#

**Time (min)**

ADP and Collagen were measured, signing an effective anti-platelet therapy.

**1 week later**

**3.1.3.1 ADP induced platelet aggregation in acute and elective groups** 

**3.1.3 Investigation of platelet aggregation in whole blood** 

\* = p<0.05,\*\*=p<.01,\*\*\*=p<0,01 vs. control # = p<0.05,\*\*=p<.01,\*\*\*=p<0,01 vs. before surgery + <0 05 \*\* < 01 \*\*\* <0 01 l ti

Fig. 4. ADP induced aggregation in Acute group, compared to Control (red line).

#

**A D P -in d u ced P L T aggregation in W h ole** 

of the week (Figure 4).

**b lood in A cu te grou p (O h m )**

**A D P in w h o le b lo o d o n E le c tiv e** 

**g ro u p ) (O h m )**

Fig. 5. Collagen induced platelet aggregation in Elective group.

Fig. 6. Collagen induced aggregation in whole blood in Acute group.

In whole blood of healthy subjects, the collagen induced platelet aggregation started with one minute delay. In the patient groups, collagen induced aggregation started without delay, mainly in Acute group, one week after the surgery. Application of the inductor in this case resulted in an immediate induction of platelet aggregation. The effect of antiplatelet therapy was detected in platelet- rich plasma was missed in whole blood.

#### **3.1.4 Investigation of antioxidants**

#### **3.1.4.1 GSH level**

Before surgery GSH levels were similar in the three groups. In patients groups a transient reduction occurred 2 hours after and one day after the surgery, but returned to the baseline level one week after the surgery.

#### **3.1.4.2 Plasma thiol groups**

Before the surgery the plasma SH-group concentration of the patient groups did not differed from each other or the healthy controls, but a transient, significant reduction was measured 2 hours and 24 hours after surgery in both patients groups, which returned to the normal level 1 week later.

Investigation of the Oxidative Stress, the Altered Function

**Before rurgery After surgery 24 hour later 1 week later**

*\*\*\**

**+++**

*+++ \*\*\* \*\*\**

**Before rurgery After surgery 24 hour later 1 week later**

than it was measured in elective group (figure 8.).

**red blood cell haemolysate** 

Control Elective Acute

**\***

**#**

**0,00**

**0,05**

**Slo pe o**  **curvem** 

**eredeksége AU/sec/10**

 **3 sejt**

**f the free radical peo**

 **ducing** 

**0,10**

**0,15**

**0,20**

**0,25**

**Control Elective Acute**

*\*\*\**

**W B C num bers(x10 3 w hite blo o d cells)**

of the free radical generation curves and the WBC numbers (Figure 8.).

**##** \*\*

*+++ +++*

*#*

**\***

\*\*

of Platelets and Neutrophils, in the Patients with Peripheral Arterial Disease 73

production corrected to the WBC numbers, 3. the Slope: the rate of rise of the ascending part

**M axim um o f R O S /W B C co unts**

Fig. 8. Upper left panel: white blood cell numbers, upper right panel: lag time, bottom left panel: slope of free radical curve, bottom right panel: maximal free radical production. \*\*\*=<0.001 vs. control, +++=p<0.001 vs. elective, #= p< 0.01 vs. before surgery).

In Acute group WBC numbers were significantly higher (upper left panel) and Lag time was significantly shorter (upper right panel) before and one week after the surgery, than the similar values in Elective and Control Groups (\*=p<0.05 vs. Control, # = p<0.05 vs. Elective groups). Slope of the free radical producing curves became steeper in acute and elective groups, signing that more and more active WBC are present before and one week after the surgery, but in Acute group this free radical generation was significantly higher, than in the other two groups. The maximum of free radical production continuously elevated both in acute and in elective groups, but these elevation in acute group was several times higher

**3.1.5.2 Investigation of lipid peroxidation due to measurement of MDA in plasma and in** 

It is a well known fact, that one of the main consequence of long-lasting atherosclerosis is the significant decrease in the polyunsaturated fatty acid (PUFA) content of the membranes, due to saturation of the membrane lipids which responsible for the rigidity of the membranes. MDA is one of the lipid peroxydation end products, generated in the course of

**A U m ax / 10 3 cells**

**\*\***\*

Control Elective Acute

> **\*\*\* \*\* # ##**

**# ++**

**Lag tim e (sec)**

**Before rurgery After surgery 24 hour later 1 week later**

**Before rurgery After surgery 24 hour later 1 week later**

**++ ++**

**\*\*\*** \*\*\* \*\*

**# \*\***

Control Elective Acute

**+**

\*\*\*

\* **+**

**# +++**

#### **3.1.4.3 SOD activity**

Before surgery SOD activity was lower in both patients groups, compared to Control, however SOD activity in Elective group was higher than in Acute group, and remained unchanged during the study. In Acute group SOD level decreased further 24 hours after the surgery. SOD levels of patients groups remained below the normal range in the course of the whole study (Figure 7.)

Fig. 7. **Changes in SOD activity.** \*= p<0.05, \*\*=p<0.01, \*\*=p<0.005, +=p<0.05 vs. elective, #=p<0,05 vs. before surgery

Before surgery SOD activity in both patients groups were lower compared to healthy volunteers. This low level decreased further significantly n Acute group until the 24th hour of reperfusion and remained in this low level during the study. In elective group SOD level was on a constant low-level during the study. I/R injury was described almost 50 years ago, but the correct mediation, the way of prevention or treatment is under investigation in nowadays, as well. The inflammatory responses can be detected following reperfusion varies greatly and depends on the time and severity of ischemia, as it was measured in our cases too. Restoration of blood flow of ischemic tissues initiates a chain reaction of complications which can categorize into two main groups: regional and systemic. In our case restoration of blood flow in lower limb, generated a systemic response, which had been detected in blood samples (in serum, haemolysate, plasma) obtained in a great distance from the intervention.

#### **3.1.5 Investigation of prooxidants**

#### **3.1.5.1 PMA induced free radical production**

Before surgery PMA induced ROS production was higher in Acute than in Elective and Control groups. This value elevated further in the course of the study. Free radical generation was characterized with four parameters: 1. the lag-time: the time elapsed between the induction and the beginning of the free radical production, 2. maximum of free radical Angioplasty, Various Techniques and Challenges in 72 Treatment of Congenital and Acquired Vascular Stenoses

Before surgery SOD activity was lower in both patients groups, compared to Control, however SOD activity in Elective group was higher than in Acute group, and remained unchanged during the study. In Acute group SOD level decreased further 24 hours after the surgery. SOD levels of patients groups remained below the normal range in the course of

**\* \*\*\***

Fig. 7. **Changes in SOD activity.** \*= p<0.05, \*\*=p<0.01, \*\*=p<0.005, +=p<0.05 vs. elective,

Before surgery SOD activity in both patients groups were lower compared to healthy volunteers. This low level decreased further significantly n Acute group until the 24th hour of reperfusion and remained in this low level during the study. In elective group SOD level was on a constant low-level during the study. I/R injury was described almost 50 years ago, but the correct mediation, the way of prevention or treatment is under investigation in nowadays, as well. The inflammatory responses can be detected following reperfusion varies greatly and depends on the time and severity of ischemia, as it was measured in our cases too. Restoration of blood flow of ischemic tissues initiates a chain reaction of complications which can categorize into two main groups: regional and systemic. In our case restoration of blood flow in lower limb, generated a systemic response, which had been detected in blood samples (in serum, haemolysate, plasma) obtained in a great distance from

Before surgery PMA induced ROS production was higher in Acute than in Elective and Control groups. This value elevated further in the course of the study. Free radical generation was characterized with four parameters: 1. the lag-time: the time elapsed between the induction and the beginning of the free radical production, 2. maximum of free radical

**\*\*\***

**After surgery 24 hours later 1 week later**

**\*\* \*\***
