[BUG]

[isabellaCPR]

I have updated PRSice from version 2.1.9 to 2.3.2. When I run the version 2.1.9 everything runs perfectly fine, but I get an "Error: Cannot read bgen file!" when running the same script using 2.3.2 instead.

Do some files or other things need to be updated when I've updated to PRSice 2.3.2? Also it can process the SNPs in the bgen files, but the problem occurs right before the clumping process. Do you know what I can do?

Error Log PRSice-2: Polygenic Risk Score Software for Biobank-Scale Data. GigaScience 8, no. 7 (July 1, 2019) 2020-08-20 08:21:56 ./2.3.2/PRSice_linux --a1 A1 --a2 A2 --bar-levels 0.001,0.05,0.1,0.2,0.3,0.4,0.5,1 --base /PRS/base.file.txt --beta --binary-target T --bp BP --chr CHR --clump-kb 250kb --clump-p 1.000000 --clump-r2 0.100000 --cov /PRS/covariate.tsv --cov-col sex,age,BMI --cov-factor sex --extract out.dosage.valid --ignore-fid --interval 5e-05 --keep PRS/keep.txt --lower 5e-08 --num-auto 22 --out out.dosage --pheno /PRS/pheno.tsv --pheno-col pheno --pvalue PVAL --seed 211324058 --snp SNP --stat Effect --target /bgen/bgen_chr#_clean --thread 28 --type bgen --upper 0.5

Initializing Genotype info from file: /bgen/bgen_chr#_clean (bgen)

Start processing base.file.txt Base file: /PRS/base.file.txt Header of file is: SNP CHR BP A1 A2 BETA SE PVAL Effect

7151746 variant(s) observed in base file, with: 1080490 total variant(s) excluded based on user input 6071256 total variant(s) included from base file

Loading Genotype info from target

---

Assume phonetype file has header line: FID IID pheno 91737 people (0 male(s), 0 female(s)) observed 56398 founder(s) included

7161939 variants(s) not found in previsou data 6 variant(s) with mismatch information 5229633 variant(s) included

Phenotype file: /PRS/pheno.tsv Column name of sample ID: FID Note: If the phenotype file does not contain a header, the column name will be displayed as the Sample ID which is expected.

There are a total of 1 phenotype to precess

Start performing clumping

terminate called after throwing a intance of 'std::runtime_error' what(): Error: Cannot read the bgen file! Error: Execution halted

[choishingwan]

Likely some bug. Will try and see if I can replicate and fix it.

[choishingwan]

Hmm, I was unable to replicate the problem using my uk biobank data. Can you check if your files are all readable?

On Mon, 24 Aug 2020 at 1:11 PM, Shing Wan Choi choishingwan@gmail.com wrote:

Likely some bug. Will try and see if I can replicate and fix it.

-- Dr Shing Wan Choi Postdoctoral Fellow Genetics and Genomic Sciences Icahn School of Medicine, Mount Sinai, NYC

[isabellaCPR]

Thanks for your fast response! It works perfectly fine with PRSice version 2.1.9. So I would assume all the files are readable? Or did something change from version 2.1.9 to 2.3.2?

[isabellaCPR]

I now made it work! Seems like it was the --keep file that was not sorted in the same order as the sample file. Though, now another error shows up:

PRSice-2: Polygenic Risk Score Software for Biobank-Scale Data. GigaScience 8, no. 7 (July 1, 2019) 2020-09-02 08:56:22 ./2.3.2/PRSice_linux --a1 A1 --a2 A2 --allow-inter --bar-levels 0.001,0.05,0.1,0.2,0.3,0.4,0.5,1 --base /PRS/base.file.txt --beta --binary-target T --bp BP --chr CHR --clump-kb 250kb --clump-p 1.000000 --clump-r2 0.100000 --cov /PRS/covariate.tsv --cov-col sex,age,BMI --cov-factor sex --extract out.dosage.valid --ignore-fid --interval 5e-05 --keep PRS/keep.txt --lower 5e-08 --num-auto 22 --out out.dosage --pheno /PRS/pheno.tsv --pheno-col pheno --pvalue PVAL --seed 1517885392 --snp SNP --stat Effect --target /bgen/bgen_chr#_clean --thread 28 --type bgen --upper 0.5

Initializing Genotype info from file: /bgen/bgen_chr#_clean (bgen)

Start processing base.file.txt

---

SNP extraction/exclusion list contains 5 columns, will assume first column contains SNP ID

Base file: /PRS/base.file.txt Header of file is: SNP CHR BP A1 A2 BETA SE PVAL Effect

7151746 variant(s) observed in base file, with: 1080490 total variant(s) excluded based on user input 6071256 total variant(s) included from base file

Loading Genotype info from target

---

Assume phonetype file has header line: FID IID pheno 91737 people (0 male(s), 0 female(s)) observed 56398 founder(s) included

7161939 variants(s) not found in previsou data 6 variant(s) with mismatch information 5229633 variant(s) included

Calculate MAF and perform filtering on target SNPs

---

5229633 variant(s) included

Phenotype file: /PRS/pheno.tsv Column name of sample ID: FID Note: If the phenotype file does not contain a header, the column name will be displayed as the Sample ID which is expected.

There are a total of 1 phenotype to precess

Start performing clumping

Number of variants after clumping: 294822

Processing the 1st phenotype 54643 control(s) 1754 case(s)

Processing the covariate file: /PRS/covariate.tsv

---

Include Covariates: Name Missing Number of levels sex 0 2 age 2055 - BMI 2721 -

After reading the covariate file, 53676 sample(s) included in the analysis

Start Processing Processing 0.02%Error: Execution halted

[choishingwan]

Yes, there was a problem with 2.3.2 related to the keep file. If you use 2.3.3, you should in theory have no problem running the command. Maybe give it a go? Also use —print-snp in your command so that when you rerun prsice, you can directly give prsice the snp file generated from the previous run and skip clumping, which for bgen can save quite a bit of time

Sam

On Thu, 3 Sep 2020 at 2:41 PM, isabellaCPR notifications@github.com wrote:

I now made it work! Seems like it was the --keep file that was not sorted in the same order as the sample file. Though, now another error shows up:

PRSice-2: Polygenic Risk Score Software for Biobank-Scale Data.

GigaScience 8, no. 7 (July 1, 2019)

2020-09-02 08:56:22

./2.3.2/PRSice_linux

--a1 A1

--a2 A2

--allow-inter

--bar-levels 0.001,0.05,0.1,0.2,0.3,0.4,0.5,1

--base /PRS/base.file.txt

--beta

--binary-target T

--bp BP

--chr CHR

--clump-kb 250kb

--clump-p 1.000000

--clump-r2 0.100000

--cov /PRS/covariate.tsv

--cov-col sex,age,BMI

--cov-factor sex

--extract out.dosage.valid

--ignore-fid

--interval 5e-05

--keep PRS/keep.txt

--lower 5e-08

--num-auto 22

--out out.dosage

--pheno /PRS/pheno.tsv

--pheno-col pheno

--pvalue PVAL

--seed 1517885392

--snp SNP

--stat Effect

--target /bgen/bgen_chr#_clean

--thread 28

--type bgen

--upper 0.5

Initializing Genotype info from file: /bgen/bgen_chr#_clean (bgen)

Start processing base.file.txt

---

SNP extraction/exclusion list contains 5 columns, will assume first column contains SNP ID

Base file: /PRS/base.file.txt

Header of file is:

SNP CHR BP A1 A2 BETA SE PVAL Effect

7151746 variant(s) observed in base file, with:

1080490 total variant(s) excluded based on user input

6071256 total variant(s) included from base file

Loading Genotype info from target

---

Assume phonetype file has header line: FID IID pheno

91737 people (0 male(s), 0 female(s)) observed

56398 founder(s) included

7161939 variants(s) not found in previsou data

6 variant(s) with mismatch information

5229633 variant(s) included

Calculate MAF and perform filtering on target SNPs

---

5229633 variant(s) included

Phenotype file: /PRS/pheno.tsv

Column name of sample ID: FID

Note: If the phenotype file does not contain a header, the column name will be displayed as the Sample ID which is expected.

There are a total of 1 phenotype to precess

Start performing clumping

Number of variants after clumping: 294822

Processing the 1st phenotype

54643 control(s)

1754 case(s)

Processing the covariate file:

/PRS/covariate.tsv

---

Include Covariates:

Name Missing Number of levels

sex 0 2

age 2055 -

BMI 2721 -

After reading the covariate file, 53676 sample(s) included in the analysis

Start Processing

Processing 0.02%Error:

Execution halted

— You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-686289004, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYQVYWFI4XIN6DMUJCDSD43BTANCNFSM4QIRLWTA .

-- Dr Shing Wan Choi Postdoctoral Fellow Genetics and Genomic Sciences Icahn School of Medicine, Mount Sinai, NYC

[isabellaCPR]

Perfect! Thanks! I'll give it a try!

[isabellaCPR]

I have installed the PRSice 2.3.3 version. Now I get this error: My R version is R/3.6.1-GCC-MKL.

Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion Plotting in quantile plot Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

[choishingwan]

Mind sending me the full log and command? It is way more difficult to figure out the cause of a problem with only the error code.

Thanks

On Fri, 4 Sep 2020 at 2:29 PM, isabellaCPR notifications@github.com wrote:

I have installed the PRSice 2.3.3 version. Now I get this error:

My R version is R/3.6.1-GCC-MKL.

Begin plotting

Current Rscript version = 2.3.3

Warning message:

In as.numeric(as.character(covariance.base[, i])) :

NAs introduced by coercion

Plotting in quantile plot

Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) :

contrasts can be applied only to factors with 2 or more levels

Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<-

Execution halted.

— You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-686942104, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYQ355ZIIOX7OCNXQN3SECCMJANCNFSM4QIRLWTA .

-- Dr Shing Wan Choi Postdoctoral Fellow Genetics and Genomic Sciences Icahn School of Medicine, Mount Sinai, NYC

[isabellaCPR]

Sorry - of course no problem.

PRSice 2.3.3 (2020-08-05) https://githup.com/choishingwan/PRSice (C) 2016-2020 Shing Wan (Sam) Choi and Paul F. O'Reilly GNU General Public License v3 If you use PRSice in any published work, please cite: Choi SW, O'Reilly PF. PRSice-2: Polygenic Risk Score Software for Biobank-Scale Data. GigaScience 8, no. 7 (July 1, 2019) 2020-09-03 13:11:56 ./2.3.3/PRSice_linux --a1 A1 --a2 A2 --allow-inter --bar-levels 0.001,0.05,0.1,0.2,0.3,0.4,0.5,1 --base /PRS/base.file.txt --beta --binary-target T --bp BP --chr CHR --clump-kb 250kb --clump-p 1.000000 --clump-r2 0.100000 --cov /PRS/covariate.tsv --cov-col sex,age,BMI --cov-factor sex --extract out.dosage.valid --ignore-fid --interval 5e-05 --keep PRS/keep.txt --lower 5e-08 --num-auto 22 --out out.dosage --pheno /PRS/pheno.tsv --pheno-col pheno -- print-snp --pvalue PVAL --seed 1931666980 --snp SNP --stat Effect --target /bgen/bgen_chr#_clean --thread 28 --type bgen --upper 0.5

Initializing Genotype info from file: /bgen/bgen_chr#_clean (bgen)

Start processing base.file.txt

SNP extraction/exclusion list contains 5 columns, will assume first column contains SNP ID

Base file: /PRS/base.file.txt Header of file is: SNP CHR BP A1 A2 BETA SE PVAL Effect

7151746 variant(s) observed in base file, with: 1080490 total variant(s) excluded based on user input 6071256 total variant(s) included from base file

Loading Genotype info from target

Assume phonetype file has header line: FID IID pheno 91737 people (0 male(s), 0 female(s)) observed 56398 founder(s) included

7161939 variants(s) not found in previsou data 6 variant(s) with mismatch information 5229633 variant(s) included

Calculate MAF and perform filtering on target SNPs

5229633 variant(s) included

Phenotype file: /PRS/pheno.tsv Column name of sample ID: FID Note: If the phenotype file does not contain a header, the column name will be displayed as the Sample ID which is expected.

There are a total of 1 phenotype to precess

Start performing clumping

Number of variants after clumping: 294822

Processing the 1st phenotype 54643 control(s) 1754 case(s)

Processing the covariate file: /PRS/covariate.tsv

Include Covariates: Name Missing Number of levels sex 0 2 age 2055 - BMI 2721 -

After reading the covariate file, 53676 sample(s) included in the analysis

Start Processing Processing 100.00% There are 1 region(s) with p-value less than 1e-5. Please note that these results are inflated due to the overfitting inherent in finding the best-fit PRS (but it's still best to find the best-fir PRS!). You can use the --perm option (see manual) to calculate an empirical P-value.

Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion Plotting in quantile plot Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

[choishingwan]

Think this might be a small bug in the Rscript.

For now, you can just ignore --cov-factor sex and rerun the script with --plot and that should in theory work.

Meanwhile, I will add this to something that I should investigate.

Thanks for reporting.

[isabellaCPR]

I have tried to run it again, but it produces the same error. I guess it starts plotting, so no log file is created.

This is the command I run: Rscript ../2.3.3/PRSice.R --dir . --prsice ../2.3.3/PRSice_linux --a1 A1 --a2 A2 --allow-inter T --base /PRS/base.file.txt --beta --binary-target T --bp BP --chr CHR --cov-file /PRS/covariate.tsv --cov-col sex,age,BMI --extract out.dosage.valid --ignore-fid --keep PRS/keep.txt --out out.dosage --pheno /PRS/pheno.tsv --pheno-col pheno --plot --print-snp --pvalue PVAL --quantile 20 --snp SNP --stat Effect --target /bgen/bgen_chr#_clean --thread 28 --type bgen

and I get the same error: Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion Plotting in quantile plot Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

[choishingwan]

I’ll have to look into it. Surprise that it seems like it is still doing something related to factor

Will have a look

On Sat, 5 Sep 2020 at 1:50 PM, isabellaCPR notifications@github.com wrote:

I have tried to run it again, but it produces the same error. I guess it starts plotting, so no log file is created.

This is the command I run:

Rscript ../2.3.3/PRSice.R --dir .

--prsice ../2.3.3/PRSice_linux

--a1 A1

--a2 A2

--allow-inter T

--base /PRS/base.file.txt

--beta

--binary-target T

--bp BP

--chr CHR

--cov-file /PRS/covariate.tsv

--cov-col sex,age,BMI

--extract out.dosage.valid

--ignore-fid

--keep PRS/keep.txt

--out out.dosage

--pheno /PRS/pheno.tsv

--pheno-col pheno

--plot

--print-snp

--pvalue PVAL

--quantile 20

--snp SNP

--stat Effect

--target /bgen/bgen_chr#_clean

--thread 28

--type bgen

and I get the same error:

Begin plotting

Current Rscript version = 2.3.3

Warning message:

In as.numeric(as.character(covariance.base[, i])) :

NAs introduced by coercion

Plotting in quantile plot

Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) :

contrasts can be applied only to factors with 2 or more levels

Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<-

Execution halted.

—

You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-687556350, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYWXT6IN6LSJ54BDJPDSEHGR3ANCNFSM4QIRLWTA .

-- Dr Shing Wan Choi Postdoctoral Fellow Genetics and Genomic Sciences Icahn School of Medicine, Mount Sinai, NYC

[choishingwan]

Oh, if you don't mind, could you please give me a small chunk of your phenotype +covariate file (without any IDs) that replicate the problem? That'd help me speed up the process.

[isabellaCPR]

I just replaced the ID with the number from 1 -10. Is this okay? Files are tab-separated.

FID IID pheno 1 1 0 2 2 0 3 3 0 4 4 0 5 5 1 6 6 0 7 7 0 8 8 0 9 9 0 10 10 0

FID IID sex age BMI 1 1 1 23 20.1 2 2 2 29.4 23.1 3 3 2 33.1 23.4 4 4 1 27.8 22.3 5 5 2 39.1 25.8 6 6 1 23.4 25.8 7 7 2 26.3 22.9 8 8 1 35.7 21.3 9 9 1 26.3 20.7 10 10 2 34.5 25.3

[choishingwan]

Do you have a fragment of your file that caused the problem? With the data you sent me, I was unable to replicate the error.

[choishingwan]

I suspect that the error might be coming from the glm model when plotting the quantile plot.

I have now attached a modified Rscript where it will generate a DEBUG file, that should give me some idea of what is the problem. (You can check if the debug file includes any sensitive information. In theory, that should have removed all the IDs). Send me the DEBUG file together with the on screen log and that should help me to pinpoint the problem.

Also, run PRSice with the --plot option to skip the time needed to re-calculate PRS.

Sam

[isabellaCPR]

Thank you very much! I checked the phenotype and covariate files. There are some NAs otherwise the rest of the files are similar to the fragment I sent you.

Great! Did you attach the R script? It seems like a cannot see it?

[choishingwan]

I wish github will notify me when they remove my attachments. Always fall for that...

Please download the script from here <https://www.dropbox.com/s/kklgepakvlz5m3n/PRSice.R?dl=0>

On Tue, Sep 8, 2020 at 4:13 PM isabellaCPR notifications@github.com wrote:

Thank you very much! I checked the phenotype and covariate files. There are some NAs otherwise the rest of the files are similar to the fragment I sent you.

Great! Did you attach the R script? It seems like a cannot see it?

— You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-688700872, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYSH4NFJ7ZYB3WPJHVLSEXRRBANCNFSM4QIRLWTA .

[isabellaCPR]

No problem. I have run the code and the error is more or less the same, but shows up after the "before glm".

It would not be possible for me to send the entire file as it still contains person sensitive information, but you can have the first lines as before. It seems like it has introduced a Cov1 that is NA for all?

"quantile" "Pheno" "Cov1" "Cov2" "Cov3" "Cov4" "1" "9" 0 NA 1 23 20.1 "2" "2" 0 NA 2 29.4 23.1 "3" "7" 0 NA 2 33.1 23.4 "4" "10" 0 NA 1 27.8 22.3 "5" "11" 1 NA 2 39.1 25.8 "6" "2" 0 NA 1 23.4 25.8 "7" "19" 0 NA 2 26.3 22.9 "8" "8" 0 NA 1 35.7 21.3 "9" "16" 0 NA 1 26.3 20.7 "10" "3" 0 NA 0 2 34.5 25.3

Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion Plotting in quantile plot [1] "Before glm" Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

[choishingwan]

I think that's where the problem is. Somehow, when reading in your file, R might have read in the file in a rather unexpected way. Unfortunately, as I can't access your files, I can only send you one modified scripts after another and ask for the output until I can pinpoint the error. if you don't mind, could you please show me the error message from this Rscript? Thanks

https://www.dropbox.com/s/kklgepakvlz5m3n/PRSice.R?dl=0

[isabellaCPR]

No problem - thank you for going through all of it!!

Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion IID Pheno 2 1 0 3 2 0 4 3 0 5 4 0 6 5 1 7 6 0 IID Cov1 Cov2 Cov3 Cov4 1 1 NA 1 23 20.1 2 2 NA 2 29.4 23.1 3 3 NA 2 33.1 23.4 4 4 NA 1 27.8 22.3 5 5 NA 2 39.1 25.8 6 6 NA 1 23.4 25.8 Plotting in quantile plot [1] "Before glm" Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

[choishingwan]

Hi sorry,

I thought I had replied to this email.

Could you please check the format of your covariance file? It seems odd that when we read in the file, it already has the NA column. That's what caused the error in the downstream analysis.

On Sat, Sep 12, 2020 at 2:15 PM isabellaCPR notifications@github.com wrote:

No problem - thank you for going through all of it!!

Begin plotting Current Rscript version = 2.3.3 Warning message: In as.numeric(as.character(covariance.base[, i])) : NAs introduced by coercion IID Pheno 2 1 0 3 2 0 4 3 0 5 4 0 6 5 1 7 6 0 IID Cov1 Cov2 Cov3 Cov4 1 1 NA 1 23 20.1 2 2 NA 2 29.4 23.1 3 3 NA 2 33.1 23.4 4 4 NA 1 27.8 22.3 5 5 NA 2 39.1 25.8 6 6 NA 1 23.4 25.8 Plotting in quantile plot [1] "Before glm" Error in 'contrasts<- '('tmp', value = contr.funs[1 + isOF[nn]]) : contrasts can be applied only to factors with 2 or more levels Calls: process_plot ... model.matrix -> model.matrix.default -> contrasts<- Execution halted.

— You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-691422705, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYTHDUX6JNMLWNWCD7DSFMGZLANCNFSM4QIRLWTA .

[isabellaCPR]

Hi, no problem. The covariance file does not have a NA column, when I read in the file. But you helped me locate the error, so I just removed the NA column in the R script and now everything works perfectly fine! Thanks :-)

[choishingwan]

I see, thanks for letting me know. I guess one of the possible reason for that is if the separator of your covariance file is different in one of the columns. But anyway, good that this is now resolved.

Sam

On Wed, Sep 30, 2020 at 2:25 PM isabellaCPR notifications@github.com wrote:

Hi, no problem. The covariance file does not have a NA column, when I read in the file. But you helped me locate the error, so I just removed the NA column in the R script and now everything works perfectly fine! Thanks :-)

— You are receiving this because you were assigned. Reply to this email directly, view it on GitHub https://github.com/choishingwan/PRSice/issues/222#issuecomment-701184156, or unsubscribe https://github.com/notifications/unsubscribe-auth/AAJTRYQDLV75XB6SWUZTX7LSILFOFANCNFSM4QIRLWTA .

[isabellaCPR]

Could easily be that some separator of the covariance file is different. I actually did not check. But it did not happen in previous versions of PRSice (2.1.9). Thank you for the help and all the time!

[stale[bot]]

This issue has been automatically marked as stale because it has not had recent activity. It will be closed if no further activity occurs. Thank you for your contributions.