**5. Results**

In this part of the article, we have a presentation of the results gained with processing, in other words, a statistic data analysis, needed for fulfilling the research aims set.

As respondents in the research, we've included 32 patients with clinically confirmed Idiopathic Parkinson's disease (IPD), diagnosed to Brain Bank Criteria, treated in the University Clinic of Neurology in Skopje – ward for extra pyramidal diseases.

The gender structure of the respondents is presented with 18 (56.25%) males and 14 (43.75%) females, table 1 and picture 1.

The average age of the respondents is 52, 7+10.3 years. The youngest respondent is 30 years and the oldest is 78 years. The median value with value of 50 years shows that 16 of the respondents (50%) are in the age group of 50-59 years, and even 23 of the patients are older than 50 years, table 2 and picture 2.


Table 1. Respondents distribution by gender

220 Neuroimaging for Clinicians – Combining Research and Practice

This is a prospective study, clinical and genetic, approved by the Ethical Committee, at Medical Faculty of Skopje. The clinical part of the study was made at University Clinic of Neurology in Skopje. Laboratory and genetic part of the study was made at the laboratory of molecular biology at Faculty of Math and Natural sciences at University of "St. Cyril and

In the study were included and were analyzed 30 patients. At each of them, Idiopathic Parkinson Disease was diagnosed, by using many diagnostic criteria such as: History of the patient's disease; Neurological examination; Neurophysiologic examinations such as EEG, ENMG, EP (VEP, SEP, BAEP); Neuroimaging methods, such as CT scan and MRI of brain;

*DNA Isolation:* under aseptic conditions, from each one of the patients, blood sample was taken, with the function of a vein. Each of those blood samples are kept invaccum pots

*Amplification:* amplification of regions of SNCA gene was made with polymerase chain

*Detection of mutations:* to detect the mutations in selected regions of SNCA gene, polymerase

Amplification of the region of egzon 3 of SNCA gene was made, using the primers 5'- GTC TCA CAC TTT GGA GGG TTT C-3' and 5' CAC CTA CCT ACA CAT ACC TCT GAC and

PCR amplification of region of egzon 4 of SNCA gene was made by using the primers 5' GCT AAT CAG CAA TTT AAG GCT AG-3' and 5' GAT ATG TTC TTA GAT GCT and

Amplificated products were digested with appropriate restrictive endonuclease under the

G88C mutation in egzon 3 of SNCA gene was detected with restricted digestion with

In this part of the article, we have a presentation of the results gained with processing, in

As respondents in the research, we've included 32 patients with clinically confirmed Idiopathic Parkinson's disease (IPD), diagnosed to Brain Bank Criteria, treated in the

The gender structure of the respondents is presented with 18 (56.25%) males and 14 (43.75%)

The average age of the respondents is 52, 7+10.3 years. The youngest respondent is 30 years and the oldest is 78 years. The median value with value of 50 years shows that 16 of the respondents (50%) are in the age group of 50-59 years, and even 23 of the patients are older

**4. Genetic analysis: Detection of mutations in gene of alpha-synuclein** 

(Vaccutainer), and anticoagulant EDTA Na2 is added in each of the pots.

chain reaction- Restriction Fragment Length Polymorphism, was used.

G209A mutation in egzon 4 of SNCA gene was detected with enzyme Mael.

other words, a statistic data analysis, needed for fulfilling the research aims set.

University Clinic of Neurology in Skopje – ward for extra pyramidal diseases.

CAG-3'. All of these amplifications have length of 215 bp.

**3. Materials and methodes** 

Methodius" in Skopje.

Neuropsychological tests.

**(SNCA)** 

reaction.

TC-3'.

optimal conditions.

females, table 1 and picture 1.

than 50 years, table 2 and picture 2.

enzyme Mval.

**5. Results** 

Fig. 1. Respondents distribution by gender


Table 2. Respondents distribution by age

In table 3 and picture 3, the representation of neurological symptoms with extra pyramidal origin, typical for diagnosed with IPD, is presented.

The objective neurogical finding, confirmed the presence of rigidity and hypertonia with extrapyramidal origin, to all 32 respondents. This also happened with tremor, which is one of the key symptoms of IPD.

Dyskinesia, as a symptom of long-lasting usage of substitution therapy (levodopa), is found in 15 patients (46.9%) and lack of these symptoms is seen in 17 patients (53.1%).


Table 3. Rigidity, tremor and dyskinesia distribution

Clinical and Genetic Aspects in Patients with Idiopathic Parkinson Disease 223

patients(6.25%) and in 4 patients(12.5%) a fully normal speech is present and the speech

Present 32 100 31 96.87 28 87.5 Absent / / 1 3.13 4 12.5 Total 32 100 32 100 32 100

Table 4. Bradykinesia, reduced postural reflexes and bradydilalia distribution

32 31

Fig. 4. Bradykinesia, reduced postural reflexes and bradydilalia distribution

bradykinesia reduced

(BAEP) and electroencephalography (EEG).

**N**

Table 5. Visual evoked potentials (VEP) in patients with IPD

The respondents gained the following neurophysiologic examinations: visual evoked potentials (VEP), somatosensory evoked potentials (SSEP), acoustic evoked potentials

postu. refl.

VEP N % Proper finding 29 90.62 Low-volted response 1 3.12 Prolonged latency 2 6.26 Total 32 100

The VEP is with proper finding in 29 respondents (90.6%), 2 of them (6.3%) have prolonged latency and the finding of one respondent shows low-volted response, table 5 and picture 5.

Number % Number % Number %

**1**

bradylalia

reflexes Bradylalia

28

**4**

present absent

itself is estimated with 0, according to UPDRS scale.

Symptoms Bradykinesia Reduced postural

Fig. 2. Respondents distribution by age

Fig. 3. Rigidity, tremor and dyskinesia distribution

Bradykinesia or stuntment in movements, as main of the list of cardinal symptoms, according to Brain Bank Criteria of UK Parkinson's Disease, was found present in all respondents.

Reduced postural reflexes are positive in dominant number of respondents, 31 (96.6%).

Bradydilalia or the changes in speech (stunt and monotonous speech), is found in 28 patients (87.5 %) and in 4 of our patients (12.5%) a fully normal speech is present, table 4 and picture 4.

Dominant patients with bradydilalia are those with minimal changes in speech, estimated by UPDRS scale, with 1 are 16 patients(50%), with 2,8 patients (25%), with 3 and 4, 2

**50**

Bradykinesia or stuntment in movements, as main of the list of cardinal symptoms, according to Brain Bank Criteria of UK Parkinson's Disease, was found present in all

Dominant patients with bradydilalia are those with minimal changes in speech, estimated by UPDRS scale, with 1 are 16 patients(50%), with 2,8 patients (25%), with 3 and 4, 2

Reduced postural reflexes are positive in dominant number of respondents, 31 (96.6%). Bradydilalia or the changes in speech (stunt and monotonous speech), is found in 28 patients (87.5 %) and in 4 of our patients (12.5%) a fully normal speech is present, table 4 and

rigidity tremor dyskinesia

**15,62**

15

17

**6,26**

30-39 40-49 50-59 60-69 70>

present absent

**12,5**

Fig. 3. Rigidity, tremor and dyskinesia distribution

32 32

Fig. 2. Respondents distribution by age

respondents.

picture 4.

**15,62**

patients(6.25%) and in 4 patients(12.5%) a fully normal speech is present and the speech itself is estimated with 0, according to UPDRS scale.


Table 4. Bradykinesia, reduced postural reflexes and bradydilalia distribution

Fig. 4. Bradykinesia, reduced postural reflexes and bradydilalia distribution

The respondents gained the following neurophysiologic examinations: visual evoked potentials (VEP), somatosensory evoked potentials (SSEP), acoustic evoked potentials (BAEP) and electroencephalography (EEG).

The VEP is with proper finding in 29 respondents (90.6%), 2 of them (6.3%) have prolonged latency and the finding of one respondent shows low-volted response, table 5 and picture 5.


Table 5. Visual evoked potentials (VEP) in patients with IPD

Proper finding

Low volted respond

Clinical and Genetic Aspects in Patients with Idiopathic Parkinson Disease 225

22 of the respondents (68.7%) have proper finding of caustic evoked potentials BAEP and 10

19 of the responders (59.4%) have proper finding of electroencephalography; the finding of 12 of them (37.5%) shows unstable basic brain activity, without pathological graph elements and in only one patient, unstable brain activity with presence of different types of

0

EEG findings N %

Proper finding 19 59.37

Total 32 100

12 37.5

Proper finding

Low volted response

1 3.13

of them (31.2%) are with low-volted responses, table 7 and picture 7.

Table 7. Caustic evoked potentials BAEP in patients with IPD

Fig. 7. Caustic evoked potentials BAEP in patients with IPD

10

22

pathological graph elements is found, table 8 and picture 8.

Unstable basic brain activity without pathological

Unstable basic brain activity, with presence of

graph elements

0

5

10

15

20

25 **N**

pathological graph elements

Table 8. EEG in patients with IPD

BAEP N %

Proper finding 22 68.75 Low-volted responses 10 31.25 Total 32 100

Prolonged latency

30 of the respondents have proper finding of somatosensory potentials SSEP and in 2 of them (6.2%), those potentials show low-volted responses, table 6 and picture 6.

Fig. 5. Visual evoked potentials (VEP) in patients with IPD


Table 6. Somatosensor evoked potentials in patients with IPD

Fig. 6. Somatosensory evoked potentials in patients with IPD

30 of the respondents have proper finding of somatosensory potentials SSEP and in 2 of

them (6.2%), those potentials show low-volted responses, table 6 and picture 6.

Fig. 5. Visual evoked potentials (VEP) in patients with IPD

90,62

0

20

40

60

80

100

**%**

Table 6. Somatosensor evoked potentials in patients with IPD

6, 25%

Fig. 6. Somatosensory evoked potentials in patients with IPD

SSEP N %

3,12 6,26

Proper finding 30 93.75 Low-volted responses 2 6.25 Total 32 100

93,75%

Proper finding

Proper finding

Low volted respond

Prolonged latency

Low volted response

22 of the respondents (68.7%) have proper finding of caustic evoked potentials BAEP and 10 of them (31.2%) are with low-volted responses, table 7 and picture 7.


Table 7. Caustic evoked potentials BAEP in patients with IPD

Fig. 7. Caustic evoked potentials BAEP in patients with IPD

19 of the responders (59.4%) have proper finding of electroencephalography; the finding of 12 of them (37.5%) shows unstable basic brain activity, without pathological graph elements and in only one patient, unstable brain activity with presence of different types of pathological graph elements is found, table 8 and picture 8.


Table 8. EEG in patients with IPD

Clinical and Genetic Aspects in Patients with Idiopathic Parkinson Disease 227

In 12 patients (37.5%), the results of CT and MRI of brain are normal; 11 (34.4%) have

Doppler color sonography of the extra cranial blood vessels is with proper finding in 20 respondents (62.5%), 7 (21.9%) are with lightly expressed intimacy and proper hemodynamic parameters; 5(15.6%) have expressed arteriosclerosical changes of carotidal

7 21.87

5 15.63

Normal finding

Lightly changes

Expressed changes

Doppler color sonography of the carotid system N % Normal finding 20 62.5

Total 32 100

Table 10. Doppler color sonography of the carotid system in patients with IPD

Fig. 10. Doppler color sonography of the carotid system in patients with IPD

**21,87 15,63**

global reduced cognitive and mnestical capacities, found in 3 respondents (9.4%).

In tables 11a and 11b, the results of the neuropsychological testing are shown. It's visible that only 3 patients (9.4%) have proper cognitive capacities. In table 11a all neuropsychological findings are examined individually, with percentage representation in our 32 respondents. Depression is found in 24 patients (84.4%); anxiety in 3 patients (9.4%); in 13 patients (40.6%), initial or global cognitive and mnestic reduced changes are found. In table 11b, a statistic analysis of different combinations of findings from neuropsychological testing is presented, so between the patients dominant are those with depression and initial reduced cognitive capacities, as well as global reduced cognitive capacities. According to the frequency of occurrence, next is the finding of depression and

The rest findings from the neuropsychological testing: depression, anxiety, initial reduced cognitive capacities, depression and anxiety together, as well as the mutual presence of

discrete cortical reductive changes and 9 (28, 1 %) have global cortical changes.

Lightly expressed intimacy with proper chemo dynamic

Expressed arteriosclerotic changes of the blood vessels of the

**62,5**

blood vessels, table 10 and picture 10.

parameters

**%**

carotid system

Fig. 8. EEG in patients with IPD


Table 9. CT and MRI of brain in patients with IPD

Fig. 9. CT and MRI of brain in patients with IPD

All patients gained neuroimaging examinations (CT and MRI of brain) and the examination results are presented in table 9 and picture 9.

Proper findings

Unstable brain activity without grapholements

Unstable brain activity with grapholements

Proper finding

Discrete cortical reduc. changes

Global cortical reductive changes

CT and MRI of brain findings N % Proper finding 12 37.5 Discrete cortical reductive changes 11 34.37 Global cortical reductive changes 9 28.13 Total 32 100

**3,1**

All patients gained neuroimaging examinations (CT and MRI of brain) and the examination

**28,13**

Fig. 8. EEG in patients with IPD

**%**

**%**

Table 9. CT and MRI of brain in patients with IPD

**37,5 34,37**

**59,3**

**37,5**

Fig. 9. CT and MRI of brain in patients with IPD

results are presented in table 9 and picture 9.

In 12 patients (37.5%), the results of CT and MRI of brain are normal; 11 (34.4%) have discrete cortical reductive changes and 9 (28, 1 %) have global cortical changes.

Doppler color sonography of the extra cranial blood vessels is with proper finding in 20 respondents (62.5%), 7 (21.9%) are with lightly expressed intimacy and proper hemodynamic parameters; 5(15.6%) have expressed arteriosclerosical changes of carotidal blood vessels, table 10 and picture 10.


Fig. 10. Doppler color sonography of the carotid system in patients with IPD

In tables 11a and 11b, the results of the neuropsychological testing are shown. It's visible that only 3 patients (9.4%) have proper cognitive capacities. In table 11a all neuropsychological findings are examined individually, with percentage representation in our 32 respondents. Depression is found in 24 patients (84.4%); anxiety in 3 patients (9.4%); in 13 patients (40.6%), initial or global cognitive and mnestic reduced changes are found.

In table 11b, a statistic analysis of different combinations of findings from neuropsychological testing is presented, so between the patients dominant are those with depression and initial reduced cognitive capacities, as well as global reduced cognitive capacities. According to the frequency of occurrence, next is the finding of depression and global reduced cognitive and mnestical capacities, found in 3 respondents (9.4%).

The rest findings from the neuropsychological testing: depression, anxiety, initial reduced cognitive capacities, depression and anxiety together, as well as the mutual presence of

Clinical and Genetic Aspects in Patients with Idiopathic Parkinson Disease 229

the distribution shown in the table. It is noticeable that half of the male respondents (9.50%), have light disease symptoms; 2 (11.1%) have minimal signs or symptoms according to UPDRS scale; 4 (22.2%) are with extremely visible symptoms. In the female respondents, the disease is often manifested with minimal symptoms; 9(64.3%) and 4(28.6%) have light symptoms and there aren't any female respondents with extremely visible symptoms.

**9**

**4**

**1**

**0**

Minimal signs

Expressed signs

Extremely expressed

Light signs

signs

Fig. 11. The immensity of the clinical state, according to the respondents gender

males females

symptoms of Parkinson's disease, is 53, 6+10.3 years.

**3**

4

**2**

**9**

insignificant (p>0.05).

The average age of the respondents, which according to UPDRS scale, have minimal

The respondents with light signs have average age of 52.8+8.1 years; the respondents with extremely expressed symptoms have average age of 45+15.3 years, table 13 and picture 12.

The connection between the age and the immensity of the clinical state of the respondents, estimated by UPDRS scale is presented in picture 13. That correlation has value for Spearmenov's correlation coefficient, R=0.039, which indicates indirect or negative correlation. Parkinson's disease has more difficult clinical state in younger patients and vice versa lighter symptoms in older patients. But, considering the value of the coefficient, we can conclude that the correlation between these two parameters is weak and statistically

UPDRS N Age (Mean) Age (SD) Minimal signs 11 53.63 10.28 Light signs 13 52.85 8.10 Expressed signs 4 57.75 11.73 Extremely expressed symptom 4 45.00 15.30 Table 13. The immensity of the clinical state (UPDRS), according to the patient's age

depression and anxiety, joined with initial reduced cognitive and mnestical capacities, are found only in one of the respondents.


Table 11.a. Neuropsychological testing of respondents with IPD


Table 11.b. Neuropsychological testing of respondents with IPD


Table 12. The immensity of the clinical state, according to the respondents' gender Mann-Whitney U=47,5 Z=2,98 p=0,0029

The immensity of the clinical state of Parkinson's disease in our respondents, of different gender, is presented in table 12 and picture 11. The male respondents have significantly tougher clinical state comparing to female respondents. This statistic comment is a result of

depression and anxiety, joined with initial reduced cognitive and mnestical capacities, are

Neuropsychological testing N % Proper finding 3 9.37 Depression 27 84.37 Anxiety 3 9.37 Initial reduced cognitive and mnestical capacities 13 40.62 Global reduced cognitive and mnestical capacities 13 40.62

Neuropsychological testing N % Proper finding 3 9.37 Depression 1 3.12 Anxiety 1 3.12 Global reduced cognitive and mnestic capacities 10 31.25 Initial reduced cognitive and mnestic capacities 1 3.12 Depression and anxiety 1 3.12 Depression and global reduced cognitive and mnestic capacities 3 9.37

Total 32 100

UPDRS Gender Total Males Females

11.11%

50.00%

16.67%

18 56.25%

Table 12. The immensity of the clinical state, according to the respondents' gender Mann-

The immensity of the clinical state of Parkinson's disease in our respondents, of different gender, is presented in table 12 and picture 11. The male respondents have significantly tougher clinical state comparing to female respondents. This statistic comment is a result of

11 34.37

1 3.12

9

4

1

14

22.22% 0 4

64.29% <sup>11</sup>

28.57% <sup>13</sup>

7.14% <sup>4</sup>

43.75% <sup>32</sup>

Table 11.a. Neuropsychological testing of respondents with IPD

Depression and initial reduced cognitive and mnestical

Depression and anxiety and initial reduced cognitive and

Table 11.b. Neuropsychological testing of respondents with IPD

Minimal signs 2

Light signs 9

Expressed symptoms 3

Extremely expressed symptoms 4

found only in one of the respondents.

capacities

Total

% of total number

Whitney U=47,5 Z=2,98 p=0,0029

mnestical capacities

the distribution shown in the table. It is noticeable that half of the male respondents (9.50%), have light disease symptoms; 2 (11.1%) have minimal signs or symptoms according to UPDRS scale; 4 (22.2%) are with extremely visible symptoms. In the female respondents, the disease is often manifested with minimal symptoms; 9(64.3%) and 4(28.6%) have light symptoms and there aren't any female respondents with extremely visible symptoms.

Fig. 11. The immensity of the clinical state, according to the respondents gender

The average age of the respondents, which according to UPDRS scale, have minimal symptoms of Parkinson's disease, is 53, 6+10.3 years.

The respondents with light signs have average age of 52.8+8.1 years; the respondents with extremely expressed symptoms have average age of 45+15.3 years, table 13 and picture 12.


Table 13. The immensity of the clinical state (UPDRS), according to the patient's age

The connection between the age and the immensity of the clinical state of the respondents, estimated by UPDRS scale is presented in picture 13. That correlation has value for Spearmenov's correlation coefficient, R=0.039, which indicates indirect or negative correlation. Parkinson's disease has more difficult clinical state in younger patients and vice versa lighter symptoms in older patients. But, considering the value of the coefficient, we can conclude that the correlation between these two parameters is weak and statistically insignificant (p>0.05).

Clinical and Genetic Aspects in Patients with Idiopathic Parkinson Disease 231

In our research, we've concluded that direct or positive statistic significant correlation between the number of hospitalizations and the immensity of the clinical state, according to

The value of the Spermen's coefficient (r=0.48), shows that these two parameters are proportionally connected, in other words, patients with IPD difficult manifestations of the disease, according to UPDRS scale, have more hospitalizations in the University clinic of

Table 14 and picture 14, show the results from the crostabulation between the CT finding and MRI of brain and the immensity of the clinical state, determined according UPDRS scale. In the respondents group with light disease symptoms, according to UPDRS scale, dominant are the findings with normal level, previously determined with CT and MRI of brain, present in 5 patients (45.4%), from this group. Between the respondents with light disease symptoms, the finding from CT and MRI of the brain shows discrete cortical reduced changes in 6 patients (46.1%). In half of the respondents, the finding from CT and MRI of brain, shows presence of discrete cortical reduced changes.75% of the respondents with highly expressed disease symptoms, have global cortical reduced changes, proved on

Immensity of the clinical state (UPDRS)

signs Light signs Expressed

5 38.46%

6 46.15%

2 15.38%

Table 14. Immensity of the clinical state (UPDRS) in relation to CT and MRI of brain

confirmed correlation is statistically insignificant, (p>0.05), picture 15.

Total 11 13 4 4 32

The connection or the correlation between the immensity of the clinical state, determined by UPDRS scale and brain CT/MRI finding, is also proved with the usage of Spearsmen's correlation coefficient. Its value (r=0.2) shows presence of direct, positive correlation between these two parameters, in other words, there is a match between the brain CT/MRI finding and the results from the UPDRS scale. In patients with difficult clinical forms of IPD, most often the brain CT/MRI finding shows global cortical reduced changes. But, the

Minimal Total

signs

1 25.00%

2

1 25.00% Extremely expressed signs

1

3

50.00% 0 11

25.00% <sup>12</sup>

75.00% <sup>9</sup>

**6. Spearman rank order correlations R=-0.039 p>0.05** 

UPDRS, indeed exists.

Neurology in Skopje.

CT and MRI of brain.

Normal finding 5

45.45%

3 27.27%

3 27.27%

CT and MRI of

brain

Discrete cortical reductive changes

Global cortical reductive changes

Fig. 12. The immensity of the clinical state (UPDRS), according to the patient's age

Fig. 13. Correlation between the immensity of the clinical state, according to UPDRS and the age of the respondents with IPD
