In a previous clinical study, a probiotic formulation (PF) consisting of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 (PF) decreased stress-induced gastrointestinal discomfort. Emerging evidence of a role for gut microbiota on central nervous system functions therefore suggests that oral intake of probiotics may have beneficial consequences on mood and psychological distress. The aim of the present study was to investigate the anxiolytic-like activity of PF in rats, and its possible effects on anxiety, depression, stress and coping strategies in healthy human volunteers. In the preclinical study, rats were daily administered PF for 2 weeks and subsequently tested in the conditioned defensive burying test, a screening model for anti-anxiety agents. In the clinical trial, volunteers participated in a double-blind, placebo-controlled, randomised parallel group study with PF administered for 30 d and assessed with the Hopkins Symptom Checklist (HSCL-90), the Hospital Anxiety and Depression Scale (HADS), the Perceived Stress Scale, the Coping Checklist (CCL) and 24 h urinary free cortisol (UFC). Daily subchronic administration of PF significantly reduced anxiety-like behaviour in rats (P,0·05) and alleviated psychological distress in volunteers, as measured particularly by the HSCL-90 scale (global severity index, P, 0·05; somatisation, P, 0·05; depression, P, 0·05; and anger-hostility, P,0·05), the HADS (HADS global score, P, 0·05; and HADSanxiety, P,0·06), and by the CCL (problem solving, P, 0·05) and the UFC level (P,0·05). L. helveticus R0052 and B. longum R0175 taken in combination display anxiolytic-like activity in rats and beneficial psychological effects in healthy human volunteers.

I n a recent clinical study, we demonstrated in the general population that Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 (PF) taken in combination for 30 days decreased the global scores of hospital anxiety and depression scale (HADs), and the global severity index of the Hopkins symptoms checklist (HSCL-90), due to the decrease of the sub-scores of somatization, depression and angerhostility spheres. Therefore, oral intake of PF showed beneficial effects on anxiety and depression related behaviors in human volunteers. From there, it is interesting to focus on the role of this probiotic formulation in the subjects with the lowest urinary free cortisol levels at baseline. This addendum presents a secondary analysis of the effects of PF in a subpopulation of 25 subjects with urinary free cortisol (UFC) levels less than 50 ng/ml at baseline, on psychological distress based on the percentage of change of the perceived stress scale (PSs), the HADs and the HSCL-90 scores between baseline and follow-up. The data show that PF improves the same scores as in the general population (the HADs global score, the global severity index of the HSCL-90 and three of its sub-scores, i.e., somatization, depression and anger-hostility), as well as the PSs score and three other subscores of the HSCL-90, i.e., "obsessive compulsive," "anxiety" and "paranoidideation." Moreover, in the HSCL-90, Beneficial psychological effects of a probiotic formulation (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) in healthy human volunteers

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Treatment of stroke, especially during the first hours or days, is still lacking. S-nitrosoglutathione (GSNO), a cerebroprotective agent with short life time, may help if administered early with a sustain delivery while avoiding intensive reduction in blood pressure. We developed in situ forming implants (biocompatible biodegradable copolymer) and microparticles (same polymer and solvent emulsified with an external oily phase) of GSNO to lengthen its effects and allow cerebroprotection after a single subcutaneous administration to Wistar rats. Arterial pressure was recorded for 3 days (telemetry, n = 14), whole-blood platelet aggregation up to 13 days (aggregometry, n = 58), and neurological score, cerebral infarct size and edema volume for 2 days after obstruction of the middle cerebral artery by autologous blood clots (n = 30). GSNO-loaded formulations (30 mg/kg) induced a slighter and longer hypotension (-10 vs. -56 ± 6 mmHg mean arterial pressure, 18 h vs. 40 min) than free GSNO at the same dose. The change in pulse pressure (-50%) lasted even up to 42 h for microparticles. GSNO-loaded formulations (30 mg/kg) prevented the transient 24 h hyper-aggregability observed with free GSNO and 7.5 mg/kg-loaded formulations. When injected 2 h after stroke, GSNO-loaded microparticles (30 mg/kg) reduced neurological score at 24 (-62%) and 48 h (-75%) vs. empty microparticles and free GSNO 7.5 mg/kg and, compared to free GSNO, divided infarct size by 10 and edema volume by 8 at 48 h. Corresponding implants reduced infarct size and edema volume by 2.5 to 3 times. The longer (at least 2 days) but slight effects on arterial pressures show sustained delivery of GSNO-loaded formulations (30 mg/kg), which prevent transient platelet hyper-responsiveness and afford cerebroprotection against the consequences of stroke. In conclusion, in situ GSNO-loaded formulations are promising candidates for the treatment of stroke.