Pilot Study In Healthy Subjects Research Articles

MRI of brachial plexus using diffusion tensor imaging: a pilot study for the use of resolve sequence surgical and radiologic anatomy.

Clinical exam is the goldstandard for surgical indication. ENMG and conventional MRI are insufficient to understand the highly variable clinical presentation of brachial plexus (BP) lesions. DTI is based on motion of water molecules and can explore nerve function. This pilot study of healthy subjects aimed to develop RESOLVE sequence for BP exploration using diffusion MRI. The main objective was to provide complete precise information from DTI cartography associated with anatomical data. Six healthy volunteers were scanned using 3T PRISMA scanner with anatomic 3D STIR SPACE and RESOLVE diffusion sequences. Diffusion parametric maps of fractional anisotropy (FA) were extracted from RESOLVE acquisitions. A reproducible method for roots volumes and angles measurements was created using 3DSlicer. ROI were segmented on Mean B0 sequences. FA measurements were obtained with ROI on Mean B0 sequences. RESOLVE sequence was adapted to the BP. Mean FA was 0.30. Angles measurements on 3D STIR SPACE sequences showed increasing values from proximal to distal roots with an 0.6 ICC. Volume measurements on anatomic sequences varied widely from one root to another but did not show any significant difference on laterality. A new and reproducible method for BP exploration was developed, using MRI RESOLVE DTI sequences. Complete mapping was obtained but a low resolution of track density imaging did not allow to exploit distal nerves. Deterministic tractography principal limit was the lack of resolution. Extraction of diffusion, volumetric and angular parameters of the plexus roots, and scripts creation for image processing was adapted to the healthy BP.

Evaluation of a new mobile virtual reality setup to alter pain perception: A pilot study in healthy subjects (Preprint)

Evaluation of a new mobile virtual reality setup to alter pain perception: A pilot study in healthy subjects (Preprint)

Transcranial Direct Current Stimulation Over the Left Dorsolateral Prefrontal Cortex Reduced Attention Bias Toward Negative Facial Expression: A Pilot Study in Healthy Subjects.

Research in the cognitive neuroscience field has shown that individuals with a stronger attention bias for negative information had higher depression risk, which may be the underlying pathogenesis of depression. This dysfunction of affect-biased attention also represents a decline in emotion regulation ability. Clinical studies have suggested that transcranial direct current stimulation (tDCS) treatment can improve the symptoms of depression, yet the neural mechanism behind this improvement is still veiled. This study aims to investigate the effects of tDCS on affect-biased attention. A sample of healthy participants received 20 min active (n = 22) or sham tDCS (n = 19) over the left dorsolateral prefrontal cortex (DLPFC) for 7 consecutive days. Electroencephalographic (EEG) signals were recorded while performing the rest task and emotional oddball task. The oddball task required response to pictures of the target (positive or negative) emotional facial stimuli and neglecting distracter (negative or positive) or standard (neutral) stimuli. Welch power spectrum estimation algorithm was applied to calculate frontal alpha asymmetry (FAA) in the rest task, and the overlapping averaging method was used to extract event-related potentials (ERP) components in the oddball task. Compared to sham tDCS, active tDCS caused an obvious increment in FAA in connection with emotion regulation (p < 0.05). Also, participants in the active tDCS group show greater P3 amplitudes following positive targets (p < 0.05) and greater N2 amplitudes following negative distracters (p < 0.05), reflecting emotion-related attention biases. These results offer valuable insights into the relationship between affect-biased attention and the effects of tDCS, which may be of assistance in exploring the neuropathological mechanism of depression and anxiety and new treatment strategies for tDCS.

Immediate Effects of Electrical Stimulation on Oropharyngeal Structure and Laryngeal Vestibular Closure: A Pilot Study in Healthy Subjects.

ABSTRACTObjectives:This study examined the immediate effects of neuromuscular electrical stimulation (NMES) on the dynamics of oropharyngeal structure and laryngeal vestibular closure (LVC) in healthy subjects.Methods:Ten healthy male volunteers participated in this controlled, before-and-after, videofluoroscopic swallowing pilot study. The study was conducted in four phases (each performed twice): (1) saliva swallow (SS) before evaluation (BEFORE), (2) NMES while at rest with no SS (NMES AT REST), (3) SS during NMES (DURING NMES), and (4) SS to examine the aftereffects of NMES (AFTER). We measured distances that oropharyngeal structures moved in the NMES AT REST phase, and we analyzed the kinematics of saliva swallowing primarily in the BEFORE and AFTER phases.Results:Four changes in the morphology of the oropharyngeal structure caused by NMES AT REST were statistically significant: anterior–upward displacement of the hyoid bone and larynx, stretch of the laryngeal vestibule, and posterior ridge of the tongue root. Regarding the kinematics measured during SS, although there was no significant change in LVC reaction times, LVC duration in the AFTER phase was significantly longer than BEFORE. Regarding maximal displacement of the hyoid bone, there was significantly greater movement AFTER than BEFORE. As additional exploratory outcomes, the velocity of hyoid bone movement was significantly slower, and the hyoid-to-larynx approximation was significantly smaller, DURING NMES than AFTER.Conclusions:Longer duration of LVC might be caused by adaptive learning with NMES-induced structural changes in the oropharynx. Further clinical studies are warranted to determine whether this approach improves dysphagia, which impairs LVC.

Dark blood cardiovascular magnetic resonance of the heart, great vessels, and lungs using electrocardiographic-gated three-dimensional unbalanced steady-state free precession

BackgroundRecently, we reported a novel neuroimaging technique, unbalanced T1 Relaxation-Enhanced Steady-State (uT1RESS), which uses a tailored 3D unbalanced steady-state free precession (3D uSSFP) acquisition to suppress the blood pool signal while minimizing bulk motion sensitivity. In the present work, we hypothesized that 3D uSSFP might also be useful for dark blood imaging of the chest. To test the feasibility of this approach, we performed a pilot study in healthy subjects and patients undergoing cardiovascular magnetic resonance (CMR).Main bodyThe study was approved by the hospital institutional review board. Thirty-one adult subjects were imaged at 1.5 T, including 5 healthy adult subjects and 26 patients (44 to 86 years, 10 female) undergoing a clinically indicated CMR. Breath-holding was used in 29 subjects and navigator gating in 2 subjects. For breath-hold acquisitions, the 3D uSSFP pulse sequence used a high sampling bandwidth, asymmetric readout, and single-shot along the phase-encoding direction, while 3 shots were acquired for navigator-gated scans. To minimize signal dephasing from bulk motion, electrocardiographic (ECG) gating was used to synchronize the data acquisition to the diastolic phase of the cardiac cycle. To further reduce motion sensitivity, the moment of the dephasing gradient was set to one-fifth of the moment of the readout gradient. Image quality using 3D uSSFP was good-to-excellent in all subjects. The blood pool signal in the thoracic aorta was uniformly suppressed with sharp delineation of the aortic wall including two cases of ascending aortic aneurysm and two cases of aortic dissection. Compared with variable flip angle 3D turbo spin-echo, 3D uSSFP showed improved aortic wall sharpness. It was also more efficient, permitting the acquisition of 24 slices in each breath-hold versus 16 slices with 3D turbo spin-echo and a single slice with dual inversion 2D turbo spin-echo. In addition, lung and mediastinal lesions appeared highly conspicuous compared with the low blood pool signals within the heart and blood vessels. In two subjects, navigator-gated 3D uSSFP provided excellent delineation of cardiac morphology in double oblique multiplanar reformations.ConclusionIn this pilot study, we have demonstrated the feasibility of using ECG-gated 3D uSSFP for dark blood imaging of the heart, great vessels, and lungs. Further study will be required to fully optimize the technique and to assess clinical utility.

Geometric Perfusion Deficits: A Novel OCT Angiography Biomarker for Diabetic Retinopathy Based on Oxygen Diffusion

To develop geometric perfusion deficits (GPD), an optical coherence tomography angiography (OCTA) biomarker based on oxygen diffusion, and to evaluate its utility in a pilot study of healthy subjects and patients with diabetic retinopathy (DR). Retrospective cross-sectional study. Commercial spectral-domain optical coherence tomography angiography (OCTA) instruments were used to acquire repeated 3× 3-mm2 and 6× 6-mm2 motion-corrected macular OCTA volumes. En face OCTA images corresponding to the superficial capillary plexus (SCP), deep capillary plexus (DCP), and full retinal projections were obtained using automatic segmentation. For each projection, the GPD percentage and the vessel density percentage, the control metric, were computed, and their values were compared between the normal and DR eyes. The repeated OCTA acquisitions were used to assess the test-retest repeatability of the GPD and vessel density percentages. Repeated OCTA scans of 15 normal eyes and 12 DR eyes were obtained. For all en face projections, GPD percentages were significantly higher in DR eyes than in normal eyes; vessel density percentages were significantly lower in all but 1 projection (DCP). Large GPD areas were used to identify focal perfusion deficits. Test-retest analysis showed that the GPD percentage had superior repeatability than the vessel density percentage in most cases. A strong negative correlation between the GPD percentage and the vessel density percentage was also found. Geometric perfusion deficits, an OCTA biomarker based on oxygen diffusion, provides a quantitative metric of macular microvascular remodeling with a strong physiological underpinning. The GPD percentage may serve as a useful biomarker for detecting and monitoring DR.

Gene expression signature induced by grape intake in healthy subjects reveals wide-spread beneficial effects on peripheral blood mononuclear cells

Abstract Using a transcriptomic approach, we performed a pilot study in healthy subjects to evaluate the changes in gene expression induced by grape consumption. Blood from twenty subjects was collected at baseline (T0), after 21 days of grape-rich diet (T1) and after one-month washout (T2). Gene expression profiling of peripheral blood mononuclear cells from six subjects identified 930 differentially expressed transcripts. Gene functional analysis revealed changes (at T1 and/or T2) suggestive of antithrombotic and anti-inflammatory effects, confirming and extending previous finding on the same subjects. Moreover, we observed several other favourable changes in the transcription of genes involved in crucial processes such as immune response, DNA and protein repair, autophagy and mitochondrial biogenesis. Finally, we detected significant changes in many long non-coding RNAs genes, whose regulatory functions are being increasingly appreciated. Altogether, our data suggest that a grape diet may exert its beneficial effects by targeting different strategic pathways.

Gaze Fixation Patterns Correlate with Visual Attention and Memory: the Results of a Pilot Study in Healthy Subjects

Gaze Fixation Patterns Correlate with Visual Attention and Memory: the Results of a Pilot Study in Healthy Subjects

50. Inter hemispheric comparison of PAS-induced cortical plasticity: A pilot study in healthy subjects

Dystonia is considered a “network” disease with an abnormal cortical plasticity induced by by paired-associated-stimulation. In this study, we applied the PAS-TMS paradigm in healthy subjects, with the aim to collect reliable data from both hemispheres in order to put the basis to future application in focal dystonia. We studied 7 healthy subjects (4 females; mean age 32.33 ± 6.79 years). We applied electrical stimulatrion of median nerve electrical and MEPs from both abductor pollicis brevis (APB) and first dorsal interosseous (FDI) muscles (2), following Stefan et al. technique, with ISI set to 25 ms. MEPs before and after PAS protocol, evaluating first the dominant and then controlateral motor cortical area. Every subject performed the study in the same experimental session. Comparison between MEPs before and after PAS on homologous hemisphere disclosed a trend toward an increased of amplitude, but without reaching statistical significance for both ABP and FDI muscles, according to Quartarone et al. At the same time, no significant difference was detected by evaluating paired inter-hemisphere analysis. These bilateral findings in normal subjects could be a possible beginning for future application on focal movement disorders.

Onset and Offset Estimation of the Neural Inspiratory Time in Surface Diaphragm Electromyography: A Pilot Study in Healthy Subjects.

This study evaluates the onset and offset of neural inspiratory time estimated from surface diaphragm electromyographic (EMGdi) recordings. EMGdi and airflow signals were recorded in ten healthy subjects according to two respiratory protocols based on respiratory rate (RR) increments, from 15 to 40 breaths per minute (bpm), and fractional inspiratory time (Ti/Ttot) decrements, from 0.54 to 0.18. The analysis of EMGdi signal amplitude is an alternative approach for the quantification of neural respiratory drive. The EMGdi amplitude was estimated using the fixed sample entropy computed over a 250ms moving window of the EMGdi signal (EMGdifse). The neural onset was detected through a dynamic threshold over the EMGdifse using the kernel density estimation method, while neural offset was detected by finding when the EMGdifse had decreased to 70% of the peak value reached during inspiration. The Bland-Altman analysis between airflow and neural onsets showed a global bias of 46ms in the RR protocol and 22ms in the Ti /Ttot protocol. The Bland-Altman analysis between airflow and neural offsets reveals a global bias of 11ms in the RR protocol and -2ms in the Ti/T tot protocol. The relationship between pairs of RR values (Pearson's correlation coefficient of 0.99, Bland-=Altman limits of -2.39 to 2.41bpm, and mean bias of 0.01bpm) and between pairs of Ti/Ttot values (Pearson's correlation coefficient of 0.86, Bland-Altman limits of -0.11 to 0.10, and mean bias of -0.01) showed a good agreement. In conclusion, we propose a method for determining neural onset and neural offset based on noninvasive recordings of the electrical activity of the diaphragm that requires no filtering of cardiac muscle interference.

Bilateral transfer phenomenon: A functional magnetic resonance imaging pilot study of healthy subjects.

The bilateral transfer of a motor skill is a physiological phenomenon: the development of a motor skill with one hand can trigger the development of the same ability of the other hand. The purpose of this study was to verify whether bilateral transfer is associated with a specific brain activation pattern using functional magnetic resonance imaging (fMRI). The motor task was implemented as the execution of the Nine Hole Peg Test. Fifteen healthy subjects (10 right-handers and five left-handers) underwent two identical fMRI runs performing the motor task with the non-dominant hand. Between the first and the second run, each subject was intensively trained for five minutes to perform the same motor task with the dominant hand. Comparing the two functional scans across the pool of subjects, a change of the motor activation pattern was observed. In particular, we observed, in the second run, a change in the activation pattern both in the cerebellum and in the cerebral cortex. We found activations in cortical areas involved in somatosensory integration, areas involved in procedural memory. Our study shows, in a small group of healthy subjects, the modification of the fMRI activation pathway of a motor task performed by the non-dominant hand after intensive exercise performing the same task with the dominant hand.

Influence of dopamine receptor gene polymorphisms on circulating T lymphocytes: A pilot study in healthy subjects

Dopamine is a key transmitter in the neuroimmune network, acting through five dopaminergic receptors (DR): the D1-like D1 and D5 and the D2-like D2, D3 and D4. Several DR gene variants exist and may affect DR expression and activity. We assessed total lymphocytes, CD3+, CD4+ and CD8+ T lymphocytes in peripheral blood of healthy subjects and their association with selected DR gene variants (DRD1 rs4532 and rs686, DRD5 rs6283, DRD2 rs1800497 and rs6277, DRD3 rs6280 and rs1800828, DRD4 rs747302 and 7 48-base pair VNTR). DRD1 rs4532 and rs686 and DRD5 rs6283 were associated with total lymphocytes, and with CD3+ and CD4+ (but not CD8+) T lymphocytes, while none of the D2-like DR gene variants showed any association with lymphocyte counts. An arbitrary score based on the activity of D1-like vs D2-like DR correlated with total lymphocytes, CD3+ and CD4+ T cells (but not with CD8+ T cells). The association between D1-like DR gene variants and lymphocyte count, and in particular with CD4+ (but not CD8+) T lymphocytes, may imply a functional prevalence of D1-like over D2-like DR in CD4+ T cells. This is the first study showing an influence of DR gene polymorphisms on lymphocyte count, and in particular on CD4+ T cells. Future studies should address the possible association between DR gene variants and the immune function in health and disease. The relevance of these findings for the immune effects of dopaminergic agents should be also carefully examined.

Effect of Colchicine on Platelet-Platelet and Platelet-Leukocyte Interactions: a Pilot Study in Healthy Subjects.

The cardioprotective mechanisms of colchicine in patients with stable ischemic heart disease remain uncertain. We tested varying concentrations of colchicine on platelet activity in vitro and a clinically relevant 1.8-mg oral loading dose administered over 1 h in 10 healthy subjects. Data are shown as median [interquartile range]. Colchicine addition in vitro decreased light transmission platelet aggregation only at supratherapeutic concentrations but decreased monocyte- (MPA) and neutrophil-platelet aggregation (NPA) at therapeutic concentrations. Administration of 1.8 mg colchicine to healthy subjects had no significant effect on light transmission platelet aggregation but decreased the extent of MPA (28 % [22-57] to 22 % [19-31], p = 0.05) and NPA (19 % [16-59] to 15 % [11-30], p = 0.01), platelet surface expression of PAC-1 (370 mean fluorescence intensity (MFI) [328-555] to 333 MFI [232-407], p = 0.02) and P-selectin (351 MFI [269-492] to 279 [226-364], p = 0.03), and platelet adhesion to collagen (10.2 % [2.5-32.6] to 2.0 % [0.2-9.5], p = 0.09) 2 h post-administration. Thus, in clinically relevant concentrations, colchicine decreases expression of surface markers of platelet activity and inhibits leukocyte-platelet aggregation but does not inhibit homotypic platelet aggregation.

Higher visual functions in the upper and lower visual fields: A pilot study in healthy subjects.

Visual perception is not identical in the upper and lower visual hemifields. The mechanisms behind this difference can be found at the retinal, cortical, or higher attentional level. In this study, a new visual test battery, that involves real-time comparisons of complex visual stimuli, such as shape of objects, and speed of moving dot patterns, in the upper and lower visual hemifields, is presented. This study represents, to our knowledge, the first to implement such a visual test battery in an immersive environment composed of a hemisphere, in order to present visual stimuli in precise regions of the visual field. Ten healthy volunteers were tested in this pilot study. The results showed a higher accuracy in the image matching when the visual test was performed in the lower visual hemifield.

The administration of nasal drops in the "Kaiteki" position allows for delivery of the drug to the olfactory cleft: a pilot study in healthy subjects.

Systemic treatment with corticosteroids shows therapeutic effects, few patients benefit from intranasal topical drug application, probably due to limited access of the drug to the olfactory epithelium. The aim of the present study was to investigate how drops distribute within the nasal cavity when the "Kaiteki" position is performed. Thirteen healthy volunteers participated. Subjects were lying on the side with the head tilted and the chin turned upward. Blue liquid was used to visualize the intranasal distribution of the nasal drops. The investigation was carried out using photo documentation thorough nasal endoscopy; the intranasal distribution of the dye was judged by two independent observers in both a decongested state and a natural state where no decongestants had been used. With regard to the main criterion of this study, using the "Kaiteki" position, nasal drops reached the olfactory cleft in 96 % of the decongested cases and 75 % of the cases who had not been decongested. However, this difference was not statistically different. Because the "Kaiteki" maneuver is not too difficult to perform, it is more likely that topical steroids can be helpful in cases of olfactory loss.

Vitamin C supplementation modulates gene expression in peripheral blood mononuclear cells specifically upon an inflammatory stimulus: a pilot study in healthy subjects

In order to study the effects of vitamin C supplementation on gene expression and compare its action between physiological and inflammatory conditions, a pilot study was set up utilizing microarray and qPCR technologies. Five healthy volunteers were supplemented with 1 g vitamin C (Redoxon®) per day for five consecutive days. Peripheral blood mononuclear cells (PBMNC) were isolated before and just after the last supplementation, and RNA was isolated for the Affymetrix gene 1.0 ST chip analysis. PBMNC were also, ex vivo, treated with LPS, and gene expression was quantified by means of a “Human NFkB Signaling” qPCR array. Only a very moderate effect on the baseline gene expression modulation was associated with vitamin C supplementation. However, in spite of the limited number of subjects analyzed, vitamin C supplementation resulted in a markedly different modulation of gene expression upon the inflammatory stimulus, specifically at the level of the MyD88-dependent pathway and of the anti-inflammatory cytokine IL-10 synthesis. This study suggests that vitamin C supplementation in healthy subjects, not selected according to a specific genetic profile, consuming an adequate amount of vitamin C, and having a satisfactory vitamin C plasma concentration at the baseline, does not result in a significant modification of gene expression profile. Under this satisfactory micronutrient status, supplementation of vitamin C is “buffered” within a homeostatic physiological equilibrium. Differently, following a second “hit” constituted of an inflammatory stimulus such as LPS, able to trigger a critical burst to the normal physiological state, the higher availability of ascorbic acid emerges, and results in a significant modulation of cell response.

Effects of exercise with or without blueberries in the diet on cardio-metabolic risk factors: An exploratory pilot study in healthy subjects

Background. The improvement of insulin sensitivity by exercise has been shown to be inhibited by supplementation of vitamins acting as antioxidants.Objective. To examine effects of exercise with or without blueberries, containing natural antioxidants, on cardio-metabolic risk factors.Methods. Fifteen healthy men and 17 women, 27.6 ± 6.5 years old, were recruited, and 26 completed a randomized cross-over trial with 4 weeks of exercise by running/jogging 5 km five times/week and 4 weeks of minimal physical activity. Participants were also randomized to consume 150 g of blueberries, or not, on exercise days. Laboratory variables were measured before and after a 5 km running-race at maximal speed at the beginning and end of each period, i.e. there were four maximal running-races and eight samplings in total for each participant.Results. Insulin and triglyceride levels were reduced while HDL-cholesterol increased by exercise compared with minimal physical activity. Participants randomized to consume blueberries showed an increase in fasting glucose levels compared with controls, during the exercise period (blueberries: from 5.12 ± 0.49 mmol/l to 5.32 ± 0.29 mmol/l; controls: from 5.24 ± 0.27 mmol/l to 5.17 ± 0.23 mmol/l, P = 0.04 for difference in change). Triglyceride levels fell in the control group (from 1.1 ± 0.49 mmol/l to 0.93 ± 0.31 mmol/l, P = 0.02), while HDL-cholesterol increased in the blueberry group (from 1.51 ± 0.29 mmol/l to 1.64 ± 0.33 mmol/l, P = 0.006).Conclusions. Ingestion of blueberries induced differential effects on cardio-metabolic risk factors, including increased levels of both fasting glucose and HDL-cholesterol. However, since it is possible that indirect effects on food intake were induced, other than consumption of blueberries, further studies are needed to confirm the findings.

MRI Acquisition and Analysis Protocol for In Vivo Intraorbital Optic Nerve Segmentation at 3T

To present a new acquisition and analysis protocol for reliable and reproducible segmentation of the entire intraorbital optic nerve (ION) mean cross-sectional area by means of magnetic resonance imaging (MRI) at 3 tesla (T). Eight healthy volunteers (mean age 31, five were male) gave written informed consent and both of their IONs were imaged individually using a coronal-oblique T2-weighted fast multidynamic image acquisition scheme; the proposed acquisition scheme has its rationale in combining separately acquired volumes and registering them to account for motion-related artifacts commonly associated with longer acquisitions. Mean cross-sectional area of each ION was measured using a semiautomated image analysis protocol that was based on an active surface model previously described and used for spinal cord imaging. Reproducibility was assessed for repeated scans (scan-rescan) and repeated image analysis performance (intraobserver). Mean and SD values of the left ION cross-sectional area for the eight healthy volunteers were 5.0 (±0.7) mm² and for the right ION were 5.3 (±0.8) mm². Mean scan-rescan coefficient of variation (COV) for the left ION was 4.3% and for the right was 4.4%. Mean intraobserver COV for the left ION was 2.1% and for the right was 1.8%. This study presents a new MRI acquisition and analysis protocol for reliable and reproducible in vivo measurement of the entire ION mean cross-sectional area as demonstrated in a pilot study of healthy subjects. The protocol presented here can be used in future studies of the ION in disease state.

L-Arginine enriched biscuits improve endothelial function and glucose metabolism: A pilot study in healthy subjects and a cross-over study in subjects with impaired glucose tolerance and metabolic syndrome

ObjectiveThe aim of this study was to evaluate the effects of a new l-arginine-enriched biscuit on endothelial function, insulin sensitivity/secretion and body composition. Materials/MethodsThe project was composed of two studies. The first study was an acute pilot postprandial study in 7 healthy subjects that evaluated bio-availability and vascular effects of l-arginine-enriched biscuits that contained 6.6gl-arginine, 21.9g carbohydrates, 3.6g protein, 7.5g fat and 4.3g dietary fiber compared with placebo biscuits and 6.6g powdered l-arginine. Subjects underwent the tests in random order, in at least 14-day intervals. The second study was a double-blind crossover study in 15 obese subjects with IGT and MS. These subjects consumed 6.6g of l-arginine-enriched biscuits or placebo biscuits in a 1600kcal diet. Each study period lasted 2weeks with a 2-week washout in between. Endothelial function, glucose tolerance, insulin sensitivity and insulin secretion were evaluated at the end of each intervention period. ResultsIn the first study, the groups that received the l-arginine-enriched biscuits and the powdered l-arginine had similarly increased l-arginine, NOx and cGMP levels and post-ischemic blood flow (PI-BF). In both cases, these levels were significantly higher than those in the placebo biscuit recipient group. In the second study, the l-arginine-enriched biscuit recipient group displayed increased l-arginine, NOx, cGMP, PI-BF, and Matsuda index levels, whereas their circulating glucose, proinsulin/insulin ratio and fat mass were decreased compared with the placebo biscuit recipient group. Conclusionsl-Arginine-enriched biscuits with low sugar and protein content enhance endothelial function and improve glucose metabolism, insulin sensitivity and insulin secretion in subjects with IGT and MS.

Measurement of S‐methylcysteine and S‐methyl‐mercapturic acid in human urine by alkyl‐chloroformate extractive derivatization and isotope‐dilution gas chromatography–mass spectrometry

S-methylcysteine (SMC) is a minor amino acid naturally excreted in human urine, a protective agent against oxidative stress and a biotransformation product of the fumigant biocide methyl bromide and of nicotine. A metabolic source of SMC is catabolism of the repair catalytic protein MGMT (EC 2.1.1.37), which specifically removes the methyl group from the modified DNA nucleotide O-6-methyl-guanine to revert the normal GC base pairing. To assess the value of SMC and of S-methylmercapturic acid (SMMA) as candidate biomarkers of proliferative phenomena, a sensitive analytical method by GC-MS was applied in a pilot study of healthy subjects to assess their urinary elimination and the intra- and inter-individual variability. Extractive alkylation with butylchloroformate-n-butanol-pyridine (Husek technique) was employed for sample derivatization and isotope dilution GC-MS with S-[CD(3) ]-SMC and -SMMA was applied for specific and sensitive detection. To resolve the target analytes from the main coeluting interferents in the derivatized urine extract a medium-polarity stationary phase was employed. SMMA was not detected in the morning urine of three healthy fertile-age women followed for one month above the minimum detectable level of approx. 500 µg/L while SMC concentrations were in the 0.02-0.7 µg/mL range (n = 61) with large inter-day and inter-individual variations. In a young healthy male urine samples taken throughout a few days yielded concentrations in the same 90-810 µg/L range (n = 11). These preliminary results points at SMC as a candidate biomarker for the study of methylation turnover in several biochemical processes.