text:
In silico prediction of B- and T- cell epitope on Lassa virus proteins for peptide based subunit vaccine design
Sitansu Kumar Verma1, Soni Yadav1, Ajay Kumar2
1 Department of Biotechnology, Madhav Institute of Technology and Science, Gwalior, Madhya Pradesh, India
2 Department of Biotechnology, Faculty of Engineering and Technology, Rama University, Kanpur, Uttar Pradesh, India
|Date of Submission||28-Feb-2014|
|Date of Acceptance||05-Jan-2015|
|Date of Web Publication||28-Sep-2015|
Sitansu Kumar Verma
Department of Biotechnology, Madhav Insti tute of Technology and Science, Gwalior, Madhya Pradesh - 474 005
Source of Support: Nil, Conflict of Interest: None declared.
Background: Lassa fever is a severe, often-fatal and one of the most virulent disease in primates. However, the mechanism of escape of virus from the T-cell mediated immune response of the host cell is not explained in any studies yet. In our studies we had aimed to predict B- and T- cell epitope of Lassa virus protein, for impaling the futuristic approach of developing preventive measures against this disease, further we can also study its presumed viral- host mechanism.
Materials and Methods: Peptide based subunit vaccine was developed from all four protein against Lassa virus. We adopted sequence, 3D structure and fold level in silico analysis to predict B-cell and T-cell epitopes. The 3-D structure was determined for all protein by homology modeling and the modeled structure validated.
Results: One T-cell epitope from Glycoprotein (WDCIMTSYQ) and one from Nucleoprotein (WPYIASRTS) binds to maximum no of MHC class I and MHC class II alleles. They also specially bind to HLA alleles namely, A*0201, A*2705, DRB*0101 and DRB*0401.
Conclusions: Taken together, the results indicate the Glycoprotein and nucleoprotein are most suitable vaccine candidates against Lassa virus.
Keywords: B-cell, homology modeling, Lassa virus, T-cell
|How to cite this article:|
Verma SK, Yadav S, Kumar A. In silico prediction of B- and T- cell epitope on Lassa virus proteins for peptide based subunit vaccine design. Adv Biomed Res 2015;4:201
|How to cite this URL:|
Verma SK, Yadav S, Kumar A. In silico prediction of B- and T- cell epitope on Lassa virus proteins for peptide based subunit vaccine design. Adv Biomed Res [serial online] 2015 [cited 2023 Mar 26];4:201. Available from: https://www.advbiores.net/text.asp?2015/4/1/201/166137
| Introduction|| |
Lassa fever is an acute viral zoonotic illness caused by Lassa virus, a member of the Arenaviridae family and responsible for a severe hemorrhagic fever characterized by fever, sour throat, muscle pain, nausea. Lassa fever was first described in Sierra Leone in 1950s but the virus responsible for the disease was not identified until 1969 when two missionary nurses died in Nigeria, West Africa, and the cause of their illness was found to be Lassa virus, named after the town in Nigeria (Lassa in the Yedseram River valley) where the first cases were isolated., There are an estimated 300,000 to 500,000 cases of Lassa fever each year , with a mortality rate if 15–20% for hospitalized patient. Mortality rate has become high as 50% during epidemic and 90% in third month pregnancy for the expectant mother and the fetus both., Since then, a number of outbreaks of Lassa virus infection were reported in various parts of Nigeria including Jos, Zonkwua, Onitsha, Owerri, Abo Mbaise, Lafiya and Epkoma.,,,, Epidemics of Lassa fever were also documented in other West African countries including Liberia, Sierra Leone, Guinea, Mali and Senegal.,, A few cases of the importation of Lassa virus into other parts of the world for example by travelers were documented.
Morphologically, Lassa virus consists of enveloped particles that vary in diameter from approximately 60 to more than 300 nm, with mean particle size of 92 nm. The virus is approximately spherical, enveloped particles that rang in diameter from 50 to 300 nm. Lassa virus having ambisense genomic organization (two viral genes separated by an intergenic region), negative sense, bisegmented, ssRNA genome S (small, ~3.4 kb) and L (large, ~7.2 kb) segments., The small segment encodes the 75 KDa glycoprotein precursors (GPC) and the 63 KDa nucleoprotein (NP). After part translation modification GPC is cleaved into GP1 and GP2, the large segment of RNA encodes the 11KDa Z protein, which binds zinc and as Matrix protein and the 200KDa L protein, which acts as matrix protein. Replication and transcription of the genome occur in the cytoplasm of an infected cell and both take place within Rib nucleoprotein complex.,
Lassa virus is transmitted to human being from the rodent reservoir Mastomys natalensis, by direct content with infected tissue, food contaminated with excreta. Mostomys natalensis, is a common rodent in village houses, is therefore primary human infection and common (r) Lassa fever may also spread through person to person contact. Lassa virus trasmition occurs when a healthy person comes in contact with virus in the blood, secretion, tissue or excretion of any infected individual. The virus cannot be transmitting through skin to skin contact without exchange of body fluid. Lassa virus enters the cell via the alpha-dystroglucan receptor. Furthermore, ribavirin should be made available in hospitals and health centers in the endemic areas particularly in rural communities. This would help to control the disease. The aim of this study is to design the peptide based vaccines for Lassa virus.
| Materials and Methods|| |
Proteome data retrieval and analysis
Z protein (NP_694871), Nucleoprotein (NP_694869), L protein (NP_694872), and Glycoprotein (NP_694870) are the protein of Lassa Virus available in the NCBI Protein data base and hence it is used for this analysis. Bioinformatics tools were used for the analysis of proteome of Lassa virus. The expected molecular weight, isoelectric point (pI) highly repeated amino acids
(%) of repetition values were calculated using ExPaSy (http://www.expasy.org/)., All four proteins were analyzed for antigenicity using VaxiJen web server [Figure 1].
B-cell and T-cell epitope prediction
All of these targeted proteins of the Lassa virus strain found namely Z protein Nucleoprotein L protein and Glycoprotein were analyzed for the B-cell epitopes using BCPred. Identify common B-cell epitope by using both BCPred and AAP prediction method of BCPred. B-cell epitope with >0.8 BCPred and >0.4 Vaxijen score were re-elected for the identification of T cell epitope. These selected epitope further subjected to ProPred1 and ProPred analysis.,,, Both (Propred1 and Propred) are matrix-based method that allows prediction of MHC binders for various alleles based on the multiplication and additional matrices, proteosome cleavage site, simultaneously. ProPred1 allow predicting MHC class I binding peptide (CTL epitope) for 47 Allele and ProPred to predict MHC class II binding peptide (HTL peptide) for 51 alleles. Common epitope that bind both the MHC classes were selected for further analysis.
These epitopes were analyzed with VaxiJen v2.0 web server. The IC50 (inhibitory concentration 50) value of corresponding epitope was deduced from MHCpred server. Epitope having less than 1000 nM IC50 values for DRB1*0101 of MHC class II were selected. T epitope designer and MHCpred are the second screening method. T-epitope Designer is a structure based QSAR simulation method to predict HLA-peptide binding based on virtual binding pocked using X-ray crystal structure of HLA-peptide complex. In the second screening, the following selection criteria were used: i) Binding with large number of HLA alleles (>1000), (ii) must bind to A*0201, A*0204 and A*2705. Predicted epitope well further analyzed for fold level topology.
Molecular modeling and fold level analysis
The 3D structure of Glycoprotein was not available in database PDB. Query glycoprotein and template 3MKO after alignment were used as input in Modeller program, giving five models for target. Modeller objective function and dope score helped in the validation of the model of glycoprotein. The validated glycoprotein models were chosen for further studies and refinement. The most acceptable model of NDM-1 was finalized by Ramachandram Plot, providing the position of residue in particular segment based on phi (Ø) and psi (ψ) angles between N-Cα and Cα-C atoms of residue. The theoretical model generated was validated by using the programs PROCHECK , and ProSA. PROCHECK is a suite of programs to check the stereo chemical quality of protein structure. It includes parameters such as bond length, bond angle, main chain and side chain properties and residue-by residue properties to assess whether the geometry of the residues in a given protein structure is normal or unusual. ProSA (Protein Structure Analysis) program exploits interactive web- based applications to check the three-dimensional models of protein structures for potential errors by displaying scores and energy plots.
Pepitope servers were used to analyze the folding and cluster of selected epitopes in folded protein PepSurf and Mapitope algorithms used by this server to predict fold level topology. PepSurf algorithm helps to map the epitope onto the surface of the antigen. By Mapitope algorithms epitopes shared by the entire set of peptides are detected by the following steps user carried out to predict fold level topology. i) Prediction algorithm was executed and ii) 3D structure of predicted epitope are visualized using 3D structure viewer.
Characterization of epitopes
DISTELL server was used to design the 3D structures of the predicted binding peptides based on the similarity with PDB temples. After designing the structures epitopes were then validated with PROCHECK and ProSA. For prediction of domain, motif and functionality of epitope ProFunc, Motif Scan and InterProScan are used. ProteinDigest web server was used to predict pI, molecular weight and enzyme degradation site.
Molecular docking studies
After designing the epitopes structures molecular docking of selected alleles and epitopes was performed with the help of Autodock. This program used a simulated annealing approach to explore the conformation space between the ligand and target protein. The energy evaluation process is done by using grid-based molecular affinity potential, the docking was performed on the basis of Lamarckian Genetic Algorithm. The PMV (Python Molecular Viewer) was used for the visualization of Binding, position, H-bonding between the selected peptides and alleles.
| Results|| |
Physicochemical analysis and antigenicity prediction
The L protein has the highest molecular weight of about 253431.9 KDa which consists of Leucine (L) a neutral nonpolar amino acid residue has the highest percentage of repetition (11.9%). The physicochemical properties of putative proteins are given in [Table 1]. The pI value of any protein indicates the stability of protein in that particular isoelectric point. Isoelectric points of these proteins were ranged 6.17 to 8.95.
|Table 1: It comprises the data of Lassa virus proteins, molecular weight, pI and percentage of highly repeated amino acid residues in individual protein with antigenisity score|
Click here to view
Prediction of B and T-cell epitope
Epitopes which are capable to induce both type immunity (B-cell and T-cell) are known to be good vaccine candidate. For identification of these epitopes amino acid sequence of all four proteins were subjected to BCPred for B- cell epitope prediction. B-cell epitope prediction is the initial step for vaccine construct. BCPred is web based method that uses a novel method of subsequence kernel which was used to predict linear B-cell epitope from each protein. BCPed and AAP (amino acid pair) methods are used to predict fixed length epitope [Table 2]. Finally, two out of three B-cell epitope from Z protein, seven out of ten epitope from Glycoprotein, seven out of fourteen epitope from Nucleoprotein and sixteen out of twenty five epitope from L protein were selected for further analysis. These selected epitopes were analyzed for T-cell epitope identification. On the first level, sequence based 2D screening propred 1, propred and MHCPred were used to identify the potential T-cell epitope. Default ProPred1 and Propred parameters were used to determine the T-cell epitope [Table 3].
|Table 2: B-cell epitope prediction using BCPred (BCPred+AAP) and anigenicity of peptide using Vaxijen|
Click here to view
|Table 3: Common epitope (B and T-cell) along with their various parameters (selected epitope are highlighted in bold)|
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Common epitopes that bind with both MHC class I and II, and interact with DRB1*0101 are shown in [Table 4]. Epitopes with best Vaxijen and MHCPred score were selected for further analysis. T-epitope Designer was used to predict identified epitope binding abilities to >1000 MHC allele. T-epitope Designer is implemented based on a MHC-peptide prediction model described recently.
|Table 4: 3D QSAR based T-cell epitope prediction using T epitope designer|
Click here to view
Modeling and fold level analysis
Modeller 9.10 generated the 3-D model of query glycoprotein protein at resolution of 1.80 Å based on template protein 3MKO with sequence query coverage 26%. PROCHECK analysis of the model structure from Modeller 9.10 through Ramachandran plot showed that 96.1% of the residues were in favored and additional allowed regions [Figure 2]. Qualitative assessment of the model through ProSA analysis revealed that the model matched with the NMR region of the plot with Z score of -8.02. GROMOS96 which was used for energy minimization optimized the model structure from initial energy -3568.30 KJ/mol to final energy of -9434.68 KJ/mol.
|Figure 2: (a) 3D model of Glycoprotein, (b) Ramachandram plot for Glycoprotein model|
Click here to view
Positional fold level topologies of predicted epitope on theoretical models of proteins were predicted using Pepitope web server. Figure shows that the epitopes were present within the cluster on the surface of the protein. The glycoprotein is antigenic and one epitope WDCIMTSYQ from cluster I (score: 8.1608, Residue No. 8) was found to be antigenic (Vaxijen score: 1.1117) and can bind 64 MHC molecule of both MHC I and II molecule. The IC50 values of this epitope for DRB1*0101 and DRB1*0401 were 70.96 and 186.64, respectively, which indicates a good inhibition this epitope has also been found to bind all selected MHC molecules (A*0201, A*0204, B*2705) and 96.57% HLA molecule of T epitope designer shown in [Table 5]. The Nucleoprotein is also antigenic and one epitope WPYIASRTS from cluster I (Score: 10.301 Residue No. 8) was found to be antigenic (VaxiJen score: 1.3451) and can bind 64 MHC molecule of both MHC class I and II. The IC50 values of this epitope for DRB1*0101 and DRB1*0401 were 293.76 and 146.89, respectively, which indicates a good inhibition this epitope has also been found to bind all selected MHC molecules (A*0201, A*0204, B*2705) and 96.57% HLA molecule of T epitope designer [Figure 3].
|Table 5: The selected epitopes showing MHC binding and inhibition values predicted from 3D QSAR based T-epitope designer and
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|Figure 3: Fold level topology of epitope analyzed by Pepitope server (a) Glycopeotein epitope (WDCIMTSYQ) shown in red colour, (b) Nucleoprotein epitope (WPYIASRTS) shown in red color|
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After designing the 3D structures both the tools (PROCHECK and ProSA-web) are used for model validation. Furthermore no motif or domain could be assigned using ProFunc, Motif Scan and InterProScan for both proteins. Calculated M. wt. and pI of the WDCIMTSYQ from glycoprotein were 1146.30 Da and 3.08 respectively. Protein was found to be un digested with Tripsin, Clostripain, Proline Endopept, Staph Protease, Trypsin K, Trypsin R, as analyzed by ProteinDigest server for epitope WPYIASRTS from nucleoprotein, M. wt. and pI were 1080.21 and 8.75 respectively, and found to be undigested by Trypsin, Cyanogen_Bromide, Staph_Protease, Trypsin_K, AspN.
Molecular docking analysis
Some of the conserved peptides (WDCIMTSYQ, WPYIASRTS) with their interaction energies in kcal mol -1 are given in [Table 6] and shown in [Figure 4] and [Figure 5]. These peptides are promiscuous HLA binders. It will be useful to include these peptides in a chimeric constructs containing both cytotoxic and helper epitopes. It is expected that though this T-cell vaccine would not prevent Lassa virus infection, it would aid in quick clearance of the virus and prevent the severe infection.
|Table 6: The Docking results of interactions obtained with docking energy, build hydrogen bond and active site residues|
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|Figure 4: Two best docking conformation ((a). WDCIMTSYQ with, (b). WDCIMTSYQ with 1KLG) analyzed by Python Molecular Viewer (docked ligand shown by balls and sticks while hydrogen bonds shown by black sticks)|
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|Figure 5: Two best docking conformation ((a). WPYIASRTS with 1J8H, (b). WPYIASRTS with 1AOS) analyzed by Python Molecular Viewer (docked ligand shown by balls and sticks while hydrogen bonds shown by black sticks)|
Click here to view
| Discussion|| |
In the present study, three putative proteins of Lassa virus were used for the physicochemical analysis such as molecular weight, isoelectric point (pI value) and antigenic nature. Vaxijen was used to predict antigenisity, which is based on auto cross covariance (ACC) transformation of protein sequence into uniform vectors of principal aminoacid properties. The l00-CV (leave one – out cross validation) was used to identify antigenicity of protein with 91% sensitive, 82% accuracy and 72 specificity for viral sheces. The resultant protein was antigenic.
Epitopes which are capable to induce both type immunity (B-cell and T-cell) are known to be good vaccine candidate. Propred1 and ProPred are matrix-based methods that allow prediction of MHC binders for various alleles based on the multiplication and additional matrices, proteosome cleavage site, simultaneously. This is based on the observations made in previous studies which demonstrate that MHC binders having proteosome cleavage site at their C terminus have high potency to become T-cell epitopes.
MHCPred runs as a CGI server, and uses partial least square base approach for the identification of binding affinity to MHC molecule. MHCPred server generated IC50 nM values as output. A lover value of IC50 shows higher affinity with MHC molecule. The epitope bind with maximum number of alleles was selected for molecular docking analysis. The Pepitope server is a web based tool to predict discontinuous epitope based on set of peptide that have affinity against a monoclonal antibody or peptide. The server aligns a linear peptide sequence on to a 3D protein structure. The DISTILL model server was used to design the 3D structures of the predicted binding peptides. It is important to identify those peptides which are conserved across the various strains of Lassa virus and in this study that has been shown for the conserved peptides present in the constituent proteins of Lassa virus. The analysis reveals that there are number of suitable peptides from all four which may be included in the construction of poly epitopes T-cell vaccine.
| Conclusion|| |
The screening of putative epitopes using bioinformatics tools thus suggests that Glycoprotein and Nucleoprotein protein of Lassa virus could be used for preparation of immunological constructs. Molecular simulation and binding tests also suggest that the two nonameric epitopes WDCIMTSYQ and WPYIASRTS predicted and reported for the first time have considerable binding with MHC molecules and low energy minimization values providing stability to the peptide-MHC complex. These peptide construct will further undergo for wet lab studies, for the development of targeted vaccine against Lassa virus strains. Using a similar approach the short listing of candidate epitopes for vaccine design using other proteins can also be targeted that would reduce time and experimental expense.
| Acknowledgement|| |
The authors are grateful to Director, Madhav Institute of Technology and Science, Gwalior, M.P., India for providing necessary facilities and encouragement. The authors are also thankful to all faculty members of the Institute of Department of Biotechnology, Madhav Institute of Technology and Science, Gwalior, M.P., India for their generous support during the course of experimental work and manuscript preparation.
| References|| |
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If I had to be an item of jewellery it would be the Angler necklace from Abel, a jewellery line designed and manufactured by Kelsey Lim.
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WA’s croquet best shine bright in Bunbury
Competitors from across WA hit the Moorabinda Croquet Club for the men’s and women’s Golf Croquet WA Championshipson January 13 and 14.
Five players from Bunbury took the opportunity to test their skills against the best in WA, but it was the club’s Dale Jury who was the talk of the town, finishing second to Australian representative Fluer Brockway in the women’s competition.
Jury said she was surprised to come second.
“It was a very hot day and we all did our best,” she said.
“We had 14 entrants which is a good number for the annual tournament.
“I played well on and off. I think the heat got to us all.”
In the men’s it was Gosnells’ Martin Clarke who walked away with the victory, despite temperatures reaching an uncomfortable 41C.
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Local musician Marshall Mandell and friend Mark Will release a rock opera depicting a true soccer tragedy in their new album, 96.
At 96, interior design pioneer Ruth Adler Schnee is feted with a Cranbrook Art Museum exhibit featuring her acclaimed projects.
The famous rabbi visited multiple synagogues and groups in Metro Detroit to discuss African-Jewish relations and more on MLK Day.
Israeli nephrologist at Sheba Medical Center uncovers new findings about reversing kidney disease. Featured photo by Roland Metzger A groundbreaking study has shown that it is...
Israeli Prime Minister Benjamin Netanyahu has officially been indicted on charges of fraud, breach of trust and bribery.


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Nov 7, 2013, 3:00 PM EDT
Fans heading to Austin, Texas, for the 2013 United States Grand Prix will be able to attend two events ahead of the race weekend that should perfectly prepare them for the Formula One action at the Circuit of the Americas.
The Formula One Teams’ Association (FOTA) is set to hold another of its highly successful and popular fan forums later this month ahead of the second grand prix to be held at the Circuit of the Americas in Austin.
The forum sees a panel of Formula One drivers, team owners and legends face an audience of fans who can ask any burning questions, air any thoughts about the sport they may have and get a direct response. Following events in Montreal and London earlier this year, the forum will return to Austin on November 13, the Wednesday before the grand prix.
This year’s panel will be made up of Sergio Perez, Esteban Gutierrez, Adrian Sutil, Pastor Maldonado and Caterham’s American test driver, Alexander Rossi.
FOTA chairman Martin Whitmarsh expressed his happiness about returning to Austin, saying: “We are delighted to be returning to Austin again, and the enthusiasm and knowledge of the fans at last year’s fans’ forum and Grand Prix made it an easy decision to host another forum in Austin.
“Fan engagement is extremely important for all the Teams, and we have a great selection of participating drivers, and are excited to be utilising the venue of the fans’ fest which also gives us an opportunity to accommodate as many fans as possible.”
One day later, NBCSN’s very own Will Buxton will be hosting his second annual Buxton’s Big Time Bash in Austin. Last year’s event was an unmitigated success, raising over $2,000 for charity and providing a great evening of entertainment for the fans.
For this year’s event, a donation of $5 will be made on the door but in return fans will receive a ticket for a draw to win some money-can’t-buy F1 prizes, as well as entry to a great evening featuring some special guests and plenty of F1 entertainment ahead of the race weekend. All funds raised on the night will go to Meals on Wheels charity. The event will take place on November 14 at Speakeasy on Congress Avenue, Austin, and there appears to be no better way of spending an evening ahead of the on-track action at the Circuit of the Americas on Friday.
If you are lucky enough to be heading to Austin for the race, get your weekend off to the best possible start by attending both events. Failing that, be sure to watch FP2, qualifying and the race live on NBC as Sebastian Vettel goes in search of an eighth consecutive victory.
Kyle Busch roars back to win Truck race at Texas; Matt Crafton gets closer to clinching championship
Oct 31, 2014, 11:45 PM EDT
Kyle Busch rallied from ninth place with five laps left in regulation time, then roared up through the pack and “set sail” to a green-white-checker victory in the Truck Series race Friday night at Texas Motor Speedway.
Oct 31, 2014, 8:57 PM EDT
16-time NHRA Funny Car champ John Force will make a major sponsorship announcement Saturday in Las Vegas, and is expected to follow that up Tuesday with what major auto manufacturer he’ll be aligned with in 2015.
Oct 31, 2014, 8:23 PM EDT
Kenseth says the only way he’d feel a little better about it is if Harvick manages to make the Championship Race.
Tony Stewart sets fastest qualifying speed ever on 1.5-mile track (200.111 mph), but Matt Kenseth wins pole at Texas
Oct 31, 2014, 8:17 PM EDT
Matt Kenseth won the pole Friday at Texas Motor Speedway, but Tony Stewart lived up to his nickname and smoked the field with the fastest qualifying speed ever on a 1.5-mile track.
Oct 31, 2014, 8:00 PM EDT
Complete with news and analysis, here is all of the action from the COTA paddock on practice day for the United States Grand Prix.
Oct 31, 2014, 7:15 PM EDT
Speaking to NBCSN’s Will Buxton, America’s great F1 hopeful speaks about recent events at Marussia and his hopes for the future.
Oct 31, 2014, 7:00 PM EDT
An on-the-ground perspective of the speed, sound and crowd from Austin second practice.
Racing legend Dan Gurney earns one of rarest and most prestigious awards for lifetime achievements (video)
Oct 31, 2014, 6:59 PM EDT
Very few race car drivers deserve to be called “legendary.” Dan Gurney is most definitely one of those, and was honored this week with the Edison-Ford Medal for his incomparable lifetime achievements.
Oct 31, 2014, 6:30 PM EDT
17-year-old Toro Rosso driver continues to prove that age is simply a number in racing.
Oct 31, 2014, 6:10 PM EDT
Austin Dillon was the fastest driver in Friday’s final Nationwide Series practice at Texas Motor Speedway. The defending NNS champ who is now in his rookie season in Sprint Cup, Dillon is making his first NNS start this season in Saturday’s race.
Oct 31, 2014, 6:00 PM EDT
Gauging the pulse of the fans on site at Austin.
Oct 31, 2014, 5:45 PM EDT
The Outlaw attempts the “Candy Corn Challenge” – fitting 100 pieces inside his mouth.
Oct 31, 2014, 5:30 PM EDT
Is there more to Fernando’s future than just McLaren?
Oct 31, 2014, 5:00 PM EDT
Administrators still hoping to get the team racing in Abu Dhabi, but will aim for the grid in 2015 if this is not possible.
Oct 31, 2014, 4:45 PM EDT
Reddick captures his second Truck pole in the last three races.
Oct 31, 2014, 4:32 PM EDT
British driver finishes just 0.003 seconds ahead of Rosberg in FP2, equating to just 6.7 inches around the Circuit of The Americas in Austin, Texas.
Oct 31, 2014, 4:00 PM EDT
The six-time Sprint Cup champion is driving the No. 48 Lowe’s “Red Vest” Chevrolet this weekend.
Oct 31, 2014, 3:45 PM EDT
Finnish driver has endured a difficult return to Maranello in 2014 after five years away.
Oct 31, 2014, 3:00 PM EDT
Chase competitors Kevin Harvick and points leader Jeff Gordon crack the Top 5 speeds.
Oct 31, 2014, 2:45 PM EDT
Can Lewis Hamilton double up in the second practice session in Austin? Join us from 3p ET to find out.
- Kyle Busch roars back to win Truck race at Texas; Matt Crafton gets closer to clinching championship 0
- Tony Stewart sets fastest qualifying speed ever on 1.5-mile track (200.111 mph), but Matt Kenseth wins pole at Texas 0
- United States GP Paddock Notebook – Friday 0
- Hamilton inches away from Rosberg to top FP2 at COTA 0
- Sebastian Vettel to start United States GP from pit lane 0
- Hamilton quickest in first practice for United States GP 1
- 2014 United States Grand Prix Preview 0


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Designed by Naoto Fukasawa for Magis
Japanese designer Naoto Fukasawa offers a contemporary twist on tradition with his Cu-Clock for Magis. Cleverly stripping away the ornamentation, wooden gears, pendants, and chains that define the look of the traditional Black Forest cuckoo clock, Fukasawa left in its place clean lines, standard injection-molded ABS, and battery power. Cu-Clock is a minimalist interpretation of the traditional, ornate Black Forest cuckoo clock. A simple white house with a vibrantly colored roof provides shelter for a tiny, abstract bird that sings on the hour. The white Cu-Clock—available with a roof in white, orange, or green—will add a note of humor to any environment.

domain: design


2015: Bass in The Sound of Musicand Carousel. Third season. Stratford: Slim, the bass player, in Crazy for You; Fiddler on the Roof. Elsewhere: For nearly 20 years, Michael has been music director, producer, arranger and bassist for award-winning singer and broadcaster Heather Bambrick. Fourteen seasons at Stage West (over 40 productions). He is on the faculty of Toronto’s Humber College and the National Music Camp of Canada. Writing/Arranging Credits: Dance Legends (Drayton/Grand); The Wizard of Oz (Grand); The Producers, Cabaret, California Dreaming, Fever, Love Train, One Hit Wonders, et al. (Stage West); International Association of Jazz Educators (inaugural winner of the Phil Nimmons/SOCAN emerging composer award). Training: Master of Music, University of Toronto. Recordings: Can be heard on dozens of CDs as bassist, producer and/or arranger. Michael’s own CD, Sing In Me Muse, is available on iTunes, or visit michaelmcclennan.com.


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definately one of your best songs. has more structure than your other pieces in 05 that you submitted in 07. Keep doing this style. keep up the good work.
(im your 100th reviewer!)
This sounds like an improbed version of Midnight Siege, well done Dimrain ; )
Has so many downloads but too small of a rating.
u are like a song mastermind!!!!!!!
All of your songs are amazing man.
When are you going to release an album?Or maybe you already did....
Please contact me if you would like to use this in a project. We can discuss the details.


Politics in America right now is disturbing. It is becoming obvious that the resources of government were used for political purposes against a presidential candidate. Now that the candidate is in office, the ‘deep state’ continues to oppose him. During the next year and a half, we are going to be subject to endless investigations of everything Donald Trump has ever done combined with a media that wants to recapture the power they had during Watergate (the ability to drive a President from office). So what are we to do about it?
American history tells us that during the American Revolution, it is estimated that only 3% of the colonists were actively fighting in the field against British forces at any given time. These 3% were people who saw what was going on and chose to be involved. We need that 3% now. We need people who are willing to look past the lies being told in the mainstream media and do their own research. We need people who don’t believe the constant drumbeat of the major media that says “Orange man bad” and are willing to look at what the Trump administration has accomplished.
Next November there will be an election. President Trump will run again. A lot can happen between now and then, but even a casual glance shows that currently almost all of the Democrat candidates have wandered away from the mainstream of America. It’s up to voters to do their homework, decide what they want for America, and vote. The plans of the Democrat candidates will negatively impact our freedom and our economy. All of us who care about our country should fight those plans with everything we have. Study voting records of those in office, and study campaign contributions (opensecrets.org lists campaign contributions of all candidates).
Get involved. You future, your children’s future, and your grandchildren’s future depend on it.


FONTANA, CA 92335
Shift: 7am -3:30pm
Seeking a Maintenance Mechanic with background in fabrication equipment. Local manufacturing plant specializes in roll forming of Architectural heavier gauge single metal wall and roofing systems.
Must have a minimum of 3-5yrs exp. To be considered. Must have steel toes.
* Troubleshooting and repair of equipment.
* Adhere to preventative maintenance schedule.
* Strong electrical and mechanical experience.
* Critical thinking and problem solving skills.
* Ability to work unsupervised as well as with a team.
* Work in a safe manner
* Available to work weekends
Skills and Knowledge Required:
* Minimum 3-4 years experience
* Effective communication skills
* Detail oriented
* A sense of urgency


This is a pure python implementation of the NetworkTables protocol, derived from the wpilib ntcore C++ implementation. In FRC, the NetworkTables protocol is used to pass non-Driver Station data to and from the robot across the network.
Don’t understand this NetworkTables thing? Check out our basic overview of NetworkTables.
This implementation is intended to be compatible with python 2.7 and python 3.3+. All commits to the repository are automatically tested on all supported python versions using Travis-CI.
NetworkTables is a protocol used for robot communication in the FIRST Robotics Competition, and can be used to talk to SmartDashboard/SFX. It does not have any security, and should never be used on untrusted networks.


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ANGER DISORDERS & ANGER MANAGEMENT
Get Therapeutic Support to Manage Anger
Anger Management Therapy
Anger is a normal and natural response to certain circumstances. However, if someone regularly experiences outbursts, aggression, or emotional escalations, they may benefit from learning tools to manage these emotions in a healthier way. As a program, anger management is a well-researched psycho-therapeutic approach for anger control. If you feel that either yourself or someone you love may benefit from speaking to a therapist at Bhatia Psychology Group, don’t hesitate to reach out to us. We’re here to help.
Help for Anger Management
If you are worried about a family member or friend that may be struggling with anger management issues, look for the following behavioural patterns:
Becomes angry and/or violent with alcohol consumption
Refuses to compromise or come to a mutual agreement without getting angry
Difficulty expressing how they feel in a calm and collected manner
Internal aggression that may involve self-harm and isolation
External aggression that may involve swearing, yelling, violence or threats
Behaviour triggered by or connected with substance abuse/addiction
Repetitive patterns of behaviour that negatively affects the relationships
Refuses to speak to someone or entirely ignores them when angry
Treatment for Anger Management
Anger alone isn’t considered a disorder, but it can be a symptom of a mental health condition — this can range from anxiety, depression and paranoia to frustration, insomnia and even social isolation. As a result, each case requires an evaluation and assessment to review underlying issues such as social or environmental influences, mental health challenges, trauma, alcohol or substance abuse, and other factors. Treatment often includes Cognitive Behavioural Therapy (CBT), as well as other tools around communication, problem-solving, avoiding triggering environments, and even humour. The number of sessions and the length of treatment will depend on the rate of progress, stress levels, support system and severity of symptoms.
FAQs – Anger Management
What are the symptoms of anger management issues?
Anger management problems come with both emotional and physical symptoms. These can include:
- Trouble sleeping
- Social isolation
- High blood pressure
- Heart palpitations
How do you know you have an anger problem?
It’s completely normal to get angry at times. However, you may have an anger management issue if you find your emotions have begun to impact friends, family, or coworkers — either mentally or physically.
Additionally, you may have an anger problem if:
- You punch objects or walls to feel a release
- You react to problems quickly and violently
- You accuse others of having an issue
- You struggle to calm or control your anger
- You constantly argue with family, friends, or coworkers
- You feel frustrated during arguments and often feel regretful of your actions afterwards
Can anger management issues be cured?
While you can’t ‘cure’ an emotion, you can learn to manage the impact that anger has on yourself and others, as well as learn to reduce its intensity or effect.
Does anger management really work?
It can for many people. It takes practice, effort, and work, but you can learn to manage your anger and not let it control your life or the lives of the ones you love and care about. There are various techniques and therapeutic options available through Bhatia Psychology Group.
How does anger management work?
While many times these behaviours are related to other factors and mental health disorders, anger management can help you learn to recognize your triggers and understand them better. It can further help you learn to approach the situation in a calm, healthy, and constructive way.
Why is anger management important?
This is essential for your own well-being and those around you. Unmanaged anger can escalate into self-harm or harming of others.

domain: psychology


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4) Hmm... weird again. Dunno what books you've read, but that is totally false. Nothing is perfect and the "LoA" I subscribe to doesn't say what you've described. Visualizing is only one tool of LoA... one which I don't really use. The perfect relationship and perfect diet are total BS. you don't visualize those things, you either build them (a good healthy relationship, not a perfect one) or follow them (a good diet).
Law Of Attraction What Is Resistance
Take some time to ‘feel’ how if feels like getting what you want. Close your eyes and live the moments after you succeed on your goal. For example, now that you’ve got the money you asked, how is you life? Visualize speaking with the ones you love, explaining them you current reality. Pay attention to their reactions. Make this experience as real as possible. Visualization speeds up manifestation. To Manifest Anything you want, you need to ‘see’ it first with you spiritual eyes.
Law Of Attraction On Relationships
Jasmine L. Thomas, M. Ed. is a follower of Christ, lover of family, a powerful educator, an entrepreneur, and an encourager to everyone she encounters. She is a passionate advocate for educational and Christian-based empowerment amongst youth and women. To live out this calling, Jasmine owns Limitless Learning Tutoring, LLC, The Mindset Queen Coaching Program, and Thomas & Company Publishing. Jasmine believes that people should be given the proper mind enriching tools to elevate their lives to the next level.
For some people, it seems like the stars repeatedly align, as if they wrote their own destiny. Despite challenges or hiccups, these people come out of even the toughest situations unfazed, blessed in one way or another by what seems like a miracle. How do they do it? And what if you learned there was a secret to cultivating the life you want? It sounds pretty crazy, but the Law of Attraction is something that many people believe in. So what's the deal with all of it, anyway?
You claim that "positive thoughts always resulting in positive things" is not the basis of the LOA. In part you are correct. The foundational basis in science, as I alluded to in the article was that thought stuff particles travel through ether (not air) and interact with formless stuff to create whatever it is that you are thinking about. This was how "the law" started.
Listen, I get it, we live in a fast-moving environment today, but the old adage, “stop and smell the roses” really comes to mind when manifesting. You see, if your mind is focused on what you don’t have, or that things aren’t working as quickly as you’d like, you will miss out on the small victories. Celebrate those small wins! You have to acknowledge the victories if you want to continue to find trust and faith in the process. Keep your eyes, ears, and heart open to signs from the Universe that it has your back. It could be someone acknowledging a change in you, a quote on social media that was exactly what you needed to hear that day, or even a smile from a stranger that warms your heart. Stay open, be present, and take note.
Law Of Attraction For Exam Success
Success rate of 0.1%? Where'd you get that statistic? But whatever the actual number, your question is an invalid one. The law of gravity works 100% of the time, except we can fly planes, right? Yes, I know that sounds silly, but it's really to show you how silly YOUR question is because "LoA" works even "negatively" (put in quotations because human perception is what labels things as positive or negative when really, the universe is 'neutral' and things don't happen as a way to punish us... that's just ridiculous, it's not personal).
A favorite Abraham-Hicks’ quote of mine is “a belief is just a thought you keep thinking.” What does that tell us? That beliefs can and do change. But knowledge, understood as an evidence-based understanding of something, is unwavering. Well, maybe there’s more of an 80/20 rule about it. I mean, once upon a time we just knew the Earth was flat. Now, we know it’s not.
Whatever you want already exists in your imagination, doesn’t it? Therefore it is just as real as any other idea you have ever had that came to fruition. Play with the belief that what you want is already waiting for you. You’ve placed the order, now you’re on your way to pick it up. What have you got to lose? After all, you don’t “need” what you desire, it’s just for fun.
They don’t obtain the ability to find life in their life. Yes, ability. It’s not just a choice. It may start with a choice. But it’s a practice, a daily one, to produce nectar (joy) from what’s in front of you as you keep building and climbing. This is what keeps you positive and attracting. It’s like pedaling a bike. Once you are no longer able to produce joy with what you have, you are no longer attracting.
I am an energy person. I believe in attracting what you’re putting out into the Universe. It’s crucial to stay at a high vibration if you want to manifest your dreams. Limit negative self talk, or negative conversations and people around you. Pay attention to when jealousy or comparison gets you down, and switch out those tasks for things that give you confidence and inspiration. A daily mindfulness practice is essential. It’s also why I recommend starting with something like A Course In Miracles or May Cause Miracles (the “millennial” version of ACIM), to get you in the habit of a daily practice. Really there are tons of apps out their for meditation and inspiration today, that there’s no excuse to not take a mindful moment every day.
How To Do Law Of Attraction Meditation
Math says that 1 + 2 = 2. If you talk with one person who loves math and another who hates math, they’ll each have a different perception of math. One person believes math is intriguing and fun, while the other may have a belief that math is terribly frustrating. So, how you perceive and interpret your reality will also have an effect on what you are attracting in your life.
My biggest mistake in this area was with my fiance. We both knew we didn’t want to get married until we hit our 30s. For us, it was more about building in our 20s to come together later in a marriage. Although we both agreed to get married at 25, the truth is that we winded up going through a horrible break-up because we didn’t do what felt right for us individually. In essence, we hustled into an engagement.
Focus on understanding your part in this overall process, rather than feeling guilty or ashamed. Recognizing how your fears have manifested in your life does not mean that the bad things in your life are your fault. It simply means that the Law of Attraction has responded to the signal you sent out. Just acknowledge that the system works and your new awareness will guide you. No need to shame yourself or even feel responsible.
How To Enhance Law Of Attraction


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NOTTINGHAM, MD—A Nottingham man who was shot in Baltimore City on Saturday morning has died.
At around 9 a.m. on Saturday, officers responded to the 400-block of E. 21st Street for a report of a shooting.
At the scene, police found that 30-year-old Rawshad Divers, of Nottingham, had been shot while sitting in his car.
He was taken to an area hospital in grave condition.
On Monday morning, the Baltimore Police Department reported that Rawshad Divers had died from his injuries.
Anyone with information on this case is asked to contact police at 410-396-2100 or Metro Crime Stoppers at 1-866-7LOCKUP.


$400M TTF Funding Bill Includes Funding for Engineering Study
TRENTON – Senator Nicholas P. Scutari today announced the state will provide funding to advance the Tremley Point Connector Road project, which is vital to improving safety and fueling economic development in the region. Funding for the project is included in the $400 million transportation funding bill passed by the Legislature and signed into law by the governor today.
Senator Scutari has been working with the Department of Transportation to advance the project. The transportation funding legislation (S-3076) signed into law today includes funding to complete the preliminary engineering for the Tremley Point Connector Road. The project will create a key access road to remove heavy truck traffic from the local roadway and provide a connection between the Turnpike and Tremley Point area of Linden.
“We knew that getting transportation dollars out the door immediately was crucial to getting vitally-needed infrastructure projects moving across the state. This law provides critical funding to advance the Tremley Point Connector Project, which will improve safety, create jobs and fuel economic development in the region,” said Senator Scutari (D-Union, Somerset, Middlesex). “This is significant progress on a project that we have been working on for years and that will benefit the residents of Linden and Union County. I am pleased that we were able to move quickly on this legislation that will help advance this project and to continue to work in preparation for the construction phase.”
The Exit 12 access road from the Turnpike now directs traffic through the South Wood Avenue section of the city. The proposed connector will intersect with Industrial Highway in Carteret, traverse the Rahway River and connect at Tremley Point Road in Linden. The Tremley Point Connector Road will provide a direct connection to the Tremley Point section of Linden, an industrialized area that relies on commercial traffic and offers the potential for economic expansion.
The transportation funding bill received final legislative approval last Thursday, and includes $140 million for NJ Transit for technology improvements and system safety and $260 million for work on roads and bridges throughout the state.
The funding will be distributed within 100 days for projects identified by the Department of Transportation through the Transportation Trust Fund selection process. The added funding for the 2017 Fiscal Year will boost total spending to $2 billion, the same level that will be sustained over the next eight years under the plan that renewed the TTF.


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Gum disease is one of the most common dental conditions in the U.S.A. In fact, it is the number one cause of lost teeth for American adults. About 80% of Americans will have a gum disease infection at some point in their life, which is an enormous number! Gum disease does not follow geographic, socioeconomic, or educational lines. It is based purely on the bacteria in your mouth and how far is is allowed to go into your gums. Don’t join these gum disease statistics. Instead, know the three levels of battle!
1) At-Home Care
Brushing, flossing, and even an antiseptic mouthwash is the first line of battle against gum disease. Following an oral hygiene routine the best way to protect yourself from gum disease, though sometimes it’s just not enough.
2) Fighting Gingivitis with Prophylaxis
The first level of gum disease is called gingivitis. This is a surface infection of the gums, but it can be easily removed with prophylaxis, which is another word for a professional dental cleaning.
3) Fighting Periodontitis
If gingivitis isn’t caught early, the infection can work down below the surface, turning into an infection called periodontitis. This simply means that the bone structure is in harm’s way. Removing periodontitis takes much more complex procedures like scaling and root planing or possibly oral surgery.
Stop gum disease before it has a chance to damage your teeth. Don’t join the statistics of patients who have lost teeth to this easily preventable disease. Contact us today to set up your next prophylaxis and exam. We can’t wait to help you get a happier, healthier smile!

domain: biology


The WFCC brings together wildland fire management agencies, partners, and collaborators in Canada and around the world. These biennial conferences focus on wildland fire management, ecology, and science in Canada, and provide a neutral forum for the exchange of best practices. vft North America will be present leading the solutions for wildland firefighting in collaboration with our distributor in Canada Motion Industries.
What news awaits?
Discover our skid units for UTV vehicles
Robust and reliable skid units equipped with a very capable centrifugal pump.
New vft Pumps
A whole range of portable wildland fire pumps. Light, powerful, efficient and durable.
What else will you find there?
Next-generation protective equipment designed exclusively so wildland firefighters and emergency units may do their work safely and comfortably. And the most innovative line tools for building defense lines, setting backburns and for burnout operations.
Where will you find us?
Visit us at booth #12.
Do not miss these and many other news, visit our booth!


Dragon has a long history of service to the oil and gas industry. Spanning the world from the fields of the Middle East and Caspian, Dragon has what it takes to meet the growing global demand for oil and natural gas projects. We have the proven expertise to perform in all facets of the industry in getting material and equipment to the most remote locations throughout the globe. Our power and the vessels that we operate provides our customers with the right ship to meet the unique requirements of the job at hand. We, as Dragon, have demonstrated our commitment and ability to exceed customer’s expectations for reliable and safe carriage of their cargo. We offer the Oil and Gas Industry superior choices in the handling of heavy lift and over-dimensional project shipments anywhere in the World. As the global demand for effective and reliable ocean transportation increases, Dragon offers efficient and quality solutions for the challenges that lay ahead. By anticipating global market trends and frequent communication with our valued customers, we are well equipped to surpass our customers’ needs and expectations.


US pushes for extension of Syria gas attack probe
US Ambassador to the UN Nikki Haley said the Security Council "must act now" to extend an investigation of chemical weapons attacks in Syria
US Ambassador Nikki Haley told the UN Security Council on Friday that it must "act now" to extend an investigation of chemical weapons attacks in Syria, a move that could be vetoed by Russia.
The United States has circulated a draft resolution extending the mandate of the joint UN-OPCW (Organisation for the Prohibition of Chemical Weapons) panel for another year and diplomats said a vote at the council could be held as early as Monday.
The measure is facing a possible veto from Russia, Syria's ally, which wants to first see a report due on Thursday on the sarin gas attack in Khan Sheikhun before deciding on the fate of the investigation.
"We must act now and support the continuation of the JIM's important work," Haley said in a letter to the council obtained by AFP.
The Joint Investigative Mechanism (JIM) was established by the council in 2015 to determine who is responsible for chemical weapons attacks in Syria's six-year civil war. Its mandate expires on November 18.
The United States accuses Russia of linking the panel's survival to its findings on the April 4 Khan Sheikhun attack.
France, Britain, and the United States have accused Assad's forces of carrying out the attack on the opposition-held village that killed at least 87 people, including more than 30 children.
Last month, UN war crimes investigators said they had evidence that the Syrian air force was behind Khan Sheikhun, despite repeated denials from Damascus.
Haley argued that the OPCW was reviewing more than 60 cases of alleged chemical weapons use in Syria, including a recently-uncovered sarin attack on an opposition-held village on March 30.
"This is not about politics or the contents of the next report, but about ensuring accountability for those who are using these terrible weapons, whether they are UN member states or non-state actors," Haley wrote.
The JIM has already determined that Syrian government forces were responsible for chlorine attacks on three villages in 2014 and 2015, and that Islamic State jihadist group used mustard gas in 2015.
The head of Russia's non-proliferation department, Mikhail Ulyanov, told UN member states last week that Russia will study the report on Khan Sheikhun by the JIM to "judge if it deserves the extension."
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CLEVELAND – The new owners of Key Center, The Millennia Companies, have submitted their plans for a renovation of the Key Tower’s lobby, plaza and the hotel.
In a presentation to the Cleveland Landmarks Commission, Millennia stated that the renovations would hopefully begin with the outdoor plaza of the complex. The plan is to add much more greenery to the area, walking paths and an outdoor pavilion.
The outdoor renovations are estimated to cost $1 million, and are part of the overall $24 million renovation plan. The outdoor renovations will begin in May and are scheduled to be finished by early fall.
The Key Center, located on Public Square, is a 57-story tower which houses the Cleveland Marriott Downtown, offices, a parking garage and is attached to the 10-story historic Society Corporation bank building.


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Play Online Baccarat
Can you like playing baccarat? If you do, then you’re definitely not alone! Baccarat is probably the hottest 인터넷 바카라 casino games available and is played by millions of people from around the globe. Online, you can get players around the world that share exactly the same passion for playing this casino game.
Online, there are many baccarat game websites proclaiming to offer you free games, bonus offers and cash prizes. You should use these offers to improve your odds at winning while playing baccarat online, concurrently as having fun. On line, share the various ways to play baccarat for real money, and the very best online casinos for baccarat available taking players from all over the world, anywhere in the word. Actually, some countries like France, Spain, Italy, Cyprus, Spain, Turkey, Romania, Greece and Brazil are known for their top online baccarat game sites.
As well as having a baccarat game, you may also purchase baccarat accessories to enhance your online experience and enhance your chances of winning. One of these accessories is called the wenn sie baccarat. This baccarat accessory is actually a head piece which you put on your skull ahead of playing. It looks like a metal band and also imitates the sound of the true dealer making the dealer calls.
The wenn sie baccarat is made up of plastic or metal, depending upon your choice. Gleam baccarat playing disc with the logo of the baccarat handmade cards. This disk is positioned in a metal case that looks just like the original case of the baccarat cards. The complete accessory is enclosed in a carrying case that looks as being a jeweler’s box.
Also you can purchase the same online as an attachment to your online baccarat casino games. The same type of accessory can be available as a download to your personal computer. All the players can get on the online baccarat casino games and choose to download the accessory should they like. Players do not need to pay any additional charges therefore to download the same.
Once downloaded to the personal computer, the players may then use it to play the web card games in the casino. When a dealer is called, the players go through the corresponding danglers and the dealer will make another call, much just as as though the dealer made a call to a regular baccarat player in true to life. The dealer in the online baccarat games can make the same calls as he or she would in a real live casino.
The third type of accessories is the Macau baccarat game offerings from third party casinos. Alternative party gambling sites are not governed by the laws of the country where they operate their casinos. Some jurisdictions prohibit gambling and some do not. That is why the Macau baccarat emerges in jurisdictions in which it is illegal for players to gamble for real in casinos. However, the sites offer all the Macau casino baccarat games, including the online ones, so players do have the option of playing in other countries.
Players can choose to play with a number of cards as in a regular baccarat game or they can play with a wheel, just like in slot machines. Regarding the wheel the player may spin the wheel no more than seven times, at which point they need to stop and wait for the next card. In this case the player is not betting on if they will hit the ball, but on the opportunity that another card will come out from the deck.

domain: hobby


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First Sweater: Feb 12, 19 & 26, 2-4 pm
|Availability:||In stock (6)|
This classic raglan sweater works up quickly and looks great! Sweater No. 9 is a perfect place to start if you are new to making garments.
Three 2 hour sessions
- Knit and purl confidently
- Working in the round
- Increasing and decreasing
- Learn top-down raglan sweater construction
- How to make a garment that fits
- Bust size: XS(32-34”), S(34-36”), M(36-38”), L(40-42”), XL(43-45”), XXL(46-48”).
- Knitting for Olive Heavy Merino 6,6,7,8,9,10 balls and Knitting for Olive Soft Silk Mohair 3,4,4,5,5,5 balls. The sweater is knit with both strands of yarn. Or chose 822,822,959,1096,1233, 1370 yards of any bulky yarn
- US 8 circulars (16”) US 11 circulars (24” and 32”) plus US 8 and US 11 double points. (These needle sizes are a suggestion – use any needle that gets gauge)
- Stitch markers (one unique for the beginning of the round), scissors, tape measure, tapestry needle, waste yarn for holding stitches
- A purchased copy of Sweater No. 9 by My Favorite Things
Read the entire pattern, and choose a size based on your measurements. Intended sweater ease is 6”. Choose your size based on the ease you’d like. Make a gauge swatch in the round. Wash the swatch as you would the finished garment. Bring all materials to the first class.


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GSA notes that the venerable Ballarat Couriers Melissa Cunningham reports that:
July 10, 2014, midnight
THERE was public outcry in the City of Ballarats council chambers as councillors voted to defer two contentious items in Wednesday nights council meeting agenda.
All councillors except Councillor Amy Johnson voted to defer the two items including a proposal to establish free dumping points for caravans, campervans and motorhomes to dispose of their waste and another proposing free camping for recreational vehicles in Pioneer Park.
The deferral was due to the citys request to seek legal assistance on the two matters.
Campervan and Motorhome Club (of Australia) co-director Keith Moxham vented his frustration to councillors and said he had travelled from Melbourne to make his submissions for both proposals on behalf of the recreational vehicle lobby group.
I guess its lucky I didnt fly one of the directors in from Brisbane for this meeting, he said.
A bit of notice would have been appreciated. We just want this issue resolved, it is a small issue but it is becoming this enormous thing and I would like to see some rationality come back into it.
Also in the chamber was Victorian Caravan Parks Association chief executive office Elizabeth White, who had travelled from West Melbourne to make a submission in support of Ballarats existing caravan park owners.
Motorhomes at Beechworth Lake Sambell.
Ms White told The Courier she had submitted an urgent letter to the council on Tuesday on the advice of her legal advisor, outlining the concerns of the association.
I think it is important that the council take the time to go through the issues that we raised in the letter, including the significant costs we believe council will incur if this goes ahead, and the considerable risks these proposal will have that maybe the council has not yet considered, Ms White said.
Ms White said she would return to Ballarat to make her public submission at a later date.
Ballarat resident and (CMCA) member Grant Tillett said he was disappointed by the outcome.
This has been going on for two years. It is beyond a joke, Mr Tillett. It is being made into a fuss when it shouldnt be.
Cr Des Hudson said the council was awaiting legal advice on both items and a deferral was required to ensure the right outcome was met.
The council needs to be across all legalities involved in this item, Cr Hudson said.
Cr Amy Johnson opposed the deferral and encouraged the people who had planned to make public submissions to still do so.
People have put in a lot of time and effort into their submissions and made the effort to attend the meeting and I think it is ridiculous to defer it now, Cr Johnson said.
While Cr Johnson was the only councillor to vote against the deferral, Cr Peter Innes voiced his disappointment.
Jayco Optimum pops its awning.
What I find intriguing is that council officers have been working really hard and looking into this for three years but these issues havent been raised until today, he said.
Cr Morris said it was critical the council made the right decision on both items and a deferral was the only way to ensure this occurred.
Concerns have been raised about both items and we want to get the right results, Cr Morris said. We see these items as two quite separate issues and we are seeking separate legal advice on both these reports and once we have that assistance we will be better placed to make the right decision.
On May 11, 2012 Victorias Minister for Innovation, Services and Small Business, Minister for Tourism and Small Business, Louise Asher wrote to then Caravan and Motorhome Club of Australia General Manager, Richard Barwick (now CEO) after meeting with CMCA regarding the clubs proposed member stop-over-scheme.
Tourism Victoria has been working with the CMCA and the Department of Planning and Community Development to examine the Park Over Properties(POP) proposal, she said.
I am advised that there is no legal foundation for councils to oppose your proposed scheme. Under current legislation, residential property owners do not require a permit to allow RV tourist to carry out the activities in the manner that you have described in your scheme provided that no fee or other consideration is charged.
Motorhomes popular Wodonga .
She went on In relation to the other issues raised by CMCA, such as dump points and rest spots, I have asked Tourism Victoria to work with the CMCA, Regional Tourism Boards and local councils to actively address these issues in areas of high demand such as the Great Ocean Road and other key touring routes.
I wish you ongoing success in your efforts to grow the RV tourism market in Victoria, she said.
Editors Note: Ballarat, as part of Victorias high profile Goldfields Region, is a focal destination player in a key state touring route. The Goldfields touring route includes Creswick, Daylesford, Castlemaine, Maldon, Bendigo, Maryborough, Avoca, Stawell and Ararat. The CMCA claims 66,000 members. A majority are active road tourers, the CMCA says. Beechworth Lake Sambell Caravan Park has a CMCA Members: $ 5 per person per night basic campsite option available under the CMCAs Leave No Trace Scheme. GSA reported on this innovation. Please follow the hyperlink to access the details in the GoSee library.


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Normally, diabetes is of two types i.e. type 1 and type 2 diabetes. The patients who have type1 diabetes have the maximum chance of hypoglycemia. When glucose level falls too low it is called hypoglycemia. Glucose is transported to muscle cells with the help of insulin. In a few cases, hypoglycemia can be treated by eating glucose tablets, sweets and drinking fruit juices.
The main signs of hypoglycemia are confusion, unconsciousness, nausea, sweating. If hypoglycemia is left untreated it can permanently damage the nervous system. In severe cases, it leads to coma and death. According to 2018 medical news, today patients with type2 diabetes may be over treated which can lead to abuse of test and waste supplies. According to the Mayo clinic, the US is in high risk of hypoglycemia due to overtreatment of diabetes.
The researchers have found that patients in US with diabetes are treated with too many medications than their hemoglobin A1C level which leads to hypoglycemia.A1C test is a primary test which provides information regarding average blood sugar level. The A1C levels are as follows for normal the percentage is below 5.7%, for prediabetes is 5.7 to 6.4% and for diabetes 6.5% and above. During 2011-2014 research records there was a glucose lowering remedy in the US by the National Health and Nutrition Examination Survey.
Older people and other patients are considered acutely confused and are at more risk to develop hypoglycemia because of excessive treatment. The researches explain that the influential persons are attached t control hypoglycemia. The treatments should be managed by taking proper care of patients health condition, mental situation and the existence of daily life. A proper balance should be taken for treatment for patients suffering from diabetes. Undertreatment and overtreatment both are harmful to patients.

domain: medical


Brown Eyed Girls’ JeA Drops “Bad Girl” MV Featuring Jung Yeop
Brown Eyed Girls leader JeA has made a comeback!
On April 15 at midnight KST, JeA released her new ballad entitled “Bad Girl,” which features R&B singer Jung Yeop of Brown Eyed Soul. The music video features JeA looking gorgeous as she sings sadly to a lover, telling him not to forgive her for all the bad things she’s done that have hurt him.
Watch the video below!
JeA released her first solo album “Just JeA” in 2013, which makes this her first solo venture in three years. JeA produced her new single album herself, and “Bad Girl” was composed and written by Uniqnote and Jung Yeop.
What do you think of JeA’s solo comeback track?


CLEARFIELD – The Clearfield County Commissioners have announced they are soliciting requests for qualifications to select professional organizations to provide various forms of housing programs and projects in the county.
This request will act to fulfill two, separate qualifications for two housing programs in Clearfield County.
- Clearfield County Housing Assistance Program (HAP): This funding is received from the Pennsylvania Department of Human Services annually. It is typically used as rental assistance funding for consumers needing help paying rent.
- Clearfield County 2015 PHARE Rental Housing Rehab Program: This program receives funding from Pennsylvania Housing and Finance Agency Act 13 Funds. It is available to develop affordable housing units out of structurally-sound vacant and or blighted structures.
Potential applicants include not-for-profit organizations, housing and human service providers, private organizations and developers.
For a copy of the RFQ, please contact the County Planning and Community Development office at 814-765-5149.
Responses to this RFQ should be submitted no later than Feb. 26. Request for Proposals of the HAP and PHARE programs will be sent to organizations and developers as qualifications are met.


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JAMIE YOST - RYMAN AUDITORIUM
4 boneless, skinless chicken breasts
1 cup buttermilk
1 cup Panko bread crumbs
½ tsp smoked paprika
½ tsp garlic powder
½ tsp onion powder
Salt & Pepper
8 slices deli ham
3 oz bleu cheese, crumbled
1 Bunch Rainbow Chard
½ white onion, chopped
1 10.5 oz. can cream of mushroom soup
½ cup sour cream
½ cup red cooking wine
Preheat oven to 375 degrees.
Cover chicken breasts with plastic wrap - pound breasts until approximately ½" thick. Place in buttermilk and allow to sit for at least 30 minutes or as long as overnight. Combine bread crumbs, paprika, garlic & onion powders, and salt & pepper to taste. Dip each chicken breast in bread mixture, coating both sides.
Sprinkle approximately half of the bleu cheese in the chicken breasts, distributing evenly. Top each chicken breast with two pieces of ham.
Separate chard leaves from stems. Chop stems and place in 9x13 casserole dish along with the onions.
Roll up the chicken breasts so that the ham and cheese are on the inside of the roll. Secure with toothpicks and place in casserole dish on top of chard stems & onions.
Bake for approximately 10 minutes. While the dish is cooking, combine cream of mushroom soup, sour cream, and cooking wine. Chop chard leaves coarsely.
Remove dish from oven and nestle in remaining chard leaves around the chicken breasts. Sprinkle remaining bleu cheese evenly over entire dish. Pour soup mixture over casserole dish and return to oven for 20 - 25 minutes, or until chicken is cooked through.

domain: food


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There are many opportunities to engage with others in the community. Check out what’s coming up:
- This event has passed.
What’s New in Food Gardening with Brian Minter
January 15, 2021 @ 2:00 pm – 3:00 pm PST
Do you wish you could grow your own vegetables for cooking but you just don’t have the space to do so? Think again. Join Brian Minter, an experienced master gardener as he introduces us to hot new varieties of easy-to-grow food plants for 2021. His talk will also feature helpful tips and tricks for how to successfully grow these foods yourself in containers and other small spaces around your house.
Brian Minter is a Horticulturalist, Entrepreneur and Businessman. He is the President and Co-Owner of Minter Country Garden Ltd. In Chilliwack, BC, a 30-acre innovative destination garden centre and greenhouse growing operation. Brian is also a garden writer and TV personality, a Board member for the Chilliwack Economic Partners Corporation (CEPCO), the Chair for Tourism Chilliwack, the First Chancellor of the University of Fraser Valley, and the former Chair of BCAA.
Click the button below to be directed to the event.


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D.M. Bowman, Inc., named as one of the Top 200 Carriers company in the nation, operates its dedicated transportation, warehousing and logistics organization from headquarters strategically located at the crossroads of Interstates 70 & 81 in Maryland. Founded in 1959, we are privately held, and take great pride in our motto, “We Carry Through!” During these economic times, it should be noted that we remain financially sound; focusing upon developing long-term, mutually beneficial relationships based upon sound logistics solutions.
From regional van to bulk, flatbed and JIT transportation services to warehousing, inventory management, distribution and value-added services, D. M. Bowman, Inc. is ready and fully committed to be your total transportation and logistics provider. We have CTPAT cross border certification, TWICS drivers and are SmartWay Certified. With over 1 million square feet of warehouse space located within Foreign Trade Zone #255, we manage terminal and/or warehouse operations in several states.

domain: economics


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One of the amazing things about our job, is that we're often asked to photograph multiple siblings' weddings in the same family - that was the case with the Malamas family this year. In 2008, we photographed Maryann's wedding, then in 2011, it was Stephanie's turn - this summer, we photographed Nicole's beautiful day - and met her sisters' new babies! Nicole and Mike wanted an intimate wedding that reflected their fun personalities, the beautiful summer colours, and a view to show off Vancouver to their guests. Coal Harbour definitely achieved that - the area is lovely, and showcases the Vancouver skyline extremely well. The surrounding flowers and gardens gave the creative session a perfect background for the couple and their wedding party - Nicole and Mike's attendants were fun ad engaging and clearly had close relationships with the couple, which made for lots of great interaction between the four friends.
One thing that really surprised us, was the appearance of a former colleague of Randal's from his hi-tech days, Wendy Yacboski. Wendy was the officiant for the couple (she did a fabulous job!), and Randal had a moment of his "two lives merging into one"!
The reception featured Vancouver-based caterer, Savoury City - headed by the wonderful Donna Wadsworth and her team, the guests feasted on tasty-looking canapés and a buffet of fine foods. We loved the cupcake display and the colourful array of icing, which matched the colours and mood of the day. Nicole and Mike looked completely happy and joyous throughout the entire day - we really enjoyed capturing their looks, their sense of humour, and their sweetness together.
Ceremony & Reception // Coal Harbour Communnity Centre
Officiant // Wendy Yacboski Musicians //
Harpist - Mike's Aunt Wedding Gown // Isabelle's Bridal
Wedding Shoes // Nine West
Makeup // Pamper Me Pretty
Floral Design // Bridal Beginnings
Cake // Cupcakes
Decor // Bridal Beginnings/Lonsdale Event Rentals
DJ // Byron James
Photography by Randal Kurt for Randal Kurt Photography. Special thanks to Katerina Stavrakis for assisting us on Nicole and Mike's day!


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Four days before Christmas in 1973, Benny Hinn went with his friend, Jim Poynter, on a charter bus trip from Toronto to Pittsburgh to a meeting of Kathryn Kuhlman.
He knew very little about her ministry, but little did he know that It would be a day that would change his life forever!
She was going to minister in The First Presbyterian Church in downtown Pittsburgh. As at the time they got there, hundreds of people were already there, but the door was not opened as it wasn’t yet time for the program. What a wonderful thing to know multitude are panting after God.
Benny Hinn stood in line, but as he was standing, he suddenly began to shake, nothing like that had ever happened before and it also didn’t stop. He felt too embarrassed to tell Jim his friend or anyone around me.
When they got in, they were able to sit close to the front on the third row of the auditorium, but still the shaking still continued. Soon he began to enjoy it he said “The longer it continued, the more beautiful it became”.
Then Kathryn Kuhlman appeared on stage. The atmosphere in the building changed immediately. Everyone began to singing “How Great Thou Art.” Tears gushed down his face, it wasn’t just tears but it was the feeling of intense glory. He also was literally singing the words from his soul.
Benny Hinn in his young Christian experience, he has been touched severally by God but on that day it was special because the way God was touching him was not as it used to be, it was unique and powerful and he felt as if the waves of a gentle breeze were flowing around him.
As Kathryn Kuhlman began ministering to the people, he was so lost in the Spirit that all he could whisper was, “Dear Jesus, please have mercy on me.” He felt he was so unworthy and that propelled him to say those words again and again.
The service was three hours which may have been so long for some people but to Benny Hinn it seemed like a fleeting moment. He said he saw the deaf begin to hear; people rise from their wheelchairs, saw tumors vanish, and so much more. Then everything stopped.
Benny Hinn said he prayed silently, “Please, Lord, don’t ever let this meeting end.” Because he was enjoying what the Holy Spirit was doing to him and other people around.
Kathryn was burying her head in her hands, sobbing so loudly that everything came to a standstill. Then, standing just a few feet in front of Benny Hinn, he said “her eyes seemed aflame as she took on a boldness he had never seen in anyone before then she began pleading as she looked out over the audience, “Please, don’t grieve the Holy Spirit!”. But he felt as if her eyes were looking straight at him. Then she said, “Don’t you understand? He’s all I’ve got!” That statement got Benny Hinn thinking, “What is she talking about?” She continued speaking, “Please! Don’t wound Him. He’s all I’ve got. Don’t wound the One I love!”
Benny Hinn said “i will never forget those words; I can remember the intensity of her breathing when she said them. Those moments are etched permanently into my memory. Then she pointed her finger down at me and said with such powerful clarity, “He’s more real than anything in the world!” At that moment, I cried and said, “I’ve got to have this!” Then, within moments, the service was over”.
Benny Hinn was still shaking but at this time he was shaking to the core. He knew that God had used that service and His mighty evangelist to reveal His power that day.
All through his way back to Toronto, he kept thinking, “What did she mean? What was she saying when she talked about the Holy Spirit?”
On getting home he was totally exhausted, but he could not sleep.
Benny Hinn said “As I lay on my bed, I felt as if someone were pulling me off the mattress and onto my knees. It was a strange sensation, but I didn’t resist. I had never spoken to the Holy Spirit before. I didn’t know He could be addressed that way. Finally, I prayed, “Holy Spirit, Kathryn Kuhlman says You are her Friend. I don’t think I know You. Before today, I thought I did. But after that meeting, I realize I really don’t. I don’t think I know You.” Then, like a child with my hands raised, I asked, “Can I meet You? Can I really meet You?”
He said a few brief moments passed, then like a jolt of electricity his body began to vibrate all over again, exactly like at the church in Pittsburgh. But this time he was afraid to open his eyes. He also felt like a warm blanket of God’s power was wrapped all over him.
That continued until he finally slept that night. But still could not tell what really had happened to him.
The next morning, three days before Christmas, he woke up and the first words he spoke to his newfound Friend were, “Good morning, Holy Spirit!” At that instant, he knew He was there with him.
Benny Hinn went on to say “As I opened my Bible, He was there sitting beside me and from that moment, everything about the Bible, my prayer life, my worship, and my relationship with God took on an entirely new dimension. For the next eight hours I had an incredible experience with the Holy Spirit that changed the course. Tears of wonderment and joy coursed down my cheeks. I had just turned twenty-one. Christmas was just around the corner. I had received the best present ever!”
Less than a year later, God open the door for Benny Hinn to step behind a pulpit to preach for the first time, and in that moment He would simultaneously heal a lifelong stuttering problem and that gave birth to a ministry that is eventually reaching out to people around the globe.
That’s why, forty years later, I commemorate all that God has done! As I boarded the charter bus for Pittsburgh back in December 1973, who could have fathomed all that would happen as a result of that momentous little trip to see Kathryn Kuhlman? Says Benny Hinn.

domain: literature


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Eating cholesterol has gained a bad reputation. Many people believe that consuming cholesterol can increase their cholesterol levels and, in turn, negatively affect their heart health. The truth is that dietary cholesterol doesn’t have the same effect on cholesterol levels as saturated fat does. In this post, we’ll take a look at the truth about dietary cholesterol, debunk common myths, and discuss the health implications of dietary choices.
What is Dietary Cholesterol?
Dietary cholesterol is the kind of cholesterol found in food items and beverages. It is found in animal sources such as eggs, red meat, poultry, and dairy products and is also found in some shellfish and organ meats. Unlike the cholesterol your body produces, dietary cholesterol is not created by your body and must be obtained from your diet.
Do Dietary Cholesterol Levels Affect Your Health?
A diet high in saturated fat has been linked to an increase of cholesterol levels in the body. But, dietary cholesterol is not believed to have the same effect on cholesterol levels in your body. Studies indicate that increasing or decreasing dietary cholesterol in your diet has little to no impact on your cholesterol levels.
The Health Benefits Of Consuming Cholesterol
Despite its negative reputation, there are some health benefits associated with consuming dietary cholesterol. For starters, cholesterol is essential for your body to produce hormones and cell membranes. Additionally, some studies have shown that consuming cholesterol may have a positive effect on brain health and memory.
Debunking Common Myths
A common misconception is that dietary cholesterol can increase your cholesterol levels. In fact, the opposite is true – saturated fat is the main culprit when it comes to raising one’s cholesterol levels. Additionally, dietary cholesterol does not increase one’s risk for developing heart disease.
Ultimately, the truth about dietary cholesterol is that it does not have the same effect on cholesterol levels as saturated fat. While there has been a lot of conflicting information over the years, recent studies indicate that dietary cholesterol can be consumed healthily. However, it is important to keep in mind that the healthiest diet is low in saturated fat.
Dietary Cholesterol, Saturated Fat, Cholesterol Levels, Heart Disease, Brain Health, Memory, Healthy Diet

domain: health


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Iran's Esteqlal soccer team has advanced to the knock-out stage of the AFC Champions League after defeating Uzbekistan's Nasaf Qarshi 2-0 away.
Serbian striker Goran Jerkovic gave the Blues the lead ten minutes after the time on Wednesday.
Five minutes after the break, Esteqlal midfielder, Mojtaba Jabbari, doubled the score from the penalty spot.
Esteqlal’s defender Mehdi Amirabadi was sent off in the 69th minute when he received the second yellow card.
Esteqlal, which ended the AFC group stage in the second place in Group A, will face Iran's newly-crowned Premier League champion, Sepahan, next Tuesday.
On Tuesday, May 15, Sepahan finished top of Group C after a 2-1 victory over Saudi Arabian team al-Ahli in Isfahan.
Meanwhile, another Iranian team, Persepolis, secured a place in the AFC Champions League round of 16 following a 3-1 victory over UAE’s al-Shabab.
Persepolis, which finished the second in Group D behind al-Hilal, will face another Saudi side al-Ittihad in the round of 16.


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[UPDATE: 2013 Luzerne County Budget Released]
WILKES-BARRE — The 2013 Luzerne County Budget Proposal was set to be released to Luzerne County Council members today, but it hasn’t been made public yet.
The Proposed 2013 Luzerne County Budget was supposed to be released earlier this week but got delayed until today because County Manager Bob Lawton said he needed more time. Newswatch 16 spoke with Lawton off-camera earlier today, and here’s what he told us.
Initially, the county had a budget gap of $12 million.
But Lawton says every day he’s been narrowing it down bit by bit so that when it’s released. There is no gap.
He says he’s been trying to avoid a tax hike at all costs. He also says there will be less spending in 2013 than there was this year. We asked him if that meant layoffs were likely. He didn’t use the word layoffs, but said the County would be quote unquote right-sized.
Again, this budget is supposed to be emailed to County Council members and posted to the county web site at some point tonight.
A formal presentation of the budget will happen at the County Council meeting on October 30th.


WASHINGTON, D.C. - The result of Wednesday's House of Representatives debate was never in doubt. Nobody expected Congress would approve Rep. Dennis Kucinich's resolution ordering President Obama to withdraw U.S. troops from Afghanistan by year's end.
The point was to give members of Congress three hours to officially sound off on U.S. military involvement in the Middle East.
"Regardless of your support or opposition, this resolution is about ensuring meaningful and open debate," the Cleveland Democrat told colleagues on the House floor. "This is going to be the first opportunity to evaluate critically where the authorization of the use of military force has taken us in the past eight and a half years."
Backers of Kucinich's measure said the U.S. has spent close to $1 trillion on the Iraq and Afghanistan wars that could have buttressed domestic needs like housing, jobs, health care and education.
"The more troops we send to Afghanistan, the more support the Taliban gains as resisters of foreign occupation," Kucinich said.
Opponents said passing his resolution would keep U.S. troops from thwarting terrorists.
"I am keenly aware that even if we remain in Afghanistan, there is no guarantee we will prevail in this fight, but if we don't fight, we are guaranteed to fail," said California Democratic Rep. Howard Berman, who chairs the House Foreign Affairs Committee.
Alliance Democratic Rep. John Boccieri, an Air Force Reserve major who served in Afghanistan, said the U.S. must establish political stability in Afghanistan and Pakistan to keep the region's nuclear weapons from falling into terrorist hands.
"It's been said we need a foreign policy based on realism, rather than idealism and that is why I will not be supporting this effort today," said Boccieri.
Democratic backers of President Barack Obama's troop surge in Afghanistan joined with Republicans to defeat Kucinich's measure in a 65-to-356 vote. Kucinich was the only Ohio member of Congress to support the resolution.
Five Republicans joined with liberal Democrats to back Kucinich's effort. Ron Paul of Texas called the war "illegal," "immoral" and "unconstitutional." John Duncan of Tennessee said fiscal conservatives "should be the most horrified about the hundreds of billions" spent there. Rep. Walter Jones of North Carolina added: "We need to understand, we can't police the world anymore."
But most Republicans were against it. Mark Kirk of Illinois, who served as a Naval Reserves intelligence officer in Afghanistan, said the debate plays into the agenda of Taliban propagandists. California's Duncan Hunter, a U.S. Marine who served in Iraq and Afghanistan, said the House floor discussion would "make America weaker" and undermine U.S. troops and their families.
"What we are doing in Afghanistan right now is going to make it so my children won't have to go and fight the same Islamofascists we are fighting," Hunter said.