Attention Deficit Hyperactivity Disorder (ADHD)
Conditions
Keywords
methylphenidate, Attention Deficit Hyperactivity Disorder, ADHD, oxidative stress
Brief summary
The aim of this study is to investigate the effect of methylphenidate treatment on oxidative stress by measuring the levels of Total Oxidant Status (TOS), Total Antioxidant Status (TAS), Oxidative Stress Index (OSI=TOS/TAS), Malondialdehyde (MDA), Oxidized LDL (Ox-LDL), and Superoxide Dismutase (SOD) in serum samples of patients diagnosed with Attention Deficit Hyperactivity Disorder (ADHD) who have either started or are planned to start methylphenidate treatment. The measurements will be taken at baseline (0 months, before treatment initiation) and at the 3rd month (after treatment has begun) to assess the effect of methylphenidate on oxidative stress. Patients who were already planned to begin methylphenidate treatment will be invited to participate in this study. Since this is not an interventional study, no additional treatments will be administered or altered beyond the treatment the patient is already required to receive.
Detailed description
The study population will consist of patients who are diagnosed with Attention Deficit Hyperactivity Disorder (ADHD) according to DSM-5-TR, who have applied to the child and adolescent psychiatry outpatient clinics of Bakırköy Prof. Dr. Mazhar Osman Mental Health and Neurological Diseases Training and Research Hospital and have been started or planned to start methylphenidate treatment by the physician they have been examined by. After an in-clinic announcement, these patients will be directed to the researcher, Enes Faruk Altunkılıç. After being informed both verbally and in writing through the informed consent process, those who agree to participate, sign the informed consent form, and meet the inclusion and exclusion criteria will be included in the study. It will also be explained to the patients and their families that their participation or non-participation in this study will not affect the treatment they will receive. After informed consent, a structured clinical interview for DSM-5-TR will be conducted using the Mood Disorders and Schizophrenia Form for School-Aged Children - Now and Lifetime DSM-5 - Turkish Adaptation (ÇDŞG-ŞY-DSM-5-T). To obtain sociodemographic and clinical data for the participants, the Sociodemographic and Clinical Data Form created by the researchers will be completed. For the ADHD diagnosed group, the Conners Parent Rating Scale - Revised Short and Conners Teacher Rating Scale - Revised Short will be applied to determine the severity of the disorder, symptoms, and predominant subtyping. After the diagnosis is made and evaluated according to the exclusion criteria, blood samples will be taken from the patient after a 10-12 hour fasting period, between 9-12 AM, in a yellow-capped tube, before the routine methylphenidate treatment is used. After waiting for 10-20 minutes at room temperature, the sample will be centrifuged at 3000 RPM for 20 minutes, and the serum will be collected in Eppendorf tubes and stored at -80°C until analysis. After 3 months of treatment, blood samples will be taken again in the same manner and stored. Once all the samples are collected, the serum samples will be analyzed for total antioxidant status (TAS), total oxidant status (TOS), malondialdehyde (MDA), superoxide dismutase (SOD), and oxidized LDL levels according to the human ELISA kit protocols at the Biochemistry Laboratory of Bakırköy Dr. Sadi Konuk Training and Research Hospital by biochemist Dr. Hacer Eroğlu İçli. After 3 months of treatment, the Conners Parent Rating Scale - Revised Short and Conners Teacher Rating Scale - Revised Short forms will be applied again. In addition to the markers, the Oxidative Stress Index (OSI = TOS/TAS) will be calculated and included in the evaluation before and 3 months after the treatment.
Interventions
OTHERThis is not an interventional study. Patients who are planned to start methylphenidate treatment have been invited to participate in the study.
This is not an interventional study. Patients diagnosed with Attention Deficit Hyperactivity Disorder (ADHD), for whom methylphenidate treatment is planned, have been invited to participate in the study. The patients have been enrolled in follow-up, and our contact information has been provided to them. As part of routine care, patients have been called in once a month. Necessary dosage adjustments have been made in accordance with medical guidelines, without any intervention for the study. Patients with complaints or those who needed to be seen earlier have been seen in between. As a result, the patients were followed for a total of 3 months for this study. Serum samples for oxidative stress markers were collected at baseline (before treatment started) and at the 3rd month (after treatment began).
Sponsors
Saglik Bilimleri Universitesi
Study design
Observational model
COHORT
Time perspective
PROSPECTIVE
Eligibility
Sex/Gender
ALL
Age
6 Years to 11 Years
Healthy volunteers
Yes
Inclusion criteria
* \- A diagnosis of Attention Deficit Hyperactivity Disorder according to DSM-5 TR, and the initiation or planned initiation of methylphenidate treatment as part of routine care. * Being between the ages of 6 and 11. * Providing consent to participate in the study after being informed about the study.
Exclusion criteria
* \- Presence of a psychiatric disorder diagnosis other than Attention Deficit Hyperactivity Disorder. * The diagnosis of Attention Deficit Hyperactivity Disorder has been made, but methylphenidate treatment has not been planned. * Being under the age of 6 or over the age of 11. * Presence of organic brain damage, mental retardation, autism spectrum disorder, neurological diseases, or any physical illness affecting neurocognitive functions. * A history of alcohol and/or psychoactive substance use. * Presence of ongoing active infection, allergic diseases, or chronic illness. * Previous use of psychiatric medication. * Presence of a chronic illness. * Use of regular medication.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Change in Total Oxidant Status | Immediately before starting the treatment and up to the 3rd month of treatment. | Total Oxidant Status (TOS) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. TOS was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1599Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution. |
| Change in Total Antioxidant Status | Immediately before starting the treatment and up to the 3rd month of treatment. | Total antioxidant status (TAS) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment.TAS was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E4350Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution. |
| Change in Superoxide Dismutas | Immediately before starting the treatment and up to the 3rd month of treatment. | superoxide dismutas level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. Superoxide dismutas was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E0918Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution. |
| Change in Malondialdehyde | Immediately before starting the treatment and up to the 3rd month of treatment. | Malondialdehyde (MDA) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. MDA was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1371Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution. |
| Change in Ox-LDL | Immediately before starting the treatment and up to the 3rd month of treatment. | Ox-LDL level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. Ox-LDL was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1521Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution. |
| Change in Oxidative Stress Index (OSI) | Immediately before starting the treatment and up to the 3rd month of treatment. | Oxidative Stress Index (OSI) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. OSI is an index created by the ratio of TOS to TAS. OSI is a parameter that shows the direction in which the body's oxidative stress balance shifts. An increase or decrease in the OSI value indicates that the oxidative balance has changed. While an increase in OSI value indicates a change in balance in favor of oxidants; a decrease in OSI value indicates a change in balance in favor of anti-oxidants. |
Countries
Turkey (Türkiye)
Participant flow
Pre-assignment details
Initially, 39 patients were enrolled in the study, and each patient who dropped out of the study during follow-up was replaced by a new patient in accordance with the protocol. Thus, a total of 53 patients were included in the study and 14 patients left the study for various reasons.
Participants by arm
| Arm | Count |
|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity Patients diagnosed with attention deficit hyperactivity and planned to start methylphenidate treatment | 39 |
| Total | 39 |
Withdrawals & dropouts
| Period | Reason | FG000 |
|---|---|---|
| Overall Study | concerns about possible side effects | 2 |
| Overall Study | Lost to Follow-up | 5 |
| Overall Study | medication refusal | 2 |
| Overall Study | side effects (dizziness) | 1 |
| Overall Study | stigmatization concerns | 2 |
| Overall Study | wanting to seek another specialist opinion | 2 |
Baseline characteristics
| Characteristic | Patients Diagnosed With Attention Deficit Hyperactivity | — |
|---|---|---|
| Age, Continuous | 8.38 years STANDARD_DEVIATION 1.11 | — |
| body mass index (BMI) | 16.60 kilogram(kg)/height(metre)² STANDARD_DEVIATION 1.96 | — |
| Body Weight | 27.95 kilogram (kg) STANDARD_DEVIATION 7.38 | — |
| Race and Ethnicity Not Collected | — | — Participants |
| Region of Enrollment Turkey | 39 participants | — |
| Sex: Female, Male Female | 26 Participants | — |
| Sex: Female, Male Male | 13 Participants | — |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | 0 / 53 |
| other Total, other adverse events | 26 / 53 |
| serious Total, serious adverse events | 0 / 53 |
Outcome results
Primary
Change in Malondialdehyde
Malondialdehyde (MDA) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. MDA was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1371Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
| Arm | Measure | Group | Value (MEDIAN) |
|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Malondialdehyde | Malondialdehyde (before starting treatment) | 9.58 nmol/ml |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Malondialdehyde | Malondialdehyde (after starting treatment) | 8.65 nmol/ml |
p-value: <0.001Wilcoxon (Mann-Whitney)
Primary
Change in Oxidative Stress Index (OSI)
Oxidative Stress Index (OSI) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. OSI is an index created by the ratio of TOS to TAS. OSI is a parameter that shows the direction in which the body's oxidative stress balance shifts. An increase or decrease in the OSI value indicates that the oxidative balance has changed. While an increase in OSI value indicates a change in balance in favor of oxidants; a decrease in OSI value indicates a change in balance in favor of anti-oxidants.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Oxidative Stress Index (OSI) | Oxidative Stress Index (OSI) (before starting treatment) | 0.75 Index | Standard Deviation 0.12 |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Oxidative Stress Index (OSI) | Oxidative Stress Index (OSI) (after starting treatment) | 0.72 Index | Standard Deviation 0.12 |
Primary
Change in Ox-LDL
Ox-LDL level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. Ox-LDL was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1521Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
| Arm | Measure | Group | Value (MEDIAN) |
|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Ox-LDL | Ox-LDL (before starting treatment) | 34.19 ng/mL |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Ox-LDL | Ox-LDL (after starting treatment) | 29.59 ng/mL |
p-value: 0.001Wilcoxon (Mann-Whitney)
Primary
Change in Superoxide Dismutas
superoxide dismutas level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. Superoxide dismutas was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E0918Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
Population: 2 patients were excluded because there was no reaction and no color change in the measurement.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Superoxide Dismutas | superoxide dismutas (before starting treatment | 175.89 U/L | Standard Deviation 158.95 |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Superoxide Dismutas | superoxide dismutas (after starting treatment) | 160.88 U/L | Standard Deviation 143.49 |
p-value: 0.003t-test, 2 sided
Primary
Change in Total Antioxidant Status
Total antioxidant status (TAS) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment.TAS was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E4350Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Total Antioxidant Status | total antioxidant status (before starting treatment) | 7.79 U/ml | Standard Deviation 5.65 |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Total Antioxidant Status | total antioxidant status (after starting treatment) | 7.33 U/ml | Standard Deviation 5.4 |
p-value: 0.021t-test, 2 sided
Primary
Change in Total Oxidant Status
Total Oxidant Status (TOS) level was measured according to the human ELISA kit protocol. Serum samples from 39 patients were compared immediately before and after 3 months of treatment. TOS was measured according to the human ELISA kit protocol as follows (bioassay technology laboratory, Cat.No E1599Hu): Standards added to wells. 40μl of serum sample was added to the sample wells, followed by 10μl of anti-TOS antibody. Incubated at 37°C for 60 minutes. plate was washed 5 times with wash buffer. 50μl of substrate solution A was added to the wells and then 50μl of substrate solution B was added to each well. The plate was incubated for 10 minutes at 37°C in the dark. 50μl of Stop Solution was added to the wells and the blue color immediately changed to yellow. The optical density (OD value) of each well was determined using a microplate reader set to 450 nm within 10 min after adding the stop solution.
Time frame: Immediately before starting the treatment and up to the 3rd month of treatment.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Total Oxidant Status | total oxidant status (before starting treatment) | 6.29 U/ml | Standard Deviation 5.37 |
| Patients Diagnosed With Attention Deficit Hyperactivity | Change in Total Oxidant Status | total oxidant status (after starting treatment) | 5.62 U/ml | Standard Deviation 4.76 |
p-value: <0.001t-test, 2 sided