Adv Physiol Educ 34: 128–133, 2010;
Staying Current
When t-tests or Wilcoxon-Mann-Whitney tests won’t do
Fiona McElduff,1 Mario Cortina-Borja,1 Shun-Kai Chan,2 and Angie Wade1
1
Medical Research Council Centre of Epidemiology for Child Health and 2Nephro-Urology Unit, Institute of Child Health,
University College London, London, United Kingdom
Submitted 1 February 2010; accepted in final form 9 July 2010
McElduff F, Cortina-Borja M, Chan SK, Wade A. When t-tests
or Wilcoxon-Mann-Whitney tests won’t do. Adv Physiol Educ 34:
128 –133, 2010; doi:10.1152/advan.00017.2010.—t-Tests are widely
used by researchers to compare the average values of a numeric
outcome between two groups. If there are doubts about the suitability
of the data for the requirements of a t-test, most notably the distribu-
tion being non-normal, the Wilcoxon-Mann-Whitney test may be used
instead. However, although often applied, both tests may be invalid
when discrete and/or extremely skew data are analyzed. In medicine,
extremely skewed data having an excess of zeroes are often observed,
representing a numeric outcome that does not occur for a large
percentage of cases (so is often zero) but which also sometimes takes
relatively large values. For data such as this, application of the t-test
or Wilcoxon-Mann-Whitney test could lead researchers to draw in-
correct conclusions. A valid alternative is regression modeling to
quantify the characteristics of the data. The increased availability of
software has simplified the application of these more complex statis-
tical analyses and hence facilitates researchers to use them. In this
article, we illustrate the methodology applied to a comparison of cyst
counts taken from control and steroid-treated fetal mouse kidneys.
Poisson; negative binomial; zero inflation
WHEN FACED WITH THE TASK of comparing a numeric outcome
between two groups, most clinicians perform either a t-test or,
if the data set is small and/or the assumptions for parametric
testing are not met, a Wilcoxon-Mann-Whitney test (WMW;
also called the Mann-Whitney-Wilcoxon test, Wilcoxon rank-
sum test, Wilcoxon test, or Mann-Whitney U-test). Statistical
methods may be divided according to the assumptions they
demand from the data. Parametric methods suppose that the
data follow a specific probability distribution whose parame-
ters can be estimated, whereas the basic principle of nonpara-
metric tests, such as the WMW test, is that the observed pooled
values are ranked and no particular distributional assumptions
are made. Statistical analyses of highly skewed distributions
can be especially problematic, particularly when the data are
discrete and exhibit value inflation.
The t-test is a parametric test of the difference in mean
between two groups that assumes that the data are normally
distributed and the groups have equal variances. Welch’s
approximation (13) can be used when the assumption of equal
variances is not satisfied. While it is commonly believed that
the WMW test compares medians in the same way that a t-test
compares means, this does not always hold. In some cases, a
WMW test indicates a significant difference between groups
when the medians are equal (2, 3, 6), in contrast to what many
researchers would expect. There are two versions of the WMW
Address for reprint requests and other correspondence: F. McElduff, Med-
ical Research Council Centre of Epidemiology for Child Health, UCL Institute
of Child Health, 30 Guilford St., London WC1N 1EH, UK (email:
f.mcelduff@ich.ucl.ac.uk).
128 1043-4046/10 Copyright © 2010 The American Physiological Society
doi:10.1152/advan.00017.2010.
test: a general form and the location-shift model, both of which
can be problematic when skewed discrete data are analyzed.
The general form of the WMW test assesses the probability
(P) that any observation drawn at random from the first group
(X) will exceed any observation from the second group (Y) i.e.,
Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017
P (X ⬎ Y) ⫽ 0.5 against P (X ⬎ Y) ⫽ 0.5 or a one-sided
alternative, P (X ⬎ Y) ⬎ 0.5 or P (X ⬎ Y) ⬍ 0.5. It assumes
that all the observations are independent and are either contin-
uous or ordinal with no possibility of ties (6). This is often an
unrealistic assumption for ordinal and discrete data. Although
the variance of the WMW test statistic can be adjusted in the
presence of ties, this adjustment always results in an increase in
the test statistic and smaller P values, and this version of the
WMW test becomes less conservative (12) for increasing
proportions of tied observations in the pooled sample.
The location-shift form of the WMW test is more restrictive
and assumes that the distribution of values in each group differ
only on location. Using this form, the test compares the
significance of any differences in medians (6). Fagerland and
Sandvik (5) showed that the WMW test in its location-shift
form is not robust in the presence of skewed data. Hence, when
the distributions are skewed and dispersions are unequal, this
form of the WMW test is inappropriate, and, consequently,
incorrect interpretations may be made. If all necessary assump-
tions are made, then it is unlikely that the t-test of means would
be invalid (3), and, therefore, the location-shift form has very
limited applicability (6).
In this article, we analyzed counts of cysts in two groups of
mouse kidneys: one treated with a steroid and a control group.
Kidney cyst counts were often zero and had a highly skewed
distribution. When comparisons were made between the two
groups, a WMW test detected a significant difference, although
the group medians were both equal to zero. We used this data
set to illustrate alternative and valid methods for quantifying
and formally testing the significance of differences for data of
this form. The standard practice of applying WMW tests to
overcome the usual assumptions of the t-test without consid-
eration of the data’s distribution is not a panacea for every data
set where a t-test is inappropriate. In this, the assumptions of
the location-shift form of the hypothesis of the WMW test are
invalid, and the cyst data set is inappropriate for the correct
application of this test. While the general version of the WMW
test is not incorrect in this instance, it provides less information
about the difference in location than other methods. We
showed how considering several regression models offers a
viable and more informative alternative means of analysis for
data with an excess of zeroes and extreme skewness. Regard-
less of the issue of validity of the WMW test, a regression
approach has some advantages over simple tests of compari-
sons: 1) model parameters can be estimated to allow for greater
interpretation of the data and 2) goodness-of-fit statistics allow
for comparisons between regression models.

WHEN t-TESTS OR WILCOXON-MANN-WHITNEY TESTS WON’T DO
This is a training article to illustrate common misconcep-
tions about the t-test and WMW test and to propose the use of
discrete regression modeling as an alternative. The data set we
used represents data resulting from a typical experiment found
in research. It provides a simple example of a situation where
the assumptions of the commonly applied t-test and WMW test
are violated. Although these tests are clearly inappropriate for
the real-life data presented here, we include them for illustra-
tive purposes.
METHODS
Data. The example data set originated from a study (3a) investi-
gating the effect of a corticosteroid on cyst formation in mice fetuses
undertaken within the Department of Nephro-Urology at the Institute
of Child Health of University College London. Embryonic mouse
kidneys were cultured, and a random sample was subjected to steroids
(n ⫽ 111), whereas the remainder acted as controls (n ⫽ 103). Six
days later, cyst counts were compared between the groups.
Summary descriptive statistics and bar charts were used to describe
the data. Normality was formally assessed using the Shapiro-Wilks
test (12), and the control and steroid-treated groups were compared
using a t-test and WMW test. Mean and median differences are
presented with 95% confidence intervals (CIs). A ␹2-test (4) assessed
the difference in the percent zero counts between the two groups.
Regression modeling. A series of different regression models was
used to describe the relationship between cyst counts and steroid
status. First, a normal linear regression model was used to compare
the means between the two groups, which for a single binary predictor
(steroid/control) is equivalent to the t-test. The assumptions of linear
regression are the same as for the t-test, i.e., that the data are
continuously numeric and normally distributed with equal variances
in each group. The purpose of presenting this regression model is to
allow comparisons of model fit diagnostics with other, less commonly
used, models. Regression diagnostics are not available when the t-test
is used.
Second, a Poisson regression model was fitted to the data. This
model is appropriate when the outcome is a count or a rate and
quantifies the mean number of events (or counts). The mean and
variance are assumed to be equal in each of the two groups (steroid/
control) (1). If the variance of the distribution is significantly greater
than the mean, there is overdispersion, and the Poisson regression
model will correctly estimate the covariate parameters but will un-
derestimate their SEs (8). An extension of the Poisson regression
model is the negative binomial regression model (1), which incorpo-
rates an extra parameter to allow for overdispersion in the data. The
improvement in model fit obtained using the negative binomial rather
than the Poisson model can be formally tested.
Allowing for zero inflation. Zero inflation occurs where there is an
excess number of zeroes in the population, often arising from a
separate process in the population. For example, some individuals are
infertile and will never have children, and others are fertile but may be
childless; the total number of individuals with no children depends on
both processes combined.
Both the Poisson and negative binomial regression models can
be extended to incorporate an additional parameter that allows for
zero inflation in the data. The resulting models are known as the
zero-inflated Poisson (ZIP) and zero-inflated negative binomial
(ZINB) (1), and the extra parameter quantifies the excess of zeroes.
These models result in two parameter estimates for each covariate
entered into the model. For example, in our data set, one parameter
will estimate the increased probability of a kidney being unable to
produce cysts. The other parameter will estimate the difference in cyst
counts between the steroid-treated and untreated groups of those
kidneys that can/do produce cysts (which can take values of 0, 1, 2,
etc.). Note that the total number of zeroes is the sum of kidneys unable
Advances in Physiology Education • VOL
Staying Current
129
to produce cysts and those kidneys that can produce cysts but haven’t
during this experiment.
Model parameters. Parameter estimates in linear regression give
the difference in means between the groups. In contrast, the Poisson
and negative binomial models yield rate ratios (RRs), which estimate
the change in the relative (rather than absolute) mean number of
events between the groups. RRs can be expressed in different ways.
For example, RR ⫽ 1.25 indicates that the mean in one group is, on
average, 1.25 times higher or, alternatively, that there is a 25%
increase in one group compared with the other. On the other hand,
e.g., RR ⫽ 0.83 indicates a 17% decrease in one group compared with
the other.
Zero-inflated regression models additionally yield the relative odds
of the proportions of zeros in each group. The odds and probability are
calculated by dividing the number of events by the number without
Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017
the event and by the total number of observations, respectively. For
example, an odds value of 2 (for every two individuals that experience
the event, one individual does not) equates to a probability of 0.33
[equal to 2/(1⫹2) ⫽ 2/3]; odds of 0.25 (for every individual for whom
the event occurs, there are four individuals for whom it does not)
equates to a probability of 0.2 (equal to 1/5). Odds ratios are calcu-
lated as the odds in one group divided by the odds in another group.
Relative risks and odds ratios are always positive, and a value of 1
means that there is no difference between the groups. All estimates
(mean differences and rate and odds ratios) are presented with 95%
CIs.
Comparison of regression models. There are a variety of ways of
deciding which regression model best describes the data. A more
complex model, with additional parameters, generally improves the
goodness of fit (i.e., how closely the model follows the data) but
requires the estimation of extra parameters. Ideally, our chosen opti-
mal model should describe the data’s main features without losing
information or being overly complex. It is important that model
selection procedures avoid including redundant parameters (overfit-
ting); we do not want to have more parameters than necessary.
The regression models described above (linear, Poisson, negative
binomial, ZIP, and ZINB) are not all direct extensions of each other.
Hence, we need to use comparison methods that do not rely on the
models being nested, i.e., that all the parameters of the smaller model
of the two being compared are included in the larger model. There-
fore, we compared models using the Bayesian information criterion
(BIC), which is a well-established measure of goodness of fit (10) that
also applies to nonnested models. The model’s likelihood is a measure
of how well it fits the data set. The BIC is based on the model’s log
likelihood larger values indicating better goodness of fit. To account
for the fact that more complex models always lead to the likelihood
remaining the same or increasing (adding parameters cannot make the
overall fit worse), BIC makes an adjustment penalizing for the number
of estimated parameters and is more conservative against overfitting,
i.e., it leads to models with smaller number of parameters than other
measures of goodness of fit. This is particularly important when the
potential number of covariates to be adjusted for is large. Models with
lower BIC values indicate a better penalized goodness of fit; however,
its absolute value is meaningless by itself, and only differences of BIC
matter.
All analyses were performed using the R environment for statistical
computing (version 2.10.1) (11) in a Windows platform. The glm
function in the base package of R was used to fit normal, Poisson, and
negative binomial models, and the pscl library (15) was used to fit
zero-inflated models, both of which use maximum likelihood estima-
tion methods.
RESULTS
Exploratory data analysis. Bar charts of the cyst frequencies
(Fig. 1) showed that most kidneys had no cysts (74.3% over-
all), although there are a few kidneys in each group with large
34 • SEPTEMBER 2010
Staying Current
130 WHEN t-TESTS OR WILCOXON-MANN-WHITNEY TESTS WON’T DO

Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017

Fig. 1. Bar graphs of the frequencies of cysts in the kidneys for the two groups [steroid-treated group (top) and control group (bottom)].

cyst counts; hence, the distributions were highly skewed to the met, but we present the results purely for comparative pur-
right and zero inflated. The overall mean number of cysts was poses. A t-test yielded a highly significant difference between
0.87 (SD: 2.28), indicating overdispersion since the variance is the means of 1.40 [95% CI: (0.82, 1.99), P ⬍ 0.01]. The
almost four times larger than the mean. The values showed WMW test under the general null hypothesis P (X ⬎ Y) ⫽ 0.5
larger number of kidneys with low counts in the control group. produced P ⬍ 0.01, similarly indicating a significant differ-
The steroid-treated group had 1 kidney with 19 cysts, which ence. A median difference of 0 and a value of 0 for both the
was much higher than the maximum number of cysts found in upper and lower 95% CI for the median difference were also
the group of control kidneys (maximum: 3). Not surprisingly, calculated, and this contrasts with the results from the WMW
the Shapiro-Wilks test for normality gave P ⬍ 0.01, indicating test, where a significant difference was detected. A comparison
that the counts did not follow a normal distribution. Similarly, of the number of cysts with zero counts showed that there were
Bartlett’s F-test for homogeneity of variances yielded P ⬍ significantly more (␹2-statistic ⫽ 28.23, degree of freedom: 1,
0.01, which implied a significant difference in the variances
P ⬍ 0.01) cysts with zero counts in the control group than in
between the two groups. Note that this test assumes normality.
The Ansari-Bradley test for equal scale parameters (9) also
gave P ⬍ 0.01, indicating a significant difference in dispersion
between the groups. Table 1. Summary statistics for counts of cysts for the
Table 1 shows summary statistics for the steroid-treated and steroid-treated and control groups
control groups. The means were 1.55 and 0.15 for the steroid-
Steroid-Treated
treated and control groups, with SDs of 2.98 and 0.51, respec- Group Control Group
tively. The medians for the two groups were both 0, with the
Mean 1.55 0.15
steroid-treated group having an interquartile range of 2 and the SD 2.98 0.51
control group an interquartile range of 0. The mean and SD in Median 0 0
the steroid-treated group were much higher than those in the Interquartile range (quartile 1, quartile 3) 2 (0, 2) 0 (0, 0)
control group, although the medians were equal. Minimum 0 0
Tests of comparisons for the two groups. Clearly, the as- Maximum 19 3
Percent zeroes 58.56 91.26
sumptions for the application of a two-sample t-test were not

Advances in Physiology Education • VOL 34 • SEPTEMBER 2010

 Staying Current
WHEN t-TESTS OR WILCOXON-MANN-WHITNEY TESTS WON’T DO 131

the steroid-treated group [percent difference: 32.7%, 95% CI:
(21.1%, 44.3%)].
Regression models. Table 2 shows the parameter estimates,
CIs, P values, and BIC for each of the regression models
described above. As expected, linear regression gave the same
information as the t-test but additionally yielded a BIC value
that could be compared with those obtained from other regres-
sion models. The Poisson model showed that the average
number of cysts in kidneys in the steroid-treated group were
almost 11 times [RR: 10.64, 95% CI: (6.28, 18.03), P ⬍ 0.01]
higher than kidneys not treated with steroids.
The negative binomial model gave the same rate ratio as the
Poisson model but with larger SEs, resulting in a wider CI than
the Poisson model. As expected, the overdispersion parameter
the BIC values clearly indicated that the negative binomial
model resulted in an improvement in fit to the data (450.68
compared with 667.07 for the Poisson model).
For the ZIP model, the zero inflation parameter was signif-
icant (model 4: P ⫽ 0.04), and the odds of a kidney having zero
cysts was 0.20 [95% CI (0.08, 0.51), P ⬍ 0.01] times lower in
the steroid-treated group (model 5). Given that there are cysts,
the rate was over three times [RR: 3.23, 95% CI: (1.51, 6.94),
P ⬍ 0.01] higher on average for kidneys in the steroid-treated
group compared with the control group. The additional param-
eter to model the zero-inflated component in the Poisson model
reduced the BIC values greatly (507.94) and even further when
the variation between zero inflation of the steroid-treated and
control groups was included (505.98), with both providing a

Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017

in the negative binomial model was significant (P ⬍ 0.01), and much better fit than the simple Poisson model (BIC: 667.07).

Table 2. Estimates, 95% confidence, P values, and BIC values for the various models for the control and steroid-treated
groups

Estimate 95% Confidence Intervals for the Estimate P Value BIC Value

Model 1: linear (raw estimate)

Control group (constant) 0.15 ⫺0.27, 0.57 0.50
Steroid-treated group 1.40 0.82, 1.99 ⬍0.01 953.41
Model 2: Poisson (rate)
Control group (constant) 0.15 0.09, 024 ⬍0.01
Steroid-treated group 10.64 6.28, 18.03 ⬍0.01 667.07
Model 3: negative binomial (rate)
Control group (constant) 0.15 0.08, 0.27 ⬍0.01
Steroid-treated group 10.64 5.09, 22.24 ⬍0.01
Dispersion 1.31 1.17, 1.47 ⬍0.01 450.68
Model 4: zero-inflated Poisson (3 parameters)
Count model (rate)
Control group (constant) 0.40 0.21, 0.74 ⬍0.01
Steroid-treated group 9.22 4.85, 17.52 ⬍0.01
Zero inflation (odds)
Control group (constant) 1.52 1.02, 2.25 0.04 507.94
Model 5: zero-inflated Poisson (4 parameters)
Count model (rate)
Control group (constant) 1.13 0.53, 2.38 0.76
Steroid-treated group 3.23 1.51, 6.94 ⬍0.01
Zero inflation (odds)
Control group (constant) 6.73 2.87, 15.81 ⬍0.01
Steroid-treated group 0.20 0.08, 0.51 ⬍0.01 505.98
Model 6: zero-inflated negative binomial (4 parameters)
Count model (rate)
Control group (constant) 0.19 0.06, 0.62 ⬍0.01
Steroid-treated group 10.47 5.05, 21.68 ⬍0.01
Dispersion 5.49 0.96, 31.30 0.33
Zero inflation (odds)
Control group (constant) 0.31 0.01, 17.44 0.57 455.20
Model 7: zero-inflated negative binomial (5 parameters)
Count model (rate)
Control group (constant) 0.52 0.13, 2.02 0.35
Steroid-treated group 4.38 1.33, 14.40 0.02
Dispersion 1.67 0.41, 6.89 0.48
Zero inflation (odds)
Control group (constant) 2.57 0.48, 13.72 0.27
Steroid-treated group 0.18 0.03, 1.02 0.05 457.80
Parameter estimates for the control (constant) and steroid-treated groups are given as either raw estimates, rate, or odds ratios for the regression models;
dispersion parameters are also given for the negative binomial models. Upper and lower confidence intervals for all the parameter estimates are shown as well
as P values and Bayesian information criterion (BIC) values.

Advances in Physiology Education • VOL 34 • SEPTEMBER 2010

Staying Current
132 WHEN t-TESTS OR WILCOXON-MANN-WHITNEY TESTS WON’T DO

The basic negative binomial model (model 3) gave a better
fit than any of the Poisson models (BIC: 450.68), with signif-
icant overdispersion, as expected. The constant zero inflation
term (model 6) was nonsignificant, and the model that allowed
zero inflation to vary according to group showed the difference
to be almost significant [odds ratio: 0.18, 95% CI (0.03, 1.02),
P ⫽ 0.05]. However, increasing the complexity of the negative
binomial model resulted in a higher BIC value (455.20 and
457.80 for ZINB with and without zero inflation varying by
group) as it was penalized by the incorporation of the addi-
tional parameters, indicating models that do not provide a more
parsimonious fit to the data.
Comparison of regression models. The observed and pre-
dicted numbers of cysts for models 1–3, 5, and 7 are shown in
tion can result in a misleading difference between means being
detected. Value inflation (namely, zero inflation) can lead to a
high proportion of tied values in the location-shift version of
the WMW test, resulting in incorrect conclusions being drawn
from P values. In these cases, tests may lead to conflicting
results and may not agree with observations made from ex-
ploratory data analyses where the medians do not indicate a
difference between groups. While an advantage of t-tests and
WMW tests are their simplicity and ease of application, they
are greatly disadvantaged when applied to count data due to
these erroneous assumptions (14). It can be shown using
simulation that the WMW test is particularly vulnerable to zero
inflation, particularly when the probability of belonging to the
zero class is ⬎0.75 and less so to overdispersion. More details

Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017

Table 3. In the distributions for the control group, the predicted
distributions for negative binomial, ZIP, and ZINB were all
similar to the observed data. However, for the steroid-treated
group, there were differences between the distributions. The
normal distribution provided the worst fit to the data, and this
was reflected in the BIC value in Table 2. The Poisson
distribution also yielded a poor fit, as indicated by both the
fitted values and BIC value. The additional zero inflation
parameter provided a better fit to the data in the ZIP model but
did not fit as well as the negative binomial or ZINB models, in
terms of both the BIC value and fitted values shown in Table
3 (steroid-treated group). The BIC values shown in Table 2
indicate that the negative binomial model was the most parsi-
monious, and therefore preferred, model.
DISCUSSION
The most commonly used methods for comparing location in
two groups are the t-test or WMW test. The advantage of
permutation tests (7) is that they provide a distribution-free
alternative to test differences of group means; however, they
are disadvantaged as they are difficult to generalize to a
regression setup such as discrete regression modeling. The
normality assumption of the t-test is inappropriate for highly
skewed discrete data, where overdispersion and/or zero infla-
on the simulations performed are available upon request from
the corresponding author.
Here, we present a number of regression models for discrete
data as an alternative to the t-tests and WMW tests. Poisson
regression models can be used to model the mean number of
events, and the negative binomial model includes an extra
parameter to allow for overdispersion. A further extension of
these models is to model zero inflation in the data, resulting in
the ZIP and ZINB models, each with corresponding additional
parameters.
While regression analyses using distributions such as the
Poisson, negative binomial, ZIP, and ZINB models might be
more complex, they model discrete counts, allowing for correct
assumptions, and also provide more valid and useful estimates
of differences between groups. Software for fitting the models
is now widespread, for example, in R and STATA. BIC values
are provided alongside parameter estimates and SEs in R,
whereas in other packages such as STATA, the log likelihood
(ᐉ) is given instead, from which the BIC value can easily be
calculated as follows: ⫺2ᐉ⫹k ⫻ log(n), where k is the number
of parameters in the model and n is the sample size.
A further advantage is that the regression modeling approach
is very flexible. Models can be extended to quantify the
associations with additional covariates, which may be contin-

Table 3. Observed and fitted numbers of cysts for the control and steroid-treated groups

Frequency of Cysts

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19

Control group
Observed numbers 94 4 4 1
Fitted number
Model 1: normal 77 20
Model 2: poisson 89 13 1
Model 3: negative binomial 94 4 2 1 1
Model 5: zero-inflated poisson 94 5 3 1
Model 7: zero-inflated negative binomial 94 6 2 1
Steroid-treated group
Observed numbers 65 14 10 6 4 2 2 2 1 1 1 2 1
Fitted numbers
Model 1: normal 13 15 15 13 11 8 5 3 1 1
Model 2: poisson 24 37 28 15 6 2
Model 3: negative binomial 64 17 9 6 4 3 2 2 1 1 1 1
Model 5: zero-inflated poisson 65 5 8 10 9 7 4 2 1
Model 7: zero-inflated negative binomial 65 14 9 6 4 3 2 2 1 1 1 1
n ⫽ 103 mice in the control group and 111 mice in the steroid-treated group.

Advances in Physiology Education • VOL 34 • SEPTEMBER 2010


WHEN t-TESTS OR WILCOXON-MANN-WHITNEY TESTS WON’T DO
uous (as opposed to the single binary covariate, steroid/control,
used in our example data set). In this example, the number of
groups could be extended to include more than one type of
steroid or a continuous variable, e.g., the growth of the kidneys
over the number of days cultured. Interactions between covari-
ates can also be investigated in the models, e.g., we could look
at the interaction between the growth of kidneys and steroid-
treated groups. A further use of discrete regression is in
modeling rates. For counts of events measured in a certain time
period, where the time period may vary across observation,
dividing the counts of events by the length of a time period
gives a rate. In discrete regression modeling, an offset can be
included to adjust for the different time periods that has no
parameter estimates or P values.
In the example presented in this article, both the t-test and
WMW test indicated that a significant difference exists in the
number of cysts between the control and steroid-treated
groups. While exploratory data analysis revealed that there was
a difference between the means of the two groups, there was no
difference in the medians. Of the seven regression models
fitted, the BIC values indicated that the negative binomial
model provided the best fit to the data. This model showed that
the mean number of cysts was significantly higher in the
steroid-treated group than in the control group and that there
was significant overdispersion present in the data.
The cyst counts on a given day provide a snapshot of each
kidney’s ability to produce cysts. The underlying capacities of
the cells may or may not be identical. The Poisson model
assumes that they are the same, whereas the negative binomial
model allows for variation. Therefore, the data suggest that the
more complex model was appropriate. However, the prefer-
ence of the negative binomial model over the ZINB model
showed that there did not appear to be a subset of kidneys that
cannot produce cysts.
This article demonstrates how commonly used tests for the
comparison of two groups, namely, the t-test and WMW test,
can be problematic when dealing with discrete counts. Erro-
neous assumptions may need to be made for the application of
the t-test and linear regression to skewed discrete data, hence
invalidating results. Less obviously, nonparametric tests such
as the WMW test, where value inflation leads to an increased
number of ties, may also lead to incorrect conclusions. We
present the use of regression modeling as an alternative method
for comparing groups with skewed observations, using the
Poisson distribution, which models the mean number of events
and can be extended to allow for overdispersion and value
inflation in the negative binomial, ZIP, and ZINB models. The
approach outlined in this article provides a flexible tool for
Advances in Physiology Education • VOL
Staying Current
133
analysing discrete data that avoids making erroneous assump-
tions and facilitates comparisons between models.
ACKNOWLEDGMENTS
The authors are grateful to Adrian Woolf and David Long for the helpful
comments on a previous version of this article.
GRANTS
The Institute of Child Health of University College London receives a
portion of funding from the Department of Health’s National Institute of
Health Research Biomedical Research Centres funding scheme. The Centre for
Paediatric Epidemiology and Biostatistics also benefits from funding support
from the Medical Research Council (MRC) in its capacity as the MRC Centre
of Epidemiology for Child Health (G0400546). F. McElduff acknowledges the
support of an MRC Capacity Building Studentship. S.-K. Chan is funded by
Kidney Research UK.
Downloaded from http://advan.physiology.org/ by 10.220.32.247 on September 4, 2017
DISCLOSURES
No conflicts of interest, financial or otherwise, are declared by the author(s).
REFERENCES
1. Afifi AA, Kotlerman JB, Ettner SL, Cowan M. Methods for improving
regression analysis for skewed continuous or count responses. Annu Rev
Public Health 28: 95–111, 2007.
2. Bergmann R, Ludbrook J, Spooren WP. Different outcomes of the
Wilcoxon-Mann-Whitney test from different statistical packages. Am
Statistician 54: 72–77, 2000.
3. Bland M. An Introduction to Medical Statistics (3rd ed.). New York:
Oxford Univ. Press, 2000.
3a. Chan SK, Riley PR, Price KL, McElduff F, Winyard PJ, Welham SJ,
Woolf AS, Long DA. Corticosteroid-induced dysmorphogenesis is asso-
ciated with deregulated expression of known cystogenic molecules, as well
as indian hedgehog. Am J physiol Renal Physiol 298: 346 –356, 2010.
4. Chernoff H, Lehmann EL. The use of maximum likelihood estimates in
chi squared tests for goodness-of-fit. Annu Math Stat 25: 579 –586, 1954.
5. Fagerland MW, Sandvik L. The Wilcoxon-Mann-Whitney test under
scrutiny. Stat Med 28: 1487–1497, 2009.
6. Fay MP, Proschan MA. Wilcoxon-Mann-Whitney or t-test? On assump-
tions for hypothesis tests and multiple interpretations of decision rules.
Stat Surveys 4: 1–39, 2010.
7. Good PI. Permutation, Parametric, and Bootstrap Tests of Hypotheses
(3rd ed.). New York: Springer, 2005.
8. Hilbe JM. Negative Binomial Regression. Cambridge: Cambridge Univ.
Press, 2007.
9. Hollander M, Wolfe DA. Nonparametric Statistical Methods (2nd ed.).
New York: Wiley-Interscience, 1999.
10. Kuha J. AIC and BIC: comparisons of assumptions and performance.
Sociol Methods Res 33: 188 –228, 2004.
11. R Development Core Team. R: a Language and Environment for Statis-
tical Computing. Vienna: R Foundation for Statistical Computing, 2007.
12. Shapiro SS, Francia RS. An approximate analysis of variance test for
normality. J Am Stat Assoc 67: 215–216, 1972.
13. Welch BL. The generalization of “Student’s” problem when several
different population variances are involved. Biometrika 34: 28 –35, 1947.
14. Whitley E, Ball J. Statistics review 6: nonparametric methods. Crit Care
6: 509 –513, 2002.
15. Zeileis A, Kleiber C, Jackman S. Regression models for count data in R.
J Stat Software 27: 2008.
34 • SEPTEMBER 2010