Introduction to

Statistics
Colm ODushlaine

Neuropsychiatric Genetics, TCD

codushlaine@gmail.com

1
 Overview
Descriptive Statistics & Graphical Presentation of
Data
Statistical Inference
Hypothesis Tests & Confidence Intervals
T-tests (Paired/Two-sample)
Regression (SLR & Multiple Regression)
ANOVA/ANCOVA
Intended as an interview. Will provide slides after
lectures
Whats in the lectures?...
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Lecture 1 Lecture 2 Lecture 3
Lecture 4
Descriptive Statistics and Graphical
Presentation
1. Terminology of Data
2. Frequency Distributions/Histograms
3. Measures of data location
4. Measures of data spread
5. Box-plots
6. Scatter-plots
7. Clustering (Multivariate Data)

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Lecture 1 Lecture 2 Lecture 3
Lecture 4 Statistical Inference

1. Distributions & Densities

2. Normal Distribution
3. Sampling Distribution & Central Limit Theorem
4. Hypothesis Tests
5. P-values
6. Confidence Intervals
7. Two-Sample Inferences
8. Paired Data

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Lecture 1 Lecture 2 Lecture 3
Lecture 4 Sample Inferences

1. Two-Sample Inferences
Paired t-test
Two-sample t-test
2. Inferences for more than two samples
One-way ANOVA
Two-way ANOVA
Interactions in Two-way ANOVA
3. DataDesk demo

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Lecture 1 Lecture 2 Lecture 3
Lecture 4

1. Regression
2. Correlation
3. Multiple Regression
4. ANCOVA
5. Normality Checks
6. Non-parametrics
7. Sample Size Calculations
8. Useful tools and websites
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FIRST, A REALLY USEFUL SITE
Explanations of outputs
Videos with commentary
Help with deciding what test
to use with what data

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1. Terminology
Populations & Samples
Population: the complete set of individuals,
objects or scores of interest.
Often too large to sample in its entirety
It may be real or hypothetical (e.g. the results from an
experiment repeated ad infinitum)

Sample: A subset of the population.

A sample may be classified as random (each member
has equal chance of being selected from a population)
or convenience (whats available).
Random selection attempts to ensure the sample is
representative of the population.
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Variables
Variables are the quantities measured in a
sample.They may be classified as:
Quantitative i.e. numerical
Continuous (e.g. pH of a sample, patient
cholesterol levels)
Discrete (e.g. number of bacteria colonies in a
culture)
Categorical
Nominal (e.g. gender, blood group)
Ordinal (ranked e.g. mild, moderate or severe
illness). Often ordinal variables are re-coded to be
quantitative.

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Variables

Variables can be further classified as:

Dependent/Response. Variable of primary interest
(e.g. blood pressure in an antihypertensive drug trial).
Not controlled by the experimenter.
Independent/Predictor
called a Factor when controlled by experimenter. It

is often nominal (e.g. treatment)

Covariate when not controlled.

If the value of a variable cannot be predicted in

advance then the variable is referred to as a
random variable
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Parameters & Statistics
Parameters: Quantities that describe a
population characteristic. They are usually
unknown and we wish to make statistical
inferences about parameters. Different to
perimeters.

Descriptive Statistics: Quantities and

techniques used to describe a sample
characteristic or illustrate the sample data
e.g. mean, standard deviation, box-plot

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2. Frequency Distributions

An (Empirical) Frequency Distribution or

Histogram for a continuous variable presents the
counts of observations grouped within pre-
specified classes or groups

A Relative Frequency Distribution presents the

corresponding proportions of observations within
the classes

A Barchart presents the frequencies for a

categorical variable
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Example Serum CK

Blood samples taken from 36 male

volunteers as part of a study to determine the
natural variation in CK concentration.

The serum CK concentrations were

measured in (U/I) are as follows:

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Serum CK Data for 36 male
volunteers

121 82 100 151 68 58

95 145 64 201 101 163
84 57 139 60 78 94
119 104 110 113 118 203
62 83 67 93 92 110
25 123 70 48 95 42
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 Relative Frequency Table
Serum CK Frequency Relative Cumulative Rel.
(U/I) Frequency Frequency
20-39 1 0.028 0.028
40-59 4 0.111 0.139
60-79 7 0.194 0.333
80-99 8 0.222 0.555
100-119 8 0.222 0.777
120-139 3 0.083 0.860
140-159 2 0.056 0.916
160-179 1 0.028 0.944
180-199 0 0.000 0.944
200-219 2 0.056 1.000
Total 36 1.000
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 Frequency Distribution
Distributions
CK-concentration-(U/l)
Quantiles
8 100.0% maximu
99.5%
97.5%
90.0%
6 75.0% quart
50.0% media
25.0% quart

Frequency
10.0%
4 2.5%
0.5%
0.0% minimu

20 40 60 80 100 120 140 160 180 200 220

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 Relative Frequency Distribution
Distributions
CK-concentration-(U/l)
Quantiles
Mode
Shaded area is 100.0% maxim
percentage of 99.5%
males with CK 0.20 97.5%
values between 90.0%
60 and 100 U/l, 75.0% quar

Relative Frequency
i.e. 42%. 0.15 50.0% med
Right tail 25.0% quar
10.0%
(skewed) 2.5%
0.10 0.5%
0.0% minim
Left tail
0.05

20 40 60 80 100 120 140 160 180 200 220

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3. Measures of Central
Tendency (Location)
Measures of location indicate where on the number
line the data are to be found. Common measures of
location are:

(i) the Arithmetic Mean,

(ii) the Median, and
(iii) the Mode

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 The Mean

Let x1,x2,x3,,xn be the realised values of a

random variable X, from a sample of size n.
The sample arithmetic mean is defined as:

n
x 1
n xi
i 1

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Example

Example 2: The systolic blood pressure of

seven middle aged men were as follows:
151, 124, 132, 170, 146, 124 and 113.

x
151 124 132 170 146 124 113
The mean is
7
137.14

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The Median and Mode

If the sample data are arranged in increasing

order, the median is
(i) the middle value if n is an odd number, or
(ii) midway between the two middle values if n is
an even number
The mode is the most commonly occurring
value.

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Example 1 n is odd

The reordered systolic blood pressure data seen

earlier are:

113, 124, 124, 132, 146, 151, and 170.

The Median is the middle value of the ordered data,

i.e. 132.

Two individuals have systolic blood pressure = 124

mm Hg, so the Mode is 124.

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Example 2 n is even

Six men with high cholesterol participated in a study to investigate

the effects of diet on cholesterol level. At the beginning of the study,
their cholesterol levels (mg/dL) were as follows:
366, 327, 274, 292, 274 and 230.
Rearrange the data in numerical order as follows:

230, 274, 274, 292, 327 and 366.

The Median is half way between the middle two readings, i.e.
(274+292) 2 = 283.

Two men have the same cholesterol level- the Mode is 274.

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Mean versus Median

Large sample values tend to inflate the mean. This will happen if
the histogram of the data is right-skewed.

The median is not influenced by large sample values and is a better

measure of centrality if the distribution is skewed.

Note if mean=median=mode then the data are said to be

symmetrical

e.g. In the CK measurement study, the sample mean = 98.28. The

median = 94.5, i.e. mean is larger than median indicating that mean
is inflated by two large data values 201 and 203.

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4. Measures of Dispersion

Measures of dispersion characterise how

spread out the distribution is, i.e., how variable
the data are.
Commonly used measures of dispersion
include:
1. Range
2. Variance & Standard deviation
3. Coefficient of Variation (or relative standard
deviation)
4. Inter-quartile range

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Range
the sample Range is the difference
between the largest and smallest
observations in the sample
easy to calculate;
Blood pressure example: min=113 and
max=170, so the range=57 mmHg
useful for best or worst case scenarios
sensitive to extreme values

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Sample Variance

The sample variance, s2, is the arithmetic

mean of the squared deviations from the
sample mean:
n
xi x
2

s i 1
2
n 1

>

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Standard Deviation

The sample standard deviation, s, is the

square-root of the variance

n
xi x
2

i 1
s
n 1

s has the advantage of being in the same units

as the original variable x
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 Example
Data Deviation Deviation2
151 13.86 192.02
124 -13.14 172.73
132 -5.14 26.45
170 32.86 1079.59
146 8.86 78.45
124 -13.14 172.73
113 -24.14 582.88
Sum = 960.0 Sum = 0.00 Sum = 2304.86
x 137.14 29
Example (contd.)

x x
2
i 2304.86
i 1

Therefore, 2304.86
s
7 1
19.6
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 Coefficient of Variation
The coefficient of variation (CV) or relative
standard deviation (RSD) is the sample standard
deviation expressed as a percentage of the mean,
i.e.
s
CV 100%
x
The CV is not affected by multiplicative changes in
scale
Consequently, a useful way of comparing the
dispersion of variables measured on different scales
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Example

The CV of the blood pressure data is:

19.6
CV 100 %
137.1
14.3%
i.e., the standard deviation is 14.3% as large as
the mean.

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 Inter-quartile range
The Median divides a distribution into two halves.

The first and third quartiles (denoted Q1 and Q3) are

defined as follows:
25% of the data lie below Q1 (and 75% is above Q1),
25% of the data lie above Q3 (and 75% is below Q3)

The inter-quartile range (IQR) is the difference

between the first and third quartiles, i.e.
IQR = Q3- Q1

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Example

The ordered blood pressure data is:

113 124 124 132 146 151 170

Q1 Q3

Inter Quartile Range (IQR) is 151-124 = 27

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60% of slides complete!

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5. Box-plots

A box-plot is a visual description of the

distribution based on
Minimum
Q1
Median
Q3
Maximum
Useful for comparing large sets of data

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Example 1

The pulse rates of 12 individuals arranged in

increasing order are:
62, 64, 68, 70, 70, 74, 74, 76, 76, 78, 78, 80

Q1=(68+70)2 = 69, Q3=(76+78)2 = 77

IQR = (77 69) = 8

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Example 1: Box-plot

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Example 2: Box-plots of intensities
from 11 gene expression arrays

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12
10
8

AG_04659_AS.cel AG_11745_AS.cel KB_5828_AS.cel KB_8840_AS.cel

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 Outliers

An outlier is an observation which does not

appear to belong with the other data
Outliers can arise because of a measurement
or recording error or because of equipment
failure during an experiment, etc.
An outlier might be indicative of a sub-
population, e.g. an abnormally low or high
value in a medical test could indicate presence
of an illness in the patient.
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Outlier Boxplot

Re-define the upper and lower limits of the

boxplots (the whisker lines) as:
Lower limit = Q1-1.5IQR, and
Upper limit = Q3+1.5IQR
Note that the lines may not go as far as these
limits
If a data point is < lower limit or > upper limit,
the data point is considered to be an outlier.
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Example CK data

outliers

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6. Scatter-plot

Displays the relationship between two

continuous variables

Useful in the early stage of analysis when

exploring data and determining is a linear
regression analysis is appropriate

May show outliers in your data

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Example 1: Age versus Systolic
Blood Pressure in a Clinical
Trial

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Example 2: Up-regulation/Down-
regulation of gene expression across an
array (Control Cy5 versus Disease Cy3)

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Example of a Scatter-plot matrix
(multiple pair-wise plots)

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Other graphical representations

Dot-Plots, Stem-and-leaf plots

Not visually appealing
Pie-chart
Visually appealing, but hard to compare two datasets. Best
for 3 to 7 categories. A total must be specified.
Violin-plots
=boxplot+smooth density
Nice visual of data shape

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Multivariate Data

Clustering is useful for visualising multivariate

data and uncovering patterns, often reducing its
complexity

Clustering is especially useful for high-

dimensional data (p>>n): hundreds or perhaps
thousands of variables

An obvious areas of application are gel

electrophoresis and microarray experiments
where the variables are protein abundances or
gene expression ratios
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 7. Clustering
Aim: Find groups of samples or variables sharing
similiarity

Clustering requires a definition of distance between

objects, quantifying a notion of (dis)similarity
Points are grouped on the basis on minimum distance
apart (distance measures)

Once a pair are grouped, they are combined into a

single point (using a linkage method) e.g. take their
average. The process is then repeated.
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Clustering
Clustering can be applied to rows or columns of a data set
(matrix) i.e. to the samples or variables

A tree can be constructed with branch length proportional to

distances between linked clusters, called a Dendrogram

Clustering is an example of unsupervised learning: No use is

made of sample annotations i.e. treatment groups, diagnosis
groups

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UPGMA
Unweighted Pair-Group Method Average
Most commonly used clustering method
Procedure:
1. Each observation forms its own cluster
2. The two with minimum distance are grouped into a single
cluster representing a new observation- take their average
3. Repeat 2. until all data points form a single cluster

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Contrived Example
5 genes of interest on 3 replicates arrays/gels
Array1 Array2 Array3

p53 9 3 7
mdm2 10 2 9
bcl2 1 9 4
d xy ( x1 y1 ) ( x2 y2 ) ( x3 y3 )
2 2 2

cyclinE 6 5 5
caspase 8 1 10 3

Calculate distance between each pair of genes

e.g. d ( p53, mdm2) (9 10) 2 (3 2) 2 (7 9) 2 2.5

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 Example

Construct a distance matrix of all pair-wise distances

p53 mdm2 bcl2 cyclinE caspase 8

p53 0 2.5 10.44 4.12 11.75

mdm2 - 0 12.5 6.4 13.93
bcl2 - - 0 6.48 1.41
cyclinE - - - 0 7.35
caspase 8 - - - - 0

Cluster the 2 genes with smallest distance

Take their average & re-calculate distances to other genes

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 {caspase-8 &
p53 mdm2 cyclin E
bcl-2}
p53 0 2.5 4.12 10.9
mdm2 0 6.4 9.1

cyclin E 0 6.9
{caspase-8 &
0
bcl-2}

{p53 & {caspase-8 &

cyclin E
mdm2} bcl-2}
{p53 & mdm2} 0 3.7 9.2
cyclin E 0 6.9

{caspase-8 & bcl-2} 0

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Example (contd)

..and the final cluster:

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Example of a gene expression dendrogram

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 Variety of approaches to clustering
Clustering techniques
agglomerative -start with every element in its own cluster, and
iteratively join clusters together
divisive - start with one cluster and iteratively divide it into
smaller clusters
Distance Metrics
Euclidean (as-the-crow-flies)
Manhattan
Minkowski (a whole class of metrics)
Correlation (similarity in profiles: called similarity metrics)
Linkage Rules
average: Use the mean distance between cluster members
single: Use the minimum distance (gives loose clusters)
complete: Use the maximum distance (gives tight clusters)
median: Use the median distance
centroid: Use the distance between the average member or
each cluster 57
Clustering Summary

The clusters & tree topology often depend highly on

the distance measure and linkage method used

Recommended to use two distance metrics, such

as Euclidean and a correlation metric

A clustering algorithm will always yield clusters,

whether the data are organised in clusters or not!

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