APOE4 STATUS AND COGNITIVE FUNCTION
IN MIDDLE-AGED AND ELDERLY PEOPLE
Pavel N.A.1, Paun M.R.2, Matei P.V.1,2, Dutu I.1, Tudose C.1,2
*Corresponding Author: Valentin P. Matei, Associate Proffesor, MD, PhD. 2nd Ward of “Prof. Dr. Alexandru
Obregia” Hospital, Berceni Street no 10, Bucharest, Romania. E-mail: valipmatei@yahoo.com
page: 6
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MATERIAL AND METHODS
We conducted a cross-sectional study on patients
from a primary care clinic who were recruited while performing
their clinical routine checkup at their General
Practitioners (GPs).
The study was conducted in accordance with the Declaration
of Helsinki [11] and was approved by the Local
Ethics Committee (no. 11/06/03.2020). Patients included in
the study had to understand and sign the Informed Consent
Form and fulfill all the described criteria: (a) age between
50-80 years, (b) Mini-Mental State Examination (MMSE)
score over 24, (c) Functional Assessment Questionnaire
(FAQ) below 9, (d) Hamilton Depression total score below
12, (e) Hamilton Anxiety total score below 17 and (f) no
substance use in the previous 6 months other than caffeine
or tobacco. Exclusion criteria were: (a) diagnosis of
major or mild neurocognitive disorder according to DSM
5,[12] (b) presence of cerebro-vascular disease translated
as Hachinski score over 4, (c) current diagnosis of neurodevelopmental
disorder, major depressive disorder, anxiety
disorder according to DSM 5, (d) severe somatic disorders
such as epilepsy, organ failure or other disease that could
impair collection of data from the patient such as severe
hearing/seeing impairment, motor deficit).
We collected social, demographic, and clinical information
from the participants, as well as their full psychiatric
history.
For genetic assessment of APOE status, we collected
3 milliliters of venous blood using EDTA coated tubes,
which were then kept refrigerated at 4 degrees Celsius
until DNA extraction, which was conducted using DNA
extraction kits (Wizzard© Genomic DNA Purification Kit,
Promega) form a volume of 300 μL white blood cells.
The genotyping was done using the Taqman SNP Genotyping
Assay method from Thermo Fished Scientific, according
to the manufacturer’s instructions. The assay was
conducted by PCR amplification of two oligonucleotide
primers, rs429358 and rs7412. Patients with at least one
ApoE4 allele were considered ApoE4 positive while the
rest were coded as ApoE4 negative and were considered
as control group.
Cognitive examination was conducted using the
MMSE [13], Rey Auditory-Verbal Learning Test (RAVLT)
[14], Rey-Osterrieth Complex Figure Test [15], Verbal
Fluency [16] and Trail Making Test (TMT) [14].
The Mini-Mental State Examination [13] is a short
questionnaire that evaluates attention, memory, calculation,
visuo-spatial ability and executive functioning. The
maximum score is 30 and scores under 24 are considered
suggestive for cognitive impairment.
The RAVLT [14] consists of a list of 15 words, and is
designed to evaluate the ability to assess verbal memory.
The evaluator reads a list of 15 words after which the
participant is asked to recall as many as he can; this is
done for a total of 5 times (Trials 1-5). Afterwards, the
same exercise is done with a different list of 15 words for
a single trial and the participant is then asked to recall the
words from the first list (Trial 6). Trial 7 (Delay) is done
after a 5-minute pause, without the list being reread to the
patient. The last examination of RAVLT (Recognition)
consists of a text which includes the first list of words,
and the patient is asked to recognize them.
The Rey-Osterrieth Complex Figure Test [15] consists
of 2 trials which examine memory, visuo-spatial ability
and executive function. For the first trial (copying), the
patient is asked to copy a complex figure, after which the
figure is put away. Following a 3-minute break the patient
is asked to draw the figure from memory. The maximum
score is 36 with higher scores representing better cognitive
functioning.
The Verbal Fluency [16] test examines verbal ability
and executive function. We used the letter fluency variant,
consisting of 3 successive 1-minute trials, in which
the patient must produce as many words as possible that
begin with a given letter. Each trial used a different letter,
and all patients received the same letters in the same order.
In our analysis we summed all the correct words from all
trials into a single score.
The Trail Making Test [14] examines a variety of
cognitive functions such as attention, visual and spatial
ability, sequencing and shifting, psychomotor speed, abstraction,
flexibility and executive function [14]. It consists
of two timed trials (A and B). Patient is considered to be
deficient in the presented cognitive domains if he finishes
Trial A after 78 second and Trial B after 273 seconds.[17]
The ApoE4 and control groups were matched for age,
sex and education and there were no patients with current
depressive or anxiety disorder. Descriptive statistics
were used to present the sample characteristics. We used Chi-square test for the categorical data such as (ApoE4
status, demographic characteristics) and Student t-test (for
normal distributed variables) and Mann-Whitney Test (for
non-parametric variables) for continuous variables (such
as age, cognitive evaluation test scores). Results are presented
with mean (standard deviation) for normally distributed
data and median (Inter Quartile Range - IQR) for
non-parametric distribution. Statistical significance was
considered at alpha below .05.
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