**3. Results**

Clinical and biochemical characteristics of the patients with T1DM without and with diabetic complications and controls (CTRL) are reported in Table 1.


a significant difference in comparison with CTRL

b significant difference in comparison with +DC group T1DM

Table 1. Clinical and biochemical characteristics of the patients with T1DM and controls

As seen, HbA1c and FAM were significantly elevated in both diabetic groups in comparison with controls and also in +DC vs. –DC those. Serum AGEs were significantly elevated in +DC compared to –DC and also to controls, but the difference between –DC and controls was not significant. The levels of AOPP were evidently higher in +DC compared to controls, but the difference was not significant. The levels of LPO were significantly elevated in +DC vs. controls, the differences between both diabetic groups and between –DC vs. controls were not significant. The levels of TGF-beta1 similarly to s-AGEs were significantly elevated in +DC compared to –DC and also to controls, but the difference between –DC and controls was not significant (Fig. 1). In terms of the VCAM-1 values, only between +DC and controls there were found significant difference there (Fig. 2).

The levels of TGF-beta1 are significantly elevated in +DC compared to –DC (10.49 ± 4.55 vs. 5.9 ± 4.14 ng/ml, p<0.05) and also to controls (10.49 ± 4.55 vs. 3.30 ± 3.41ng/ml, p<0.05), but the difference between –DC and controls (5.9 ± 4.14 vs. 3.30 ± 3.41ng/ml, p>0.05) was not statistically significant.

#### **3.1 The relationships between clinical and biochemical parameters 3.1.1 The subgroup of patients with T1DM without diabetic complications**

The relationships characterized by Pearson´s correlation coefficient r or Spearman´s coefficient R between the parameters described within the text are reported in Table 2. As seen, we found significant linear correlations of FAM with HbA1cA (r=0.676), LPO with HbA1cP (r=-0.507) and AOPP (R=0.671). The relationship between LPO (y) and HbA1cP(x) is possible to describe by non-linear equation y=19x2-354x+1752 (R=0.632, R2=0.400, p<0.05). VCAM-1 significantly correlated with age (r=-0.478), HbA1cA (r=0.653, Fig. 3), HbA1cP (r=0.501) and with FAM (r=0.630, Fig. 4).

association between parameters described within the text, in all studied patients and in diabetic and non-diabetic subgroups. P values less than 0.05 were accepted as being statistically significant. All statistical analyses were carried out using Excel 2003, Origin 8

Clinical and biochemical characteristics of the patients with T1DM without and with

 CTRL n T1DM -DC n T1DM +DC N Age (yrs.) 9.0(6.1,14.0) 26 14.4(12.4, 17.9)ab 20 16.4(15.1, 17.6)a 26 DD (yrs.) - 0 6(5.5, 8.1)b 20 10.0(7.9, 12.9) 26 HbA1cA (%) 5.0 ± 0.3 18 8.3 ± 1.4 ab 20 10.4 ± 1.4a 26 FAM (mmol/l) 1.67 ± 0.31 24 2.64 ± 0.38 ab 20 3.06 ± 0.48a 26 s-AGEs (A.U.) 54.9 ± 9.9 22 64.4 ± 10.1 ab 20 71.8 ± 11.6 24 AOPP (micromol/l) 58.8(52.0, 71.8) 11 43.3(42.6, 60.4) 17 78.2(49.5, 114.6) 20 LPO (nmol/ml) 100(88, 110) 10 106(105, 161) 19 127(109, 152)a 17 TGF-beta1 (ng/ml) 3.30 ± 3.41 8 5.9 ± 4.14b 10 10.49 ± 4.55a 16 VCAM-1 (ng/ml) 12.6 ± 3.7 15 17.1 ± 3.1 19 17.4 ± 3.3a 26

Table 1. Clinical and biochemical characteristics of the patients with T1DM and controls

As seen, HbA1c and FAM were significantly elevated in both diabetic groups in comparison with controls and also in +DC vs. –DC those. Serum AGEs were significantly elevated in +DC compared to –DC and also to controls, but the difference between –DC and controls was not significant. The levels of AOPP were evidently higher in +DC compared to controls, but the difference was not significant. The levels of LPO were significantly elevated in +DC vs. controls, the differences between both diabetic groups and between –DC vs. controls were not significant. The levels of TGF-beta1 similarly to s-AGEs were significantly elevated in +DC compared to –DC and also to controls, but the difference between –DC and controls was not significant (Fig. 1). In terms of the VCAM-1 values, only between +DC and controls

The levels of TGF-beta1 are significantly elevated in +DC compared to –DC (10.49 ± 4.55 vs. 5.9 ± 4.14 ng/ml, p<0.05) and also to controls (10.49 ± 4.55 vs. 3.30 ± 3.41ng/ml, p<0.05), but the difference between –DC and controls (5.9 ± 4.14 vs. 3.30 ± 3.41ng/ml, p>0.05) was not

The relationships characterized by Pearson´s correlation coefficient r or Spearman´s coefficient R between the parameters described within the text are reported in Table 2. As seen, we found significant linear correlations of FAM with HbA1cA (r=0.676), LPO with HbA1cP (r=-0.507) and AOPP (R=0.671). The relationship between LPO (y) and HbA1cP(x) is possible to describe by non-linear equation y=19x2-354x+1752 (R=0.632, R2=0.400, p<0.05). VCAM-1 significantly correlated with age (r=-0.478), HbA1cA (r=0.653, Fig. 3), HbA1cP

diabetic complications and controls (CTRL) are reported in Table 1.

a significant difference in comparison with CTRL

b significant difference in comparison with +DC group T1DM

there were found significant difference there (Fig. 2).

(r=0.501) and with FAM (r=0.630, Fig. 4).

**3.1 The relationships between clinical and biochemical parameters 3.1.1 The subgroup of patients with T1DM without diabetic complications** 

statistically significant.

and BioSTAT 2009.

**3. Results** 

Fig. 1. Comparison of TGF-beta1 levels of patients with T1DM and controls

Fig. 2. Comparison of VCAM-1 levels of patients with T1DM and controls

The levels of VCAM-1 are significantly elevated in +DC compared to controls (17.4 ± 3.3 vs. 12.6 ± 3.7 ng/ml, p<0.05). The values of VCAM-1 in –DC subgroup differ obviously from those in controls, but the difference is non statistically significant (17.1 ± 3.1 vs. 12.6 ± 3.7 ng/ml, p>0.05). There are similar levels in both diabetic subgroups ((17.4 ± 3.3 vs. 17.1 ± 3.1 ng/ml, p>>0.05).

The Study of Glycative and Oxidative Stress in Type 1 Diabetes Patients

Fig. 4. The linear correlation between VCAM-1 and FAM

VCAM-1 (ng/ml)

DD

FAM

s-AGEs

AOPP

LPO

TGF-beta1

(† R, <sup>∆</sup> p<0.05)

T1DM with diabetic complications

parameters, but those were not statistically significant.

in Relation to Circulating TGF-Beta1, VCAM-1 and Diabetic Vascular Complications 11

234 FAM (mmol/l)

beta1 VCAM-1

(yrs.) HbA1cA HbA1cP FAM s-AGEs AOPP LPO TGF-

Age (yrs.) 0.448<sup>∆</sup> -0.295 -0.422∆ -0.023 -0.068 -0.165 -0.044† 0.541†∆ 0.067 DD (yrs.) N -0.070 -0.110 -0.170 -0.069 -0.034 -0.297† 0.247† 0.009 HbA1cA (%) N 0.539∆ 0.581<sup>∆</sup> 0.221 0.278 0.123† -0.429† 0.006 HbA1cP (%) N 0.3405 0.247 0.116 0.127† -0.679†∆ 0.291

(mmol/l) N 0.479∆ 0.538<sup>∆</sup> 0.471† -0.708†∆ 0.183

(A.U.) N 0.119† 0.125† -0.356† 0.432<sup>∆</sup>

(micromol/l) N 0.355† -0.545† -0.026

(nmol/ml) N -0.612† -0.174†

(ng/ml) N -0.069†

Table 3. The relationships between parameters described within the text in patients with

As seen, in +DC subgroup we found significant correlations of FAM with HbA1cA (r=0.581), s-AGEs with FAM (r=0.479) and AOPP with FAM (r=0.538). LPO correlated with FAM (r=0.471), but this relation is not statistically significant (p=0.056). TGF-beta1 correlated with age (R=0.541), HbA1cP (R=-0.679) and FAM (R=-0.708). Statistically significant moderate linear correlation was found between VCAM-1 and s-AGEs (r=0.432). Moderate relationships were found also between TGF-beta1 and oxidative stress

Fig. 3. The linear correlation between VCAM-1 and HbA1cA


Table 2. The relationships between the parameters in patients with T1DM without diabetic complications († R, <sup>∆</sup> p<0.05)

In this subgroup LPO and VCAM-1 were in association also with other parameters, but those were not statistically significant. Non linear statistically significant relationship with regression line equation y=0.33x2 – 4.10x + 28.26 was found between VCAM-1(y) and HbA1cA(x) (R=0.694, R2=0.481, p<<0.05).

#### **3.1.2 The subgroup of patients with T1DM with diabetic complications**

The relationships characterized by Pearson´s correlation coefficient r or Spearman´s coefficient R between the parameters described within the text are reported in Table 3.

Fig. 4. The linear correlation between VCAM-1 and FAM


(† R, <sup>∆</sup> p<0.05)

10 Type 1 Diabetes – Complications, Pathogenesis, and Alternative Treatments

5 6 7 8 9 10 11 12 HbA1cA (%)

beta1 VCAM-1

(yrs.) HbA1cA HbA1cP FAM s-AGEs AOPP LPO TGF-

Age (yrs.) 0.205 -0.267 -0.232 -0.328 -0.030 0.130† 0.334 0.406† -0.478<sup>∆</sup> DD (yrs.) N 0.152 -0.008 0.326 -0.256 -0.193† -0.001 0.036† 0.089 HbA1cA (%) N 0.748∆ 0.676<sup>∆</sup> 0.217 -0.298† -0.398 -0.588† 0.653<sup>∆</sup> HbA1cP (%) N 0.433 0.302 -0.400† -0.507<sup>∆</sup> -0.042† 0.501<sup>∆</sup>

(mmol/l) N 0.134 0.081† -0.260 -0.224† 0.630<sup>∆</sup>

(A.U.) N 0.197† 0.270 0.006 0.068

(micromol/l) N 0.671†<sup>∆</sup> 0.477† -0.447†

(nmol/ml) N 0.200† -0.404

(ng/ml) N -0.578†

Table 2. The relationships between the parameters in patients with T1DM without diabetic

In this subgroup LPO and VCAM-1 were in association also with other parameters, but those were not statistically significant. Non linear statistically significant relationship with regression line equation y=0.33x2 – 4.10x + 28.26 was found between VCAM-1(y) and

The relationships characterized by Pearson´s correlation coefficient r or Spearman´s coefficient R between the parameters described within the text are reported in Table 3.

**3.1.2 The subgroup of patients with T1DM with diabetic complications** 

Fig. 3. The linear correlation between VCAM-1 and HbA1cA

VCAM-1 (ng/ml)

DD

FAM

s-AGEs

AOPP

LPO

TGF-beta1

complications († R, <sup>∆</sup> p<0.05)

HbA1cA(x) (R=0.694, R2=0.481, p<<0.05).

Table 3. The relationships between parameters described within the text in patients with T1DM with diabetic complications

As seen, in +DC subgroup we found significant correlations of FAM with HbA1cA (r=0.581), s-AGEs with FAM (r=0.479) and AOPP with FAM (r=0.538). LPO correlated with FAM (r=0.471), but this relation is not statistically significant (p=0.056). TGF-beta1 correlated with age (R=0.541), HbA1cP (R=-0.679) and FAM (R=-0.708). Statistically significant moderate linear correlation was found between VCAM-1 and s-AGEs (r=0.432). Moderate relationships were found also between TGF-beta1 and oxidative stress parameters, but those were not statistically significant.

The Study of Glycative and Oxidative Stress in Type 1 Diabetes Patients

complications.

in Relation to Circulating TGF-Beta1, VCAM-1 and Diabetic Vascular Complications 13

FAM were significantly elevated in patients having diabetic complications only other than diabetic retinopathy compared to –DC (3.10(2.93, 3.54) vs. 2.54(2.42, 2.91) mmol/l, p<0.05, Fig. 5). HbA1c levels are elevated in patients having diabetic retinopathy against to –DC (9.8(9.6, 10.2) vs. (7.9(7.4, 9.1)%, p<0.05), in subgroup of patients having diabetic retinopathy with other complication/s compared to -DC (10.4(8.6, 11.2) vs. (7.9(7.4, 9.1) %, p<0.05, Fig.6) and also the subgroup of patients having diabetic complications only other than diabetic retinopathy compared to –DC (10.5(10.0, 11.1 vs. (7.9(7.4, 9.1) %, p<0.05, Fig. 6). Serum AGEs were significantly higher in subgroup with only other diabetic complications than diabetic retinopathy compared to –DC one (74.8(71.2, 76.5) vs. 61.9(58.9, 71.0) A.U., Fig. 7), and non-significantly higher in patients with retinopathy only than in those with others DC and also in patients with DR and another DC compared to ODC group, however, p-values were only slightly higher than 0.05 (Fig. 7). The values of LPO were significantly elevated in patients with complications other than retinopathy compared to those with retinopathy only (138(129, 165) vs. 101(93, 109) nmol/ml, Fig. 8). No significant differences were found between others in LPO. There were the significant differences between patients having only diabetic retinopathy vs. –DC in TGF-beta1 levels (14.17(13.32, 15.52) vs. 5.7(2.23, 8.71) ng/ml, p<0.05, Fig. 9) and also between subgroup of patients having only diabetic retinopathy and those having diabetic complications other than diabetic retinopathy (14.17(13.32, 15.52) vs. 9.05(5.29, 10.39) ng/ml, p=0.05, Fig. 9). Neither AOPP parameters nor VCAM-1 showed any significant differences between subgroups with regard to presence/absence diabetic retinopathy or other diabetic

Fig. 5. The values of FAM in subgroups of patients with T1DM with regard to diabetic retinopathy presence/absence (DR – having diabetic retinopathy only, DR+ODC – having

diabetic retinopathy and another complications, ODC – having only other diabetic

complications except diabetic retinopathy, -DC – having no complications)
