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high peak density sites and high-frequency SD chi-square test #9

Closed rmflight closed 2 years ago

rmflight commented 2 years ago

I created a modified version of the high-peak-density detector that works in frequency space, and converted the xcalibur peak lists to frequency space.

Then I can find our own peaks that would have been part of HPD sites using the ranges from the HPD detector.

I also have the peaks that we decided had unusually high standard deviations (HighSD) from the underlying frequency level peaks.

It is trivial to find the peaks that are either HPD or HighSD, and generate a contingency table of them.

              HPD
HighSD  FALSE TRUE
  FALSE     0   29
  TRUE     58   49

Of course, with a contingency table, we can easily do a Chi-square test. In this case, we get a really significant p-value:

    Pearson's Chi-squared test with Yates'
    continuity correction
data:  cont_table
X-squared = 25.237, 
df = 1, 
p-value = 5.069e-07

@hunter-moseley, does this make sense as a way to evaluate the overlap of the HighSD and HPD? We can plot them as well obviously.

hunter-moseley commented 2 years ago

Robert,

Yes, the chi-squared test shows an association between high SD and HPD sites. I bet if you quantified the width of HPD sites against SD, you will see the wide HPD sites have high SD. If this is the case, then this description would be more informative.

Warm regards, Hunter

On Thu, Mar 17, 2022 at 3:53 PM Robert M Flight @.***> wrote:

I created a modified version of the high-peak-density detector that works in frequency space, and converted the xcalibur peak lists to frequency space.

Then I can find our own peaks that would have been part of HPD sites using the ranges from the HPD detector.

I also have the peaks that we decided had unusually high standard deviations (HighSD) from the underlying frequency level peaks.

It is trivial to find the peaks that are either HPD or HighSD, and generate a contingency table of them.

          HPD

HighSD FALSE TRUE FALSE 0 29 TRUE 58 49

Of course, with a contingency table, we can easily do a Chi-square test. In this case, we get a really significant p-value:

Pearson's Chi-squared test with Yates' continuity correction data: cont_table X-squared = 25.237, df = 1, p-value = 5.069e-07

@hunter-moseley https://github.com/hunter-moseley, does this make sense as a way to evaluate the overlap of the HighSD and HPD? We can plot them as well obviously.

— Reply to this email directly, view it on GitHub https://github.com/MoseleyBioinformaticsLab/manuscript.peakCharacterization/issues/9, or unsubscribe https://github.com/notifications/unsubscribe-auth/ADEP7BZUIWAVY3RN25CAAP3VAOET5ANCNFSM5Q73W5MA . Triage notifications on the go with GitHub Mobile for iOS https://apps.apple.com/app/apple-store/id1477376905?ct=notification-email&mt=8&pt=524675 or Android https://play.google.com/store/apps/details?id=com.github.android&referrer=utm_campaign%3Dnotification-email%26utm_medium%3Demail%26utm_source%3Dgithub.

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

hunter-moseley commented 2 years ago

Robert,

Just thought about this some more.

HPD peak widths may be narrow at the scan level. But I do think mean/median scan-level peak widths for HPD and high SD peaks may be informative.

Warm regards, Hunter

On Thu, Mar 17, 2022 at 4:22 PM Hunter Moseley @.***> wrote:

Robert,

Yes, the chi-squared test shows an association between high SD and HPD sites. I bet if you quantified the width of HPD sites against SD, you will see the wide HPD sites have high SD. If this is the case, then this description would be more informative.

Warm regards, Hunter

On Thu, Mar 17, 2022 at 3:53 PM Robert M Flight @.***> wrote:

I created a modified version of the high-peak-density detector that works in frequency space, and converted the xcalibur peak lists to frequency space.

Then I can find our own peaks that would have been part of HPD sites using the ranges from the HPD detector.

I also have the peaks that we decided had unusually high standard deviations (HighSD) from the underlying frequency level peaks.

It is trivial to find the peaks that are either HPD or HighSD, and generate a contingency table of them.

          HPD

HighSD FALSE TRUE FALSE 0 29 TRUE 58 49

Of course, with a contingency table, we can easily do a Chi-square test. In this case, we get a really significant p-value:

Pearson's Chi-squared test with Yates' continuity correction data: cont_table X-squared = 25.237, df = 1, p-value = 5.069e-07

@hunter-moseley https://github.com/hunter-moseley, does this make sense as a way to evaluate the overlap of the HighSD and HPD? We can plot them as well obviously.

— Reply to this email directly, view it on GitHub https://github.com/MoseleyBioinformaticsLab/manuscript.peakCharacterization/issues/9, or unsubscribe https://github.com/notifications/unsubscribe-auth/ADEP7BZUIWAVY3RN25CAAP3VAOET5ANCNFSM5Q73W5MA . Triage notifications on the go with GitHub Mobile for iOS https://apps.apple.com/app/apple-store/id1477376905?ct=notification-email&mt=8&pt=524675 or Android https://play.google.com/store/apps/details?id=com.github.android&referrer=utm_campaign%3Dnotification-email%26utm_medium%3Demail%26utm_source%3Dgithub.

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-- Hunter Moseley, Ph.D. -- Univ. of Kentucky Associate Professor, Dept. of Molec. & Cell. Biochemistry / Markey Cancer Center / Institute for Biomedical Informatics / UK Superfund Research Center Not just a scientist, but a fencer as well. My foil is sharp, but my mind sharper still.

Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

-- Hunter Moseley, Ph.D. -- Univ. of Kentucky Associate Professor, Dept. of Molec. & Cell. Biochemistry / Markey Cancer Center / Institute for Biomedical Informatics / UK Superfund Research Center Not just a scientist, but a fencer as well. My foil is sharp, but my mind sharper still.

Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

rmflight commented 2 years ago

So I grabbed the HPD site width in frequency space, and then calculated the median Frequency SD for HighSD peaks if they were present in the HPD site, the other peaks in the HPD site, and all peaks in the HPD site for the 4 samples to see if there was any "trend" of HPD width and Frequency SD of the peaks within them.

I don't see any trend here.

split_sd_wdith

I also tried aggregating them across samples and plotting again, and still don't see any trend here, unfortunately.

width_sd_type

rmflight commented 2 years ago

Three more things:

  1. The new graph that I just generated.
  2. Do 98th percentile and compare to the 99th percentile.
  3. scatter plot of # of peaks in each HPD by:

    • xcalibur
    • scan-level characterization 99th
    • scan-level characterization 98th
    • MSnbase
rmflight commented 2 years ago

And here is the plot for the 49lipid sample

example_comparison_hpd

I tried a scatter plot of the counts of peaks, but it didn't seem to tell us much

scatterplot_1sample

, so I calculated the fraction of peaks in the method compared to xcalibur.

  scanlevel_99 scanlevel_98 msnbase
         <dbl>        <dbl>   <dbl>
1      0.177        0.177    0.189 
2      0.0197       0.0197   0.0132
3      0.0288       0.0288   0.0791
4      0.0380       0.0380   0.0253
5      0.00593      0.00593  0.0237
6      0.0294       0.0294   0.147 
7      0.0278       0.0278   0.111 
8     NA           NA        0.0870
9     NA           NA        0.14  
hunter-moseley commented 2 years ago

Well the 98% point filter was no different than the 99% point filter. No need to include it.

Would include the number of peaks after highSD filter in the scatter plot instead of 98% point filter. Should be pretty dramatic visually.

On Fri, Mar 18, 2022 at 10:12 PM Robert M Flight @.***> wrote:

And here is the plot for the 49lipid sample

[image: example_comparison_hpd] https://user-images.githubusercontent.com/1509626/159102598-46c08ab9-a19d-4966-82a4-9668fefed20d.png

I tried a scatter plot of the counts of peaks, but it didn't seem to tell us much

[image: scatterplot_1sample] https://user-images.githubusercontent.com/1509626/159102821-93bad525-ea92-4c58-92e2-5d8221eb9838.png

, so I calculated the fraction of peaks in the method compared to xcalibur.

scanlevel_99 scanlevel_98 msnbase

1 0.177 0.177 0.189 2 0.0197 0.0197 0.0132 3 0.0288 0.0288 0.0791 4 0.0380 0.0380 0.0253 5 0.00593 0.00593 0.0237 6 0.0294 0.0294 0.147 7 0.0278 0.0278 0.111 8 NA NA 0.0870 9 NA NA 0.14 — Reply to this email directly, view it on GitHub , or unsubscribe . Triage notifications on the go with GitHub Mobile for iOS or Android . You are receiving this because you were mentioned.Message ID:

-- Hunter Moseley, Ph.D. -- Univ. of Kentucky Associate Professor, Dept. of Molec. & Cell. Biochemistry / Markey Cancer Center / Institute for Biomedical Informatics / UK Superfund Research Center Not just a scientist, but a fencer as well. My foil is sharp, but my mind sharper still.

Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

rmflight commented 2 years ago

I just realized one we could include is the scan-level where we didn't do any point filtering except having counts > 0.

Relatedly, we could also do an analysis of the number of starting regions we have using different percentile cutoffs, and not do full characterization. (Will make another issue).

On Fri, Mar 18, 2022, 10:22 PM Hunter Moseley @.***> wrote:

Well the 98% point filter was no different than the 99% point filter. No need to include it.

Would include the number of peaks after highSD filter in the scatter plot instead of 98% point filter. Should be pretty dramatic visually.

On Fri, Mar 18, 2022 at 10:12 PM Robert M Flight @.***> wrote:

And here is the plot for the 49lipid sample

[image: example_comparison_hpd] < https://user-images.githubusercontent.com/1509626/159102598-46c08ab9-a19d-4966-82a4-9668fefed20d.png

I tried a scatter plot of the counts of peaks, but it didn't seem to tell us much

[image: scatterplot_1sample] < https://user-images.githubusercontent.com/1509626/159102821-93bad525-ea92-4c58-92e2-5d8221eb9838.png

, so I calculated the fraction of peaks in the method compared to xcalibur.

scanlevel_99 scanlevel_98 msnbase

1 0.177 0.177 0.189 2 0.0197 0.0197 0.0132 3 0.0288 0.0288 0.0791 4 0.0380 0.0380 0.0253 5 0.00593 0.00593 0.0237 6 0.0294 0.0294 0.147 7 0.0278 0.0278 0.111 8 NA NA 0.0870 9 NA NA 0.14 — Reply to this email directly, view it on GitHub < https://github.com/MoseleyBioinformaticsLab/manuscript.peakCharacterization/issues/9#issuecomment-1072919608 , or unsubscribe < https://github.com/notifications/unsubscribe-auth/ADEP7BYXCK6F5TU7RF46DDTVAUZYNANCNFSM5Q73W5MA . Triage notifications on the go with GitHub Mobile for iOS < https://apps.apple.com/app/apple-store/id1477376905?ct=notification-email&mt=8&pt=524675 or Android < https://play.google.com/store/apps/details?id=com.github.android&referrer=utm_campaign%3Dnotification-email%26utm_medium%3Demail%26utm_source%3Dgithub . You are receiving this because you were mentioned.Message ID:

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

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hunter-moseley commented 2 years ago

Good idea on count>0 point filtering. Will likely show the HPD effect.

As an aside, would be interesting to see when count filtering has an effect on the number of peaks detected. 98% point filtering likely did not changed the count cutoff, since this is an integer cutoff value.

On Sat, Mar 19, 2022 at 11:54 AM Robert M Flight @.***> wrote:

I just realized one we could include is the scan-level where we didn't do any point filtering except having counts > 0.

Relatedly, we could also do an analysis of the number of starting regions we have using different percentile cutoffs, and not do full characterization. (Will make another issue).

On Fri, Mar 18, 2022, 10:22 PM Hunter Moseley @.***> wrote:

Well the 98% point filter was no different than the 99% point filter. No need to include it.

Would include the number of peaks after highSD filter in the scatter plot instead of 98% point filter. Should be pretty dramatic visually.

On Fri, Mar 18, 2022 at 10:12 PM Robert M Flight @.***> wrote:

And here is the plot for the 49lipid sample

[image: example_comparison_hpd] <

https://user-images.githubusercontent.com/1509626/159102598-46c08ab9-a19d-4966-82a4-9668fefed20d.png

I tried a scatter plot of the counts of peaks, but it didn't seem to tell us much

[image: scatterplot_1sample] <

https://user-images.githubusercontent.com/1509626/159102821-93bad525-ea92-4c58-92e2-5d8221eb9838.png

, so I calculated the fraction of peaks in the method compared to xcalibur.

scanlevel_99 scanlevel_98 msnbase

1 0.177 0.177 0.189 2 0.0197 0.0197 0.0132 3 0.0288 0.0288 0.0791 4 0.0380 0.0380 0.0253 5 0.00593 0.00593 0.0237 6 0.0294 0.0294 0.147 7 0.0278 0.0278 0.111 8 NA NA 0.0870 9 NA NA 0.14 — Reply to this email directly, view it on GitHub <

https://github.com/MoseleyBioinformaticsLab/manuscript.peakCharacterization/issues/9#issuecomment-1072919608

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

rmflight commented 2 years ago

Updated to use scanlevel_00 (really noperc_nonorm), and include the whole count and lowsd peak count. And the HPD site is different b/c it's a different sample, one of the AA samples. hpd_2ecf

n_peak_diffs_hpd

hunter-moseley commented 2 years ago

If possible, we should use spectra of the same sample.

Also in the panels of spectra, The scanlevel_00 should not highly high SD in red. And the scanlevel_99 should have one with high SD included and one below it with high SD removed. It will visually make the scanlevel_99 with high SD removed visually comparable to the MSnBase.

On Sat, Mar 19, 2022 at 3:09 PM Robert M Flight @.***> wrote:

Updated to use scanlevel_00 (really noperc_nonorm), and include the whole count and lowsd peak count. And the HPD site is different b/c it's a different sample, one of the AA samples. [image: hpd_2ecf] https://user-images.githubusercontent.com/1509626/159135118-80d2754e-588e-4c34-ad53-eb28cb7c0929.png

[image: n_peak_diffs_hpd] https://user-images.githubusercontent.com/1509626/159135121-588bfdd7-ef32-4096-b62f-40a736362929.png

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

rmflight commented 2 years ago

Just to be clear, all the spectra are the same sample in a given set of graphs. I just wanted to let you know why these plots looked different than the one I had added previously.

I added the indicate_high_sd() to all of the different scan-level processing we are doing (why I didn't have it in there before, not sure), so those will be running a good chunk of the weekend. I have it set to text me when they are done (or error), so I'll check on it to make sure it actually all ran OK.

I will update the HPD plotting code on Monday to take your comments into account.

So overall, the spectra plot would be:

  1. xcalibur
  2. scanlevel_00
  3. scanlevel_99 all peaks
  4. scanlevel_99 highsd removed
  5. msnbase

And then the scatterplot would be:

  1. msnbase
  2. scanlevel_00
  3. scanlevel_99
  4. scanlevel_99_lowsd
hunter-moseley commented 2 years ago

Sounds good.

Would also change the scatter plot axes labels to "peak count" and "Xcalibur peak count".

On Sat, Mar 19, 2022 at 3:33 PM Robert M Flight @.***> wrote:

Just to be clear, all the spectra are the same sample in a given set of graphs. I just wanted to let you know why these plots looked different than the one I had added previously.

I added the indicate_high_sd() to all of the different scan-level processing we are doing (why I didn't have it in there before, not sure), so those will be running a good chunk of the weekend. I have it set to text me when they are done (or error), so I'll check on it to make sure it actually all ran OK.

I will update the HPD plotting code on Monday to take your comments into account.

So overall, the spectra plot would be:

  1. xcalibur
  2. scanlevel_00
  3. scanlevel_99 all peaks
  4. scanlevel_99 highsd removed
  5. msnbase

And then the scatterplot would be:

  1. msnbase
  2. scanlevel_00
  3. scanlevel_99
  4. scanlevel_99_lowsd

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

rmflight commented 2 years ago

Will do.

On Sat, Mar 19, 2022, 3:39 PM Hunter Moseley @.***> wrote:

Sounds good.

Would also change the scatter plot axes labels to "peak count" and "Xcalibur peak count".

On Sat, Mar 19, 2022 at 3:33 PM Robert M Flight @.***> wrote:

Just to be clear, all the spectra are the same sample in a given set of graphs. I just wanted to let you know why these plots looked different than the one I had added previously.

I added the indicate_high_sd() to all of the different scan-level processing we are doing (why I didn't have it in there before, not sure), so those will be running a good chunk of the weekend. I have it set to text me when they are done (or error), so I'll check on it to make sure it actually all ran OK.

I will update the HPD plotting code on Monday to take your comments into account.

So overall, the spectra plot would be:

  1. xcalibur
  2. scanlevel_00
  3. scanlevel_99 all peaks
  4. scanlevel_99 highsd removed
  5. msnbase

And then the scatterplot would be:

  1. msnbase
  2. scanlevel_00
  3. scanlevel_99
  4. scanlevel_99_lowsd

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Email: @. (work) @. (personal) Phone: 859-218-2964 (office) 859-218-2965 (lab) 859-257-7715 (fax) Web: http://bioinformatics.cesb.uky.edu/ Address: CC434 Roach Building, 800 Rose Street, Lexington, KY 40536-0093

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rmflight commented 2 years ago

OK, here is the figure based on everything brought up above

manuscript_hpd_figure

hunter-moseley commented 2 years ago

This looks really good!

One question that will come up is the overlap of peaks between our method and MSnBase. May be good to find a place in the manuscript to put a Venn Diagram showing overlap of peaks between our method and MSnBase.

On Mon, Mar 21, 2022 at 8:03 AM Robert M Flight @.***> wrote:

OK, here is the figure based on everything brought up above

[image: manuscript_hpd_figure] https://user-images.githubusercontent.com/1509626/159257244-4cf9b9e4-08d0-49c8-9fc2-f0e6981738d8.png

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rmflight commented 2 years ago

OK. will put it in another issue.

hunter-moseley commented 2 years ago

Throw these results into supplemental material.

We can talk about them in the main text.

On Fri, Mar 18, 2022, 10:00 AM Robert M Flight @.***> wrote:

So I grabbed the HPD site width in frequency space, and then calculated the median Frequency SD for HighSD peaks if they were present in the HPD site, the other peaks in the HPD site, and all peaks in the HPD site for the 4 samples to see if there was any "trend" of HPD width and Frequency SD of the peaks within them.

I don't see any trend here.

[image: split_sd_wdith] https://user-images.githubusercontent.com/1509626/159016326-3e2f8b88-30b5-47a4-94e7-b94e9d88afa2.png

I also tried aggregating them across samples and plotting again, and still don't see any trend here, unfortunately.

[image: width_sd_type] https://user-images.githubusercontent.com/1509626/159016369-6463c7a3-488d-455f-a83d-cfa3062dc600.png

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