2
.
2
. Analysis The sera were obtained after a 12-h fast. Serum total
cholesterol levels were measured by the enzymatic method [42]. The total concentration of HDL choles-
terol and the cholesterol subfractions were measured by the dextran sulfate method [43] as previously described
[44].
The precipitation reagent for total HDL cholesterol contained 10 gl Dextralipid 50 Sochibo, Boulogne,
France and 101.6 gl MgCl
2
– 6H
2
O and for HDL
3
19.1 gl Dextralipid 50 and 397.4 gl MgCl
2
– 6H
2
O. Then 250 ml serum was added to 25 ml of the appropriate
reagent and after a 15-min incubation at ambient tem- perature, supernatants were separated by centrifugation
for 20 min at 12 000 × g Heraeus Biofuge 15, os- terodeamtlarz, Germany. The HDL cholesterol con-
tents of the supernatants were analyzed enzymatically on a Monarch 2000 Analyzer Instrumentation Labora-
tory, Lexington, USA using Chod-Pap cholesterol reagents Cat No 237574; Boehringer Mannheim, Ger-
many, and a primary cholesterol standard Cat No 530, Orion, Espoo, Finland. The HDL
2
subfraction was calculated by subtraction of HDL
3
from total HDL. Triglyceride levels were measured by the enzy-
matic, kinetic method. Apo A1 and B levels were determined immunoturbidimetrically [45]. Serum LDL
levels were calculated as cholesterol – HDL choles- terol – 0.45 × triglycerides [46]. None of the women had
triglyceride values \ 4.0 mmoll and thus none of them were excluded from this calculation. The percentages of
HDL
2
and HDL
3
of total cholesterol and HDL choles- terol were calculated, and the values were compared
between the groups.
2
.
3
. Statistical methods BMDP software programs were used in the statistical
analyses of the material [47]. The correlations of self-re- ported alcohol consumption and lipid values were stud-
ied using bivariate plots for linear regression. The lipid values were compared between the groups, by using
analyses of variance. When variances were different, the Welch approximation was used. To compare the results
with earlier studies, the women were divided also into five groups according to self-reported alcohol consump-
tion. In both cases the lipid values that were signifi- cantly different using P
ANOVA
were also compared using pairwise t-test between the groups; separate,
when the variances between the groups differed and pooled when they did not differ. Nonnumeric parame-
ters between the groups were compared using cross- tabulations.
3. Results
Among 274 women all lipid values except LDL cholesterol significantly correlated with self-reported al-
cohol consumption Table 2. When alcoholics were excluded, only Apo A1, HDL cholesterol and HDL
3
cholesterol correlated significantly with the self-re- ported alcohol consumption Table 2. A strong signifi-
cant positive correlation was found also between Apo A1 and HDL
2
r = 0.62, P B 0.001 and Apo A1 and HDL
3
r = 0.71, P B 0.001. Lipid values were compared between different groups
based on self-report and results from two question- naires Table 3. P
ANOVA
was significant for total cholesterol, HDL
3
cholesterol, triglycerides, Apo AI and Apo B. Differences in lipid values using t-tests
were found between the moderated drinkers and the alcoholics and between heavy drinkers and alcoholics.
No differences were found between the groups in the percentage of abnormally low HDL cholesteroltotal
cholesterol B 0.20, ApoA1 B 0.86 gl, HDL
3
cholesterol B 0.8 mmoll, and HDL
2
cholesterol B 0.3 mmoll.
Table 1 Characteristics of the women divided into different groups according to the self-reported alcohol consumption and the results from structured
alcohol questionnaires Moderate drinkers n = 139
P
ANOVA a
Alcoholics n = 73 Heavy drinkers n = 62
Alcohol consumption gweek 39.0
B 0.001
160.6 934.3
38.3 42.4
B 0.001
Age years 42.3
24.3 25.3
BMI
b
kgm
2
23.5 0.062
Smokers 33.3
46.8 85.0
B 0.001
7 Anxiolytes
B 0.001
33 1
Gonadal hormones 6
6 1
\ 0.05
12 16
Other medication 16
\ 0.05
a
ANOVA, analysisis of variance.
b
BMI, body mass index.
Table 2 Correlations between the self-reported alcohol consumption and dif-
ferent lipid measures among women Serum lipids
All women All except alcoholics
n = 201 n = 274
r r
P P
0.215 Cholesterol
B 0.001
0.008 0.906
0.018 0.166
0.206 B
0.001 HDL
cholesterol 0.088
HDL
2
B 0.001
0.209 0.107
cholesterol HDL
3
B 0.001
0.227 0.284
B 0.001
cholesterol 0.272
LDL-cholester 0.060 NS
− 0.077
ol 0.392
B 0.001
0.032 0.647
Triglycerides 0.353
B 0.001
Apo A1 0.289
B 0.001
Apo B 0.240
B 0.001
− 0.005
0.946
\ 40 gday P B 0.01, P-value not shown in table and
those drinking 0 – 10 gday. The relative changes of total HDL cholesterol, its
density based subtractions and apolipoprotein A1 in the group drinking \ 10 – 20 gday compared to those
drinking 0 – 10 gday indicate constant increasing trend for HDL
3
but significant reduction of HDL
2
resulting in decreasing or plateau trend in total HDL cholesterol
and apolipoprotein A1 Fig. 1. In-groups with higher consumption the trend for both HDL subfractions was
increasing, resulting in increase in total HDL choles- terol and apolipoprotein A1.
Because Apo A1, HDL cholesterol and HDL
3
sub- fraction correlated significantly with self-reported alco-
hol consumption even when the alcoholics were excluded, we studied these values further. The mean
HDL cholesterol value among alcoholics was 0.17 mmoll higher than among moderated drinkers 95
confidence interval from 0.02 to 0.32 mmoll, and Apo A1 values 0.19 mmoll higher 95 confidence interval
from 0.12 to 0.26 mmoll. HDL
3
cholesterol values were 0.11 mmoll higher among alcoholics than in
moderated drinkers 95 confidence interval from 0.05 to 0.17 mmoll. The concentrations of HDL choles-
terol, HDL
3
cholesterol, and Apo A1 in alcoholic women were 12 P B 0.05, 11 P B 0.001, and 13
P B 0.001 higher than in the moderated drinkers, respectively.
Among the 12 alcoholic females who were followed for 2 weeks during abstinence, HDL
3
cholesterol de- creased P = 0.004, whereas LDL cholesterol P =
0.02, and Apo B P = 0.02 significantly increased. To further confirm the above findings, we studied
lipid values based only on the self-reported alcohol consumption Table 4. Here P
ANOVA
was significant concerning total HDL, Apo A1, HDL
3
cholesterol, HDL
2
, HDL
2
total cholesterol, HDL
3
total cholesterol, HDL
2
HDL cholesterol, and HDL
3
HDL cholesterol. An increasing trend of HDL cholesterol P B 0.05, P-
value not shown in table, HDL
3
cholesterol and Apo A1 P B 0.05, P-value not shown in table was first seen
among those women drinking \ 20 – 40 gday as com- pared to those drinking \ 10 – 20 gday. HDL
2
choles- terol values were significantly lower among those
drinking \ 10 – 20 gday as compared to those drinking
Table 3 Lipid values mean 9 SD of the women divided into different groups according to the self-reported alcohol consumption and the results from two
structured alcohol questionnaires
a
Moderate drinkers n = 139 Heavy drinkers n = 62
Alcoholics n = 73 P
A B
C 1234
5.10 9 0.87 Chol
5.10 9 0.95 5.80 9 2.18
c 0.983 1.55 9 0.35
1.57 9 0.45 1.72 9 0.58
0.089 HDL
0.56 9 0.26 0.54 9 0.31
HDL
2
0.61 9 0.46 0.610
HDL
3
1.00 9 0.16 1.02 9 0.19
1.11 9 0.24 c 0.375
0.31 9 0.12 0.31 9 0.09
0.31 9 0.08 HDLChol
0.959 HDL
2
Chol 0.11 9 0.06
0.11 9 0.05 0.11 9 0.08
0.939 HDL
3
Chol 0.20 9 0.04
0.21 9 0.05 0.21 9 0.06
0.615 HDL
2
HDL 0.34 9 0.10
0.32 9 0.11 0.32 9 0.13
0.384 0.66 9 0.10
0.68 9 0.11 HDL
3
HDL 0.68 9 0.14
0.399 3.10 9 0.80
3.10 9 0.88 LDL
3.30 9 2.15 0.730
1.00 9 0.51 1.00 9 0.47
Tg 1.70 9 0.93
c c
0.541 c ApoA
1
1.40 9 0.18 1.50 9 0.23
1.60 9 0.30 c
c 0.070
Apo
B
0.70 9 0.18 0.70 9 0.18
0.90 9 0.32 c 0.496
a
P
1
ANOVA, P
2
C vs. A, P
3
B vs. A, P
4
C vs. B. Chol, Cholesterol; LDL, LDL-cholesterol; Tg, Triglycerides. PB0.05, PB0.01,
c
PB0.001.
Table 4 Lipid values mean 9 SD of the women divided into different groups A–E based on self-reported alcohol consumption
a
Lipid values P n = 76
Alcohol consumption per day g \
10–20 n = 49 \
20–40 n = 33 \
40 1234
0–10 n = 106 B
C D
A 5.10 9 0.95
5.00 9 0.91 Chol
5.70 9 2.11 5.10 9 0.86
0.6120.764 HDL
1.54 9 0.34 1.47 9 0.41
1.65 9 0.42 1.74 9 0.58
c 0.1580.268 0.46 9 0.23
0.58 9 0.31 0.56 9 0.27
0.63 9 0.46 HDL
2
0.2560.788 1.01 9 0.22
1.07 9 0.17 HDL
3
1.11 9 0.24 0.98 9 0.14
c c
0.33 0.29 9 0.08
0.34 9 0.08 0.31 9 0.08
0.32 9 0.11 HDLChol
0.101 0.11 9 0.06
HDL
2
Chol 0.09 9 0.04
0.12 9 0.06 0.11 9 0.08
0.9390.729 0.20 9 0.05
0.22 9 0.04 0.20 9 0.04
0.21 9 0.06 HDL
3
Chol 0.1170.254
0.35 9 0.11 HDL
2
HDL 0.30 9 0.08
0.33 9 0.11 0.33 9 0.13
0.3600.441 HDL
3
HDL 0.70 9 0.08
0.65 9 0.10 0.67 9 0.11
0.67 9 0.13 0.3710.451
3.10 9 0.84 2.90 9 0.76
3.18 9 0.82 3.30 9 2.08
LDL 0.524
0.95 9 0.53 Tg
1.10 9 0.49 1.10 9 0.56
1.60 9 0.92 c
c 0.3380.166
1.40 9 0.24 1.50 9 0.19
1.60 9 0.30 c
c 0.868
Apo A1 1.40 9 0.16
0.70 9 0.18 0.70 9 0.18
0.80 9 0.32 c 0.6430.491
Apo B 0.70 9 0.19
a
P
1
P
Anova
, P
2
D vs. A, P
3
C vs. A, P
4
B vs A. Chol, cholesterol; LDL, LDL-cholesterol; Tg, triglycerides,
c
PB0.001; PB0.01,
PB0.05.
4. Discussion
