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diff --git a/docs/articles/FOCUS_Z.tex b/docs/articles/FOCUS_Z.tex new file mode 100644 index 00000000..f1886acd --- /dev/null +++ b/docs/articles/FOCUS_Z.tex @@ -0,0 +1,566 @@ +%\VignetteIndexEntry{Example evaluation of FOCUS dataset Z} +%\VignetteEngine{knitr::knitr} +\documentclass[12pt,a4paper]{article}\usepackage[]{graphicx}\usepackage[]{color} +%% maxwidth is the original width if it is less than linewidth +%% otherwise use linewidth (to make sure the graphics do not exceed the margin) +\makeatletter +\def\maxwidth{ % + \ifdim\Gin@nat@width>\linewidth + \linewidth + \else + \Gin@nat@width + \fi +} +\makeatother + +\definecolor{fgcolor}{rgb}{0.345, 0.345, 0.345} +\newcommand{\hlnum}[1]{\textcolor[rgb]{0.686,0.059,0.569}{#1}}% +\newcommand{\hlstr}[1]{\textcolor[rgb]{0.192,0.494,0.8}{#1}}% +\newcommand{\hlcom}[1]{\textcolor[rgb]{0.678,0.584,0.686}{\textit{#1}}}% +\newcommand{\hlopt}[1]{\textcolor[rgb]{0,0,0}{#1}}% +\newcommand{\hlstd}[1]{\textcolor[rgb]{0.345,0.345,0.345}{#1}}% +\newcommand{\hlkwa}[1]{\textcolor[rgb]{0.161,0.373,0.58}{\textbf{#1}}}% +\newcommand{\hlkwb}[1]{\textcolor[rgb]{0.69,0.353,0.396}{#1}}% +\newcommand{\hlkwc}[1]{\textcolor[rgb]{0.333,0.667,0.333}{#1}}% +\newcommand{\hlkwd}[1]{\textcolor[rgb]{0.737,0.353,0.396}{\textbf{#1}}}% +\let\hlipl\hlkwb + +\usepackage{framed} +\makeatletter +\newenvironment{kframe}{% + \def\at@end@of@kframe{}% + \ifinner\ifhmode% + \def\at@end@of@kframe{\end{minipage}}% + \begin{minipage}{\columnwidth}% + \fi\fi% + \def\FrameCommand##1{\hskip\@totalleftmargin \hskip-\fboxsep + \colorbox{shadecolor}{##1}\hskip-\fboxsep + % There is no \\@totalrightmargin, so: + \hskip-\linewidth \hskip-\@totalleftmargin \hskip\columnwidth}% + \MakeFramed {\advance\hsize-\width + \@totalleftmargin\z@ \linewidth\hsize + \@setminipage}}% + {\par\unskip\endMakeFramed% + \at@end@of@kframe} +\makeatother + +\definecolor{shadecolor}{rgb}{.97, .97, .97} +\definecolor{messagecolor}{rgb}{0, 0, 0} +\definecolor{warningcolor}{rgb}{1, 0, 1} +\definecolor{errorcolor}{rgb}{1, 0, 0} +\newenvironment{knitrout}{}{} % an empty environment to be redefined in TeX + +\usepackage{alltt} +\usepackage{a4wide} +\input{header} +\hypersetup{ + pdftitle = {Example evaluation of FOCUS dataset Z}, + pdfsubject = {Manuscript}, + pdfauthor = {Johannes Ranke}, + colorlinks = {true}, + linkcolor = {blue}, + citecolor = {blue}, + urlcolor = {red}, + hyperindex = {true}, + linktocpage = {true}, +} +\IfFileExists{upquote.sty}{\usepackage{upquote}}{} +\begin{document} + + + +\title{Example evaluation of FOCUS dataset Z} +\author{\textbf{Johannes Ranke} \\[0.5cm] +%EndAName +Wissenschaftlicher Berater\\ +Kronacher Str. 8, 79639 Grenzach-Wyhlen, Germany\\[0.5cm] +and\\[0.5cm] +University of Bremen\\ +} +\maketitle + +\thispagestyle{empty} \setcounter{page}{0} + +\clearpage + +\tableofcontents + +\textbf{Key words}: Kinetics, FOCUS, nonlinear optimisation + +\section{The data} + +The following code defines the example dataset from Appendix 7 to the FOCUS kinetics +report \citep{FOCUSkinetics2011}, p.350. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlkwd{require}\hlstd{(mkin)} +\end{alltt} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: mkin}} + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: minpack.lm}} + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: rootSolve}} + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: inline}} + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: methods}} + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Loading required package: parallel}}\begin{alltt} +\hlstd{LOD} \hlkwb{=} \hlnum{0.5} +\hlstd{FOCUS_2006_Z} \hlkwb{=} \hlkwd{data.frame}\hlstd{(} + \hlkwc{t} \hlstd{=} \hlkwd{c}\hlstd{(}\hlnum{0}\hlstd{,} \hlnum{0.04}\hlstd{,} \hlnum{0.125}\hlstd{,} \hlnum{0.29}\hlstd{,} \hlnum{0.54}\hlstd{,} \hlnum{1}\hlstd{,} \hlnum{2}\hlstd{,} \hlnum{3}\hlstd{,} \hlnum{4}\hlstd{,} \hlnum{7}\hlstd{,} \hlnum{10}\hlstd{,} \hlnum{14}\hlstd{,} \hlnum{21}\hlstd{,} + \hlnum{42}\hlstd{,} \hlnum{61}\hlstd{,} \hlnum{96}\hlstd{,} \hlnum{124}\hlstd{),} + \hlkwc{Z0} \hlstd{=} \hlkwd{c}\hlstd{(}\hlnum{100}\hlstd{,} \hlnum{81.7}\hlstd{,} \hlnum{70.4}\hlstd{,} \hlnum{51.1}\hlstd{,} \hlnum{41.2}\hlstd{,} \hlnum{6.6}\hlstd{,} \hlnum{4.6}\hlstd{,} \hlnum{3.9}\hlstd{,} \hlnum{4.6}\hlstd{,} \hlnum{4.3}\hlstd{,} \hlnum{6.8}\hlstd{,} + \hlnum{2.9}\hlstd{,} \hlnum{3.5}\hlstd{,} \hlnum{5.3}\hlstd{,} \hlnum{4.4}\hlstd{,} \hlnum{1.2}\hlstd{,} \hlnum{0.7}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{c}\hlstd{(}\hlnum{0}\hlstd{,} \hlnum{18.3}\hlstd{,} \hlnum{29.6}\hlstd{,} \hlnum{46.3}\hlstd{,} \hlnum{55.1}\hlstd{,} \hlnum{65.7}\hlstd{,} \hlnum{39.1}\hlstd{,} \hlnum{36}\hlstd{,} \hlnum{15.3}\hlstd{,} \hlnum{5.6}\hlstd{,} \hlnum{1.1}\hlstd{,} + \hlnum{1.6}\hlstd{,} \hlnum{0.6}\hlstd{,} \hlnum{0.5} \hlopt{*} \hlstd{LOD,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{c}\hlstd{(}\hlnum{0}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{0.5} \hlopt{*} \hlstd{LOD,} \hlnum{2.6}\hlstd{,} \hlnum{3.8}\hlstd{,} \hlnum{15.3}\hlstd{,} \hlnum{37.2}\hlstd{,} \hlnum{31.7}\hlstd{,} \hlnum{35.6}\hlstd{,} \hlnum{14.5}\hlstd{,} + \hlnum{0.8}\hlstd{,} \hlnum{2.1}\hlstd{,} \hlnum{1.9}\hlstd{,} \hlnum{0.5} \hlopt{*} \hlstd{LOD,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{),} + \hlkwc{Z3} \hlstd{=} \hlkwd{c}\hlstd{(}\hlnum{0}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{NA}\hlstd{,} \hlnum{0.5} \hlopt{*} \hlstd{LOD,} \hlnum{9.2}\hlstd{,} \hlnum{13.1}\hlstd{,} \hlnum{22.3}\hlstd{,} \hlnum{28.4}\hlstd{,} \hlnum{32.5}\hlstd{,} + \hlnum{25.2}\hlstd{,} \hlnum{17.2}\hlstd{,} \hlnum{4.8}\hlstd{,} \hlnum{4.5}\hlstd{,} \hlnum{2.8}\hlstd{,} \hlnum{4.4}\hlstd{))} + +\hlstd{FOCUS_2006_Z_mkin} \hlkwb{<-} \hlkwd{mkin_wide_to_long}\hlstd{(FOCUS_2006_Z)} +\end{alltt} +\end{kframe} +\end{knitrout} + +\section{Parent compound and one metabolite} + +The next step is to set up the models used for the kinetic analysis. As the +simultaneous fit of parent and the first metabolite is usually straightforward, +Step 1 (SFO for parent only) is skipped here. We start with the model 2a, +with formation and decline of metabolite Z1 and the pathway from parent +directly to sink included (default in mkin). + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.2a} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{))} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.2a} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.2a, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.2a)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_1-1} +\begin{kframe}\begin{alltt} +\hlkwd{summary}\hlstd{(m.Z.2a,} \hlkwc{data} \hlstd{=} \hlnum{FALSE}\hlstd{)}\hlopt{$}\hlstd{bpar} +\end{alltt} +\begin{verbatim} +## Estimate se_notrans t value Pr(>t) +## Z0_0 9.701488e+01 3.55313691 2.730401e+01 1.679214e-21 +## k_Z0_sink 1.281376e-11 0.22689470 5.647447e-11 5.000000e-01 +## k_Z0_Z1 2.236006e+00 0.16507604 1.354531e+01 7.396594e-14 +## k_Z1_sink 4.821248e-01 0.06585369 7.321150e+00 3.552015e-08 +## Lower Upper +## Z0_0 91.4058170 102.6239462 +## k_Z0_sink 0.0000000 Inf +## k_Z0_Z1 1.8419826 2.7143172 +## k_Z1_sink 0.4005856 0.5802613 +\end{verbatim} +\end{kframe} +\end{knitrout} + +As obvious from the parameter summary (the \texttt{bpar} component of the +summary), the kinetic rate constant from parent compound Z to sink +is negligible. Accordingly, the exact magnitude of the fitted parameter +\texttt{log k\_Z0\_sink} is ill-defined and the covariance matrix is not +returned (not shown, would be visible in the complete summary). +This suggests, in agreement with the analysis in the FOCUS kinetics report, to +simplify the model by removing the pathway to sink. + +A similar result can be obtained when formation fractions are used in the model +formulation: + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.2a.ff} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{),} + \hlkwc{use_of_ff} \hlstd{=} \hlstr{"max"}\hlstd{)} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.2a.ff} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.2a.ff, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.2a.ff)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_2-1} +\begin{kframe}\begin{alltt} +\hlkwd{summary}\hlstd{(m.Z.2a.ff,} \hlkwc{data} \hlstd{=} \hlnum{FALSE}\hlstd{)}\hlopt{$}\hlstd{bpar} +\end{alltt} +\begin{verbatim} +## Estimate se_notrans t value Pr(>t) Lower +## Z0_0 97.0148812 3.55314944 27.303912 1.679369e-21 91.3287912 +## k_Z0 2.2360064 0.21684747 10.311425 3.661846e-11 1.8052909 +## k_Z1 0.4821248 0.06585372 7.321147 3.552045e-08 0.3996826 +## f_Z0_to_Z1 1.0000000 0.10147342 9.854798 9.707056e-11 0.0000000 +## Upper +## Z0_0 102.7009713 +## k_Z0 2.7694843 +## k_Z1 0.5815722 +## f_Z0_to_Z1 1.0000000 +\end{verbatim} +\end{kframe} +\end{knitrout} + +Here, the ilr transformed formation fraction fitted in the model takes a very +large value, and the backtransformed formation fraction from parent Z to Z1 is +practically unity. Again, the covariance matrix is not returned as the model is +overparameterised. + +The simplified model is obtained by setting the list component \texttt{sink} to +\texttt{FALSE}.\footnote{If the model formulation without formation fractions +is used, the same effect can be obtained by fixing the parameter \texttt{k\_Z\_sink} +to a value of zero.} + +In the following, we use the parameterisation with formation fractions in order +to be able to compare with the results in the FOCUS guidance, and as it +makes it easier to use parameters obtained in a previous fit when adding a further +metabolite. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.3} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{),} \hlkwc{use_of_ff} \hlstd{=} \hlstr{"max"}\hlstd{)} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.3} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.3, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.3)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_3-1} +\begin{kframe}\begin{alltt} +\hlkwd{summary}\hlstd{(m.Z.3,} \hlkwc{data} \hlstd{=} \hlnum{FALSE}\hlstd{)}\hlopt{$}\hlstd{bpar} +\end{alltt} +\begin{verbatim} +## Estimate se_notrans t value Pr(>t) Lower +## Z0_0 97.0148815 2.68177093 36.17568 2.363587e-25 91.5215237 +## k_Z0 2.2360064 0.14686110 15.22531 2.246514e-15 1.9545310 +## k_Z1 0.4821248 0.04268712 11.29439 3.068581e-12 0.4021552 +## Upper +## Z0_0 102.5082392 +## k_Z0 2.5580177 +## k_Z1 0.5779966 +\end{verbatim} +\end{kframe} +\end{knitrout} + +As there is only one transformation product for Z0 and no pathway +to sink, the formation fraction is internally fixed to unity. + +\section{Including metabolites Z2 and Z3} + +As suggested in the FOCUS report, the pathway to sink was removed for metabolite Z1 as +well in the next step. While this step appears questionable on the basis of the above results, it +is followed here for the purpose of comparison. Also, in the FOCUS report, it is +assumed that there is additional empirical evidence that Z1 quickly and exclusively +hydrolyses to Z2. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.5} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{),} \hlkwc{use_of_ff} \hlstd{=} \hlstr{"max"}\hlstd{)} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.5} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.5, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.5)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_5-1} + +\end{knitrout} + +Finally, metabolite Z3 is added to the model. We use the optimised +differential equation parameter values from the previous fit in order to +accelerate the optimization. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.FOCUS} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z3"}\hlstd{),} + \hlkwc{Z3} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{),} + \hlkwc{use_of_ff} \hlstd{=} \hlstr{"max"}\hlstd{)} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.FOCUS} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.FOCUS, FOCUS_2006_Z_mkin,} + \hlkwc{parms.ini} \hlstd{= m.Z.5}\hlopt{$}\hlstd{bparms.ode,} + \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.FOCUS)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_6-1} +\begin{kframe}\begin{alltt} +\hlkwd{summary}\hlstd{(m.Z.FOCUS,} \hlkwc{data} \hlstd{=} \hlnum{FALSE}\hlstd{)}\hlopt{$}\hlstd{bpar} +\end{alltt} +\begin{verbatim} +## Estimate se_notrans t value Pr(>t) Lower +## Z0_0 96.83849566 2.05886133 47.034977 5.583742e-44 92.70517559 +## k_Z0 2.21540897 0.11813046 18.753919 7.744034e-25 1.99051134 +## k_Z1 0.47829896 0.02928763 16.331089 3.333376e-22 0.42297603 +## k_Z2 0.45161653 0.04421268 10.214638 3.110936e-14 0.37103495 +## k_Z3 0.05869343 0.01429485 4.105914 7.285393e-05 0.03599566 +## f_Z2_to_Z3 0.47150952 0.05705233 8.264510 2.809889e-11 0.36038193 +## Upper +## Z0_0 100.97181573 +## k_Z0 2.46571661 +## k_Z1 0.54085782 +## k_Z2 0.54969886 +## k_Z3 0.09570371 +## f_Z2_to_Z3 0.58553462 +\end{verbatim} +\begin{alltt} +\hlkwd{endpoints}\hlstd{(m.Z.FOCUS)} +\end{alltt} +\begin{verbatim} +## $ff +## Z2_Z3 Z2_sink +## 0.4715095 0.5284905 +## +## $SFORB +## logical(0) +## +## $distimes +## DT50 DT90 +## Z0 0.3128755 1.039350 +## Z1 1.4491923 4.814113 +## Z2 1.5348136 5.098540 +## Z3 11.8096217 39.230714 +\end{verbatim} +\end{kframe} +\end{knitrout} + +This fit corresponds to the final result chosen in Appendix 7 of the FOCUS +report. Confidence intervals returned by mkin are based on internally +transformed parameters, however. + +\section{Using the SFORB model for parent and metabolites} + +As the FOCUS report states, there is a certain tailing of the time course of metabolite +Z3. Also, the time course of the parent compound is not fitted very well using the +SFO model, as residues at a certain low level remain. + +Therefore, an additional model is offered here, using the single first-order +reversible binding (SFORB) model for metabolite Z3. As expected, the $\chi^2$ +error level is lower for metabolite Z3 using this model and the graphical +fit for Z3 is improved. However, the covariance matrix is not returned. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.mkin.1} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z3"}\hlstd{),} + \hlkwc{Z3} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFORB"}\hlstd{))} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.mkin.1} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.mkin.1, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.mkin.1)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_7-1} +\begin{kframe}\begin{alltt} +\hlkwd{summary}\hlstd{(m.Z.mkin.1,} \hlkwc{data} \hlstd{=} \hlnum{FALSE}\hlstd{)}\hlopt{$}\hlstd{cov.unscaled} +\end{alltt} +\begin{verbatim} +## NULL +\end{verbatim} +\end{kframe} +\end{knitrout} + +Therefore, a further stepwise model building is performed starting from the +stage of parent and two metabolites, starting from the assumption that the model +fit for the parent compound can be improved by using the SFORB model. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.mkin.3} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFORB"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{))} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.mkin.3} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.mkin.3, FOCUS_2006_Z_mkin,} \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.mkin.3)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_9-1} + +\end{knitrout} + +This results in a much better representation of the behaviour of the parent +compound Z0. + +Finally, Z3 is added as well. These models appear overparameterised (no +covariance matrix returned) if the sink for Z1 is left in the models. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.mkin.4} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFORB"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z3"}\hlstd{),} + \hlkwc{Z3} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{))} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.mkin.4} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.mkin.4, FOCUS_2006_Z_mkin,} + \hlkwc{parms.ini} \hlstd{= m.Z.mkin.3}\hlopt{$}\hlstd{bparms.ode,} + \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.mkin.4)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_10-1} + +\end{knitrout} + +The error level of the fit, but especially of metabolite Z3, can be improved if +the SFORB model is chosen for this metabolite, as this model is capable of +representing the tailing of the metabolite decline phase. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{Z.mkin.5} \hlkwb{<-} \hlkwd{mkinmod}\hlstd{(}\hlkwc{Z0} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFORB"}\hlstd{,} \hlstr{"Z1"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z1} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z2"}\hlstd{,} \hlkwc{sink} \hlstd{=} \hlnum{FALSE}\hlstd{),} + \hlkwc{Z2} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFO"}\hlstd{,} \hlstr{"Z3"}\hlstd{),} + \hlkwc{Z3} \hlstd{=} \hlkwd{mkinsub}\hlstd{(}\hlstr{"SFORB"}\hlstd{))} +\end{alltt} +\begin{verbatim} +## make[1]: warning: jobserver unavailable: using -j1. Add '+' to parent make rule. +\end{verbatim} + + +{\ttfamily\noindent\itshape\color{messagecolor}{\#\# Successfully compiled differential equation model from auto-generated C code.}}\begin{alltt} +\hlstd{m.Z.mkin.5} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.mkin.5, FOCUS_2006_Z_mkin,} + \hlkwc{parms.ini} \hlstd{= m.Z.mkin.4}\hlopt{$}\hlstd{bparms.ode[}\hlnum{1}\hlopt{:}\hlnum{4}\hlstd{],} + \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.mkin.5)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_11-1} + +\end{knitrout} + +The summary view of the backtransformed parameters shows that we get no +confidence intervals due to overparameterisation. As the optimized +\texttt{k\_Z3\_bound\_free} is excessively small, it seems reasonable to fix it to +zero. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlstd{m.Z.mkin.5a} \hlkwb{<-} \hlkwd{mkinfit}\hlstd{(Z.mkin.5, FOCUS_2006_Z_mkin,} + \hlkwc{parms.ini} \hlstd{=} \hlkwd{c}\hlstd{(m.Z.mkin.5}\hlopt{$}\hlstd{bparms.ode[}\hlnum{1}\hlopt{:}\hlnum{7}\hlstd{],} + \hlkwc{k_Z3_bound_free} \hlstd{=} \hlnum{0}\hlstd{),} + \hlkwc{fixed_parms} \hlstd{=} \hlstr{"k_Z3_bound_free"}\hlstd{,} + \hlkwc{quiet} \hlstd{=} \hlnum{TRUE}\hlstd{)} +\hlkwd{plot_sep}\hlstd{(m.Z.mkin.5a)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_11a-1} + +\end{knitrout} + +As expected, the residual plots for Z0 and Z3 are more random than in the case of the +all SFO model for which they were shown above. In conclusion, the model +\texttt{Z.mkin.5a} is proposed as the best-fit model for the dataset from +Appendix 7 of the FOCUS report. + +A graphical representation of the confidence intervals can finally be obtained. + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlkwd{mkinparplot}\hlstd{(m.Z.mkin.5a)} +\end{alltt} +\end{kframe} +\includegraphics[width=\maxwidth]{figure/FOCUS_2006_Z_fits_11b-1} + +\end{knitrout} + +The endpoints obtained with this model are + +\begin{knitrout} +\definecolor{shadecolor}{rgb}{0.969, 0.969, 0.969}\color{fgcolor}\begin{kframe} +\begin{alltt} +\hlkwd{endpoints}\hlstd{(m.Z.mkin.5a)} +\end{alltt} +\begin{verbatim} +## $ff +## Z0_free_Z1 Z1_Z2 Z2_sink Z2_Z3_free Z3_free_sink +## 1.0000000 1.0000000 0.4634431 0.5365569 1.0000000 +## +## $SFORB +## Z0_b1 Z0_b2 Z3_b1 Z3_b2 +## 2.447138640 0.007512589 0.080007099 0.000000000 +## +## $distimes +## DT50 DT90 DT50_Z0_b1 DT50_Z0_b2 DT50_Z3_b1 DT50_Z3_b2 +## Z0 0.3042972 1.184811 0.283248 92.26476 NA NA +## Z1 1.5147787 5.031986 NA NA NA NA +## Z2 1.6413857 5.452565 NA NA NA NA +## Z3 NA NA NA NA 8.663571 Inf +\end{verbatim} +\end{kframe} +\end{knitrout} + +It is clear the degradation rate of Z3 towards the end of the experiment +is very low as DT50\_Z3\_b2 (the second Eigenvalue of the system of two differential +equations representing the SFORB system for Z3, corresponding to the slower rate +constant of the DFOP model) is reported to be infinity. However, this appears +to be a feature of the data. + +\bibliographystyle{plainnat} +\bibliography{references} + +\end{document} +% vim: set foldmethod=syntax: |