Amyotrophic Lateral SclerosisIMPACT-ALSNCT07178067
Longitudinal Evaluation of Short-Chain Fatty Acid Profiles During the Natural Disease Course and a Mediterranean Diet Intervention in Amyotrophic Lateral Sclerosis Patients
An observational study of Amyotrophic Lateral Sclerosis, run by George Emil Palade University of Medicine, Pharmacy, Sciences and Technology of Targu Mures at 1 site in Romania.
Trial summary
This observational study explored the connection between the gut microbiota and the brain in patients with amyotrophic lateral sclerosis (ALS), specifically the modulation of short-chain fatty acids during disease progression and after following a Mediterranean diet for 6 months. Read more about this study
- Who is it for?
- Ages 18 years and olderHealthy volunteers can take part
- Conditions the study is for
- Amyotrophic Lateral Sclerosis
- What is being tested?
- Mediterranean diet
- How many people
- 44 participants
- Where
- 1 siteRomania
- Study dates
- 21 Jul 2022to 30 Nov 2023
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Study population: This prospective, single-center study enrolled adults from Romania diagnosed with ALS at Neurology Clinic 1, Mureș County Clinical Emergency Hospital , with an ALS diagnosis within the past 12 months, disease duration under 1 year, El Escorial (1996) criteria-clinically probable or clinically definite ALS. ALS cohort: N=44 at baseline (T0, Aug-Oct 2022); mean age 58.4 ± 12.5 years; 28 men / 16 women. Follow-up: T1 (6 months): N=36 (8 deaths by T1). T2 (12 months): N=30 (6 additional deaths between T1 and T2). Healthy controls: N=40, age- and sex-matched community volunteers without known neurodegenerative or major systemic disease. The patients were recruited through consecutive recruitment at the Neurology Clinic 1, Mureș County Clinical Emergency Hospital (Târgu Mureș, Romania) over a 3-month window (01 Aug-31 Oct 2022).
Who may be able to join
- Patients diagnosed with amyotrophic lateral sclerosis (ALS) in the past 12 months.
- Disease duration of less than 1 year.
- Fulfillment of the El Escorial diagnostic criteria (1996) for clinically probable or clinically definite ALS.
Who cannot join
- Patients with an expected survival <18 months.
- Presence of other neurodegenerative diseases.
- Major comorbidities, such as: heart failure, cancer, autoimmune diseases, systemic diseases.
- Gastrointestinal diseases that could influence gut microbiota.
- Recent or ongoing treatment with antibiotics, probiotics, or prebiotics.
- Low compliance with study procedures.
Trial locations and contacts
1 site in 1 country.
Romania1 site
University of Medicine, Pharmacy, Science and Technology of Târgu Mureș 'George Emil Palade'
Târgu Mureş, Mureș County, 540142
About this study
Official title: Short Chain Fatty Acids Profiles in Amyotrophic Lateral Sclerosis: Longitudinal Effects of Disease and Mediterranean Diet Intervention
This observational study explored the connection between the gut microbiota and the brain in patients with amyotrophic lateral sclerosis (ALS), specifically the modulation of short-chain fatty acids during disease progression and after following a Mediterranean diet for 6 months. Recent research suggests that the gut microbiome-the community of bacteria and other microorganisms living in our intestines-may influence how ALS develops and progresses. The hypothesis was that changes in the gut microbiome and the substances it produces, such as short-chain fatty acids (SCFAs), may play an important role in ALS progression. Additionally, the effect of the Mediterranean diet on circulating short-chain fatty acid concentrations was assessed.
Read the detailed description
This study investigates the role of the microbiome-gut-brain axis in the progression of amyotrophic lateral sclerosis (ALS), with a focus on short-chain fatty acids (SCFAs) and their associated biomarker panel (SCFAGGYC-PROF). The investigators hypothesized that alterations in the gut microbiota composition and SCFA metabolism contribute to the heterogeneity of ALS phenotypes, influencing whether the disease follows a rapidly progressive or more slowly progressive course.
Participants with ALS and healthy controls undergo blood sampling and stool collection at baseline and follow-up. Serum analyses include: measurement of circulating SCFA levels using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
In a subset of ALS patients, dietary interventions consistent with a Mediterranean dietary pattern are introduced to evaluate the impact of nutrition on gut microbiota composition, SCFA production, and clinical disease progression. This 12-month study (6 months natural course + 6 months dietary intervention) evaluated Mediterranean diet effects on ALS progression rate and circulating levels of SCFA. The diet-known for neuroprotective and anti-inflammatory properties-was associated with significant changes in SCFA plasma levels.
A key process stage involved developing and applying an LC-MS/MS method for plasma quantification of acetic (C2), propionic (C3), butyric (C4), 3-hydroxybutyric (3-OH-C4), hexanoic (C6), and iso-hexanoic acid (Iso-C6; 4-methyl-valeric), using acetic acid-C13, butyric-d8 and hexanoic-d11 as internal standards.
After extraction and derivatization of analytes from plasma, samples were analyzed by reversed-phase LC and detected via MS/MS in MRM mode with negative ESI. The method enabled quantitative determination of free plasma carboxylic acids.
Analytical development and performance verification were carried out for simultaneous analysis of 6 short-chain carboxylic acids plus 3 homologous internal standards via LC-MS/MS with prior derivatization using EDC, 3-NPH, and pyridine. Validation covered LOD/LOQ, linear range and signal-concentration dependence (R>0.9900), accuracy (85-115%), inter-series precision (<15%), and carryover (none), ensuring confidence in the method.
Working solutions were prepared from STOCK/STD solutions fresh each day for method/derivatization checks and for spiking blank plasma at defined levels to build daily calibration curves for routine analysis. Solvent water:methanol 3:7 (v/v) was freshly prepared each analysis day.
Samples were prepared fresh on analysis day and immediately loaded in the autosampler. Samples were grouped into 4 batches: Control, T0, T1, T2. Plasmas were thawed once and equilibrated to room temperature. Sample preparation mirrored that of calibration/control plasma, spiked only with internal standards.
Quantification of SCFAs in Control (n=40), T0 (n=44), T1 (n=36), T2 (n=30). Each batch was run independently with fresh daily calibration curves from pooled healthy plasma spiked at method concentrations. Blanks (unspiked plasma and ISTD-only plasma) were also run to assess endogenous signal. Curves were 6-point linear (R>0.9900), used for quantification with internal-standard peak area correction: acetate C2, propionate C3, and 3-OH-C4 with acetate-C13 ISTD; butyrate C4 with butyrate-d8 ISTD; iso-hexanoic (Iso-C6) and hexanoic C6 with hexanoic-d11 ISTD.
Statistical interpretation using descriptive statistics per analyte and batch: means, SD, %CV (RSD), min, max. Chromatographic peaks processed in MultiQuant (Sciex) linked to Analyst 1.7.1; data/statistics in Microsoft Excel 2019. Method validated for sensitivity, selectivity, linearity, LLOQ, carryover, within-run/between-run precision and accuracy, and recovery.
- Conditions
- Amyotrophic Lateral Sclerosis
- Being tested
- Mediterranean diet
- Keywords
- short-chain fatty acid, amyotrophic lateral sclerosis, microbiota-gut-brain axis, LC-MS/MS
- Medical subject headings
- Amyotrophic Lateral Sclerosis
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Mediterranean dietOther
The Mediterranean diet, characterised by a high intake of fruits, vegetables, whole grains, legumes, nuts, and olive oil, along with moderate consumption of fish and poultry, has been associated with anti-inflammatory and neuroprotective effects. Notably, this dietary pattern has the potential to enhance the production of short-chain fatty acids (SCFAs) and support gut microbial diversity, representing a promising strategy for nutritional intervention in patients with ALS.
| Study group | Role | What they receive |
|---|---|---|
| ALS patients | 44 patients diagnosed with amyotrophic lateral sclerosis (ALS) in the past 12 months. Receives: Other: Mediterranean diet |
What is being measured
The primary outcome is the main question the study is designed to answer.
Analysis of serum SCFA levels in ALS patients and healthy controls, and evaluating the change in circulating SCFA concentrations associated with Mediterranean diet intervention
This study compares plasma levels of short-chain fatty acids (SCFAs): acetic acid (ng/mL), propanoic acid (ng/mL), butyric acid (ng/mL), 3-OH butyric acid, 4-methyl-valeric acid (ng/mL) between adults with amyotrophic lateral sclerosis (ALS) and age/sex-matched healthy controls at baseline (T0) and then tracks how these levels change after 6 months of natural disease progression (T1) and 6 months after introducing a Mediterranean diet (T2) in ALS patients. Outcomes: Mean SCFA concentrations (±SD) (ng/mL) at T0 between-group (ALS vs control) differences at baseline, and within-patient changes over time to assess whether the Mediterranean diet is associated with favorable SCFA shifts (T0-T1, T1-T2).
Measured: 12 months
Secondary outcomes (1)
Association of Serum SCFA Levels With ALS Progression Measured by ALSFRS-R at T0, T1, and T2
The investigators analysed whether mean plasma levels of short-chain fatty acids (SCFAs)-acetate, propionate, butyrate, 3-hydroxybutyrate, hexanoate, and iso-hexanoate (ng/mL)-are associated with clinical progression of amyotrophic lateral sclerosis (ALS) (ALSFRS-R progression rate (ΔPR).
Measured: 12 months
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted17 Sep 2025
- Study start21 Jul 2022
- Primary completion30 Nov 2023
- Study completion30 Nov 2023
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (1)Papers the sponsor has linked to this study.
- Other trials for Amyotrophic Lateral SclerosisStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Prospective
- Enrolment
- 44 participants (actual)
Sponsor and investigators
- Lead sponsor
- George Emil Palade University of Medicine, Pharmacy, Sciences and Technology of Targu Mures
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Motataianu Anca, George Emil Palade University of Medicine, Pharmacy, Sciences and Technology of Targu Mures
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07178067
- Sponsor's protocol ID
- TE 74/2022
- Other IDs
- PN-III-P1-1.1-TE-2021-0960
- Acronym
- IMPACT-ALS
- Status last verified
- Sep 2025
- First submitted
- 3 Sep 2025
- Last updated
- 17 Sep 2025
- Results posted
- No
- Data monitoring committee
- Yes
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No: Given the small, regionally identifiable ALS cohort and the rarity of the condition, the risk of re-identification remains high even after de-identification.
Publications
Motataianu A, Ion V, Dumitreasa M, Ormenisan I, Farczadi L, Andone S, Balasa R, Roman MM. Short-Chain Fatty Acid Profiles in Amyotrophic Lateral Sclerosis: Longitudinal Effects of Disease and Mediterranean Diet Intervention. Biomolecules. 2025 Sep 28;15(10):1380. doi: 10.3390/biom15101380.
PubMed 41154611
Study at a glance
From the registry record, last updated by the sponsor on 17 Sep 2025. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Short Chain Fatty Acids Profiles in Amyotrophic Lateral Sclerosis: Longitudinal Effects of Disease and Mediterranean Diet Intervention
- ClinicalTrials.gov ID
- NCT07178067
- Acronym / sponsor ID
- IMPACT-ALS / TE 74/2022
- Conditions
- Amyotrophic Lateral Sclerosis
- Keywords
- short-chain fatty acid, amyotrophic lateral sclerosis, microbiota-gut-brain axis, LC-MS/MS
- Study type
- Observational
- Recruitment status
- Completed
- Enrolment
- 44 participants (actual)
- Lead sponsor
- George Emil Palade University of Medicine, Pharmacy, Sciences and Technology of Targu MuresUniversity, hospital or other
- Study start
- 21 Jul 2022 (actual)
- Primary completion
- 30 Nov 2023 (actual)
- Study completion
- 30 Nov 2023 (actual)
- First posted
- 17 Sep 2025 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
This observational study explored the connection between the gut microbiota and the brain in patients with amyotrophic lateral sclerosis (ALS), specifically the modulation of short-chain fatty acids during disease progression and after following a Mediterranean diet for 6 months. Recent research suggests that the gut microbiome-the community of bacteria and other microorganisms living in our intestines-may influence how ALS develops and progresses. The hypothesis was that changes in the gut microbiome and the substances it produces, such as short-chain fatty acids (SCFAs), may play an important role in ALS progression. Additionally, the effect of the Mediterranean diet on circulating short-chain fatty acid concentrations was assessed.
Detailed description
This study investigates the role of the microbiome-gut-brain axis in the progression of amyotrophic lateral sclerosis (ALS), with a focus on short-chain fatty acids (SCFAs) and their associated biomarker panel (SCFAGGYC-PROF). The investigators hypothesized that alterations in the gut microbiota composition and SCFA metabolism contribute to the heterogeneity of ALS phenotypes, influencing whether the disease follows a rapidly progressive or more slowly progressive course.
Participants with ALS and healthy controls undergo blood sampling and stool collection at baseline and follow-up. Serum analyses include: measurement of circulating SCFA levels using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
In a subset of ALS patients, dietary interventions consistent with a Mediterranean dietary pattern are introduced to evaluate the impact of nutrition on gut microbiota composition, SCFA production, and clinical disease progression. This 12-month study (6 months natural course + 6 months dietary intervention) evaluated Mediterranean diet effects on ALS progression rate and circulating levels of SCFA. The diet-known for neuroprotective and anti-inflammatory properties-was associated with significant changes in SCFA plasma levels.
A key process stage involved developing and applying an LC-MS/MS method for plasma quantification of acetic (C2), propionic (C3), butyric (C4), 3-hydroxybutyric (3-OH-C4), hexanoic (C6), and iso-hexanoic acid (Iso-C6; 4-methyl-valeric), using acetic acid-C13, butyric-d8 and hexanoic-d11 as internal standards.
After extraction and derivatization of analytes from plasma, samples were analyzed by reversed-phase LC and detected via MS/MS in MRM mode with negative ESI. The method enabled quantitative determination of free plasma carboxylic acids.
Analytical development and performance verification were carried out for simultaneous analysis of 6 short-chain carboxylic acids plus 3 homologous internal standards via LC-MS/MS with prior derivatization using EDC, 3-NPH, and pyridine. Validation covered LOD/LOQ, linear range and signal-concentration dependence (R>0.9900), accuracy (85-115%), inter-series precision (<15%), and carryover (none), ensuring confidence in the method.
Working solutions were prepared from STOCK/STD solutions fresh each day for method/derivatization checks and for spiking blank plasma at defined levels to build daily calibration curves for routine analysis. Solvent water:methanol 3:7 (v/v) was freshly prepared each analysis day.
Samples were prepared fresh on analysis day and immediately loaded in the autosampler. Samples were grouped into 4 batches: Control, T0, T1, T2. Plasmas were thawed once and equilibrated to room temperature. Sample preparation mirrored that of calibration/control plasma, spiked only with internal standards.
Quantification of SCFAs in Control (n=40), T0 (n=44), T1 (n=36), T2 (n=30). Each batch was run independently with fresh daily calibration curves from pooled healthy plasma spiked at method concentrations. Blanks (unspiked plasma and ISTD-only plasma) were also run to assess endogenous signal. Curves were 6-point linear (R>0.9900), used for quantification with internal-standard peak area correction: acetate C2, propionate C3, and 3-OH-C4 with acetate-C13 ISTD; butyrate C4 with butyrate-d8 ISTD; iso-hexanoic (Iso-C6) and hexanoic C6 with hexanoic-d11 ISTD.
Statistical interpretation using descriptive statistics per analyte and batch: means, SD, %CV (RSD), min, max. Chromatographic peaks processed in MultiQuant (Sciex) linked to Analyst 1.7.1; data/statistics in Microsoft Excel 2019. Method validated for sensitivity, selectivity, linearity, LLOQ, carryover, within-run/between-run precision and accuracy, and recovery.
Study design
- Observational model
- Cohort
- Time perspective
- Prospective
- Number of arms
- 1
| Arm | Type | Receives |
|---|---|---|
| ALS patients | – | Other: Mediterranean diet44 patients diagnosed with amyotrophic lateral sclerosis (ALS) in the past 12 months. |
Who may qualify
- Ages
- 18 years and older
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult, Older adult
- Study population
- This prospective, single-center study enrolled adults from Romania diagnosed with ALS at Neurology Clinic 1, Mureș County Clinical Emergency Hospital , with an ALS diagnosis within the past 12 months, disease duration under 1 year, El Escorial (1996) criteria-clinically probable or clinically definite ALS. ALS cohort: N=44 at baseline (T0, Aug-Oct 2022); mean age 58.4 ± 12.5 years; 28 men / 16 women. Follow-up: T1 (6 months): N=36 (8 deaths by T1). T2 (12 months): N=30 (6 additional deaths between T1 and T2). Healthy controls: N=40, age- and sex-matched community volunteers without known neurodegenerative or major systemic disease. The patients were recruited through consecutive recruitment at the Neurology Clinic 1, Mureș County Clinical Emergency Hospital (Târgu Mureș, Romania) over a 3-month window (01 Aug-31 Oct 2022).
Key inclusion criteria (3 listed)
- Patients diagnosed with amyotrophic lateral sclerosis (ALS) in the past 12 months.
- Disease duration of less than 1 year.
- Fulfillment of the El Escorial diagnostic criteria (1996) for clinically probable or clinically definite ALS.
Key exclusion criteria (6 listed)
- Patients with an expected survival <18 months.
- Presence of other neurodegenerative diseases.
- Major comorbidities, such as: heart failure, cancer, autoimmune diseases, systemic diseases.
- Gastrointestinal diseases that could influence gut microbiota.
- Recent or ongoing treatment with antibiotics, probiotics, or prebiotics.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Mediterranean dietThe Mediterranean diet, characterised by a high intake of fruits, vegetables, whole grains, legumes, nuts, and olive oil, along with moderate consumption of fish and poultry, has been associated with anti-inflammatory and neuroprotective effects. Notably, this dietary pattern has the potential to enhance the production of short-chain fatty acids (SCFAs) and support gut microbial diversity, representing a promising strategy for nutritional intervention in patients with ALS. | Other | ALS patients |
Primary outcome
Analysis of serum SCFA levels in ALS patients and healthy controls, and evaluating the change in circulating SCFA concentrations associated with Mediterranean diet intervention
Time frame: 12 months
This study compares plasma levels of short-chain fatty acids (SCFAs): acetic acid (ng/mL), propanoic acid (ng/mL), butyric acid (ng/mL), 3-OH butyric acid, 4-methyl-valeric acid (ng/mL) between adults with amyotrophic lateral sclerosis (ALS) and age/sex-matched healthy controls at baseline (T0) and then tracks how these levels change after 6 months of natural disease progression (T1) and 6 months after introducing a Mediterranean diet (T2) in ALS patients. Outcomes: Mean SCFA concentrations (±SD) (ng/mL) at T0 between-group (ALS vs control) differences at baseline, and within-patient changes over time to assess whether the Mediterranean diet is associated with favorable SCFA shifts (T0-T1, T1-T2).
1 secondary outcome
Association of Serum SCFA Levels With ALS Progression Measured by ALSFRS-R at T0, T1, and T2
Time frame: 12 months
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 17 Sep 2025.
- Derived from this study: PubMed 41154611Motataianu A, Ion V, Dumitreasa M, Ormenisan I, Farczadi L, Andone S, Balasa R, Roman MM. Short-Chain Fatty Acid Profiles in Amyotrophic Lateral Sclerosis: Longitudinal Effects of Disease and Mediterranean Diet Intervention. Biomolecules. 2025 Sep 28;15(10):1380. doi: 10.3390/biom15101380.
- Individual participant dataSharing planned: No. Given the small, regionally identifiable ALS cohort and the rarity of the condition, the risk of re-identification remains high even after de-identification.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
The record lists no contact. Use the official record to reach the sponsor.
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Amyotrophic Lateral Sclerosis, run by George Emil Palade University of Medicine, Pharmacy, Sciences and Technology of Targu Mures at 1 site in Romania.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 3 inclusion and 6 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 17 Sep 2025. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Romania. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Analysis of serum SCFA levels in ALS patients and healthy controls, and evaluating the change in circulating SCFA concentrations associated with Mediterranean diet intervention (time frame: 12 months). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT07178067, which also lists 1 linked publication.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 17 Sep 2025.