Stimulated Growth and Innate Immunity in Brook Charr (Salvelinus fontinalis) Treated with a General Probiotic (Bactocell®) and Two Endogenous Probiotics That Inhibit Aeromonas salmonicida In Vitro.

Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium causing furunculosis, an opportunistic infection of farmed salmonid fish. Current treatment methods against furunculosis rely heavily on antibiotherapy. However, strains of this opportunistic fish pathogen were found to possess genes that confer resistance to major antibiotics including those used to cure furunculosis. Therefore, dispensing bacterial symbionts as probiotics to susceptible hosts appears to be a promising alternative. Here, we present the genomic characterization and in vivo safety assessment of two brook charr (Salvelinus fontinalis) bacterial symbionts that inhibited A. salmonicida subsp. salmonicida growth in vitro (Pseudomonas fluorescens ML11A and Aeromonas sobria TM18) as well as a commercialized probiotic, Pediococcus acidilactici MA18/5M (Bactocell®). The genomic sequences of ML11A and TM18 obtained by whole-genome shotgun sequencing lack key virulence factor genes found in related pathogenic strains. Their genomic sequences are also devoid of genes involved in the inactivation (or target modification of) several key antimicrobial compounds used in salmonid aquaculture. Finally, when administered daily to live brook charr fingerlings, ML11A, TM18 and Bactocell® helped improve several physiological condition metrics such as mean body weight, Fulton's condition factor and blood plasma lysozyme activity (an indicator for innate immune activity).

Polar lipids from wheat extract oil improve skin damages induced by aging: Evidence from a randomized, placebo-controlled clinical trial in women and an ex vivo study on human skin explant.

BACKGROUND: Polar lipids from wheat (Triticum vulgare/aestivum) extract oil (WEO) are known to improve skin hydration. AIMS: These studies aimed to assess WEO benefits on the skin appearance of middle-aged women. METHODS: A double-blind, randomized, placebo-controlled clinical study was carried out on 64 healthy women, aged from 45 to 60 years, to investigate antiaging effects and benefits for the skin. The study lasted 20 weeks including 12 weeks of oral supplementation with WEO or placebo and 8 weeks of follow-up. Wrinkles in the "crow's-feet" area were evaluated by the Lemperle score. Skin hydration was measured using a corneometer, while roughness and radiance were determined by clinical scoring. Collagen content was quantified in human skin explants exposed to ultraviolet (UV) irradiations and treated with WEO or vehicle control. RESULTS: Compared to the placebo group, the Lemperle score was significantly reduced in the WEO group between W0 and W8 to reach a clinically significant 1 grade at W12. Facial hydration was significantly improved in the WEO group from W0 to W12, whereas leg hydration was significantly increased after 4 weeks and lasted throughout the supplementation period. Skin roughness and radiance were also significantly improved from W0 to W8 in the WEO group compared to placebo group. A higher collagen content was measured in the UV-irradiated skin explants treated with WEO compared to the untreated ones. CONCLUSION: These results confirmed the moisturizing effect of WEO and, for the first time, revealed its potential antiaging properties.

Strain rate hardening: a hidden but critical mechanism for biological composites?

Natural materials such as nacre, bone, collagen and spider silk boast unusual combinations of stiffness, strength and toughness. Behind this performance is a staggered microstructure, which consists of stiff and elongated inclusions embedded in a softer and more deformable matrix. The micromechanics of deformation and failure associated with this microstructure are now well understood at the "unit cell" level, the smallest representative volume for this type of material. However, these mechanisms only translate to high performance if they propagate throughout large volumes, an important condition which is often overlooked. Here we present, for the first time, a model which captures the conditions for either spreading of deformations or localization, which determines whether a staggered composite is brittle or deformable at the macroscale. The macroscopic failure strain for the material was calculated as function of the viscoplastic properties of the interfaces and the severity of the defect. As expected, larger strains at failure can be achieved when smaller defects are present within the material, or with more strain hardening at the interface. The model also shows that strain rate hardening is a powerful source of large deformations for the material as well, a result we confirmed and validated with tensile experiments on glass-polydimethylsiloxane (PDMS) nacre-like staggered composites. An important implication is that natural materials, largely made of rate-dependent materials, could rely on strain rate hardening to tolerate initial defects and damage to maintain their functionality. Strain rate hardening could also be harnessed and optimized in bio-inspired composites in order to maximize their overall performance.

Antihypertensive treatment in patients with class 3 obesity.

BACKGROUND: Even though patients with class 3 obesity (body mass index ≥ 40 kg/m(2)) are prone to arterial hypertension and respond less to antihypertensive drugs, they are not considered in hypertension treatment guidelines and data from prospective clinical trials are lacking. METHODS: In a post hoc analysis of a clinical trial, we compared patients with class 3 obesity with patients with class 1/2 obesity. RESULTS AND CONCLUSIONS: Blood pressure control in class 3 obesity was less likely to be achieved with hydrochlorothiazide monotherapy. While addition of amlodipine, irbesartan, or aliskiren to hydrochlorothiazide improved the blood pressure response, amlodipine was less effective and induced peripheral edema in 19% of patients with class 3 obesity.

Complement dependent cytotoxicity activity of therapeutic antibody fragments is acquired by immunogenic glycan coupling

Oligosaccharides are implicated in the development of the immune response notably in complement activation. Anti-tumoural immunotherapy by monoclonal antibodies (mAbs) offers some advantages to chemotherapy including cell targeting but some of them are inefficient to generate cytotoxicity dependent complement (CDC) known to be important in the antibody’s efficacy. The aim of this study is to give a CDC activity of mAb by linkage of a complement activating oligosaccharide to this antibody via a hetero-bifunctional linker allowing control of the conjugation reaction. We worked on non Hodgkin Burkitt’s lymphoma as cancer source, Fab fragments of rituximab devoid of complement activity as mAb and the trisaccharide Gal alpha(1→3)Gal beta(1→4)GlcNAc as immunogenic glycan. The bioconjugate Fab-Gal was characterized by biochemical methods and we demonstrated that the α-Gal epitope was recognized by seric immunoglobulins. After checking the recognition capacity of the Fab-Gal conjugate for the CD20 epitope, in vitro assays were performed to evaluate the activation of the complement cascade by the Fab-Gal conjugate. The effect of this bioconjugate was confirmed by the evaluation of the proliferation response of Burkitt’s cell line. The relative facility realization of this strategy represents new approaches to increase activities of mAbs.

Efficacy, safety, and tolerability of aliskiren monotherapy administered with a light meal in elderly hypertensive patients: a randomized, double-blind, placebo-controlled, dose-response evaluation study.

This randomized, double-blind, placebo-controlled study assessed the efficacy, safety, and tolerability of aliskiren 75, 150, and 300 mg to clarify the dose-response relationship and characterize the optimum aliskiren dose when given with a light meal to elderly hypertensive patients. After washout, 754 patients aged ≥65 years with hypertension (mean sitting systolic blood pressure [msSBP] ≥150 and <180 mm Hg; mean sitting diastolic blood pressure [msDBP] <110 mm Hg) were randomized to aliskiren 75, 150, or 300 mg or placebo for 8 weeks; medication was taken each morning with a light meal. The primary efficacy variable was change in msSBP from baseline to week 8 end point. Change from baseline in msDBP and dose-response curves for aliskiren 75, 150, and 300 mg were also assessed. At week 8 end point, all 3 aliskiren doses provided significantly greater least squares mean reductions in msSBP/msDBP (75 mg, 13/5 mm Hg; 150 mg, 15/6 mm Hg; 300 mg, 14/7 mm Hg) compared with placebo (8/4 mm Hg; P < .05). Aliskiren was generally well tolerated at all doses. There was a significant dose-response relationship for aliskiren, with an estimated minimum effective dose of 81.9 mg. In conclusion, aliskiren 150 and 300 mg provided effective blood pressure control in elderly patients when given with a light meal.

Efficacy and safety of aliskiren in Japanese hypertensive patients with renal dysfunction.

This 12-week, multicenter, open-label study assessed the efficacy, pharmacokinetics and safety of a once-daily aliskiren in Japanese hypertensive patients with renal dysfunction. Patients (n=40, aged 20-80 years) with mean sitting diastolic blood pressure (msDBP) >or=95 and <110 mm Hg and serum creatinine between >or=1.3 and <3.0 mg per 100 ml in males or between >or=1.2 and <3.0 mg per 100 ml in females were eligible. Patients began therapy with a once-daily morning oral dose of 75 mg of aliskiren. In patients with inadequate blood pressure control (msDBP >or=90 or mean sitting systolic blood pressure [msSBP] >or=140 mm Hg) and without safety concerns (serum potassium >5.5 mEq l(-1) or an increase in serum creatinine >or=20%), the aliskiren dose was increased to 150 mg and then to 300 mg in sequential steps starting from Week 2. Efficacy was assessed as change in msSBP/msDBP from baseline to the Week 8 endpoint (with the last observation carried forward). The mean reduction from baseline to Week 8 endpoint was 13.9+/-16.6 and 11.6+/-9.7 mm Hg for msSBP and msDBP, respectively. At the Week 8 endpoint, 65% patients had achieved blood pressure response (msDBP <90 or a 10 mm Hg decrease or msSBP <140 or a 20 mm Hg decrease) and 30% had achieved blood pressure control (msSBP <140 mm Hg and msDBP <90 mm Hg). Aliskiren was well tolerated with no new safety concerns in Japanese hypertensive patients with renal dysfunction.

Combination therapy with valsartan/hydrochlorothiazide at doses up to 320/25 mg improves blood pressure levels in patients with hypertension inadequately controlled by valsartan 320 mg monotherapy.

OBJECTIVES: To investigate the efficacy and tolerability of valsartan (Val) 320 mg once daily (o.d.), Val/hydrochlorothiazide (HCTZ) 320/12.5 mg o.d. and Val/HCTZ 320/25 mg o.d. in patients with hypertension not adequately controlled by Val monotherapy. METHODS: This double-blind, active-controlled, parallel-group, randomized trial recruited patients > or =18 years with mild-to-moderate essential hypertension, defined as mean sitting diastolic blood pressure (MSDBP) of > or =95 mmHg and <110 mmHg without treatment. After washout, 3805 eligible patients received Val 320 mg o.d. single-blind for 4 weeks. Subsequently, patients with MSDBP > or =90 and <110 mmHg (n=2702) were randomized to double-blind treatment with Val 320 mg, Val/HCTZ 320/12.5 mg or Val/HCTZ 320/25 mg for 8 weeks. Mean changes in MSDBP and mean sitting systolic BP (MSSBP) from the start of the single-blind period were analysed, as well as the proportion of responders (MSDBP <90 mmHg or > or =10 mmHg decrease from the start of the double-blind period). Tolerability and safety were also assessed. RESULTS: Reductions in MSDBP and MSSBP were observed in all groups. Both combinations were associated with significantly greater reductions than monotherapy for MSDBP and MSSBP at Weeks 8 and 12 (all p<0.0001). Both combinations also resulted in significantly greater proportion of responders at study end (74.9% and 68.8% for Val/HCTZ 320/25 mg and Val/HCTZ 320/12.5 mg, respectively) than monotherapy (52.7%; both p < 0.0001). In addition, a dose-response was observed with increasing dose of HCTZ with respect to MSSBP. All treatments were well tolerated. CONCLUSIONS: The combination ofVal and HCTZ at doses of 320/12.5 mg and 320/25 mg increases antihypertensive efficacy in patients with mild-to-moderate hypertension inadequately controlled with Val 320 mg monotherapy, without compromising tolerability.

Direct Renin inhibition with aliskiren in obese patients with arterial hypertension.

Current guidelines from the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure recommend first-line treatment with a thiazide diuretic but do not provide specific guidance for obese patients. The renin system is activated in obesity-associated arterial hypertension. Therefore, we tested the hypothesis that the oral direct renin inhibitor aliskiren could provide additive blood pressure lowering in obese patients with hypertension (body mass index >or=30 kg/m(2); mean sitting diastolic blood pressure: 95 to 109 mm Hg) who had not responded to 4 weeks of treatment with hydrochlorothiazide (HCTZ) 25 mg. After a 2- to 4-week washout, 560 patients received single-blind HCTZ (25 mg) for 4 weeks; 489 nonresponders were randomly assigned to double-blind aliskiren (150 mg), irbesartan (150 mg), amlodipine (5 mg), or placebo for 4 weeks added to HCTZ (25 mg), followed by 8 weeks on double the initial doses of aliskiren, irbesartan, or amlodipine. After 8 weeks of double-blind treatment (4 weeks on the higher dose), aliskiren/HCTZ lowered blood pressure by 15.8/11.9 mm Hg, significantly more (P<0.0001) than placebo/HCTZ (8.6/7.9 mm Hg). Aliskiren/HCTZ provided blood pressure reductions similar to those with irbesartan/HCTZ and amlodipine/HCTZ (15.4/11.3 and 13.6/10.3 mm Hg, respectively), with similar tolerability to placebo/HCTZ. Adverse event rates were highest with amlodipine/HCTZ because of a higher incidence of peripheral edema (11.1% versus 0.8% to 1.6% in other groups). In conclusion, combination treatment with aliskiren is a highly effective and well-tolerated therapeutic option for obese patients with hypertension who fail to achieve blood pressure control with first-line thiazide diuretic treatment.

Norfloxacin-poly(L-lysine citramide imide) conjugates and structure-dependence of the drug release.

Norfloxacin (Nflx), an antibiotic which is active against some intracellular bacteria, was coupled to a polymeric carrier, namely poly(L-lysine citramide) via a lysine or an ethylcarbamate spacer to obtain a macromolecular prodrug. The carrier, which derived from the two metabolites citric acid and L-lysine, is known to be biocompatible and slowly degradable under slight acidic conditions. Conjugates were characterised by UV, 1H and 13C NMR and SEC. The presence of Norfloxacin and the lysine type spacer caused chain aggregation, due to a probable physical cure. The release of Norfloxacin from these prodrugs and from a prodrug where Norfloxacin is bound to the carrier backbone without spacer arm was investigated comparatively in vitro. Conjugation via a carbamate-type linkage appeared as a method to achieve the release of Norfloxacin from a PLCA-type conjugate at neutral.

Preference of insomniac patients between a single dose of zolpidem 10 mg versus zaleplon 10 mg.

We assessed the preference of insomniac patients between a single dose of 10 mg zolpidem or zaleplon, respectively, administered in random order on two consecutive nights. Fifty-three patients (mean age 52.2 years, 51% females) with a history of recurrent episodes of insomnia and currently complaining of difficulties in falling asleep were included into a randomized, double-blind, cross-over study by 12 general practitioners. After each night, the patients were asked to fill in a sleep questionnaire and visual analogue scales (VAS) to subjectively assess both the quality of sleep (in the morning) and the quality of the day (in the evening). After the second study night, patients' self-assessed preference was established through a drug preference questionnaire. 62% of patients preferred zolpidem, while 38% preferred zaleplon (p = 0.08). The quality of sleep items getting to sleep and quality of sleep were significantly more improved after zolpidem (p = 0.03 and p < 0.0001, respectively). On the VAS, subjective sleep quality was significantly better after zolpidem (p < 0.0001). Diurnal awakeness and quality of day life were satisfying in both groups, without significant difference. The subjective total duration of sleep was 8.0 h for zolpidem and 8.1 h for zaleplon (n.s.). Safety was good and similar between the two drugs. Insomniac patients tended to prefer zolpidem to zaleplon on both nocturnal and diurnal assessments. These results provide additional information for the physician's choice, based on the patient's preference for a given drug.

Improved brain uptake and pharmacological activity of dalargin using a peptide-vector-mediated strategy.

The blood-brain barrier restricts the passage of substances into the brain. Neuropeptides, such as enkephalins, cannot be delivered into the brain when given systemically because of this barrier. Therefore, there is a need to develop efficient transport systems to deliver these drugs to the brain. Recently, we have demonstrated that conjugation of doxorubicin or penicillin to peptide vectors significantly enhances their brain uptake. In this study, we have conjugated the enkephalin analog dalargin with two different peptide vectors, SynB1 and SynB3, to improve its brain delivery and its pharmacological effect. We show by in situ brain perfusion that vectorization markedly enhances the brain uptake of dalargin. We also show using the hot-plate model that this enhancement in brain uptake results in a significant improvement in the observed antinociceptive effect of dalargin. These results support the usefulness of peptide-mediated strategies for improving the availability and efficacy of central nervous system drugs.