text:
Decentral gene expression analysis: analytical validation of the Endopredict genomic multianalyte breast cancer prognosis test
Genetics, genomics and epigenetics
EndoPredict (EP) is a clinically validated multianalyte gene expression test to predict distant metastasis in ER-positive, HER2-negative breast cancer treated with endocrine therapy alone. The test is based on the combined analysis of 12 genes in formalin-fixed, paraffin-embedded (FFPE) tissue by reverse transcription-quantitative real-time PCR (RT-qPCR). Recently, it was shown that EP is feasible for reliable decentralized assessment of gene expression. The aim of this study was the analytical validation of the performance characteristics of the assay and its verification in a molecular-pathological routine laboratory.
Gene expression values to calculate the EP score were assayed by one-step RT-qPCR using RNA from FFPE tumor tissue. Limit of blank, limit of detection, linear range, and PCR efficiency were assessed for each of the 12 PCR assays using serial samples dilutions. Different breast cancer samples were used to evaluate RNA input range, precision and inter-laboratory variability.
PCR assays were linear up to Cq values between 35.1 and 37.2. Amplification efficiencies ranged from 75% to 101%. The RNA input range without considerable change of the EP score was between 0.16 and 18.5 ng/μl. Analysis of precision (variation of day, day time, instrument, operator, reagent lots) resulted in a total noise (standard deviation) of 0.16 EP score units on a scale from 0 to 15. The major part of the total noise (SD 0.14) was caused by the replicate-to-replicate noise of the PCR assays (repeatability) and was not associated with different operating conditions (reproducibility). Performance characteristics established in the manufacturer's laboratory were verified in a routine molecular pathology laboratory. Comparison of 10 tumor samples analyzed in two different laboratories showed a Pearson coefficient of 0.995 and a mean deviation of 0.15 score units.
The EP test showed reproducible performance characteristics with good precision and negligible laboratory-to-laboratory variation. This study provides further evidence that the EP test is suitable for decentralized testing in specialized molecular pathological laboratories instead of a reference laboratory. This is a unique feature and a technical advance in comparison with existing RNA-based prognostic multigene expression tests.
EndoPredict (EP) is a multigene assay which predicts the risk of distant metastasis in ER+/HER2- breast cancer and identifies a subgroup of patients who have an excellent prognosis if treated with endocrine therapy alone
Translation of the EP test from research laboratory to clinical practice covered the necessary steps for development of a laboratory test (Figure
Figure 1
Translation of the EndoPredict multigene expression test from research laboratory to clinical practice
Translation of the EndoPredict multigene expression test from research laboratory to clinical practice. Workflow of sequential discovery and clinical as well as analytical validation is shown.
Besides EndoPredict, other prognostic multigene expression tests for patients with breast cancer like MammaPrint
The aim of this study was a comprehensive analytical validation of the EP test to complete development before implementation in clinical practice. Analytical validation of multianalyte assays is still a challenge, as these types of assays require a more complex evaluation of the performance characteristics compared with single analyte assays in order to assure reliable performance in the clinical routine. Adequate performance evaluation includes the control of the process from the acquisition of the tumor samples and isolation of the RNA to the assessment of each single analyte as well as the combination of the single results to a comprehensive score by an algorithm. Moreover, guidelines for analytical validation of multianalyte genomic assays are rare. Here, we analytically validated the EndoPredict multianalyte gene expression assay according to the adapted guideline MM17-A of the Clinical and Laboratory Standards Institute (CLSI) addressing the analytical validation of nucleic acid-based qualitative and semiquantitative multiplex assays
Reference and Testing Materials
Nucleic acids for test development and validation were selected based on the specific purpose of the respective analytical performance characteristics to be tested. Sample material was comparable to the specimen used in the clinical testing, i.e. DNA-free total RNA or genomic DNA from FFPE tissue which is fragmented to nucleic acid pieces by formalin-fixation
As positive controls of RT-qPCR assays a standardized reference RNA (Stratagene qPCR Human Reference Total RNA, Agilent Technologies, Böblingen, Germany) and Human Genomic DNA (Roche Applied Bioscience, Mannheim, Germany) were tested on each plate.
Isolation of RNA and DNA
Total RNA and DNA was extracted from FFPE tissue sections (10 μm) using a fully-automated silica-coated magnetic bead-based method in combination with a liquid handling robot (VERSANT Tissue Preparation System, Siemens Healthcare Diagnostics, Eschborn, Germany) as published previously
The mean of Cq (quantification cycle) values of the EP reference genes
Gene expression analysis using RT-qPCR
Expression of 8 genes-of-interest (
For calculation of the EP score genes were measured in triplicates. This is mandatory to control for PCR imprecision and to enable outlier removal
Assessment of limit of blank (LoB), limit of detection (LoD), linear dynamic range and PCR efficiency
LoB, defined as the 5%-percentile of the distribution of Cq-values measured in a blank sample without analyte, was calculated as described in supplemental data (see Additional file 1)
Assessment of precision
The precision experiment was designed according to CLSI guidelines
For verification in the laboratory of Charité as a representative routine laboratory an abbreviated precision experiment with fewer variables was performed: day (n=5), day time (n=2), position of sample on 96-well EndoPredict plate (n=2), lot of enzyme/master mix (n=2), and operator (n=2).
Variable noise, replicate noise and total noise was calculated using univariate N-way analysis of variance (ANOVA) and indicated as standard deviations
For EP scores 95% confidence intervals (CI) were calculated as described
Limit of blank, limit of detection, linear dynamic range and PCR efficiency
For each of the 12 genes the analytical performance of the RT-qPCR assays was assessed. For a type I error of 5% the LoB was at a Cq value of 40 for all genes (Table
Additional file 1
Supplemental Data: File contains detailed descriptions of methods as well as supplemental tables and figures which could not be included in the main manuscript.
Click here for file
All 11 RNA-specific assays were linear up to dilutions between 2-9 and 2-16 corresponding to Cq values between 35.1 and 37.2 (Table
Figure 2
RNA input range and reproducibility of EndoPredict
RNA input range and reproducibility of EndoPredict. (A) EP scores depending on amount of input RNA. RNAs from 6 different FFPE samples were diluted and EP scores were assessed dependent on RNA input (Cq-ARG as surrogate marker). 95% confidence intervals (CI) of EP scores calculated from the noise model are indicated. (B) Correlation between Cq-ARG and total RNA concentration assessed by RIBOGREEN assay. Lower RNA input limit is indicated by dotted lines. (C) Reproducibility of 160 EP scores assessed in three different RNA samples (low risk, close to the decision point, high risk) over time (11 different working days distributed over 21 calendar days). Individual EP measurement results are indicated by dots.
Input range
For a multigene expression test it is essential to determine the acceptable range of input RNA within which the assay yields accurate results for all variants tested
Precision of the multigene expression assay was evaluated under various stipulated operating conditions including day, day time, PCR instrument, position of the sample on the EndoPredict plate, plate lot, reagent lots, and operators and using three different test RNA samples from FFPE breast cancer tissue with a low EP score (2.4), a high EP score (13.5), and an EP score close to the decision point (4.9). In total, 160 EndoPredict tests (Figure
Verification of performance characteristics in an independent laboratory
The performance characteristics of the EndoPredict assay were verified in a routine laboratory at the Charité in Berlin to confirm that the test performs to specifications also in a routine diagnostic laboratory. The parameters verified were efficiency of the single PCR assays, precision, input range, and analytical accuracy with respect to reference values.
For assessment of linear range and efficiency two independent series of seven 1:2 dilutions of the reference RNA pools and DNA from FFPE tissue were generated. Each nucleic acid concentration was assessed four times. The 11 RNA-specific assays were linear over the whole range of concentrations analyzed (dilutions up to 2-7), the
Figure 3
Verification of performance data at Charité
Verification of performance data at Charité. (A) EP scores depending on amount of input RNA assessed as Cq-ARG. Pre-specified input limit is indicated by a dotted line. 95% CI are given. (B) Correlation of EP scores of 10 different breast cancer FFPE samples assessed at the molecular-pathological laboratory at the Charité compared to reference values assessed at Sividon. The cutoff value between low and high risk for distant metastasis is indicated by dotted lines.
Moreover, precision of the EndoPredict test was verified assessing the impact of the day, day time, the position of the sample on the 96-well EndoPredict plate, the reagent lot, and the operator on the reproducibility of the assay. The two RNAs from the tumors with the low and the high EP score were analyzed using the EndoPredict test resulting in 659 Cq values, 160 ΔCq values and 20 EP scores. The total variation (standard deviation) of the EP scores was 0.18 (Table
Finally, the analytical accuracy of the EndoPredict assay performed in the Charité laboratory was examined. For that purpose, ten breast tumor samples were selected and the EP scores were determined at Sividon. These pre-determined scores ranging from 3.3 to 11.0 were used as reference values. Five of the cases were very close to the predefined cutoff of the EP score. The pre-specified aim for the verification study at Charité was that the difference between the EP score at Charité and the reference EP score was below 1.0 EP units for at least 9 of 10 samples. Charité received a 10 μm tissue section of each of the ten tumors, isolated RNA and performed the EndoPredict test. The aim of this verification study was achieved as the largest deviation from the reference value was 0.36 score units with a mean deviation of 0.15 (Figure
In this study, we showed by means of a defined analytical validation and verification process developed according to the CLSI guidelines that the RT-qPCR-based EndoPredict multianalyte gene expression test is a robust test that can be performed reproducibly and accurately. The resulting performance characteristics therefore meet the requirements needed for a diagnostic test. Moreover, we verified that a comparable performance with respect to assay efficiency, precision, and accuracy can also be achieved in a routine molecular diagnostic laboratory. In addition, this study provides the specifications for analytical verification of EndoPredict in molecular pathological laboratories.
Successful clinical validation of the EndoPredict score in two large clinical trials was published previously
The difficulty of a comprehensive analytical validation of a highly complex genomic multianalyte assay such as EndoPredict is that there are no evaluation guidelines to cover all relevant aspects. Therefore, we specified a validation program for both, the 12 single analyte assays and the combined multigene assay. Where applicable, established guidelines of the CLSI as well as the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines were followed
Besides EndoPredict, other prognostic multigene expression tests for patients with breast or other cancers like MammaPrint
Robust performance of the EndoPredict test in a decentralized setting was also shown in a successful prospective proficiency testing study including seven different molecular pathology laboratories
A recent study assessed the variability of Ki-67 immunohistochemistry, which is a standard antibody-based diagnostic test in pathology used for treatment decision making in luminal breast cancer
Although our results might suggest a higher reproducibility of the PCR-based test using standardized instruments and reagents, it is important to know that the pathological laboratories involved in this technical verification study as well in the proficiency testing of EndoPredict were highly experienced in molecular work. Therefore, the results might be different in laboratories with less molecular diagnostic experience and ongoing quality control by periodical round robin tests might be reasonable.
A critical issue for accurate results from any diagnostic assay is the use of optimal sample material. For this validation study optimal FFPE tumor material was selected by experienced pathologists on the basis of adjacent HE-stained tissue slides. Hence, in order to obtain high-quality EndoPredict results an expert pathological evaluation of the tissue specimens is mandatory. Moreover, a highly standardized method for RNA isolation as used in this validation study and in the EndoPredict proficiency testing is necessary. In this study, the validation of the RNA extraction method was not a primary aim. The silica-coated magnetic beads-based method used and recommended for EndoPredict was thoroughly validated in previous studies showing a reliable analysis of RNA transcript levels by RT-qPCR in FFPE tissue
In conclusion, this study provides the analytical performance characteristics of the EndoPredict breast cancer prognosis multigene expression assay. They can be used as a reference for analytical verification of the test in molecular pathological laboratories. Moreover, the study shows a robust and reliable performance of the test and provides conclusive evidence that RT-qPCR-based quantitative multigene expression analysis of FFPE tissue samples is feasible in a decentral setting in molecular pathological laboratories. This is a major technical advance in comparison with existing prognostic multigene expression tests which are performed in central reference laboratories. Together with the clinical validation studies and the prospective analytical proficiency testing program with seven different molecular pathological laboratories these results provide the basis for the application of EndoPredict as a test to assess prognosis under endocrine therapy in clinical decision making in a decentralized environment.
EP: Endopredict; ER: Estrogen receptor; FFPE: Formalin-fixed, paraffin-embedded; RT-qPCR: Reverse transcription quantitative real-time PCR; LoD: Limit of detection; Cq: Quantification cycle; GOI: Gene-of-interest; LoB: Limit of blank; Cq-ARG: Average Cq value of reference genes; CI: Confidence interval.
Competing interests
RK, KB, FH, CR, MA, TR, CW, JCB, KEW, KF, CP are employees of Sividon Diagnostics GmbH, RK, KB, CR, MA, TR, CW, KEW, CP, MD, CD hold shares of Sividon Diagnostics GmbH. MS, MF, PD received speakers' honoraria from Sividon Diagnostics. Sividon Diagnostics has applied for a patent relating to the content of the manuscript.
Author's contributions
RK, KB, JCB, KEW, MS, MF, PD, CP, MD, CD were involved in conception and design of the study as well as in analysis or interpretation of data. JP, BVS, FH, CR, MA, TR, CW, KF, BMM acquired, analyzed and interpreted data. All authors have been involved in drafting or critically revising the manuscript and approved the manuscript.
We would like to thank Britta Bayer, Petra Wachs, and Ines Koch for their excellent technical assistance.
Part of this work was funded by the German Ministry of Science and Technology (BMBF) in the NEOpredict project.
A new molecular predictor of distant recurrence in ER-positive, HER2-negative breast cancer adds independent information to conventional clinical risk factors
Decentral gene expression analysis for ER+/Her2- breast cancer: results of a proficiency testing program for the EndoPredict assay
Comparison of the RNA-based EndoPredict multigene test between core biopsies and corresponding surgical breast cancer sections
RNA extraction from archival formalin-fixed paraffin-embedded tissue: a comparison of manual, semiautomated, and fully automated purification methods
Quantitative determination of estrogen receptor, progesterone receptor, and HER2 mRNA in formalin-fixed paraffin-embedded tissue–a new option for predictive biomarker assessment in breast cancer
Automated extraction of DNA and RNA from a single formalin-fixed paraffin-embedded tissue section for analysis of both single-nucleotide polymorphisms and mRNA expression
Tumor-associated lymphocytes as an independent predictor of response to neoadjuvant chemotherapy in breast cancer
Gene expression profiling predicts clinical outcome of breast cancer
A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer
Validation study of a quantitative multigene reverse transcriptase-polymerase chain reaction assay for assessment of recurrence risk in patients with stage II colon cancer
Supervised risk predictor of breast cancer based on intrinsic subtypes
A comparison of PAM50 intrinsic subtyping with immunohistochemistry and clinical prognostic factors in tamoxifen-treated estrogen receptor-positive breast cancer
Effect of formalin, acetone, and RNAlater fixatives on tissue preservation and different size amplicons by real-time PCR from paraffin-embedded tissue
The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments
Statistical methods for assessing agreement between two methods of clinical measurement
Use of archived specimens in evaluation of prognostic and predictive biomarkers
Converting a breast cancer microarray signature into a high-throughput diagnostic test
Analytical validation of the Oncotype DX genomic diagnostic test for recurrence prognosis and therapeutic response prediction in node-negative, estrogen receptor-positive breast cancer
Translating tumor biology into personalized treatment planning: analytical performance characteristics of the Oncotype DX Colon Cancer Assay
How reliable is Ki-67 immunohistochemistry in grade 2 breast carcinomas? A QA study of the Swiss Working Group of Breast- and Gynecopathologists
Pre-publication history
The pre-publication history for this paper can be accessed here:


text:
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text: Google seems intent on pushing UGC (user generated content) down in the search results. There are some active conversations in their product forums where Google reps have suggested that sites de-index UGC that is light (zero replies, thin content). As a forum admin, this sounds sort of insane. 88k discussions and almost 1 million messages, this is impossible to do without some level of automation. It would also have to tie in with whatever sitemap tool you are using. Google's algorithms could change for the better, but I'm not so sure. I think as forum admins, we need to spoon feed our best content and leave the thin stuff as no-index. I'd welcome ideas on how to best accomplish this going forward with XenForo. Source, Discussion on Google's Product Forums


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Interviewing is such an important part of the job process, and it's important to present yourself well. No matter how qualified you are, you're unlikely to get the job if you flub the interview.
There are lots of things that are out of your control including plain old chemistry. But don't sabotage your chances of landing a job with things you can control. Here are some interview tips on to handle the worst mistakes you can make in an interview.
Not Knowing Why Are You Applying For This Job
If your job-searching process is already long and exhausting it's easy to become a victim of this routine and apply automaticly to every job that fits you. Ofcourse, when your resume is polished relevant you will most likely get an unterview, but will you get an offer?
When you are going for an interview or applying for a job you need to give the interview compelling reasons for giving you the job and that starts with the basic question: Why do you want it? Why are you - a perfect candidate? You will put yourself at a disadvantage before you even walk into an interviewer's office if you fail to prepare for the discussion in advance.
We will never get tired to repeat that researching is a must.
READ ALSO: What You Should Know About a Company Before You Join And How To Do That
Background information including company history, locations, divisions, and a mission statement are available in an "About Us" section on most company websites. Review it ahead of time, then print it out and read it over just before your interview to refresh your memory. Also check the company's LinkedIn page and Facebook page, if they have one. Think about what you like about this job and why you want it.
Lack Of Enthusiasm
In a tightly run job race, when the two top candidates have equal qualifications, the job offer will always go to the most intelligently enthusiastic candidate. The problem for most of us is that interviews are stressful events, and under stress our defenses go up and any natural enthusiasm for our work is buried in a wall of stiff professionalism. This can get in the way of winning job offers.
READ ALSO: Job Interview: Words You Should Never Use
You must show energy and enthusiasm if you want to be seriously considered for any opportunity. If a company doesn't see your desire or enthusiasm for a position during an interview, you will likely be disqualified from consideration for the position.
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Not Establishing Common Ground
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READ ALSO: 3 Things Every Interviewer Wants to Hear
Being good at small talk isn't something everyone is automatically good at, but it's really important for getting people to like you. Develop a genuine interest in people–this is a skill that will never fail you your entire life. Be a good listener, support the conversation instead of just sitting there, staring blankly at your interviewer and waiting for them to pounce on you with the first question.
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text: This event will host three physician speakers to include Helen Hui-Chou, MD Plastic Surgeon and two of our general breast surgeons. Activities include education, stress management, health screenings to include annual mammogram if they are due, meditation, massage, healing touch, make overs, hair styling and a healthy food cooking demonstration. We are planning our fund raising activities with philanthropy and our vendor sponsors for this event.


text:
[INFO]Rooting the Nexus S 4G – Windows Version (XP/Vista/Windows 7)
Post 2 : Download Required Files
Post 3 : Windows SDK Rooting Version
Post 4 : One Click Root Version
Post 5 : Credits
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[INFO]Rooting the Nexus S 4G – Windows Version (XP/Vista/Windows 7)
Post 2 : Download Required Files
Post 3 : Windows SDK Rooting Version
Post 4 : One Click Root Version
Post 5 : Credits
[INFO]Required Files to Download
:: SDK Method Required Files ::
android-sdk_r16-windows.zip (Android SDK) Download & Extract to C:/ of your hard drive
Superuser-3.1.3-arm-signed.zip (Superuser/Root) Place on root level of sdcard on device
recovery-clockwork-220.127.116.11-crespo4g.img (ClockWorkMod recovery image) Place in your C:/android-sdk/platform-tools/
PDAnet (Aid in connection & driver installation) Download, don't install until instructed.
Optional Windows Samsung Drivers (Download) Click on file to install to computer
:: One Click Root Method Required File ::
One Click Root v5.0 (Download ONLY if your using the One Click Root method)
[NOTE]Android SDK Package - Windows
[WARN]Performing the following procedure will completely wipe your device. Make sure you contacts are synced before moving forward and backup any pictures, music, etc. from your internal sdcard location. You have been warned ![/WARN]
1. Assuming you have downloaded the required files from previous post. Make sure android-sdk setup has been extracted to your C:/ location of your hard drive. This location when extracted to C:/ will actually be named a folder called " android-sdk-windows ".
[TIP]To get your platform-tools folder properly installed inside your C:/android-sdk-windows folder, please see this POST[/TIP]
2. Place the recovery-clockwork-126.96.36.199-crespo4g.img in the C:/android-sdk-windows/platform-tools/ location if you haven't already.
3. Download the following file, Superuser-3.1.3-arm-signed.zip and place it on your root level internal storage/sdcard.
4. Starting with your device booted and everything ready to go proceed to Step #5
5. On your device go to Settings/Applications/Development/USB Debugging. Go ahead and enable this option.
6. Now plug your device via USB cable into your computer and allow the drivers to install.
7. Go back to your computer and run the PDAnet setup file.
Follow the prompts and when it gets to the following picture below to install the drivers, pick Samsung and follow up by doing as it instructs and plug in your device once again with debugging enabled to finish installing the drivers.
8. Now you will need to shut down your device and then press and hold Volume UP + Power button and you will be booted into your bootloader.
9. Go back to your computer, press the Windows Icon/Start in the lower left hand corner task bar and either from RUN or the small search bar type CMD and press ENTER. A Windows DOS window will open for you.
10. Type the following from the command line and press ENTER.
11. Now your command prompt should look like the screenshot below.Code:
cd c:\android-sdk-windows\platform-tools\
Microsoft Windows [Version 6.1.7601]
Copyright (c) 2009 Microsoft Corporation. All rights reserved.
C:\Users\PaulSR>cd c:\android-sdk-windows\platform-tools\
12. Now type in the following command prompt and press ENTER.
You should then be presented with a screen on your device like the screenshot below.Code:
fastboot oem unlock
13. Now after reading the speech on the screen about unlocking your bootloader and taking note that this will completely WIPE YOUR DEVICE, go ahead and select YES.
14. Now from your command prompt type the following command and press ENTER. This will install ClockWorkMod recovery to your device.
15. Once this is complete you can then select RECOVERY from the list on your bootloader screen. (Navigate by using Volume UP/Down and make selections by pressing Power). This will then boot you into ClockWorkMod (CWM) 22.214.171.124.Code:
fastboot flash recovery recovery-clockwork-22.214.171.124-crespo4g.img
16. Now again using the same buttons to navigate go down to Install from SDCARD. Now follow the prompts to select a file, look for the Superuser-3.0.7-efghi-signed.zip you placed on your sdcard and process to install.
17. Now you have completed the rooting process :) You may reboot your device.
Now you can go searching for your favorite ROMs, kernels, mods, etc. and enjoy a new way of using your device :)
** I took the liberty of adding the code so when you flash the Superuser file it also makes the CWM "stick".
[INFO]Version #2 � One Click Root Package
[warn]Performing the following procedure will completely wipe your device. Make sure you contacts are synced before moving forward and backup any pictures, music, etc. from your internal sdcard location. You have been warned ![/warn]
Now we all have heard the goods and bads of One Click Root packages. 99% time they work and the other 1% of the time they cause problems.
This One Click Root package works, I tested it myself and if you follow the prompts & instructions its a painless process.
1. Download the following package and extract it's contents to your C:/ drive.
One Click Root v5.0
2. Open the One Click Root folder and with your device booted, place it in debugging mode (Settings/Applications/Development/USB Debugging) and then click on the run.bat file to begin the process.
3. From there the program basically walks you through the process. It will do a little, then ask you to participate by verifying certain things.
If your unrooted, then select the unrooted/stock device option and follow its instructions. Make sure when you get to the prompt about device you select the proper device (Nexus S 4G).
Later in the installation it will give you the option of two recoveries to choose from. The common ClockWorkMod (CWM) recovery and the new TWRP (Touch Screen) recovery. Both do basically the same thing, TWRP is newer and offers a little better layout.
[TIP]I have gotten a PM or two concerning the root.zip file that is supposed to be transfered in the process to provide Superuser access not working. To fix this situation see this POST[/TIP]
There really isn't much more to this package to explain because it basically holds your hand through the process and ShabbyPenguin who designed it did an excellent job with it. [/NOTE]
[INFO]Credits & Resources
This guide was written up with use of various resources and credit to different people...
Android Central
Joshua Munoz
Any one who I didn't mention...
One more just in case....
You said seem to be having a problem where during One Click Root it didn't transfer the root file over?
1. If you boot into your bootloader and select RECOVERY what happens?
2. If you boot into ClockWorkMod or TWRP then all your are missing is the root.zip file is Superuser (root access). So in this situation then you can grab the Superuser file from the second post in this thread and flash in recovery to gain Superuser/root access. Use the Install from SDCARD option.
Please report back with your results.
when i boot into recovery it does nothing and brings up an android with a exclamation inside a triangle. only way out was to pull the battery. phone restarted normally after that
I attempted again after battery pull. it shows a picture of a box open with and arrow out for a second or so, then goes to triangle and exclamation image, I am guessing that the recovery file was not transferred correctly?
i may be getting ahead of myself but i attempted to run the one click root option again, this time i chose option 2 for have unlocked bootloader need recovery. went through steps chose CWM recovery option and get <waiting for device> on screen.
Option to try...
I guess go back and act if you are doing a fresh install...
Have that Superuser file ready on your computer. When and if you get booted into CWM recovery you have an option called Mounts & Storage from the main list.
Choose that option, then choose MOUNT USB STORAGE.
That will then connect your device to your computer and you can then move the Superuser file over to your device via Windows Explorer and then exit the USB MOUNT and then flash/install the Superuser apk.
Then you can reboot, and you should have root access and you CWM should stick instead of being erased.
I believe that worked. thanks for your help. Now onto installing ICS!!!!
thanks again.
Good deal...
I will have to amend my installation notes to take this possible situation into consideration.
Thats for pointing it out.
Okay there is a mistake probably on my part... When you extract the contents of the android-sdk_r16.zip file the folder inside that you place on your C:/ drive location is actually called.Hi Paul,
I'm about to embark on the rooting process! One initial confusion. Your instructions say:
android-sdk_r16-windows.zip (Android SDK) Download & Extract to C:/ of your hard drive
Superuser-3.0.7-efghi-signed.zip (Superuser/Root) Place on root level of sdcard on device
recovery-clockwork-184.108.40.206-crespo4g.img (ClockWorkMod recovery image) Place in your C:/android-sdk/platform-tools/
On this last step, am I supposed to create a folder on my C drive "android-sdk" and then create a folder within called "platform-tools"?
I couldn't find this location after I downloaded and extracted the android-sdk_r16 zip file above to my C drive.
I forgot the "windows" part of the folder name. I will fix my OP to reflect this.
So search for the folder android-sdk-windows on your C:/ location and you should be fine.
As for your other part of the question, inside the android-sdk-windows folder there will already be the required platform-tools. So no need to create that.
On mine, these are the internal folders within "android-sdk-windows" folder:
I couldn't find a platform-tools folder.
maybe I spoke to soon. the one click installer finished and stated you phone is not rooted, however, I installed titanium back up and busybox and they are telling me that my phone is not rooted and cannot get SU access.
I also tried to go into recovery mode again after this and it gave me the same exclamation point in triangle screen and had to pull batter to reset.
I am guessing I am doing something wrong, just need to figure out what.
Obviously some problems have crept up here... I did run through and follow my own steps that I did in the guides so I know it works. The One Click Root should be working if you follow the amended tip I placed in the installation instructions guide. I ran into this problem too with the root.zip file and thought it was just me so I didn't initially mention it.
To fix the problem of not having the platform-tools folder in your android-sdk-windows here is what you need to do.
1. Open the android-sdk-windows folder on your hard drive location.
2. Click on/run the SDK Manager file located inside.
3. This will open a SDK Manager window and you need to look on the left hand side for an option to install SDK Platform Tools. This is the only option you need to worry about. Check the box to the left of the SDK Platform Tools option and then near the bottom right hand side of the window you will see an option " Install Package(s) " .
4. Allow this to complete, shouldn't take long. Then when completed you can go back into the android-sdk-windows folder and see if you now have a platform-tools, folder.
Here is a helpful guide for this directly from Google's SDK website.
Installing the SDK | Android Developers
Look under Available Components.
On this step -- "6. Now plug your device via USB cable into your computer and allow the drivers to install."
It failed on my computer -- said it couldn't find any device drivers. I had already installed that optional samnsung drivers file that you mentioned. Should I just proceed to the next step and run PdaNet file?
Yes... PDAnet should handle the rest.
Sent from my Nexus S 4g using Tapatalk
Thank you very much, Paul!
I followed the instructions for manual rooting and I'm now rooted. Time to install a custom ROM. Just now downloaded Titantiam Backup Pro.
Thank you for being patient and helping me get a few bugs worked out of the guide.
Enjoy your new world of rooting :)
Sent from my Nexus S 4g using Tapatalk
question for you paul...still having same issue that when recovery is run i locks up and a battery pull is required. when I go into recovery folder (TWRP) via usb connection to my phone I see it is empty. is this normal? it makes me believe that the recovery is being deleted and therefore final root cannot be achieved even though the program stated I was rooted?
There has been some issues with TWRP getting it to stick. I'm not sure what the deal is really. I obtained TWRP by flashing the standard CWM recovery and then there is an available TWRP recovery flashable that you can flash in CWM to switch over to TWRP... Sorry alot of TWRP talk here.. LOL...
I would try running it again and go with CWM and here is the TWRP recovery flashable thread.
[ACS] TWRP 2.0 Recovery flashable zip
finally, looks like that did, using CWM now. Appreciate all your help.
many many thanks
The PdaNet that was posted was for the 32 bit. Is there a 64 bit exe file?
Google PDAnet and go to their site and check.
Fyi this is the same PDAnet I use on my own computer and I have/run windows 7/64 bit.
Sent from my Nexus S 4g using Tapatalk


text:
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Por: ShemarooPúblicado: 2 anos atrás
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Karan, a dreaded gangster, falls in love with Kajal, the daughter of his nemesis Prof. Siddharth. After initial hesitation Kajal reciprocates Karans advances and tries to reform him with her love. However, Prof. Siddharth forces Kajal to marry Raju, the son of his childhood friend, Ramakanth. Thus, Karan returns to his violent old days after suffering the heartbreak. Watch to know what transpires when Karan finds out that Raju is his next target. Will Karan kill Raju or stand guard for him against the odds?Watch to know more. Watch to know more.
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Hurrying Through The Street by bigbraader ()
Vue Fantasy posted on Apr 04, 2013
Report this image. Open Full Image
Members remain the original copyright holder in all their materials here at Renderosity. Use of any of their material inconsistent with the terms and conditions set forth is prohibited and is considered an infringement of the copyrights of the respective holders unless specially stated otherwise.
"The Prince of the Villains" is hurrying to a meeting nearby, escorted by his big guard "Oaktree". Things may seem quiet in the village, but the tension is just below the surface. Two local guardsmen are following the unlikely visitors with some attention - not exactly keeping a low profile on their homeground, they can't resist mocking the "dandy" looking prince a bit..."
- This is of course not mainstream Vue work, but I really like making these historical/fantasy scenes with characters and hints of a story.
Here I was also interested to see if Vue was able to pull this off - on my old 32 bit system Vue would struggle with the (great) village model alone, adding characters could be done only with caution and a great deal of planning and composite renders.
On my 64 bit system this potentially heavy scene was swift to make and render, once the character setup was done (great deal of work in that dept. though). Many huge and detailed texture maps, lots of details - no problem :)
Toolbox: Vue render, DAZ Studio, Photoshop.
Items from DAZ, Cornucopia 3D and myself.
Thanks for viewing, comments etc. welcome and appreciated.
Have a nice day,


text:
Discussion in 'NFL Zone' started by WoodysGirl, May 19, 2014.
The Los Angeles Eagles. yes! we need only classy fans representing the powerful NFC East.
And maybe move the bills from western ny to Canada.
oR as an alternate to the first nomination...
if Toronto can't handle the Bills, and the Egals won't relocate, move the Bills to Los Angeles and we trade out the filthadelphia egals to the AFC north so they have natural rivals with Pittsburgh and we take on the Cleveland "Johny Football" Browns....much better.
Screw it, all 32 teams will compete in the playoffs next year. The playoffs will run through like March and the Super Bowl will be in April.
Screw it, let's just get rid of the whole tournament thing. We'll have 16 bowl games each year.
Screw it, we don't need playoffs. Just automatically give everyone a trophy at the end of the season, just for trying.
I thought about it ..liked it for a minute then came to my senses. I really think 2 teams should be rewarded by making the playoffs and winning the regular season. A best of 7 super bowl from January to march
That would be very interesting to say the least. I think that might tend to get boring though. One of the reasons I'm not a big fan of the playoffs of other major sports is that the playoffs seem to drag on forever... which might not be a bad thing after all.
yeah we have this silliness going on here in this thread while in the draft zone we are discussing our young group of WRs. Its all good.
here's how I see this affecting the cowboys...
1) Jones pushes and pushes to get that extra team in there (and the league adds it)
2) Dallas has a great season, gets the #2 seed
3) The #7 seed is a 7-9 team
4) Dallas loses to them in round 1
5) Jones goes back to the owners meetings to remove the extra team each year
LOL. Yeah and we can stop counting the score, too. That way everyone is a winner.
Ian RapoportVerified account @RapSheet
#Cowboys owner Jerry Jones tells @KimJonesSports he's definitely for playoffs expansion. One reason? "Would help my Cowboys." #oh
It's no big surprise the owners favor the expension of playoff teams -- that was to be expected from those focused on profits far more than player safety or even paying heed to what the fans prefer. Far too many league decisions are getting away from fan preferences only to serve the almighty dollar. I'm hoping that eventually good reason prevails and the players and fans will merit more and better consideration in the future rather than lining the owners' pocketbooks. The future of the game is at stake. Many of those in positions of power seem to be turning their backs to that fact, IMHO. It certainly wouldn't be in their best interest to kill the goose that laid the golden egg; that's a given. The league and the owners had best take a deep breath and stifle the greed before it's too late. They've already succeeded in excessively watering down team rosters with their expansionism and lust for the greenbacks. Enough, already, before it's too late. Just my two cents on that . . .
Except we still wouldn't have made the playoffs last year. lol
Expanding the playoffs by only eliminating the bye would not add an extra weekend.
And while everyone wants to wallow in failure, it should be noted that under such a system we would have been in the playoffs the last few years. People would have to come up with a new meme to whinge about not winning Super Bowls.
I should note that the above statement was in no way directed at you or anyone in particular. I just don't want to make an extra post.
As a percentage of total teams, less teams get in now than did before 2000 when the NFL expanded and reset the divisions.
I was talking about a full 4 rounds so one year we would have gotten in. I still don't see a downside.
This guy is a ******.
Are you sure Dallas wouldve been in the last 3 years with said format? Im not sure but i think there were other teams with a better record who also didnt make it.
Being the last team limping into the playoffs in said format would be nothing to brag about. It actually makes the playoffs less credible by adding teams that aren't good or didnt earn it during the season. On the other hand, 10wins should put you in the playoffs. The cardinals got snubbed last year.
Of course there's wide-spread support, it will make more money for the teams.
Apparently we were 8th in 2008 but were 10th the other years so no. And getting in is getting in. 'Earn' speaks of worth and cost which is completely arbitrary.


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Fri 13th Jun 2014 10:00amRapturecamps's Uluwatu Surf Report
3ft waves coming in frequently and breaking comsistent at Racetracks Uluwutu this morning at 10 pm. 40 people out. Medium offshore wind.
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text:
Apple announced the launch of a bug bounty program, offering up to $200,000 for some discoveries.
Apple's head of security engineering and architecture, Ivan Krstic, made the announcement at Black Hat. The bug bounty program will get underway in September.
It has taken Apple some time to embrace this type of initiative, which sees researchers and hackers rewarded for their discoveries and submissions of security vulnerabilities. In fact, Apple was criticised during its battle with the FBI when the latter was trying to break into an iPhone used by Syed Farook, one of the individuals involved in the San Bernardino shooting last December. It was believed that Apple's unwillingness to reward researchers to share information of vulnerabilities resulted in the FBI being able to find a third party to exploit a flaw.
Krstic told attendees at Black Hat, "Feedback that we've heard pretty consistently both from my team at Apple and also from researchers directly is that it's getting increasingly more difficult to find some of those most critical types of security vulnerabilities. So the Apple Security Bounty Program is going to reward researchers who actually share critical vulnerabilities with Apple."
There will be five categories of risk and reward:
Interestingly, Apple is hoping that researchers who receive a reward will do some good with it. Apple is encouraging them to donate their reward to charity, and if Apple approves of a researcher's selected institution, it will match their donation.


text:
Benjamin Colton Barnes
January 2, 2012 by staff
Benjamin Colton Barnes, Benjamin Colton Barnes is a "strong person of interest" in the shooting of park ranger Magaret Anderson at Mount Rainier on Sunday. He is suspected to be armed and at large in Mount Rainier National Park.
Who is Barnes? in 2011, Barnes has had two restraining orders filed against him by the mother of his one-year-old child. In November, a court ruled that he could only have two supervised visits a week with his daughter until he completed a domestic violence and mental health investigation.
-The restraining orders indicate that Barnes is National Guard member or Reservist.
-Barnes has been named by an anonymous law enforcement source as a suspect in the Skyway shooting of four people on New Year's Eve.
Benjamin is considered to be armed and dangerous. If you see him, law enforcement urges you to not contact him and instead dial 9-1-1.
Please feel free to send if you have any questions regarding this post , you can contact on
Disclaimer: The views expressed on this site are that of the authors and not necessarily that of U.S.S.POST.


text:
Shooter headlines
PlayerUnknown's Battlegrounds: why the UI and HUD should absolutely stay the way they are
PlayerUnknown's Battlegrounds doesn't need pretty, informative UI, because it's not that kind of game.
Destiny 2 beta guide: All Exotics, weapons and armour, and how to get your hands on them
A guide to all of the weapons, Exotics, and armour – and how to get them – in the Destiny 2 beta.
Destiny 2 beta guide: how to trigger the infinite Super glitch
Supers charge extra-slow in the Destiny 2 beta, but there's a trick you can use to always have an active Super – at least in the strike.
Destiny 2 beta guide: secrets, mysteries and Easter eggs from the Tower and Nessus
If the Destiny 2 beta is hiding something, we're determined to find it. Spoilers ahead, arguably.
Which class should you roll for the Destiny 2 beta? A quick guide
The Destiny 2 beta is upon us. Which class should you roll to make sure you have the best possible time while the doors are still open?
PlayerUnknown's Battlegrounds has plans to add a ton of character customisation options so you can design them "to a tee"
PlayerUnknown's Battlegrounds has plans to expand on face and clothes customisation.
Pre-load the Destiny 2 beta today on PS4 and Xbox One, exclusive to players with pre-order early access codes
Sometimes it pays to pre-order. Especially when there are Destiny 2 beta codes at stake.
With the apparent death of season passes and $15 DLC, did we actually end up with a worse deal?
For better or worse, we're paying differently for DLC in 2017.
Overwatch: Doomfist can be devastating in the PTR, but his kit desperately needs tightening up
Overwatch's newest hero has finally punched his way onto the PTR.
Mass Effect Andromeda update ruins Cora, Jaal love triangles, brings back the Kishock Harpoon Gun - full patch notes
Mass Effect Andromeda has squashed another log full of bugs.
PlayerUnknown's Battlegrounds will be finished in 6 months, but he has nothing but sympathy for the DayZ and H1Z1 teams
PlayerUnknown's Battlegrounds will not hang around Early Access for yonks, unlike other games we might mention and in fact already did.
PlayerUnknown's Battlegrounds desert map preview is great but hey, DayZ fans: can we talk about the fact that there's a bicycle in it
PlayerUnknown's Battlegrounds is getting bikes and DayZ fans are justifiably miffed.
The story of Destiny 2, part 2: what Bungie learned over three years of hits and misses
Destiny had a rough start, narratively, and Bungie spent three years trying to turn it around. Did it succeed, and what does it mean for Destiny 2?
The story of Destiny 2, part 1: why Destiny's alien-walloping story disappointed you so much
Destiny had a rubbish story. Destiny's follow-up content mostly improved on it. Destiny 2 has a good chance of doing even better. Let's discuss.
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text: 'Buy American' hits free-trade wall By on October 23, 2009 By Leslie Jorgensen THE COLORADO STATESMAN On the face of it, "Buy American" seems to be a logical, patriotic and economically sound message. However, the few bills backing that concept that have emerged recently from the Colorado Legislature have had a hard time making it out of committee and on to the House and Senate […]This content is only available to subscribers. Login or Subscribe Share this:EmailTwitterFacebook Current News, Default


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We've all been there. That dreaded moment when you look up at the travel board and see that your flight has been delayed. Even though you were productive and arrived the suggested hour and a half early, now you're stuck for half the day. You're tired and ready to go home and now you just have to sit and wait. It is one of the most unappealing feelings in the world and is enough to wipe that vacation buzz right out of you as you feel reality set back in.
Fortunately enough for us, there are a few bright sides to being trapped at the airport. If you are able to look past the inconvenience of your flight being delayed you
The airport is truly the epitome of a melting pot. It is one of the best places in the world to expose yourself to various different cultures. If you take out your headphones and put your cell phone down you will hear languages you didn't know existed and see wardrobe choices that will make you smile from ear to ear. Open your eyes and see how many characters are wandering around the airport it is worth a double-take. For some extra fun try and observe why they may be traveling. Does it seem like a happy occasion, a sad trip, did someone just get a great new career and is anxiously awaiting the flight to their new home, or did Nana pass away and you need to travel home to say goodbye. The airport is a giant grouping of short stories. Have a little fun on your next delay and see if you can decipher a few.
Airlines are lined with outlets and charging stations but that really confines you to one space while you're trapped at the airport. If you are anything like me it helps to be up and about. If you feel the same GRACESHIP offers beautiful power wallets that allow you to stay charged on the go! Never sit in the charging station again, instead charge your phone with a stylish power wallet and cut the shackles to the outlet that has you sitting on the carpet in a corner.
Bars and booze...every airport has them. When your plane is delayed and there is nothing you can do about it what better way to pass the time than with a cocktail. Forget about the delay and drink responsibly among your fellow inconvenienced passengers. Whether you are anxious about flying or you are annoyed by the delay, a nice cold beverage may be just what you need to enjoy your extra couple hours at the airport. The time will go by much faster if you small talk at the airport bar than if you sit by your gate and exhaust your social media accounts. This is also the perfect place to learn the stories of your fellow travelers and continue to people watch.
GRACESHIP has beautiful sleek trolley bags that are small, lightweight and fashionable. Not only is it simple to roll the beautiful trolley bags from spot to spot but I guarantee you'll receive some compliments as you catch fellow people-watcher's eyes! Don't let big bulky carry-ons keep you from bar hopping around the airport!
You know what they say, "Good things come to those who wait".