Probiotic Impact on Cognitive Performance, and Metabolic Outcomes in Overweight Young Adults With Impaired Glucose Regulation

Assessed using the Cambridge Neuropsychological Test Automated Battery (CANTAB) Verbal Recognition Memory task (VRM) a computerized word list recall task, which measures accuracy in recalling previously presented verbal stimuli to evaluate verbal memory function.

Measured using the Paired Associates Learning (PAL) task (CANTAB), which assesses the ability to remember the locations of visual patterns and form new associations.

All adverse events (AEs) and serious adverse events (SAEs) will be monitored through participant self-reports and case report forms. Events will be categorized by severity and relatedness to the study product.

Measured using the Spatial Working Memory (SWM) task (CANTAB), which assesses the ability to temporarily retain and manipulate spatial information and use effective search strategies during problem-solving.

Measured using the Stop Signal Task (SST) (CANTAB), which assesses the ability to inhibit a dominant or preplanned motor response when signaled.

Measured using the Intra-Extra Dimensional Set Shift (IED) task (CANTAB), which assesses the ability to shift attention between different stimulus dimensions and adapt to changing rules. This evaluates cognitive flexibility and executive function.

Measured using the Five Choice Reaction Time (RTI) task (CANTAB), which assesses how quickly and accurately participants respond to visual stimuli.

CVR reflects the ability of cerebral blood vessels to dilate or constrict in response to changes in carbon dioxide, indicating the health of the brain's vascular system. Measured via transcranial Doppler ultrasound of the middle cerebral artery in response to hypercapnia/hypocapnia challenge.

Measured using ultrasound imaging of the brachial artery before and after a period of blood flow restriction using a pressure cuff. The artery's ability to dilate in response to the return of blood flow (reactive hyperemia) reflects endothelial function, an early indicator of vascular health and cardiovascular risk.

Measured using the Pittsburgh Sleep Quality Index (PSQI) global score, a validated 19-item self-report instrument assessing subjective sleep quality over the past month.

Assessed as a PSQI subcomponent, defined as the self-reported time (in minutes) taken to fall asleep on most nights.

Assessed as a PSQI subcomponent, reflecting the average number of hours of sleep per night.

Calculated as a PSQI subcomponent, defined as the ratio of total sleep time to time spent in bed, expressed as a percentage.

Derived from PSQI items capturing the frequency of nocturnal awakenings and other sleep interruptions.

Based on PSQI responses indicating frequency of pharmacological sleep aid use over the past month.

Evaluated using PSQI items assessing difficulties staying awake during the day and maintaining enthusiasm for activities.

HbA1c reflects average blood glucose levels over the previous 2 to 3 months and is used to assess long-term blood sugar control. Measured from capillary blood using the Afinion 2 Analyzer (Abbott).

This measure reflects short-term blood sugar levels and is used to assess baseline glucose regulation. Measured from capillary blood using an over-the-counter, CE-marked glucometer.

Assessed via a 2-hour oral glucose tolerance test (OGTT), with capillary blood glucose measured at 0, 30, 60, and 120 minutes post-glucose ingestion. These values will be used to calculate the area under the curve (AUC), which reflects how effectively the body regulates blood glucose over time.

Measured using an automated oscillometric device (Omron). This measure reflects the pressure in the arteries when the heart contracts and is a key indicator of cardiovascular health.

Measured using an automated oscillometric device (Omron). This measure reflects the pressure in the arteries when the heart is at rest between beats and is an important marker of vascular function and cardiovascular risk.

Measured from a fasting capillary blood sample using the Cholestech LDX Analyzer (Abbott). This test provides total blood cholesterol levels, which are used to assess cardiovascular risk.

Measured from a fasting capillary blood sample using the Cholestech LDX Analyzer (Abbott). LDL-C contributes to plaque buildup in the arteries and is a key cardiovascular risk marker.

Measured from a fasting capillary blood sample using the Cholestech LDX Analyzer (Abbott). HDL-C helps remove excess cholesterol from the bloodstream and is protective against heart disease.

Measured from a fasting capillary blood sample using the Cholestech LDX Analyzer (Abbott). Triglycerides are a type of fat found in the blood and are used to assess metabolic health and cardiovascular risk.

Calculated as weight in kilograms divided by height in meters squared (kg/m²). Weight is measured using a calibrated digital scale and height with a stadiometer. BMI is a gold standard measure used to classify underweight, normal weight, overweight, and obesity.

Calculated by dividing waist circumference by hip circumference. This ratio reflects fat distribution and is a marker of cardiometabolic risk.

Measured in kilograms using dual-energy X-ray absorptiometry (DXA). This measure reflects the total weight of muscle and other non-fat, non-bone tissues in the body.

Measured in kilograms using dual-energy X-ray absorptiometry (DXA). This reflects the total amount of fat tissue in the body.

Calculated from DXA-derived fat mass as a percentage of total body mass. This measure is used to evaluate body composition and excess adiposity-related risk.

Quantified from venous plasma samples using enzyme-linked immunosorbent assay (ELISA) kits. IL-6 is a pro-inflammatory cytokine associated with immune response and chronic inflammation.

Measured from venous plasma samples using ELISA kits. TNF-α is a cytokine involved in systemic inflammation and plays a role in metabolic and vascular dysfunction.

Assessed by 16S rRNA gene sequencing of DNA extracted from stool samples. Diversity will be measured using alpha diversity indices (e.g., Shannon Index), reflecting the richness and evenness of microbial communities.

Assessed using beta diversity metrics such as Bray-Curtis dissimilarity and principal coordinates analysis (PCoA). This reflects variability in microbial composition between participants and time points.

Determined from 16S rRNA gene sequencing data by calculating the proportion of individual bacterial taxa present in stool samples. This measure assesses how specific microbial populations change over time.

Assessed by capsule counts at the Week 12 visit and participant self-reports. The number of capsules taken will be calculated as a percentage of the total expected capsules over the 12-week intervention period.

Assessed using a structured, self-administered questionnaire. Participants will indicate whether they experienced specific physical symptoms during the past 14 days, and on how many days each symptom occurred. Symptom categories include gastrointestinal (e.g., constipation, diarrhoea, bloating), systemic (e.g., sore throat, headache, muscle ache, cold symptoms), and medication or menstrual-related events (e.g., use of antibiotics, vaginal treatments, menstruation).

Circulating markers of platelet activity will be quantified from venous blood samples using validated biochemical assays. These markers reflect platelet activation status and provide an index of platelet function in response to the intervention.