A cosinor model aims to model the amplitude (\(A\)), acrophase (\(\phi\)), and MESOR (\(M\)) of a rhythmic dataset.

MESOR (\(M\)) is the Midline Estimating Statistic of Rhythm, and may also be referred to as the equilibrium point.

Amplitude (\(A\)) is the difference between the MESOR and the maximum height of the rhythm.

Acrophase (\(\phi\)) is the phase at which the maximal response occurs.

These could be modelled using a cosine function:

\[Y(t) = M + Acos(\frac{2\pi t}{\tau} - \phi) + e(t)\] where \(e(t)\) is the error term.

However, these cannot be estimated using a linear modelling
framework! Other packages, including `{circacompare}`

(Parsons et al. 2020), fit this exact
nonlinear model but most packages (including this one) decomposes this
into linear terms, creating the cosinor model:

\[Y(t) = M + \beta x + \gamma z + e(t)\]

Where \(x =cos(\frac{2\pi t}{τ})\), \(z =sin(\frac{2\pi t}{τ})\), \(\beta = A cos(\phi)\), \(\gamma = A sin(\phi)\)

This linear model is passed to the package `{glmmTMB}`

(Brooks et al. 2017) in `lme4`

syntax. If the model has no random effects, `glmmTMB`

uses
maximum likelihood estimation to estimate the linear coefficients of the
model. For models with random effects, a Laplace approximation is used
to integrate over the random effects. This approximation is handled by
the `{TMB}`

(Kristensen et al. 2016) package which uses
automatic differentiation of the joint likelihood function to provide
fast computations of parameter estimates. A detailed explanation of this
process is described here (Kristensen et al. 2016)

`glmmTMB`

returns the estimates of the linear coefficients
from the linear model. To recover the estimates of the original
parameters for amplitude (\(A\)) and
acrophase (\(\phi\)), the estimates for
\(\hat\beta\) and \(\hat\gamma\) must be transformed as per the
following equations:

\[\hat\phi = \arctan(\frac{\hat\gamma}{\hat\beta}) \]

\[\hat A = \sqrt{\hat\beta ^2 + \hat\gamma
^ 2}\] These transformed parameters for acrophase and amplitude,
along with MESOR are returned as part of the `cglmm`

output.
For a more thorough introduction to cosinor modelling, see here (Cornelissen 2014).

`{GLMMcosinor}`

allows the user to fit generalised linear
models based on rhythmic data with a cosinor model. It allows users to
summarise, predict, and plot these models too. Existing packages have
focused primarily on Gaussian data. Some circadian regression modelling
packages have allowed users to specify generalised linear models, but
with limited flexibility. `{GLMMcosinor}`

takes a
comprehensive approach to modelling by harnessing the
`{glmmTMB}`

package, that has a wide range of available link
functions, allowing users to model rhythmic data from a wide range of
distributions (for full list - see `?family`

and
`?glmmTMB::family_glmmTMB`

) including:

- Binomial
- Guassian
- Inverse Gaussian
- Gamma
- Poisson
- Negative Binomial

The table below shows what features are available within
`{GLMMcosinor}`

and other methods.

Software | Language | Multicomponent | Dispersion model | Zero-inflated model | Differential rhythmicity | Parameter estimates for differences | Family (available link functions) | Estimated parameters | Reference |
---|---|---|---|---|---|---|---|---|---|

GLMMcosinor | R | ✓ | ✓ | ✓ | ✗ | ✓ | n > 15 Any family avilable in {glmmTMB}: Gaussian, gamma, binomial, Poisson, … | Amplitude, acrophase, MESOR | Parsons, 2023 |

CircaCompare | R | ✗ | ✗ | ✗ | ✗ | ✓ | n = 1 Gaussian | Amplitude, acrophase, MESOR, and exponential decay of any of these characteristics or the differences in them between groups | Parsons, 2020 |

Cosinor | R | ✗ | ✗ | ✗ | ✓ | ✓ | n = 1 Gaussian | Amplitude, acrophase, MESOR | Sachs, 2014 |

Cosinor2 | R | ✗ | ✗ | ✗ | ✓ | ✓ | n = 1 Gaussian | Amplitude, acrophase, MESOR | Mutak, 2018 |

CosinorPy | Python | ✓ | ✗ | ✗ | ✓ | ✓ | n = 3 Gaussian, Poisson, negative-binomial | Amplitude, acrophase, MESOR | Moskon, 2020 |

DiscoRhythm | R | ✗ | ✗ | ✗ | ✗ | ✗ | n = 1 Gaussian | Amplitude, acrophase | Carlucci, 2019 |

FMM | R | ✓ | ✗ | ✗ | ✗ | ✓ | n = 1 Gaussian | Amplitude, FMM phase angle parameters: alpha, beta, gamma | Fernández, 2022 |

Kronos | R | ✓ | ✗ | ✗ | ✓ | ✓ | n = 1 Gaussian | Amplitude, acrophase, MESOR | Bastiaanssen, 2023 |

LimoRhyde | R | ✗ | ✗ | ✗ | ✓ | ✓ | n = 1 Gaussian | Amplitude, acrophase, MESOR, period | Singer, 2019 |

RhythmCount | Python | ✓ | ✗ | ✓ | ✗ | ✓ | n = 6 Poisson, generalised-Poisson, zero-inflated Poisson, negative binomial, zero-inflated negative-binomial | Amplitude, acrophase, MESOR, Zenith | Velikajne, 2022 |

`cglmm()`

`cglmm()`

wrangles the data appropriately to fit the
cosinor model given the formula specified by the user. It returns a
model, providing estimates of amplitude, acrophase, and MESOR (Midline
Statistic Of Rhythm).

The formula argument for `cglmm()`

is specified using the
`{lme4}`

style (for details see
`vignette("lmer", package = "lme4")`

). The only difference is
that it allows for use of `amp_acro()`

within the formula
that is used to identify the cosinor (rhythmic) components and relevant
variables in the provided data. Any other combination of covariates can
also be included in the formula as well as random effects. Additionally,
zero-inflation (`ziformula`

) and dispersion
(`dispformula`

) formulae can be incorporated if required. For
detailed examples of how to specify these types of models, see the mixed-models,
model-specification
and multiple-components
vignettes.

For example, consider the following model and its output:

```
library(GLMMcosinor)
cosinor_model <- cglmm(
vit_d ~ X + amp_acro(time, period = 12, group = "X"),
data = vitamind
)
```

Notice how both the raw and transformed coefficients are provided as
output. The adapted `data.frame`

that was used to fit the raw
model can be accessed from the model and includes `main_rrr1`

and `main_sss1`

columns of data:

```
head(cosinor_model$newdata)
#> vit_d time X main_rrr1 main_sss1
#> 1 16.12091 11.439525 0 0.9572476 -0.2892699
#> 2 29.90624 5.807104 0 -0.9949038 0.1008285
#> 3 39.17572 1.045492 1 0.8538711 0.5204846
#> 4 35.15403 4.082983 1 -0.5371451 0.8434899
#> 5 43.67065 10.606247 1 0.7453295 -0.6666963
#> 6 31.20360 5.126054 0 -0.8971168 0.4417935
```

In this example, the `main`

prefix indicates that this is
the data for the conditional model, as opposed to (potential) dispersion
or zero-inflation models, which have the prefixes `disp`

and
`zi`

, respectively. The numeric suffix, indicates that this
is the data for the first (and only) cosinor component. If there are
multiple components, the columns of data will be named accordingly.

`cglmm()`

The `cglmm()`

function is used to fit cosinor models to a
variety of distributions using the `glmmTMB()`

function.

```
cglmm(
formula = vit_d ~ amp_acro(time, period = 12),
data = vitamind,
family = gaussian
)
#>
#> Conditional Model
#>
#> Raw formula:
#> vit_d ~ main_rrr1 + main_sss1
#>
#> Raw Coefficients:
#> Estimate
#> (Intercept) 30.25467
#> main_rrr1 2.59418
#> main_sss1 5.75079
#>
#> Transformed Coefficients:
#> Estimate
#> (Intercept) 30.25467
#> amp 6.30883
#> acr 1.14703
```

`formula`

: A formula specifying the model structure, including the response variable and the cosinor components (using`amp_acro()`

).`data`

: The`data.frame`

containing the variables used in the formula.`family`

: The family of the distribution for the response variable (e.g., poisson, gaussian, or any family found in`?family`

and`?glmmTMB::family_glmmTMB`

)

The `amp_acro()`

function is used within the formula to
specify the cosinor components. It allows you to specify the period of
the rhythm and, if necessary, the grouping structure and the number of
components. The arguments of `amp_acro()`

are:

`group`

: The name of the grouping variable in the dataset.`time_col`

: The name of the time column.`n_components`

: The number of components in the cosinor model.`period`

: The period(s) of the rhythm.

The most relevant output from the `cglmm()`

function is
likely to be the parameter estimates for MESOR, amplitude, and acrophase
under the ‘Transformed Coefficients’ heading. These are the recovered
estimates mentioned at the beginning of this vignette: the amplitude and
phase. The ‘Raw Coefficients’ are the coefficients from the cosinor
model. In this example, the `main_rrr1`

and
`main_sss1`

correspond to \(\hat\beta\) and \(\hat\gamma\) in the first section,
respectively.

The following example fits a grouped single-component model with a Guassian distribution (the default).

```
cglmm(
vit_d ~ X + amp_acro(time, period = 12, group = "X"),
data = vitamind
)
#>
#> Conditional Model
#>
#> Raw formula:
#> vit_d ~ X + X:main_rrr1 + X:main_sss1
#>
#> Raw Coefficients:
#> Estimate
#> (Intercept) 29.68980
#> X1 1.90186
#> X0:main_rrr1 0.93079
#> X1:main_rrr1 6.51029
#> X0:main_sss1 6.20099
#> X1:main_sss1 4.81846
#>
#> Transformed Coefficients:
#> Estimate
#> (Intercept) 29.68980
#> [X=1] 1.90186
#> [X=0]:amp 6.27046
#> [X=1]:amp 8.09947
#> [X=0]:acr 1.42181
#> [X=1]:acr 0.63715
```

Under the ‘Transformed Coefficients’ heading:

`(Intercept) = 29.6898`

is the MESOR estimate of group 0`[X=1] = 1.90186`

is the difference between the MESOR estimates of group 1 and 2 *`[X=0]:amp = 6.27046`

is the amplitude estimate for group 0`[X=1]:amp = 8.09947`

is the amplitude estimate for group 1`[X=0]:acr = 1.42181`

is the acrophase estimate in radians for group 0 **`[X=1]:acr = 0.63715`

is the acrophase estimate in radians for group 1

* Hence, the MESOR estimate for group 1 would be
`29.6898 + 1.90186 = 31.59166`

. This is due to the behaviour
of the `glmmTMB()`

function. This can be adjusted by adding a
`0 +`

to the beginning of the formula:

```
cglmm(
vit_d ~ 0 + X + amp_acro(time,
period = 12,
group = "X"
),
data = vitamind
)
#>
#> Conditional Model
#>
#> Raw formula:
#> vit_d ~ X + X:main_rrr1 + X:main_sss1 - 1
#>
#> Raw Coefficients:
#> Estimate
#> X0 29.68980
#> X1 31.59165
#> X0:main_rrr1 0.93079
#> X1:main_rrr1 6.51029
#> X0:main_sss1 6.20101
#> X1:main_sss1 4.81847
#>
#> Transformed Coefficients:
#> Estimate
#> [X=0] 29.68980
#> [X=1] 31.59165
#> [X=0]:amp 6.27048
#> [X=1]:amp 8.09948
#> [X=0]:acr 1.42181
#> [X=1]:acr 0.63716
```

Note how now, `[X=1] = 31.59165`

and this represents the
estimate for the MESOR for group 1, rather than the difference.

** Note how the acrophase is provided in units of radians. Since the
period is 12, an acrophase of 1.42181 radians corresponds to a time of
\(\frac{1.42181}{2 \pi} \times 12 =
2.715457\). This means the maximum response occurs at 2.715 time
units. We can check this visually using the `autoplot()`

function, looking at the `[X=0]`

level (red line)

`cglmm()`

model specificationThe `cglmm()`

function allows you to specify different
types of cosinor models with or without grouping variables. The function
can also generate dispersion models and zero-inflation models. For more
detailed explanations and examples, see the model-specification
article.

Additionally, the `cglmm()`

function provides more
advanced functionality for multi-component models, and detailed
explanations can be found in the multiple-components
article.

The `cglmm()`

function also allows mixed model
specification. See the mixed-models
article for more details.

`summary()`

and testing for differences between
estimatesThe `summary()`

method for the outputs from
`cglmm()`

provides a more detailed summary of the model and
its parameter estimates and uncertainty. It outputs the estimates,
standard errors, confidence intervals, and p-values for both the raw
model parameters and the transformed parameters. The summary statistics
do not represent a comparison between any groups for the cosinor
components - that is the role of the
`test_cosinor_components()`

and
`test_cosinor_levels()`

functions.

Here is an example of how to use `summary()`

with some
simulated data:

```
testdata_simple <- simulate_cosinor(
1000,
n_period = 2,
mesor = 5,
amp = 2,
acro = 1,
beta.mesor = 4,
beta.amp = 1,
beta.acro = 0.5,
family = "poisson",
period = 12,
n_components = 1,
beta.group = TRUE
)
```

```
object <- cglmm(
Y ~ group + amp_acro(times, period = 12, group = "group"),
data = testdata_simple, family = poisson()
)
summary(object)
#>
#> Conditional Model
#> Raw model coefficients:
#> estimate standard.error lower.CI upper.CI p.value
#> (Intercept) 4.998454142 0.003463730 4.991665356 5.00524 < 2.22e-16
#> group1 -1.002150001 0.005937109 -1.013786521 -0.99051 < 2.22e-16
#> group0:main_rrr1 1.082281784 0.003347565 1.075720677 1.08884 < 2.22e-16
#> group1:main_rrr1 0.876651963 0.006198710 0.864502714 0.88880 < 2.22e-16
#> group0:main_sss1 1.682350718 0.003919418 1.674668800 1.69003 < 2.22e-16
#> group1:main_sss1 0.481951763 0.005936670 0.470316104 0.49359 < 2.22e-16
#>
#> (Intercept) ***
#> group1 ***
#> group0:main_rrr1 ***
#> group1:main_rrr1 ***
#> group0:main_sss1 ***
#> group1:main_sss1 ***
#> ---
#> Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
#>
#> Transformed coefficients:
#> estimate standard.error lower.CI upper.CI p.value
#> (Intercept) 4.998454142 0.003463730 4.991665356 5.00524 < 2.22e-16 ***
#> [group=1] -1.002150001 0.005937109 -1.013786521 -0.99051 < 2.22e-16 ***
#> [group=0]:amp1 2.000409408 0.004275553 1.992029478 2.00879 < 2.22e-16 ***
#> [group=1]:amp1 1.000398004 0.006379631 0.987894156 1.01290 < 2.22e-16 ***
#> [group=0]:acr1 0.999134804 0.001439122 0.996314177 1.00196 < 2.22e-16 ***
#> [group=1]:acr1 0.502662067 0.005739524 0.491412807 0.51391 < 2.22e-16 ***
#> ---
#> Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
```

If we wanted to test the difference between the amplitude estimate
for component 1 between `group 1`

and `group 2`

,
we can use the `test_cosinor_levels()`

function:

```
test_cosinor_levels(object, x_str = "group", param = "amp")
#> Test Details:
#> Parameter being tested:
#> Amplitude
#>
#> Comparison type:
#> levels
#>
#> Grouping variable used for comparison between groups: group
#> Reference group: 0
#> Comparator group: 1
#>
#> cglmm model only has a single component and to compare
#> between groups.
#>
#>
#>
#> Global test:
#> Statistic:
#> 16955.27
#>
#> P-value:
#> 0
#>
#>
#> Individual tests:
#> Statistic:
#> -130.21
#>
#> P-value:
#> 0
#>
#> Estimate and 95% confidence interval:
#> -1 (-1.02 to -0.98)
```

The estimate here is the estimate of the difference between the
inputted values, along with its confidence interval. The real parameters
for `amp`

in the first component were 2 and 1 for groups 0
and 1 respectively, and so the difference is approximately -1.

Now, consider an example where the difference is not so clear.

```
testdata_poisson <- simulate_cosinor(100,
n_period = 2,
mesor = 7,
amp = c(0.1, 0.5),
acro = c(1, 1),
beta.mesor = 4.4,
beta.amp = c(0.1, 0.46),
beta.acro = c(0.5, -1.5),
family = "poisson",
period = c(12, 6),
n_components = 2,
beta.group = TRUE
)
```

```
cosinor_model <- cglmm(
Y ~ group + amp_acro(times,
period = c(12, 6),
n_components = 2,
group = "group"
),
data = testdata_poisson,
family = poisson()
)
test_cosinor_levels(
cosinor_model,
x_str = "group",
param = "amp",
component_index = 1
)
#> Test Details:
#> Parameter being tested:
#> Amplitude
#>
#> Comparison type:
#> levels
#>
#> Grouping variable used for comparison between groups: group
#> Reference group: 0
#> Comparator group: 1
#>
#> cglmm model has2 components. Component 1 is being used for comparison between groups.
#>
#>
#>
#> Global test:
#> Statistic:
#> 0.05
#>
#> P-value:
#> 0.8245
#>
#>
#> Individual tests:
#> Statistic:
#> -0.22
#>
#> P-value:
#> 0.8245
#>
#> Estimate and 95% confidence interval:
#> 0 (-0.04 to 0.03)
```

In this example, there is no significant difference in the estimate
of `amp`

for the first component between the reference group
and the comparator group. Also notice how if we are comparing between
levels, we should keep the component the same, and that is what
`component_index`

sets. Likewise, when we test between
components using `test_cosinor_components()`

, we can indicate
which level this comparison occurs using `level_index`

. There
may be multiple `groups`

, in which case we can fix the
`group`

using the `x_str`

argument.

As an example of testing the difference between components for the same level:

```
test_cosinor_components(
cosinor_model,
x_str = "group",
param = "acr",
level_index = 1
)
#> Test Details:
#> Parameter being tested:
#> Acrophase
#>
#> Comparison type:
#> components
#>
#> Component indices used for comparison between groups: group
#> Reference component: 1
#> Comparator component: 2
#>
#>
#> Global test:
#> Statistic:
#> 154.29
#>
#> P-value:
#> 0
#>
#>
#> Individual tests:
#> Statistic:
#> -12.42
#>
#> P-value:
#> 0
#>
#> Estimate and 95% confidence interval:
#> -1.89 (-2.19 to -1.59)
```

In this situation, there is a significant difference between the acrophase for the comparator group between its two components.

`predict()`

The `predict()`

method allows users to get predicted
values from the model on either the existing or new data.

```
#> predictions Y times group
#> 1 865.8332 871 17.009450 0
#> 2 701.2750 714 10.503837 0
#> 3 798.4445 861 4.800118 0
#> 4 1551.0482 1541 18.409584 0
#> 5 1733.9702 1699 12.315885 0
#> 6 1972.1517 1951 1.072893 0
```

`cglmm`

objectsThe `{GLMMcosinor}`

package includes two ways to visualise
`cglmm()`

objects. Firstly, the `autoplot()`

method creates a time-response plot of the fitted model for all
groups:

This function also allows users to superimpose the data (that was
used to fit the model) over the fitted model, using the
`superimpose.data = TRUE`

, as demonstrated above. By default,
the generated plot will have x-limits corresponding to the minimum and
maximum values of the time-vector in the original dataframe, although
the x-limits can be manually defined by the user using the
`xlims`

argument. The details of using the
`autoplot`

function are found in the model-visualisations
vignette.

Brooks, Mollie E., Kasper Kristensen, Koen J. van Benthem, Arni
Magnusson, Casper W. Berg, Anders Nielsen, Hans J. Skaug, Martin
Maechler, and Benjamin M. Bolker. 2017. “glmmTMB Balances Speed and Flexibility Among
Packages for Zero-Inflated Generalized Linear Mixed Modeling.”
*The R Journal* 9 (2): 378–400. https://doi.org/10.32614/RJ-2017-066.

Cornelissen, Germaine. 2014. “Cosinor-Based Rhythmometry.”
*Theoretical Biology and Medical Modelling* 11 (1): 1–24.

Kristensen, Kasper, Anders Nielsen, Casper W. Berg, Hans Skaug, and
Bradley M. Bell. 2016. “TMB: Automatic Differentiation and Laplace
Approximation.” *Journal of Statistical Software* 70 (5):
1–21. https://doi.org/10.18637/jss.v070.i05.

Parsons, Rex, Richard Parsons, Nicholas Garner, Henrik Oster, and Oliver
Rawashdeh. 2020. “CircaCompare: A Method to Estimate and
Statistically Support Differences in Mesor, Amplitude and Phase, Between
Circadian Rhythms.” *Bioinformatics* 36 (4): 1208–12.