Title: Report on “Advanced Laboratory Skills in Field Biology” Course
Author: Negin Eslamibidgoli
Affiliation: Wildlife Research Center, Kyoto University
Date of Submission: 2023/11/10

Table of Contents
Abstract.................................................................................................................................1
Introduction.......................................................................................................................... 2
Methods................................................................................................................................ 3● Androgen Receptor Genotyping................................................................................... 3● ALDH2 Genotyping....................................................................................................... 6
Results.................................................................................................................................. 8
Discussion.......................................................................................................................... 12
References..........................................................................................................................13

Abstract
The androgen receptor (AR) gene, plays a critical role in male phenotypes andbehavioral patterns, particularly aggression. This study investigates the polymorphic
variants of the AR gene in six participants, correlating genetic data with a chimpanzee
personality test and a human personality test (TCI). The experiment also explores
alcohol tolerance of the participants by genotyping the SNP types of ALDH2 gene andconducting a patch test. During this laboratory course, we collected buccal cell samples
from 6 participants and genetically analyzed them using various methods including
DNA extraction, PCR, Agarose gel electrophoresis, and sequencing. Additionally, werequested the participants to take multiple personality tests designed for human andnon-human primates to evaluate the reliability of our genetic data. Our results indicatethat the male participants, possessing one X chromosome, were homozygous inglutamine and glycine regions of the androgen receptor gene. Females on the other
hand, with two X chromosomes, exhibited heterozygosity. Also, among the participants,
shorter repeat numbers correlated with lower dominance and harm avoidance, while
acquiring a higher neuroticism score in the Chinpanzee personality trait assessment.
Alcohol tolerance, assessed by patch test and SNP detection of ALDH2 gene in buccal
cell samples using PCR and agarose gel electrophoresis revealed that all participants,
except one male individual, exhibited strong alcohol tolerance.

Introduction
The androgen receptor (AR) is a nuclear hormone receptor that is expressed in almost
every tissue of the body [1], and has a significant function in the development of malephenotypes [2,3]. The gene encoding this receptor, which is also known to have an
influence on behavioral patterns in terms of aggression [2], has eight exons in human[5] one of which contains microsatellite repeat regions that code for poly-glutamine andpoly-glycine (hereafter referred to as poly-Q and poly-G respectively). These tworegions are reported to be polymorphic in their length in both human and non-humanprimates [1]. Multiple studies have been done on the correlation between thepolymorphism in poly-Q and poly-G sequence repeat number and personality traits
including agression in human and other animal species; For instance, an study carriedout by Konno and colleagues (2011) on Japanese Akita Inu dogs [6] suggests theexistence of a link between the length of AR gene and aggression. According to theresults of this study, male dogs with higher score in aggressiveness (based on aquestionnaire taken by the dog’s owners) had a shorter allele of poly-Q (CAG repeat
region), which results in more activity of AR.
In this experiment, we investigated the AR gene polymorphism in six participants, ans
compared the genetic data collected with the result of a personality test designed for
chimpanzees [7,8], which is titled “Chimpanzee Personality Trait Assessment”, topredict the participants’ personality traits. We also asked them to take the TCI test [9] inorder to analyze the reliability of our evaluations of personality their personality traits.
It has been reported that approximately 50% of East Asian people [10,11] experience
severe reactions in case of alcohol consumption [12], due to a deficiency in their class 2
mitochondrial aldehyde dehydrogenase (ALDH2) activity. ALDH2 is an enzyme
consisting of four sub-units, which is responsible for metabolyzing the acetaldehydethat is formed as a result of ethanol oxidation in the liver [10]. A single point mutationin exon 12 of ALDH2 coding region can lead to a dramatic reduction in its activity. As aresult of this mutation, one code changes from G to A and consequently, glutamine
substitutes lysine [13]. Deficiency in this enzyme not only causes unpleasant reactions
to consuming alcoholic drinks, it also carries a significant risk of drinking-related
cancers [11]. This defective variant could be genotyped by detecting the single
nucleotide polymorphism (SNP) types in human genome [14]. During this laboratory
course, and in addition to investigating the relationship between emotions and AR
coding sequence variations, we also checked the alcohol tolerancy of the participants
through a combination of genotyping the ALDH2 gene and performing patch test.

Methods● Androgen Receptor Genotyping
Preparations Before and After the Experiment: Before and after each experimental
session, the surface of the tables, tube stands, and other equipments were cleaned using
first H2O and then ethanol.
Sampling: To conduct this experiment, buccal cells of the participants was collected byrubbing a cotton bud on the buccal mucous membrane of the inside of the cheeks. To
collect enough cells for this experiment, the cotton bud was rubbed 10 times on theinside of each cheek. Then, the cutton bud was washed in 1000μl saline (0.9% NaCl)
inside a 1.5ml tube. In the next step, the solution containing buccal cells wascentrifuged at 14000rpm for 1 minutes and 30 seconds, 800μl of supernatant was
discarded, and 100-200μl of the remaining solution was used for DNA extraction. In
total, 6 samples were prepared.
DNA Extraction: Students were divided into two groups of three, therefore, themeasurements were prepared for three samples. 110μl saline and 22μl protease persample were mixed. Then, 120μl of the solution containing cells was added to the mix
and everything was vortexed together. Next, 200μl of Buffer AL was added and mixed
by vortexing. The mix was then incubated at 56°C for 10min, vortexed, and spinned
down. After adding 200μl of ethanol and mixing, 700µl of the solution was added to aspin column and centrifuged at 14000rpm for 1min. Then, the spin column was placedin a new collection tube, 500µl of Buffer AW1 was added and the whole mix wascentrifuged at 8000rpm for 1min. The spin column was placed in a new collection tube,
500µl of Buffer AW2 was added and the whole mixture was centrifuged at 14000rpm
for 4min. This longer period of centrifuging removes the remaining ethanol that wasadded previously, which could later inhibit the PCR process. After that, the spincolumn, now containing the DNA, was placed in a new 1.5ml tube, 50µl of H2O(previously incubated at 56°C) was added directly to the center of the column and thetube was incubated at 56°C for 5min. Next, the mixture was centrifuged at 8000rpm for1min to separate the DNA from the filter inside the spin column, the column was
discarded and the DNA was stored at 4°C.
Measuring DNA Concentration: To Nanodrop Spectrophotometer to measure theextracted DNA’s concentration first, the device was calibrated by putting 1µl of either
H2O or buffer on the device’s sensor and running the analysis. Then, the surface waslightly wiped dry. Before puting the target DNA on the sensor for measurement, we
tapped the tube to mix the DNA and then it was spinned down. Next, 1µl of DNA wasplaced on the nano drop sensor and the concentration was measured. After finishingmeasuring the DNA concentration of all 6 samples, the sensor was cleaned in the sameway described previously.

PCR: To create the PCR mixture, the following materials were mixed in a 1.5ml tube
under a clean bench, and 9µl of it was distributed into PCR tubes:

Table 1: PCR mixture measurements per sample.

Material

Measurement

2x GCI Buffer

5.0µl

H2O

1.8µl

sNTP

1.6µl

*Primer1 (20µl)

0.25µl**Primer2 (20µl)

0.25µlLA Taq0.1µl

Template DNA

1.0µl

PCR mixture + Template DNA

10.0µl*Primer1 sequence: [ARhFF: 5'-TCCAGAATCTGTTCCAGAGCGTGC-3' (FAM labeled); ARhR:
5'-GCTGTGAGGGTTGCTGTTCCTCAT-3' ; Fragment size: around 280bp]; This primer is designed forglutamine (Q) repeat.
**Primer2 sequence: [ARGFH: 5'-CAGTGCCGCTATGGGGACCTGGCGA-3' (HEX labeled); ARGR:
5'-GGACTGGGATAGGGCACTCTGCTCACC-3' ; Fragment size: around 320bp]; This primer isdesigned for glycine (G) repeat.

1µl of template DNA was later added to the tubes under a clean bench, and we ran the
PCR reaction with the following condition: PCR reaction (LA60C30): 95°C 2min
(95°C 30s, 60/65°C 30s, 74°C 1min) x30/35, 74°C 10min, 10°C.
Genotyping: Firstly, we diluted the PCR product by adding 100μl water and 1μl PCR
product per well in a new 96 well plate, and then we spinned down the plate. In the next
step, we made a mixture of Hi-Di Formamide and size standard (For 16 samples: 160μl
Hi-Di Formamide and 1μl size standard). Hi-Di Formamide is a buffer that prevents
H2O from evaporating when it is exposed to electricity during the sequencing process.
Next, 10 μl of the mixture was added to a new 96 well plate, and 1μl of the mixture ofwater and PCR product that was prepared in the beginning was added to each well in
the plate. The samples were heated 95°C for 5 min in order to denature thedouble-stranded amplicons and separate the strands, and then they were immediately
placed on an ice box for 5 min. Before sunning the sequencer, we input samples’ data tothe plate manager to associate them with the results of the sequencing, and next, we run
the sequencer.
To have an efficient data to further investigations, we filtered the data output bychoosing specific peaks to be printed out. In other words, if there were two peaks next
to each other, one shorter than the other, we removed the shorter one if it was as high as
50% of the other.
Repeat Number Calculation: Based on Table 2, the repeat numbers for each region
were calculated. Firstly, the sequence lengths were rounded up/down (considering thefact that 3bp is required to create an amino acid code). Then, Since each amino acidcode consists of 3bp, for every additional 3bp in the allele sequence, 1 number wasadded to the repeat number.

Table 2: The repeat number of ARG and ARQ based on their sequence length

Name of The Sequence

Sequence Length

Repeat No.

ARQ

273

23.00

ARG

314

17.00

ARQ: Poly-Glutamine; ARG: Poly-Glycine

Dominance, Neuroticism: To calculate these factors, we used the data reported by
Weiss et al. (2009), which is provided in Figure 1, as a reference and first selected all
the items related to each factor:
1. Dominance: Dominant, Persistent, Stable, Stingy/Greedy, Timid, Bullying,
Aggressive, Manipulative, Inquisitive, Submissive, Dependent/ Follower,
Decisive, Depressed, Defiant, Intelligent, Disorganized, Independent
2. Neuroticism: Fearful, Stable, Autistic, Excitable, Depressed, Clumsy, Erratic,
Unemotional, Imitative)
Then, based on Figure 1, we changed the chimpanzees’ and our scores of the test to
positive and negative scores and summed the numbers of all the items in each category,
divided them by the number of the items, and reported the final number in Tbale 6.

Figure 1: Structure of Chimpanzees in Japan Rotated to Structure from King and Figueredo (1997) [7]● ALDH2 Genotyping
Sampling: DNA amplicons of the PCR reaction for the previous experiment was usedin ALDH2 genotyping.
Patch Test: Two pieces of 1.5cm square cut paper towels were soaked in water and
70% ethanol separately. The patches were next placed on the inside of participants’arms (Figure 2) for 10min, and 10min after their removal, the skin was observed forredness in the point of touch with the alcohol patch. A red skin would indicate lower
alcohol tolerancy. The H2O patches were used as control.

Figure 2: H2O and ethanol patches on the inside surface of arms.

PCR: PCR: To create the PCR mixture, the following materials were mixed in a 1.5ml
tube outside of a clean bench, and 8µl of it was distributed into PCR tubes:

Table 1: PCR mixture measurements per sample.

Material

Measurement

AmpliTaq Gold 360 Master Mix

5.0µl

H2O

2.5µl

*F Primer

0.25µl

**R/***R2 Primer

0.25µlTemplate DNA

2.0µl

PCR mixture + Template DNA

10.0µl*F Primer sequence: (ALDH F): CAAATTACAGGGTCAACTGCT; This primer is designed for bothGlu and Lys.
**R Primer sequence: (ALDH R): CCACACTCACAGTTTTCACTTC; This primer is designed for
Glu.
**R Primer sequence: (ALDH R): CCACACTCACAGTTTTCACTTC; This primer is designed for Lys.

2µl of template DNA was later added to the tubes under a clean bench, mixed withvortex, and spinned down before running the PCR reaction with the following
condition: PCR reaction (GT65C35): 95°C, 9:00 (94°C, 0:30; 65°C, 1:00; 72°C,
1:00) 35 cycles 72°C, 5:00. In this thermal cycle, in addition to the 6 samples
previously collected, 2 control DNA templates were also amplified.

Agarose Gel Electrophoresis: We first prepared a 1.5% agarose gel by measuring 60ml
of 1xTBE, and adding 0.9g of agarose powder to it until the mixture weighed 60g.
Then, we dissolved the powder in 1xTBE by heating it in the microwave for 1 to 2min.
While heating, some of the mixture evaporates, therefore, we added H2O to both restore
the original volum and cool down the liquid to 60°C. We then placed the comb in thegel tray and poured the mixtures in it to harden for 30min. Before removing the comb
and running the PCR amplicons in the gel, we added a smal amount of 1xTBE to avoid
damages to the wells and gel. In the next step, we filled the electrophoresis tank with
400ml of 1xTBE, and put the gel in it. To load the samples in the gel, we put 1μl of dye
to a piece of parafilm stuck on the table, added 6μl of PCR products to it and pipeted
them together before loading everything to the wells. We also loaded a mixture of 1μl
dye with 3μl size marker in the middle well. We ran the electrophoresis at 100V for
20min until the dye moves to the middle of the gel. Later, to preserve the data, we tooka photo of the gel on the UV Transilluminator (Figure 2).

Results
DNA Concentration: Nanodrop Spectrophotometer device was used to check whether
each sample's DNA concentration was desirable. The DNA concentration for all of thesamples is shown in Table 3. For all of the samples, an increase was observed between
250nm and 280nm. Moreover, the 260/280 factor for sample number 3 was 1.91.

Table 3: DNA concentration measured by nanodrop spectrophotometer

Sample No.

DNA Concentration

1

38.1

2

17.1

3

9.1

4

13.7

5

19.0

6

21.7

Genotyping: The androgen receptor gene was sequenced in the regions encoding
glutamine and glycine amino acids to check their hetrezygocity/homozygocity. Since
this gene is placed on the X chromosom, it only had one allele for each of the amino

acids in the male participants. In other words, all of the male samples were homozygote
for both glutamine and glycine. On the other hand, three out of four female samples
were heterozygote in glutamine region, while one of them was homozygote. The resultsof the female samples for the glycine region were the opposite, meaning that only one
female was heterozygote and the four other female individuals were homozygote (Table4). Based on Table 2, repeat numbers of each sample in the Glutamine and Glycine
alleles were calculated, and the results are shown in Table 5.

Table 4: Genotyping results for glutamine and glycine region in androgen receptor gene

Sample
No.

Sex

ARQ1

ARQ2

Heterozygote/Homozygote

ARG1

ARG2

Heterozygote/
Homozygote

1F

279.11284.86

Hetro

323.20331.11

Hetro

2

M

276.42

Homo

331.203

F276.71276.71

Homo

331.27331.27Homo4

F

279.40282.20

Hetero331.16331.16Homo5

F

276.61288.00Hetro

331.26331.26Homo

6M

276.29Homo

331.33

Homo

Homo

F: Female; M: Male; ARQ: Glutamine; ARG: Glycine

Table 5: Repeat numbers of ARQ and ARG for 6 samples

ARQ1

ARQ2

ARG1

ARG2

ARQ1
RepeatNo.

1279.00285.00323.00332.00

25.00

27.00

26.00

20.00

23.00

21.50

2

276.00－332.00

－

24.00

－

24.00

23.00

－

23.00

3

276.00276.00332.00332.00

24.00

24.00

24.00

23.00

23.00

23.00

4

279.00282.00332.00332.00

25.00

26.00

25.50

23.00

23.00

23.00

5

276.00288.00332.00332.00

24.00

28.00

26.00

23.00

23.00

23.00

6

276.00－332.00

－

24.00

-

24.00

23.00

－

23.00

Sampl
e No.

ARQ2
Average
Repeat Repeat No. for
No.
ARQ

ARG1
RepeatNo.

ARG2
RepeatNo.

Average
Repeat No.
for ARG

Additionally, the average number between ARG 1 and 2, and ARQ 1 and 2 was
calculated (Table 5). This number was later used to put the samples into two groups ofshort and long sequence length. Based on this result, samples 2, 3, and 6 were reported
short, whereas samples 1, 4, and 5 were reported to be in the long group (Table 7).
Moreover, the dominance, neuroticism, and harm avoidance indices were calculated forthe 6 participants of this experiment, using the Chimpanzee Personality Assesment(Table 6). Then, the average number of these indices in each group were calculated andtheir relevance to the sequence length was investigated (Table 6). Based on the results,
individuals with a shorter sequence length are less dominant and avoidant of harmful
situation, and have a higher neuroticism index compared to the individuals with a higher
sequence length.

Table 6: Dominance, Neuroticism, and Harm avoidance index of human samples based on the
Chimpanzee Personality Assessment and TCI test.

Sample No.

Dominance

Neuroticism

Harm Avoidance

1

2.00

-0.33

80.00

2

2.18

-0.33

Not reported

3

1.35

0.44

49.00

4

2.53

-0.11

40.00

5

2.12

-0.22

40.00

6

2.00

-0.44

68.60

Table 7: The average number of Dominance, Neuroticism, and Harm avoidance and their relevance to thesequence length.

Sequence
lengthSample No.

Dominance

Neuroticism

Harm
avoidance

Short

2,3,6

1.84

-0.11

39.20

Long

1,4,5

2.22

-0.22

60.00

To investigate the alcohol tolerance of the participants, two seperate experiments weredone and their results were aligned to determine whether the participant have low orhigh alcohol tolerancy. Firstly, we surveied the participants’ alcohol tolerance using
patch test; 2 male and 3 females. For this stage, we only tested 5 individuals since oneof the participants was absent for this laboratory session. Among the five individuals,
only one male participant developed redness in part of the skin in contact with alcohol
patch (Table 8). The rest of the participants showed no change in their skin color after
being exposed to 70% ethanol. Among the people with negative results in the patch test,
25% were male (one individual) and 75% were female (three individuals).

Table 8: Patch test results and the electrophoresis results of the presence/absence of Glu and Lys coding
sequences for investigating alcohol tolerance.

Sample No.

patch test

Glu

Lys

Alcohol tolerance

1

－

＋

－

Storong

2

－

＋

－

Storong

3

－

＋

－

Storong＋－

Storong

4
5

－

＋

－

Storong

6

+

＋

＋

Medium

In the next step, DNA samples extracted from participants’ buccal cells were genotyped
in the ALDH2 gene, looking for SNP in the Glu and Lys coding regions. The result of
running the DNA samples on Agarose Gel Electrophoresis (Figure 3) shows that all of
the samples contained the Glu coding sequence, while only sample No. 6 had the coding
sequence of Lys. Based on this result, All of the participants had strong alcohol
tolerance, except for sample 6 (Male participant) who had a medium alcohol tolerance(Table 8).
TCI Test: Since the TCI (Temperament and Character Inventory) test was available inJapanese at the time of experiment, I took the IPIP equivalent to TCI test [15]. The test
results are shown on Table 9.

Figure 3: The result of gel electrophoresis detecting the presence of Glu and Lys.

A: Amplicons of Glu (From left to right: samples 1, 2, 3, 4, 5, 6, *Control 1, **Control 2); B: Ampliconsof Lys (From left to right: samples 1, 2, 3, 4, 5, 6, Control 1, Control 2); C: Size marker.
*The combined results of control 1 indicates a person with strong alcohol tolerance (Glu: +/ Lys: –).
**The combined results of control 2 indicates a person with medium alcohol tolerance (Glu: +/ Lys: +).

Table 9: IPIP equivalent to TCI test results for participant No.3.

Personality Domain

Score

Idealistic

32

Novelty seeking

63

Harm avoidance

49

Reward dependence

47

Persistence

56

Self-directedness

70

Cooperativeness61

Discussion
In this experiment, all of the DNA samples extracted from male participant were
homozygote in both glutamine and glycine regions. This happens as a result of theandrogen receptor gene being located on the X chromosome [4], and since maleindividuals are XY, they only possess one copy of this gene. Therefore, they can only behomozygote for our target regions on the androgen receptor gene. On the other hand,samples collected from female individuals could be either homozygote or heterozygote
in the glutamine and glycine regions, because they are XX and have two copies of theandrogen receptor gene. In our study, 75% of the female samples were heterozygote inglutamine, while only 25% of them were heterozygoete in glycine region. Moreover,individuals with shorter repeat numbers in the glutamine and glycine regions, had lower
dominance factor and harm avoidance index, with a lower neuroticism. On the other
hand, individuals with a longer repeat number in the target regions were more dominant
and were expected to avoid harm more than the other group, while being more neurotic(based on the Chimpanzee Personality Assessment). In previous studies[8], it ismentioned that the shorter sequence length of Poly-Q is associated with moreaggression in male dogs, and they also reported no evidence of a correlation between
AR sequence length and aggression in female dogs. However, in our experiment, thegroup with short AR coding sequence consisted of both male and female individuals,
and according to their average dominance factor, they are less aggressive compared to
the group with a longer sequence length(Table 7). For future experiments, I suggest theuse of a personality assessment test designed for humans. Since the personality test we
used in this study was originally designed for chimpanzees, it could decrease theaccuracy of the experiment when comparing the test results with genotyping data. For
instance, while taking the test myself, I had difficulty interpreting some of the questions
from how a chimpanzee would behave to their human counterpart.
In the second part of our experiment, we investigated the alcohol tolerance of theparticipants by combining the results of two methods; Firstly, we conducted the patch
test to check the participants’ alcohol tolerance through observing their skins’ reaction
to being exposed to alcohol. The result of this test aligned perfectly with what wasdetermined through gel electrophoresis, showing that 17% of the participants (1 male)
had medium tolerancy, while 83% of them (4 female, 1 male) were strongly tolerant toalcohol.

References
1. Hiramatsu, C., Paukner, A., Kuroshima, H., Fujita, K., Suomi, S.J. andInoue-Murayama, M., 2017. Short poly-glutamine repeat in the androgen
receptor in New World monkeys. Meta gene, 14, pp.105-113.
2. Li, J., Al-Azzawi, F., 2009. Mechanism of androgen receptor action. Maturitas
63, 142–148. http://dx.doi.org/10.1016/j.maturitas.2009.03.008.
3. Gelmann, E.P., 2002. Molecular biology of the androgen receptor. Journal ofclinical oncology, 20(13), pp.3001-3015.
4. Brown, C.J., Goss, S.J., Lubahn, D.B., Joseph, D.R., Wilson, E.M., French, F.S.
and Willard, H.F., 1989. Androgen receptor locus on the human X chromosome:regional localization to Xq11-12 and description of a DNA polymorphism.

American journal of human genetics, 44(2), p.264.
5. Kuiper, G.G.J.M., Faber, P.W., Vanrooij, H.C.J., Vanderkorput, J.A.C.M.,
Risstalpers, C., Klaassen, P., Trapman, J., Brinkmann, A.O., 1989. Structuralorganization of the human androgen receptor gene. J. Mol. Endocrinol. 2,
R1–R4.
6. Konno, A., Inoue-Murayama, M. and Hasegawa, T., 2011. Androgen receptor
gene polymorphisms are associated with aggression in Japanese Akita Inu.
Biology Letters, 7(5), pp.658-660.
7. Weiss, A., Inoue‐Murayama, M., Hong, K.W., Inoue, E., Udono, T., Ochiai, T.,
Matsuzawa, T., Hirata, S. and King, J.E., 2009. Assessing chimpanzee
personality and subjective well‐being in Japan. American Journal ofPrimatology: Official Journal of the American Society of Primatologists, 71(4),pp.283-292.
8. Inoue-Murayama, M., Hibino, E., Matsuzawa, T., Hirata, S., Takenaka, O.,
Hayasaka, I., Ito, S.I. and Murayama, Y., 2006. The application of a humanpersonality test to chimpanzees and survey of polymorphism in genes relating toneurotransmitters and hormones. Cognitive development in chimpanzees,
pp.113-124.
9. Cloninger, C. R., Przybeck, T. R., Svrakic, D. M., & Wetzel, R. D. (1994). The
temperament and character inventory (TCI): A guide to its development and use.
St. Louis: Washington University Center for Psychobiology of Personality.
10. Farrés, J., Wang, X., Takahashi, K., Cunningham, S.J., Wang, T.T. and Weiner,
H., 1994. Effects of changing glutamate 487 to lysine in rat and human liver
mitochondrial aldehyde dehydrogenase. A model to study human (Oriental type)class 2 aldehyde dehydrogenase. Journal of Biological Chemistry, 269(19),pp.13854-13860.
11. Matsumoto, A., 2018. Importance of an aldehyde dehydrogenase 2
polymorphism in preventive medicine. Nihon Eiseigaku zasshi. Japanese Journalof Hygiene, 73(1), pp.9-20.
12. Mizoi, Y., Ijiri, I., Tatsuno, J., Kijima, T., Fujiwara, S., Adachi, J., and Hishida,S.
(1979) Pharmacol. Biochem. Behau. 10,303-311
13. Yoshida, A., Huang, I.Y. and Ikawa, M., 1984. Molecular abnormality of an
inactive aldehyde dehydrogenase variant commonly found in Orientals.
Proceedings of the National Academy of Sciences, 81(1), pp.258-261.
14. Chang, J.S., Hsiao, J.R. and Chen, C.H., 2017. ALDH2 polymorphism andalcohol-related cancers in Asians: a public health perspective. Journal ofbiomedical science, 24(1), pp.1-10.
15. https://psytests.org/multi/itcien.html