you and we were away from baba for an extended period of time, the first time you met your great grandparents. It was a lot of firsts for us both and it was wonderful. I just wish your baba was there too. Inshallah next time. Also, this month, all three of us prayed together! inshallah we continue to pray together and you keep on top of it for your own sake.
I am sorry for the long post. I love you with all my heart, my darling. May Allah always keep you healthy, safe and happy. May you always be guided to the right path, and may you always use Muhammad PBUH to guide your life and actions.
All my love,


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1st Step Arch Supports
1st Step Arch Supports are perfect for those that want incredible support with a slightly forgiving arch support. Start with the 1st Step Arch Supports.
1st Step Arch Supports
1st Step Arch Supports are the ideal orthotic for those wanting to see if orthotics are for them. The 1st Step Arch Support is a very dense foam rubber capable of molding to the foot over time while supporting the foot. 1st Step Arch Supports are used to treat plantar fasciitis, heel pain, tired feet, back pain, arthritis in the feet, bunions, and foot pain. The gentle arch from the 1st Step Arch Support is easily tolerated by most.
Supplied as a pair 1 for each foot.
Select the correct size from the drop down menu.
Note From the Podiatrist:
These arch supports are 1 of a few arch supports that I recommend first to my patients because they are easily tolerated. Patients slip these into their shoes under the insoles and they are off to the races. There is a minimal break in period because the insoles are so gentle on the bottom of the foot.
Additional information
X-Small (W 4-6)(M 3-5), Small (W 6-8)(M 5-7), Medium (W 8-10)(M 7-9), Large (W 10-12) (M 9-11), X-Large (W 12-14)(M 11-13)


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Jasmine Waltz - Exclusive Jasmine Waltz video!
Celeb BB's hottest housemate poses for the camera!
This video was filmed during Jasmine Waltz's world exclusive shoot with Nuts - and what a shoot it was! The Celebrity Big Brother babe knows her poses and loved slipping into that sexy lingerie. Who wouldn't with a body like that!
Filename: septimiu29-Jasmine Waltz - Exclusive Jasmine Waltz video!.mp4
Filesize: 29.2 MiB
Format: MPEG-4
PlayTime: 00:02:11.644
1 video stream(s), 1 audio stream(s), 0 subtitle stream(s).
Video Stream 0:
Length: 00:02:11.566
Frames: 3947, 30.000 fps, 1664 Kbps CBR
Codec: MPEG-4V (Family: )
Dimensions: 640x360, AR: 16/9
Encoder: Lavc55.39.100
Audio Stream 0:
Codec: AAC LC, A_AAC/MPEG4/LC, AAC Low Complexity
Bitrate: 192 Kbps
Bitrate Mode: CBR
Sampling Rate: 48 KHz
Channel(s): 2 channel
Resolution: 16 bits per sample


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It's said that living isn't a life but healthy lifestyle is a life which is real. Wellness is actually much significant not merely stay content for very long time and to stay healthy. In reality, health could be the basic state of someone in many factors.
24 Hour Fitness – Is it a Scam?
Facts, Fiction. 24 hour fitness centres have begun to flourish.
[Read more…] about 24 Hour Fitness – Is it a Scam?
How you can Get the Flow State
Once we're in flow, we experience enormous satisfaction and whatever it's that generates movement becomes it's very own benefit.
This is certainly a direction of the way that it seems to be in a state of movement
How to Increase Breast Size
Exactly what are choices to increase breast size without getting surgery. Who else really wants to read about How Exactly To Increase Breast Size? Here's the things I know about just how to increase breast size.


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Works much like the de-abstraction of the word "now" as in the context that now is all we really have and is where change must manifest. Therefore i try to inhabit the vision of that which i wish to manifest as often as possible. .
Framed by a quantum window, sending this TED talk forward makes for a meaningful now and that counts.
Create intentionally, now.
".... reimagine conventions in design .....create safe objects of long-term value, eliminate waste, and recognize the interdependence of humans and nature as well as the right of each to co-exist. "
As taken from YES magazine -William McDonough and Michael Braungart.


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The process of producing methane from manure is fairly straight forward – seal manure in an airtight container at a favorable temperature and it will break down into biogas: a mixture of Methane (CH4), Carbon dioxide (CO2), and trace amounts of other gases. Behind the apparent simplicity, though, lie complicated interactions involving several communities of microorganisms.
Anaerobic digestion is a multi-stage process (Figure 1) involving two to four steps, depending on where you want to draw lines in the process.
More than 100 different anaerobic microbes are known to contribute to the production of biogas. These microbes are organized into a number of interlinked communities. Communities of hydrolytic bacteria break complex organic matter down to simpler compounds. Acid-forming bacteria convert the simple compounds to volatile fatty acids – principally acetic acid (vinegar). Hydrolysis and acidogenisis are commonly lumped together and called anaerobic fermentation. Some microbiologists also distinguish between formation of mixed volatile fatty acids (acidosis) and reduction to Acetic acid (acetogenesis).
Methane forming Archaeans (very simple, single cell organisms similar to bacteria), called methanogens, take the end products of fermentation – volatile fatty acids, hydrogen gas (H2), CO2, and water (H2O) – and use them to form methane. Methanogens fall into two main camps depending on the pathway they use to produce methane (Figure 2). All methanogens can reduce CO2 and H2 into CH4 and H2O; those that use this pathway exclusively are called hydrotrophic methanogens. Methanogens that convert volatile fatty acids (and a number of other simple organic compounds) to CH4 and CO2 are called acetotrophic methanogens.
Anaerobic digestion is a living process, and like all biological communities, the microorganisms that carry out anaerobic fermentation and methanogensis do so under certain operating conditions.
Given an ample food supply, sufficient room to expand, the absence of predators or competing organisms, all communities of organisms – be they methanogens or human beings – grow in a pattern similar to the one shown in Figure 3. The Lag Time occurs as the organisms acclimate to their environment.
The inhibitory elements that cause endogenous growth are often the end products of a community's metabolism.
The beauty of anaerobic digestion is that it is the work of a mixed community of organisms. The toxic end product of one community is the food supply of another. Acid forming bacteria consume the simple sugars that might inhibit hydrolytic communities. Methanogens use the acids formed in fermentation to produce CH4 and CO2. And in the end, CH4 and CO2 leave the digester as biogas.
Most anaerobic digesters are designed so that the microbial communities remain in exponential growth. An important concept to grasp is the doubling rate or reproduction time of an organism. This is the time needed for a population to double in size during exponential growth. In simple terms it is the time required for the organisms to replace themselves.
Anaerobic digestion generally takes place in a liquid, continuous flow reactor (Figure 4). If the reactor volume does not change, then flow out of the reactor equals flow into the reactor, and the average time that liquid remains in the reactor is its volume divided by the flow rate. This is called the hydraulic retention time of the reactor (HRT).
If the reactor is completely mixed, that is, the microorganisms are completely suspended in the reactor, then the time the cells remain in the reactor equals the HRT. If the HRT of a completely mixed reactor equals the reproduction time of the organisms living in the rector, a new cell is formed to replace each cell leaving the reactor, and the population within the reactor remains stable. If the HRT is shorter than the reproduction time, a new cell will not be there to replace the one leaving, and the population will decline, or "wash out."
Reducing HRT decreases reactor size, and smaller reactors reduce costs; therefore, many digestion systems are designed so that microorganisms remain in the reactor longer than its HRT. For instance, we could put a screen on the outlet of the reactor in Figure 4 so that some of the microorganisms are trapped inside. We can calculate cell retention time by dividing the mass of microorganisms trapped in the reactor by the mass of organisms leaving the reactor. The microbial population is kept stable by setting cell retention time equal to reproduction time.
It is easier to measure the total mass of solid particles suspended in liquid rather than the mass of viable cells; therefore, cell retention time is approximated by Solids Retention Time (SRT), or the mass of solids in the reactor divided by the mass of solids leaving.
Microorganisms need food to reproduce and grow. Methanogens have the uncanny ability to go dormant during periods of food shortage. This is good because you can easily restart an anaerobic digester after long periods of inactivity. On the other hand, sudden increases in feeding can cause bursts of gas production, which lead to foaming and scum formation in the digester.
Methanogens thrive in two temperature ranges. Thermophyllic (heat loving) methanogens are fast growing (reproduction time 10 to 15 days), but they operate in a fairly narrow band of temperature centered on 55 degrees C. Mesophyllic methanogens are slower growing (reproduction time up to 30 days), but they tolerate a wider range of temperatures. 35 degrees C. is optimal for mesophyllic methanogens, but they can tolerate much lower temperatures. Provided they have sufficiently long SRT, digesters operated at 20 degrees C. do not show substantial loses in gas production when compared to those operated at 35 degrees C.
Methanogens are strict anaerobes, meaning the least amount of oxygen is poison to them. Acid forming bacteria are more tolerant of oxygen. So, if oxygen gets into an anaerobic digester, methane concentration will drop and carbon dioxide concentration will increase in the biogas.
pH is an major indicator of reactor health. A complex set of naturally occurring buffers control pH within an anaerobic digester, with organic acid-ammonia and carbonate-bicarbonate being the most important buffers. The optimum pH range for anaerobic digestion is neutral to slightly basic (pH 6.6 to 7.6).
To ensure that there is a living population of methane producing microbes in a digester, materials that contain methanogens are commonly added. When these materials come from an outside source, such as sludge from an active manure treatment lagoon or biosolids from a sewage treatment plant, it is called a "hot start." Sometimes, digesters are brought on line with a "cold start," meaning manure is slowly added to a liquid filled digester until gas begins to form. Hot starts are generally quicker than cold starts. Depending on digester type, a hot start can take between one and six months to bring a digester on line. It may take six months to a year for a digester to reach steady-state under a cold start.
Inhibiting elements are often called toxic, but toxicity is a misnomer. Low doses of most inhibitory agents actually stimulate biological activity. The following sets of chemicals are the major inhibitory agents found in animal manures.
Antibiotics given therapeutically or fed sub-therapeutically to animals have the potential to alter the communities of microorganisms digesting manure from treated animals. The inhibitory effect of antibiotics on gas production has been shown to be minimal if high SRT is maintained and the microorganisms are given time to acclimate.
Disinfectants and cleaning agents can also cause digester imbalance, although reports of upsets have not been reported in the literature. Dilution of cleaning water or bypassing the digester after cleaning is the safest tactic for handling disinfectants. Some doses may prove to have minimal effect on performance if high SRT is maintained.
Ammonium ion (NH4+) and its gaseous relative, Ammonia (NH3), are byproducts of protein digestion and reduction of urea. Concentrations at which Ammonical Nitrogen (NH4+NH3-N) is beneficial, inhibitory, or toxic to anaerobic digestion are given in Table 1. The toxicity of ammonia is highly dependent on pH. NH3, which is the predominant form at higher pH, is more toxic than NH4+. Usually, ammonia is not a problem in manure digesters, except for reactors treating highly nitrogenous materials such as poultry manure; and some poultry manure digesters have been able to tolerate levels as high as 6,000 mg/l if the microorganisms are given time to acclimate.
|Effect on Anaerobic Treatment
|NH4+NH3-N mg/l
|S- mg/l
|No Adverse Effect
|Inhibitory at higher pH values
Sulfate (SO4-) is not an inhibitory substance, per se. Its presence can reduce CH4 production because a group of bacteria called sulfate reducers can out-compete hydrotrophic methanogens for available H2. Competition with sulfate reducers does not usually reduce production from manure digesters since there is plenty of acetic acid around for acetotrophic methanogens to produce gas. The end product of sulfate reduction, Sulfide (S-), can be quite toxic to anaerobic digestion. Like ammonia, the toxicity of S- is dependent on digester pH. Stimulatory and inhibitory concentrations of sulfide are given in Table 1. Also, like ammonia, S- exists in a gaseous form, Hydrogen sulfide (H2S), so its control is a balance between source reduction, gas production, and pH.
The higher the soluble salt content in a digester, the harder microorganisms have to work in order to transport water in and out of their cells. Research has shown that it is mainly the cations (positive ions in solution) that inhibit microbial activity. Stimulatory and inhibitory concentrations of base cations are given in Table 2.
|Stimulatory (mg/l)
|Moderately Inhibitory (mg/l)
|Strongly Inhibitory (mg/l)
|Sodium, Na+
|Potassium, K+
|Calcium, Ca+
|Magnesium, Mg+
It should be noted that most inhibitory substances must be in solution to reduce biological activity in digesters. Therefore, the inhibitory effect of S- is reduced through precipitation of insoluble metal sulfides. Soluble Mg and NH3 are reduced by precipitation of Struvite (MgNH4PO4).
Fermentative bacteria are generally more robust and faster growing than methanogens. The first indication that something is wrong with a digester occurs when the acid formers start to overpower the methane formers. This may show up as a drop in biogas production. First, however, the CO2 concentration in the biogas will increase, and the organic acid concentration of the reactor liquid increases. Both of these will cause a drop in pH. So, daily measurement of pH is a good method to monitor digester health. The relationship between fermentative and methanogenic communities can become so unbalanced, that even the acid formers can no longer tolerate the low pH conditions. At this point we say the digester is "stuck," or it has "soured" or become "pickled." Basic steps to follow in case of digester upset are:
This document was adapted from Factsheet BAE-1747, Oklahoma State University, authored by
Peer Reviewers


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Student Placements
As a recognized leader in childrens mental health, the York Centre provides field placement opportunities for students pursuing a career in childrens mental health. While there may be exceptions, generally the Centre will provide field placement for:
- third year Child and Youth Work students - with the day treatment programs
- Master of Social Work students - with the family / social work team
- Bachelor of Social Work students - with the zero to six program
The educational facility will make recommendations and refer perspective students to the Centre. Candidates will submit resumes and avail themselves for an interview . Successful candidates must obtain a recent Vulnerable Sector Screening Criminal Reference Check and be in good standing with their respective educational institution.
Students accepted into placement at the Centre will be assigned a staff supervisor who will have similar educational background and training (ie. MSW) as the student in placement. The student will meet with their supervisor to establish the goals and objectives of the placement and to determine the evaluation requirements of the student. The supervisor will maintain required documentation and conduct evaluation as per the placement agreement and in accordance with the policies and procedures of the York Centre.
For further information please contact us.


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Litigation Legal Practice Guide 2014: British Virgin Islands
2014年 10月 15日
This article first appeared in the Chambers Legal Practice Guide to Litigation 2014, published by Chambers & Partners.
Arabella di Iorio, Ben Mays and Brian Lacy provide an overview of Litigation Law & Practice in the British Virgin Islands. Their chapter formed part of the Chambers Legal Practice Guide to Litigation 2014.


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The Association of Resident Doctors (ARD), University of Calabar Teaching Hospital (UCTH), on Friday returned to work after a three - month strike.
A News Agency of Nigeria (NAN) investigation showed that activities in the hospital were in full swing as doctors were seen attending to patients.
A communiqué signed by Dr Ukweh Ikechukwu, President of ARD, UCTH branch said the strike was suspended on passionate ground.
According to the communiqué, "the strike is hereby suspended on compassionate ground because of the suffering of patients and work is resuming on Friday, Sept. 11, 2015''.
The doctors had demanded payment of skipping allowances, among others and resolved to reconvene on Oct. 10 to re - appraise the level of implementation by management.
It also demanded that no member of ARD should be victimised on account of the strike.
Meanwhile, some residents of Calabar have expressed relief over the suspension of the strike by the doctors.
A cross section of the residents who bared their minds in separate interviews with the News Agency of Nigeria (NAN) on Friday in Calabar, said the suspension was a welcome development.
Mr Aniete Ukoh, an engineer told NAN that the suspension of the strike was long overdue and urged relevant authorities to meet the demands of the doctors to avoid future occurrences.
"I advise relevant authorities to do everything possible to meet their demands to avoid them returning to strike, '' Ukoh said.
A pregnant woman, Mrs Irene Uwakwe expressed her happiness, adding that pregnant women who registered in the hospital had been receiving skeletal services since the strike began.


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As Canadian hockey fans, we all exhibit symptoms of schizophrenia, don't we? Primarily we:
- hear or see things that aren't real (hallucinations)
- hold strong beliefs that are not real (delusions)
I don't mean to make light of a serious mental disorder that affects many Canadians and their families, but rather to illuminate the tendency of fans to lose touch with reality when observing their favourite team.
Look no further that the acquisition by the Jets of Eric Tangradi.
Should this really be a news? A trade, where the Jets gave up a 7th round pick for a player who has played in 45 NHL games and managed 1 goal and six points is not news. If you are reading this means you are breathing and you would have had an outside chance of picking up one more points than Tangradi playing on a line with Evgeni Malkin and James Neal: you would have only needed to pick up one second assist.
If you are imagining or wondering if Tangradi is some sort of answer for the Jets, you need to breath deeply and and try to focus on thinking rationally.
This post isn't intended as a veiled shot against any of my fellow bloggers or those in the MSM who chose to cover this 'story.' It would be more than a little hypocritical on my part as by writing this post I'm doing just the same myself.
My frustration lies at what is now considered news. From my point of view, it would be considered news if after 5 or 10 games Tangradi actually put up some numbers or created an imapct- that would be a story.
However, as fellow Jets Nation candidate Brett Martin pointed out in an excellent piece called ,"Jets Acquire Eric Tangradi Cheveldayoff Still Boring," when you follow a mediocre team that plays it safe and doesn't make big splashy player moves, you have to find news somewhere.
Wandering away into the comfy confines of reality created by the mind of a fan is not the sole domain of supporters of the Jets.
In Calgary, fans and management are in denial that they only have two players left from the 'glory days' of 3 months in 2004 and are nowhere close to being a contending team.
In Vancouver their window is starting to close and their fans seem intent on proving to other fans why their team is better even though they have never won anything. They also think Burrows isn't a diver and is basically a saint.
In Edmonton, fans want to sell you on what they are seeing their mind's eye rather that what is actually on display.
Do I really need to go into Toronto or Montreal?
Ottawa gets a pass to cry a little. They have just lost their best player and they also lost their top centreman a few weeks ago.
Fans everywhere have to be a little 'touched' to even be showing up after the lockout. Imagine a group of people willing to have their lives orbit around a group of whiny entitled millionaires and their even more whiny and entitled billionaire bosses. Well thats you and me, isn't it?
We all have to be a little crazy, but unfortunately it is that kind of crazy that feels good.


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Hey folks. Sorry for the long absence. I had to have surgery but am doing fine now and am back to work.
A LOT has happened since the last time I posted, so I'm going to kind of do some quick blurbs of what's going on, nothing too detailed. I also recommend visiting Billy Go Boy's website and Facebook page to learn more. "BGB" is an industry insider who has been reporting about the abuse and what goes on in the TWH industry since 1996. Mostly he was posting on the Walking Horse Chat, but due to being kicked off several times (some people can't handle the truth), he started a website and Facebook page in the past couple of months to continue to get the word out. So be sure to visit to learn more!
Wheelon Case
So we've been on pins and needles lately worrying about the Wheelon Case. Wheelon's attorney Rob White tried to get the entire case dismissed stating that the search warrant was invalid due to "woefully lacking" specific details. More information here. Today in court, White also argued that the search warrant did not state that Wheelon was running a business.
However, smarter heads prevailed, and the judge dismissed the motion to suppress (click here to read it) and the preliminary hearing is underway, with the indictment being decided tomorrow.
In the meantime, Jack G. Heffington, a lawyer and industry supporter, has released a motion for return of the horses seized from Wheelon's place to their owners. The motion states the horses were "illegally seized and held amends this motion and would show as basis for the return of the horses to the owners." Whatever that means, since clearly Heffington can't write. Click here for the entire motion as posted on the Walking Horse Chat.
How do we know he's an industry supporter? From the Walking Horse Foundation website:
Jack Heffington was born into the industry. His father and grandfather owned world champion quality horses as early as the 1930s and Jack's love for the breed began when he was a young child. Jack graduated from the University of Memphis and the University of Arkansas School of Law. He has been a practicing attorney since 1971. Jack owns The Scoop Walking Horse Publication and Tan Oak Farms, a well-known breeding operation. He is a member of TWHBEA, Charter Member of WHOA, and owns and shows many Tennessee Walking Horses including the current World Grand Champion Weanling, Bambini. Jack would like the Foundation to serve as a perpetual institution to promote and support the Tennessee Walking Horse.
And his current violations, including one- and two-year suspensions. Conflict of Interest much?
P Jack Heffington, Trainer, Shelbyville TN, The Ritz Blitz, Not Specified, tckt 28871, 7-17-10, 10/05/2010 10/04/2011, SHOW
P Jack Heffington, Trainer, Shelbyville TN, The Ritz Blitz, Other, Define In Next Field, ticket 28871, 7/17/10, 10/05/2010-02/01/2012, SHOW
P Jack Heffington, Shelbyville TN, Bilateral Sore, 10/05/2010-10/04/2011, 1000, SHOW
P Jack Heffington, Shelbyville TN, 37160, Illegal Chains, 06/25/2009-07/08/2009, SHOW
Jack Heffington, Shelbyville TN 37160, Scar Rule, 10/27/2007-11/09/2007, KWHA
Jack Heffington, Murfreesboro TN, Bilateral Sore, 10/03/2002-06/03/2003, SSHBEA
Jack Heffington, Christiana TN 37037, Unilateral Sore, 10/04/2004-10/17/2004, NHSC
Jack Heffington, Shelbyville TN, Equipment Violation, 08/06/2003-08/19/2003, KWHA
Jack Heffington, Christiana TN 37037, Unilateral Sore, 05/06/2002-05/19/2002, NHSC
PSHA vs. Everyone Else
The PSHA has continued to show their true colors. Now that the judge ruled in favor of the USDA concerning the SHOW/USDA lawsuit, SHOW is facing decertification before Black Week, which starts next week on August 21.
To get back at TWHBEA, PSHA is now telling them that they won't give them the show results to include in the iPEDS database. This literally made me laugh out loud, and I'm not kidding. Seriously, do they really think that people aren't going to find out who the winners are? Are they just going to keep it a cute little secret and not announce in the Voice or other publications who won? It's like a bunch of children on a playground with these guys. PSHA's attacks are getting weaker and weaker.
Many sound horse advocates have countered with a petition: TWHBEA (Tennessee Walking Horse Breeders and Exhibitors Association): Erase all references to Celebration awards on their registration and Ipeds. I signed this because I believe the general public no longer wants a WGC that was on stacks and chains. Anyone who spends any time studying TWHs will discover that most WGCs are one-trick ponies that can't even gait correctly without being stacked and sored. The true, natural, four-beat gait is being bred out of the TWH because of the current "need" for a crippled spider in the show ring, and TWHBEA could help by no longer recognizing it.
PSHA has also hired Purple Strategies yet again (how the heck do they even have any money left?) who expressed their opposition to S 1406 by publicly saying this. My thoughts are in green text.
Performance Show Horse Association Announces Opposition to PAST Act
Monday, August 05, 2013
Today, the Performance Show Horse Association (PSHA) shared its opposition to the Prevent All Soring Tactics (PAST) Act, being offered by U.S. Senators Kelly Ayotte (R-NH) and Mark Warner (D-VA). The House version – in amendment form – was introduced earlier this year by Congressman Ed Whitfield (R-KY).
"PSHA strongly opposes the PAST Act because it would destroy an entire equine division, costs tens of thousands of jobs, eliminate hundreds of millions of dollars in economic activity and millions in charitable giving, devastating communities and people's lives throughout the country," said Terry Dotson, PSHA Chair.
The PAST Act would:
• Eliminate a total division of the equine breed, impacting more than 10,000 horses that would be deemed no longer fit for their intended purpose; As far as I understand, there aren't even 10,000 living TWHs that are registered. From what the Industry itself has said, only 10% of all registered TWHs are stacked horses. As far as I understand, 10% of 10,000 is not 10,000.
• Take from hard-working taxpayers hundreds of millions of dollars of investments and income without cause or any scientific evidence; Where are these numbers coming from? What's their basis for this assumption? I want to see their evidence of this.
• Eliminate the Horse Protection Act's self-regulation, which is far superior to that of the federal government and which does not need the hundreds of new federal employees required by Whitfield's amendment; I seriously laugh every single time I read this. The OIG reported in September 2010 that the HIOs are not doing an adequate job and need to be eliminated. It is proven time and time again that more horses are found sore when the USDA is there than when they're not. And what happened to that public swabbing initiative that TWHSO (who has disappeared, by the way) was doing for the Celebration last year? The USDA found lots of foreign substances on a lot of horses' legs, but the TWHSO swab results are nowhere in sight. Where's that transparency now?
• Create a new federal bureaucracy that will result in a cost of tens of millions of dollars for taxpayers. No, it won't. VMOs are already on staff at USDA. All they have to do is train them to inspect horses. Not that hard to do. And again, where is this "millions of dollars" amount coming from?
Quite frankly, I thought the Horse Protection Act was just that: the HORSE PROTECTION Act. Not the TWH SORE HORSE INDUSTRY PROTECTION Act. I'm pretty sure I'm correct on that.
Black Week
Black Week, aka the National Celebration, begins soon, and the entries are down again. From the Walking Horse Report.
The entry deadline for the 75th Anniversary Celebration closed on August 6, 2013. Preliminary class entries were down 22% from 2,671 entries in 2012 to 2,081 this year. The total number of entries will grow with the championship entries on the last weekend. Preliminary classes begin August 21st and continue through August 28th.
The total number of horses that make up those entries also declined. In 2012 there were 1,469 horses while this year that number slipped to 1,112 horses, a decline of 24%. Continued economic pressure along with industry turmoil and unrest have resulted in a decline of 1,336 preliminary entries since 2009, a decline of 39%.
We also have to remember that just because there are this many entries, sometimes horses don't show for various reasons. People can scratch classes for whatever reason (horse is tired, injured, etc.), not necessarily because their horse is sored.
It has also been confirmed that 11 classes will be split. A split class is usually done so because of a large number of entries. But after finding this information on the Walking Horse Chat regarding the Aged Stallion class split, seems like the classes weren't that big... Just another way for PSHA/SHOW/TWHNC to get more money, I guess.
Aged Stallion:
A Division:
Moody Star - Bobby Hugh [HPA violator]
He's Vida Blue- Chad Wiliams [HPA violator]
Seige - Scott Beaty [HPA violator]
Lucky Grand Slam - Charlie Witherspoon [HPA violator]
He's Slim Shady - Jimmy McConnell [HPA violator and brother to Jackie McConnell]
Swing Batter Batter Swing- Danny Hughes [HPA violator]
I'm Deuce - Casey Wright [HPA violator]
B Division:
Lampshade - Bill Cantrell [HPA violator]
The Metalist - Herbert Derickson
The Golden Sovergin - Chad Williams [HPA violator]
The Thug - Dick Peebles [HPA violator]
A Bruce Pearl - Jimmy McConnell [HPA violator]
Putting Cash on the Line - Jimmy McConnell [HPA violator]
Honors - Larry Edwards [HPA violator]
Jose's Crystal Lite - Charlie Green
In a twist of good news, two of the largest TWH shows before the Celebration, Belfast and Wartrace, were extremely disappointing. Belfast was completely canceled, and Wartrace had minimal exhibitors and small turnout in the stands. There used to be five or six people deep around the arena to watch, and the stands were full. Now take a look:
|From Walking Horse Report Facebook page
Three belong to the HSUS, as pictured below.
This one has been posted anonymously by what I've gathered are basically concerned citizens who are worried about the horse. No other information has been found.
These billboards are hard to ignore and clear reminders to the City of Shelbyville and those who continue to support this nonsense of why the Celebration needs to end.
There is a rumor going about that SHOW was going to add the mandatory penalties into their rulebook on August 7, 2013, which is after the August 6, 2013 deadline to turn in entries. So far I haven't gotten confirmation of this.
Finally, so far the USDA has not moved on decertifying SHOW before the Celebration, so we ask that you send in your emails and call them to ask them to do so. Look up Dr. Chester Gipson, Dr. Rachel Cezar, APHIS Administrator Kevin Shea, and USDA Secretary Tom Vilsack for contact information.
In the meantime, the USDA has announced they will be pulling shoes at the Celebration, which is a massive change and would expose more and more of these monsters.
Information from Billy Go Boy's page tells us this, which is also good news:
According to high level USDA sources, THERE IS PRESENTLY NO 30 DAY GRACE PERIOD BEING AFFORDED TO SHOW HIO REGARDING ENFORCEMENT OF THE MANDATORY PENALTIES. SHOW HIO DECERTIFICATION WAS IN PROCESS BEFORE JUDGE MEANS RULING. More updates as there are developments.
I'll keep you posted as I learn more.
From The Commercial Appeal: "State prosecutors are asking that four Tennessee Walking Horses identified as abused in the criminal case against horse trainer Jackie McConnell be taken from their owners."
I can't tell it any better than the article itself. Below is copied and pasted from The Commercial Appeal's article written by Lela Garlington and posted August 13, 2013. Click here for more information from The Republic in Indiana.
Tennessee wants abused Tennessee Walking Horses taken from owners
State seeks abused horses in forfeiture case
State prosecutors are asking that four Tennessee Walking Horses identified as abused in the criminal case against horse trainer Jackie McConnell be taken from their owners.
Similar to a Witness Protection Program, the horses are at an undisclosed location in Tennessee but outside of Fayette County. Since the horses were seized in March 2012, the Humane Society of the United States has paid for the cost of security, boarding, treatment and care.
The state in the lawsuit filed in Somerville claims the owners either knew or should have known that McConnell, who lives in Collierville, was barred by the USDA from training or exhibiting horses for shows. The suit said McConnell was widely known in the Tennessee walking horse exhibition community and was inducted into the Tennessee walking horse Hall of Fame before his name was later stricken because of horse soring.
"The criminal state statue specifically allows for the forfeiture of abused animals," said Atty. General Mike Dunavant, who filed the lawsuit Friday in Fayette County Circuit Court. "The issue now is they knew or should have known. That's the heart of the matter."
McConnell and two others pleaded guilty in state and federal courts on charges they beat a horse with a wooden club, used an electric cattle prod on the nose and hindquarters, and painted chemical compounds on the horses legs solely for training them to compete in horse shows. The horses were under his care at Whittler Stables in Fayette County.
The Humane Society secretly shot video of the abuse.
Last month, McConnell was placed under house arrest and fined $25,000 after pleading guilty to 12 misdemeanor counts of animal cruelty. In September, McConnell received three years probation and $75,000 fine after pleading guilty to felony charges of violating the Horse Protection Act in Chattanooga.
The owners were not charged with abusing the horses either in state or federal court.
When reached at her and her husband's White Oak Farm in Atoka, horse owner Lisa Marbry said she couldn't comment about the forfeiture proceedings regarding her horse, "I'm Nancy Lopez."
"My attorney has instructed me not to talk about this," she said. Her attorney, Steve Farese, was in trial and could not be reached for comment.
The other horse owners are Joe Privett of Gates, Tenn., who owns "Taj Mahal," and Beverly Sherman of Dallas and her daughter, Kelly, of Boulder, Colo., who own "Paroled in the Night" and "Mucho Bueno."
Beverly Sherman's husband is Floyd Sherman, who is the CEO of Builders FirstSource and have been profiled in both Forbes and Business Week. Their attorney, J. Houston Gordon of Covington, said his clients just received the complaint.
"We don't think the state has complied with the forfeiture proceedings," he said. "We also do not think that Humane Society of the United State has any standing to hold these horses. They and the state have essentially taken these horses unconstitutionally."
As long as the federal and state criminal charges were pending, Gordon said his clients "had no problem using the horses as evidence." But now that McConnell has pleaded guilty and the horses are no longer needed as evidence, Gordon added, "Neither the state nor anyone acting with or on behalf of the state has the right to take anybody's property without due process."
Regarding the lawsuit's contention that the horse owner knew or should have know, Gordon said, "Obviously people wouldn't have allowed their horses to be abused if they knew it. My clients and I don't approve of any animal abuse period."
Beverly and Kelly Sherman sued the state and the Humane Society in May, 2012 demanding that their horses be returned. The case has not been heard and likely will be combined with the latest lawsuit.
Circuit Judge Weber McCraw will oversee the forfeiture proceedings. No hearing date has been set for the matter.
While there were other abused horses that were named in the state animal cruelty indictment, Dunavant said not all of the horse owners are disputing the forfeiture. McConnell also forfeited ownership of his horse named in the indictment.
"All of these owners have had the opportunity to relinquish these horses and give them a chance of a life free of cruelty," said Keith Dane, equine protection director of the Washington-based Humane Society. "They have all suffered abuse."
If the judge rules in favor of the state, the suit asked that ownership be turned over to the Humane Society and any cost associated with the care and upkeep be charged to the former horse owners during the time of their seizure.
© 2013 Memphis Commercial Appeal. All rights reserved.
And More News...
AVMA-AAEP Soring Discussion Draws Overflowing Crowd on Capitol Hill Seems that soring is getting more and more popular. Looks like a bigger crowd than the audience at the Wartrace show!
L&H Distributing From Tullahoma, Tennessee Will Be Serving Beer At the Celebration. Click here for article from Walking Horse Report. Seems that the Coke distributor from Tullahoma that sponsored last year isn't going to be back this year...hmm, I wonder why?
Roy Exum: Budweiser is Not a Sponsor It's important to note that Anheiser-Busch and Budweiser are not the new sponsors of the Celebration, and that they were shocked to learn that L&H Distributing was sponsoring. Be sure to read this article completely to learn more.
That's about it for now...Stay tuned to FTTWH and Billy Go Boy for more information!


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Monday, August 17, 2009
Join our community!
Become a Facebook fan of The Ansel Adams Gallery
Posted by The Ansel Adams Gallery at 11:11 AM
2 comments:
Beautiful work, as always! Congrats on your show!
Carolina Glaubitz
- August 20, 2009 at 9:34 AM
- professional retouching said...
A stunning landscape photo enhanced by the perspective and the reflection in the lake!
- August 23, 2009 at 6:46 AM
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|Your Compass.
photo and text by: Aria E. Appleford (http://neoscii.blogspot.com/)
Everything in this world is energy. Some of the energy vibrates at a much slower rate and gives the appearance of something solid while others vibrate at such a high rate as to be invisible. Life itself is motion – we are constantly in a state of flux and change. The fuel for that motion are the feelings we have and the ensuing thoughts that follow. These then create our emotions.
To navigate our lives we do have help. There is a sense within all of us of what feels right for us. If we have lived an authentic life we are very attuned to that, if our life has been filled with fear and guilt we might be miles away from it and need to do some work clearing the debris from our lives that keeps us from our authenticity. This sense is our compass and it can guide us towards the right things to do in times of chaos. SIlva Ultramind can help us find our compass. The problem many of us have, is that the world has taught us to worry about what everyone else is doing and that can often pull us off track and we lose the connection within ourselves. Our compasses can speak to us of things and actions that make no logical sense to others – however they feel completely right to us.
Think of how you use a compass – you look at it, you begin to walk in the direction it points, after walking a while you hold it again and walk in the slightly corrected direction. The point here is to simply become immersed in what your compass tells you right now. You may find that next check of the compass readjusts you a bit however, in doing what is right here, right now, you ARE moving forward and bridging the chasm of chaos. You may not see the whole picture or understand all the little reasons of how this will fit together – but once safely across and on the other side you will be amazed how perfectly your inner compass works … when you let it!!
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Could you be a volunteer for the Samaritans?
That is the question being asked of Ribble Valley residents as the charity that helps people in need launches a recruitment drive.
An appeal has gone out for volunteers to sign up to support people who may not have anywhere else to turn. Anyone with a few hours to spare each week would be welcome to help the charity that has a wide range of roles available to suit people with different skills, from taking calls to the helpline, providing administrative and IT support or organising fund raising activities for the Blackburn, Hyndburn and Ribble Valley branch.
The Samaritans offers round the clock emotional support for anyone feeling down, depressed, anxious or unhappy and the confidential service is offer by phone, email, text, letter and face to face.
If you would like more information o becoming a samaritan the next information session is on Monday, November 12th from 7pm until 9pm at the branch at 105, New Park Street, Blackburn. Alternatively ring 01254 682266 or email firstname.lastname@example.com.


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Welcome to the 200th 1NightStand Celebration featuring author Jessica E. Subject!!!
Guest Post
If I ever met an alien...
by Jessica E. Subject
There was a time in my life where if I had met an alien, I would have run away screaming, afraid they would want to abduct me into their spaceship, perform experiments, or worse. This belief had much to do with the movies SIGNS, and living in a farmhouse surrounded by corn fields at the same time. And the fear traveled with me when we moved back to the city.
It was a fear I had to get over and fast, with children in the house. Except for a fictional movie, I had no basis for this fear. I've always believed in aliens, but why do they have to be scary? Why can't they be sexy? My favorite superhero is Superman, and he's a sexy alien.
In some of my 1Night Stand stories, a character is meeting an alien for the first time, set up with them by the magical Madame Eve. Though the French woman knows what each applicant can handle, it stills comes as a surprise to the character when they learn their date comes from worlds away.
In both Celestial Seduction and Satin Sheets in Space, the women are shocked and scared when they first find out. But then curiosity takes over, followed by lust. This is exactly how I think I would react in their position. And it would be an experience I would never forget.
Now, I welcome the chance to meet an alien, especially if they are as hunky as in the books I read and write. If the space visitors happen to resemble those in INDEPENDENCE DAY, WAR OF THE WORLDS, or even SIGNS, I'd stare in awe for a few moments before I turned and ran. LOL
About Jessica E. Subject
Jessica Subject is the author of contemporary and science fiction romance, ranging from sweet to erotica. In her stories, you could meet clones, or a sexy alien or two. You may even be transported to another planet for a romantic rendezvous.
When Jessica isn't reading, writing, or doing dreaded housework, she likes to get out and walk. Fast. But she just may slow down if there is a waterfall nearby.
Jessica lives in Ontario, Canada with her husband and two energetic children. And she loves to hear from her readers.
Jessica is a member of RWA and the Toronto Romance Writers chapter, EPIC, and Savvy Authors. She blogs weekly at Backward Momentum, and monthly at Paranormal Romantics. You can find her at jessicasubject.com and on twitter @jsubject.
Website/Blog | Newsletter | Twitter | Facebook | Goodreads | Amazon | Authorgraph
Check out Jessica E. Subject's works:
Celestial Seduction
Erotic Sci-Fi Romance
At the end of his obligated commitment to the Space Service, Frey Berger decides to stay on Earth to further experience human emotions rather than return to Ginnun where his intended has already mated with another. Looking for real love and someone to accept him for what he is, Frey enlists in Madame Evangeline's popular dating service. Although he does not understand the concept of a one-night stand, he hopes to find his perfect mate.
Carrie Cooper's husband divorced her for someone who could provide him with children, leaving her with little self-confidence and no desire to let a man hurt her again. Her best friend convinces her to join a dating service, claiming Carrie needs to get laid, while not winding up with a broken heart since her date requires no commitment after the night is over.
Although several misunderstandings nearly derail them before the date has even begun, Carrie and Frey end up becoming closer than they ever believed possible. Still, Frey will need to convince Carrie that her perfect mate isn't human at all, and that one night can lead to a forever love.
Available From:
Beneath the Starry Sky
Erotic Contemporary Romance
Left at the altar with bald patches on her head and her self-confidence stripped away, Tamara Johnson must pick up the pieces of her life and face her Alopecia Areata, the autoimmune disease causing her hair to fall out. She is convinced no man will date a bald woman and arranges a one-night stand through Madame Evangeline's popular service, 1Night Stand. With her new wig held firmly in place, Tamara arrives at the Castillo Hotel in Las Vegas for a night of passion and pleasure. Anything more than one night would mean exposing her bald head, and she will never do that to any man again.
Running into the Castillo Hotel, away from the paparazzi constantly at his heels, Josh Summers wants to spend his next few days locked away from the flashing lights and screaming fans. When he finds an incredibly sexy woman lying on his hotel room bed in nothing but lingerie, he thinks management has made a mistake. Leaving her behind, he heads for the lobby to get the situation rectified. But an unexpected text from Madame Evangeline sends him back to his room. Though unsure whether the woman on the bed is really his perfect match or just another fan believing him to be like his character on television, he knows he must find out, even if it means he must risk telling her his own secrets. If Tamara can handle his imperfections, she just might be his ideal woman.
Available From:
Unknown Futures
Erotic Lesbian Sci-Fi Romance
Attacked as a young adult, Jewel is left scarred and alone. She applies to the 1 Night Stand service, believing no one can see past her appearance to love her. But she doesn't expect to be sent to a space station where her date could be any number of unusual alien species.
Flavia left everything behind to be one of five humans working for the Space Service space station. Extremely homesick, she is excited to learn she will be a guide to a woman from Earth visiting the station for a couple days. She is immediately drawn to the petite woman, seeing past her scars to her true inner beauty, stirring unforeseen desires.
Unfortunately, Jewel is there for a date which Flavia hasn't been told about. Can she contain her lust and simply show her a good time? Or will she let her desire lead them into an unknown future?
Available From:
Amazon (US/UK) | Decadent Publishing | All Romance | Barnes&Noble | more
Satin Sheets in Space
Erotic Sci-Fi MFM Ménage Romance
Danyka's unusual height and radiant red hair earn her the cold shoulder from any guy she approaches, unless he wants to experiment with a giant. So when her boss sets up on a one night stand, she hopes for a prince but expects the usual toad.
As a captivating stranger whisks her away from the ball and introduces her to his equally gorgeous brother, she fears her fantasies have become a nightmare. Has she finally given into impulse only to become a lab rat in an unknown world?
Available From:
Sudden Breakaway
an Erotic Interracial Sci-Fi Military Romance with light bondage
Dressed in her black designer suit, Paige Brown recruits men and women for the Space Service. After the end of her marriage, and several failed recruiting missions, she yearns to find a way out of her lifelong contract to live a normal life. To find love again.
Jared Barnes's life had been flipped upside down by two little kids. When his former Commander's wife shows up to give him a reprieve, he jumps at the chance for a break, and to visit his former squad. But it is not his buddies he meets in the private suite at a hockey game, instead a woman who has been the star of all of his recent fantasies, fulfills his wildest dreams except the one that binds his heart.
Has Madame Eve made a mistake or can two lonely hearts overcome seemingly insurmountable obstacles?
Available from:
Erotic Sci-Fi Romance
Limited by status…
Ava is a housekeeper aboard Star Spirit, a luxury cruise ship travelling between the Milky Way and the Belvarian System. Coming from a middle-class family on Earth, she is considered by passengers as nothing more than an object to be tossed away when they're done with her.
Shunned because of his lineage…
Banned from several planets across the universe, Nate struggles to run his interstellar shipping business. But the DNA of an exterminated race runs through his blood, giving him a diluted form of their powers, and leaving him labeled as a terrorist.
Both searching for the one…
Through her 1Night Stand dating service, Madame Evangeline matches these two together for a celestial escape on the pleasure planet, Elatia.
Will Ava and Nate find what they're looking for, or will their date be just another night on another planet?
Available from
Decadent Publishing | Amazon US / UK | All Romance | Barnes & Noble | more
Intergalactic Heat
5-story 1Night Stand anthology
A date on a space station? Alien lovers? Madame Evangeline, owner of the exclusive 1Night Stand dating service is known to be magical in fulfilling the ideal dates of her clients, but she's outdone herself this time, bring together beings from Earth and beyond. As long as applicants are willing to let go of their pasts and open their hearts and minds, she will find their perfect match, whether they be a former sci-fi television star, a nurse, an employee of the Space Service, a former Marine, or even an extraterrestrial. Because everyone in the universe deserves a happily ever after…
Coming Soon!!
His Alien Virgin
New Adult Science Fiction Romance
She is desperately seeking freedom...
On the eve before her eighteenth birthday, Skylar steals a ship and escapes from the planet Rebed. She refuses to be another wife to the slimy space pirate that is her betrothed. After hearing of her brothers' own success, she applies to the 1Night Stand service on her way to Earth, hoping Madame Eve can find a man to be her hero, to make her feel special, even if for only one night.
He is trying to fit in...
Daran traveled to Earth years ago, and he still struggles to fit in on the planet. But when his boss suggests a blind date, he reluctantly agrees. What would happen if anyone found out about his alien ancestry?
And they are not at all meant to be together...
As best friend to Skylar's brothers, Daran has always forced himself to fight his attraction to her. He cannot see her as anything more than a little sister, someone to protect. So, why did Madame Eve match them together?
Has Madame Eve made a huge mistake, or will Daran tread into forbidden territory to be with the one woman he's always wanted, and become the hero Skylar craves?
WIN a signed print edition of Intergalactic Heat by Jessica E. Subject and book swag (Canada and US only) or an Intergalactic Heat eBook Bundle (International). Simply enter using the rafflecopter form. No purchase necessary. Must be 18 or over to enter. Good luck!
Grand Prize Giveaway!!
1Night Out on 1Night Stand's Madame Eve!
Grand prize is a $25 Fandango gift card plus 5 1NS titles of the winner's choice
2nd prize is a paperback from the vault plus swag
3rd is 2 1NS ebooks of winner's choice (5 winners)
*Must be 18 or older to enter.


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Friday, October 2, 2015
Red Leaf Tea, Acai Berry Matcha
Our fabulous Matcha Tea cannot get any better than this! Its high-quality Green Tea leaves are young and processed in a special way that maintains all of their amazing health benefits. Matcha Tea features multiple healthful qualities that keep your body and mind strong and alert. Matcha Tea helps detoxify the blood, has cancer-fighting properties and also kills free radicals.
So, what could we possibly do to make our Matcha Tea any better? We have taken our amazing, healthy Matcha and supplemented it with Raw Organic Freeze-dried Acai Berry powder. If you have followed any health buzz for the past few years, you know about Acai. If you haven't been following it, let's take a look at what Acai can do. Acai has ten times the antioxidants of blueberries or grapes. Antioxidants, like Matcha, help fight free radicals and keep your body healthy and strong all year round.
Pure Matcha powder from green leaves, Brazilian Acai Natural Flavor
My Review:
Today I am reviewing Acai Berry Matcha from Red Leaf Tea. I debated with myself for quite a while, trying to decide how I would prepare this one. More on that in a moment.
I ordered a sample size to try out something new and different. The sample bags are small and not resealable. Once opened I fold the top over and close it with a paper clip. I usually prepare the rest in a day or two.
This sample uses the basic level matcha. It is a little darker than most of these blends have been, apparently from the powder acai.
The scent is pretty intensely berry. My first thought was to make a cup with hot water. I normally don't drink my matcha that way. I like a cold milk latte. I really wasn't sure milk and acai would be a good mix, but, I decided I would do it for science! If it turns out badly, you'll never see this and I will be writing about a hot cup today :)
Next I added an additional 4 oz of cold milk and lightly mixed. As you can see in the picture, the color is kind of light. The foam is a much smaller amount with bigger bubbles than before adding more milk. Normally I use a big tumbler for extra blending room, but it is hard to get a good picture through the graphics.
The taste is very berry! I have never had an actual acai berry. I have had a few flavored acai teas. This one is the most intense - in a good way. It is kind of its own flavor, somewhere between blueberry and blackberry, with floral notes that remind me of a light rose.
I needn't have worried about adding this to milk. It definitely holds its own and with out a taste conflict. I added no sweetener to the mug as I often do with matcha. Seriously, this does not need sweetened. The natural sweetness is more than enough. I kind of expected some tartness. I did not notice any in the cup.
The only downside I can comment on is this did not mix thoroughly. Some sediment remained in the bottom of the cup. It is possible this would not occur in a hot preparation. I was just really in the mood for a refreshing cold mug of joy. Mission accomplished.
You can find Red Leaf Tea Acai Berry Matcha here.
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Monthly Archives: June 2015
Our 2015 Billionaire Mailing List Update Is Ready For Purchase!
June 23, 2015 by Billionaire Addresses
Thank you for waiting for the update to our 2015 Billionaire Mailing List. There are 536 billionaires available to contact …
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"Queen of Versailles" Daughter Found Dead In Mansion
June 8, 2015 by Billionaire Addresses
The 18-year-old daughter of David and Jackie Siegel, who were featured in the 2012 documentary "Queen of Versailles", was found …
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See The House McDonalds Built – The Former Joan Kroc Mega Mansion
June 3, 2015 by Billionaire Addresses
A prominent local businessman and his wife bought the home of the late McDonald's philanthropist from the Salvation Army way …
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*** udpate 1.5.2013
We have a few new entries. Yes, we allow entries after the deadline. They only get 8 names, and really who the hell cares? Approx half our list is totally new folks to our Dead Pool challenge. So, welcome aboard, you sick fuckers!
the rules? Well, there are many. Really, a lot. However, it is mostly this simple. Whomever has the most people on their list die in 2013 wins. If there is a tie, that is where the rules come in play, and I thaw the lawyers. Pray not for the lawyers, friends. They are in a billing stasis not unlike what Jabba did with Han Solo. Once they are loosened on the world, the pain comes forth like Austin Powers' urine stream after being frozen for 40 years.
and in the event that can't be worked out, it is simple. The failsafe is that I win.
Derron M
Patrick S
Grace Slick
Bob Dole
Fats Domino
Oprah Winfrey
Ozzy Osbourne
Johnny Winters
Leslie West
Donald Sutherland
E-Murph/ Erin M/ Lilly von Schtupp
Chris Michas aka Mr von Scthupp
* normally, I would simply list names. However, this ranty gem of an email must be published in its glorious entirety
Nancy Reagan
Mickey Rooney
Carol Channing
David Rockefeller
Kirk Douglas
Abe Vigoda
Vera Lynn
George Herbert Walker Bush
Olivia de Havilland
Sid Caesar
Hillary Clinton
A note on my alternate, I don't think she is actually sick or injured right now despite what is being reported. I think its a political ploy and it would not hurt my feelings any if she croaked in the next few hours.
Another note to any government agency that is likely reading our emails. This email contains no threats intended or implied toward any celebrities or political figures and is merely speculation.
Kevin/ Lono/ Master of Death Emeritus
1. Kat Williams
2. Lindsay Lohan
3. George Bush (sr)
4. Zsa Zsa Gabor
5. Fidel Castro
6. Don Pardo
7. Nancy Reagan
10. BB King
Jamie Guerin aka the Arizona Monkey Boy (really, that is his email)
1. Honey Boo Boo
2. Snooki's baby
3. Angus Jones
4. Lance Armstrong
5. Bob Dole
6. Glenn Campbell
7. Bart Starr
9. The voice of Elmo guy (* editors note: Kevin Clash)
10. Lindsay Lohan will have at least 2 abortions. (* ed note - do you mean on top of the 3 she had this weekend?)
I can't think of anyone GOOD. ok. So here is my (unresearched) list:
Leann Rhimes
Aretha Franklin
Dick Cheney
HW Bush
Zsa Zsa Gabor
Kirk Douglas
Muhammad Ali
Billy Graham
Kim Kardashian
Malibu Julz
Lindsay Lohan
Zsa Zsa Gabor
Stephen Hawking
Ariel Sharon
Jan Michael Vincent
Fidel Castro
Rose Marie
Jake La Motta
Shirley Temple Black
Alternate: Aretha Franklin
Roy (aka Majikwah)
1. George W H Bush
3. Zsa Zsa Gabor
4. Stephen Hawking
5. Nancy Reagan
6. Billy Graham
7. Lawrence Walsh
8. Fidel Castro
9. Patrick Macnee
10. Helmut Schmidt