[Barriers and enablers to pharmacist-led medication review in community pharmacy: A prospective study in Grand Est, France]. / Freins et leviers à la réalisation des bilans partagés de médication en officine : retour sur une expérimentation dans la région Grand Est.

Since 2018, French community pharmacists are involved in pharmaceutical care program performing medication review (MR). Near graduated pharmacy students from two faculties of pharmacy were assigned to implement and perform 3 MR in order to identify enablers and barriers of the implementation of MR by community pharmacists. Among 179 MR performed by 117 pharmacists during 5 months, they reported 3 main barriers: the time spending to initiate and perform all steps in MR (lack of time), patients recruiting, and compensation by health care system. Communications initiatives to patients and health professionals in primary care could facilitate patient MR adhesion. Simplification of administrative approach and optimization of software will be welcome and useful in order to reinforced MR implementation and leading.

[Comparison of Lasik with femtosecond laser versus Lasik with mechanical microkeratome: predictability of flap depth, corneal biomechanical effects and optical aberrations]. / Comparaison entre le Lasik au laser femtoseconde et le Lasik au microkératome mécanique: prédictibilité des découpes, biomécanique cornéenne et aberrations optiques.

OBJECTIVES: To compare the predictability of flap thickness, high-order optic aberrations (HOAs), and biomechanical properties of cornea between patients treated by Lasik with mechanical microkeratome versus patients treated by FemtoLasik. SETTING: Department of ophthalmology, Pellegrin University Hospital, Bordeaux, France. PATIENTS AND METHODS: We conducted a retrospective study on 53 myopic patients who underwent Lasik with either mechanical microkeratome (MK group) or femtosecond laser (FS group). Refraction, central corneal thickness, high-order optic aberrations (HOAs), corneal hysteresis (CH), and corneal resistance factor (CRF), were analysed pre- and postoperatively in both groups. The central corneal thickness was measured with OCT-Visante(®) (Carl-Zeiss, Meditec), biomechanical parameters with ORA(®) (Reichert), and optical aberrations with the Wave Scan(®) (AMO) aberrometer. RESULTS: We included 44 eyes of 22 patients in the MK group and 62 eyes of 31 patients in the FS group. Preoperatively, the mean best-corrected visual acuity was 0.95 in both groups. In the MK group, the flap was significantly thicker than expected (162/130 µm), but in the FS group, there was no significant difference (117/120 µm). The biomechanical properties of the cornea were lower in both groups independently of the flap cutting technique. The HOAs increased after Lasik and were not influenced by the flap cutting technique. CONCLUSION: Neither mechanical microkeratome, nor femtosecond laser for flap creation, increases HOAs and the biomechanical changes of the cornea, according to ORA(®), significantly after Lasik.

[Comparison of the depth predictability of intra corneal ring segment implantation by mechanical versus femtosecond laser-assisted techniques using optical coherence tomography (OCT Visante(®))]. / Comparaison en tomographie en cohérence optique (OCT Visante(®)) de la prédictibilité de la profondeur d'implantation des anneaux intracornéens après tunnélisation mécanique versus tunnélisation assistée au laser femtoseconde.

UNLABELLED: Comparison of the depth predictability of intracorneal ring segment implantation by mechanical versus femtosecond laser-assisted techniques using optical coherence tomography (OCT Visante(®)). PURPOSE: To compare the depth predictability of intracorneal ring segment implantation by mechanical versus femtosecond laser-assisted techniques using OCT Visante(®). METHODS: This observational prospective study included, after patients' consent, 76 keratoconic eyes, of which 31 eyes (group 1) were operated using the mechanical technique and 45 eyes (group 2) with the femtosecond laser-assisted technique. The target depth was two-thirds of the peripheral corneal thickness, ranging from 5 to 7 mm in diameter. Every patient underwent high-resolution anterior segment OCT (OCT Visante(®)) measurement preoperatively and for the implant depth, 1 month postoperatively. Then two different sites were used to determine the segment depth, at the segment site and tangentially, at a distance of 700 µm central to the segment's inner edge. RESULTS: Both measurement techniques demonstrated that targeted implantation depth of 66% of the corneal thickness was not observed in either group. The mean difference between the preoperative expected depth and final segment implantation was 76.64 ± 48.76 µm in the manual technique and 85.85 ± 33.02 µm in the femtosecond-assisted technique, with no statistically significant difference between the groups. Comparison between the measurement sites showed implantation depth to be shallower, 54.93±6.03%, at the tangential site compared to 55.14 ± 8.08% at the implant site in group 1 and 56.17 ± 5.82% versus 58.88 ± 6.06%, respectively, in group 2. CONCLUSION: Both mechanical and femtosecond laser-assisted techniques showed a more superficial Intacs(®) placement than predicted. No statistically significant difference was observed in implantation depth between the two groups.

[Detection of keratoconus using wavefront analysis]. / Dépistage aberrométrique du kératocône.

PURPOSE: To determine the value of wavefront analysis in the detection of keratoconus. PATIENTS AND METHODS: A prospective study was conducted from May 2008 to May 2010. Four groups were formed: patients with a "healthy" cornea (group 0; n=48), patients with keratoconus "suspect" (group 1; n=43), patients with keratoconus "forme fruste" (group 2; n=31), and patients with "beginning" keratoconus (group 3; n=47). Total and corneal aberrations were analysed. The Fisher test and Student t-test were used to compare the different groups. Receiver operating characteristic (ROC) curves were plotted when aberration differentiated groups 0 and 1. RESULTS: Total and corneal coma (Z 3), the corneal trefoil, (Z 3) and the corneal secondary astigmatism (Z 4) differentiated groups 0 and 1. Sensitivities and specificities of total and corneal coma, corneal trefoil, and corneal secondary astigmatism were, respectively, 67.4%, 56.3% (Youden=0.237), 60.5%, 72.9% (Youden=0.334), 83.7%, 39.6% (Youden=0.233), and 65.1%, 58.3% (Youden=0.234). Total, corneal coma, and corneal trefoil differentiated all severity groups. The third high-order aberrations differentiated groups 1 and 2. DISCUSSION: High-order aberrations are good indicators for grading keratoconus. Total high-order aberrations are less discriminating than corneal high-order aberrations. Corneal coma (Z 3) is the most discriminating aberration to differentiate the healthy cornea from the suspect cornea. Nevertheless, it does not optimally detect keratoconus suspect (Youden=0.334 and sensitivity=60.5%). The statistical analysis suggests that keratoconus suspect and forme fruste are different entities. CONCLUSION: Wavefront analysis can be used for the detection of keratoconus suspect, forme fruste, and beginning keratoconus but must be associated with videotopography and pachymetry during consultation of refractive surgery.

Genome-wide expression analyses establish dendritic cells as a new osteoclast precursor able to generate bone-resorbing cells more efficiently than monocytes.

Dendritic cells (DCs), mononuclear cells that initiate immune responses, and osteoclasts (OCs), multinucleated bone-resorbing cells, are hematopoietic cells derived from monocytic precursor cells. Using in vitro generated dendritic cells, we previously showed that human and murine DCs could transdifferentiate into resorbing osteoclasts in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL). In this study we globally compared by transcriptomic profiling this new osteoclast differentiation pathway from DCs with the canonical differentiation pathway from monocytes. DNA chip data revealed that starting from two very distinct cell types, treatment with M-CSF and RANKL generated two highly similar types of osteoclast. In particular, DC-derived osteoclasts expressed all the characteristic marker genes of monocyte-derived osteoclasts. Two major molecular events could be observed during osteoclastogenesis: downregulation of a large set of monocyte or DC specific markers, together with upregulation of characteristic osteoclast marker genes. Most interestingly, our transcriptomic data showed a closer molecular profile between DCs and OCs than between monocytes and OCs. Our data establish DCs as a new osteoclast precursor able to generate OCs more efficiently than monocytes.

Salmonella pathogenicity island 2-encoded type III secretion system mediates exclusion of NADPH oxidase assembly from the phagosomal membrane.

Salmonella typhimurium requires a type III secretion system encoded by pathogenicity island (SPI)-2 to survive and proliferate within macrophages. This survival implies that S. typhimurium avoids or withstands bactericidal events targeted to the microbe-containing vacuole, which include intraphagosomal production of reactive oxygen species (ROS), phagosomal acidification, and delivery of hydrolytic enzymes to the phagosome via fusion with lysosomes. Recent evidence suggests that S. typhimurium alters ROS production by murine macrophages in an SPI-2-dependent manner. To gain insights into the mechanism by which S. typhimurium inhibits intraphagosomal ROS production, we analyzed the subcellular distribution of NADPH oxidase components during infection of human monocyte-derived macrophages by wild-type (WT) or several SPI-2 mutant strains of S. typhimurium. We found that the membrane component of the NADPH oxidase, flavocytochrome b(558), was actively excluded or rapidly removed from the phagosomal membrane of WT-infected monocyte-derived macrophages, thereby preventing assembly of the NADPH oxidase complex and intraphagosomal production of superoxide anion. In contrast, the NADPH oxidase assembled on and generated ROS in phagosomes containing SPI-2 mutant S. typhimurium. Subversion of NADPH oxidase assembly by S. typhimurium was accompanied by increased bacterial replication relative to that of SPI-2 mutant strains, suggesting that the ability of WT S. typhimurium to prevent NADPH oxidase assembly at the phagosomal membrane represents an important virulence factor influencing its intracellular survival.

Transient association of the nicotinamide adenine dinucleotide phosphate oxidase subunits p47phox and p67phox with phagosomes in neutrophils from patients with X-linked chronic granulomatous disease.

Optimal microbicidal activity of polymorphonuclear leukocytes (PMNs) requires recruitment of a functional nicotinamide adenine dinucleotide phosphate (NADPH) oxidase to the phagosome. In this study, we used a synchronized phagocytosis assay and immunofluorescence microscopy (IFM) to examine the association of cytosolic NADPH oxidase subunits with phagosomes containing opsonized zymosan (OpZ). Ingestion of OpZ began within 30 seconds of particle binding and forming phagosomes were enriched for both F-actin and the actin-binding protein p57. NADPH oxidase subunits p47phox and p67phox were also recruited to forming phagosomes and were retained on mature phagosomes for at least 15 minutes. Colocalization of F-actin, p57, and p47phox on phagosomes was confirmed by immunoblotting. Translocation of p67phox, but not p57, to forming phagosomes was deficient in PMNs lacking p47phox. Surprisingly, we found that in PMNs from six individuals with X-linked chronic granulomatous disease (CGD), p47phox and p67phox accumulated in the periphagosomal area during ingestion of OpZ. However, in marked contrast to normal PMNs, p47phox and p67phox were shed from nascent phagosomes along with F-actin and p57 once OpZ was internalized (approximately 5 minutes). These data support a model in which flavocytochrome b is required for stable membrane binding of p47phox and p67phox, but not their association with the cytoskeleton or transport to the cell periphery.

Effect of SK&F 96365 on extracellular Ca2+ -dependent O2- production in neutrophil-like HL-60 cells.

Store-operated Ca2+ entry is referred to a capacitative current activated by Ca2+ -stores depletion in various non-excitable cells. Neutrophil-like HL-60 cells responded to N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine (fMLP) by an early O2- production preceded by a [Ca2+]i rise. Cell stimulation in the absence of extracellular Ca2+ resulted in a major reduction of [Ca2+]i rise and O2- production. A purported inhibitor of store-operated Ca2+ entry, SK&F 96365 (1-(beta-(3-(4-methoxy-phenyl)propoxyl)-4-methoxy-phenetyl)- 1H-imidazole hydrochloride), inhibited extracellular Ca2+ -dependent [Ca2+]i rise by 30% but did not alter O2- production. In conclusion, SK&F 96365 did not modify extracellular Ca2+ -dependent O2- production, despite a significant but limited reduction in fMLP-activated membrane Ca2+ fluxes which can be ascribed to store-operated Ca2+ entry. Furthermore, Ca2+ influx is necessary for a full induction and maintenance of the biological response.

Human umbilical cord blood-derived eosinophils cultured in the presence of IL-3 and IL-5 respond to fMLP with [Ca2+]i variation and O2- production.

In the presence of interleukin-3 and interleukin-5, eosinophil precursors from human umbilical cord blood mononuclear cells were regularly differentiated into mature eosinophil-like cells expressing normal morphology and cyanide-resistant peroxidase. O2- production and [Ca2+]i rise were measured in these in vitro differentiated eosinophils after fMLP stimulation; with dihydrorhodamine-123 and fura-2, respectively. Umbilical cord blood-derived eosinophils responded to fMLP (0.01 nM to 3 microM) with a concentration-dependent production of O2- (EC50 = 63.1 +/- 17.2 nM; Emax = 71.0 +/- 6.2 pmol/min/10(6) cells). O2- production was correlated with an fMLP concentration-dependent increase in [Ca2+]i (EC50 = 32.5 +/- 14.9 nM; Emax = 200.0 +/- 23.9 nM). These results indicate that human umbilical cord blood-derived eosinophils demonstrate functional characteristics similar to adult human peripheral blood eosinophils after activation by fMLP. Therefore, the large numbers of eosinophils (2-3 x 10(6)/ml cord blood) which can be obtained by culture of human cord blood mononuclear cells may serve as a useful model for future studies which will provide insight into the pathogenesis of diseases associated with eosinophils.

Endothelin-1 does not modulate O2.release and [Ca(2+)]i variations in resting or differentiated HL-60 cells.

Endothelin-1 (ET-1) by itself was not an effective stimulus for inducing superoxide (O2.) generation in human resting or DMSO-differentiated neutrophil-like HL-60 cells. ET-1 (0.01-100 nM) was not able to modulate O2. generation stimulated by the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP, EC50 = 4.24 +/- 1.63 nM in the absence and 3.16 +/- 1.95 nM in the presence of ET-1). Neither did ET-1 (0.01-100 nM) promote the mobilization of intracellular calcium ions or modulate fMLP-induced [Ca(2+)]i increase in this model of human neutrophils. Phosphoramidon, a neutral endopeptidase inhibitor, was not able to reveal any biological (O2.) or biochemical ([Ca(2+)]i response to ET-1 in the absence or in the presence of fMLP in these cells. These results indicate that DMSO-differentiated neutrophil-like HL-60 cells are not sensitive to ET-1 in terms of O2. generation or [Ca(2+)]i variations.

A double-labelling fluorescent assay for concomitant measurements of [Ca2+]i and O2. production in human macrophages.

To measure intracellular free Ca2+ concentration ([Ca2+]i) and superoxide (O2) production in human alveolar macrophages, we used the fluorescent Ca2+ indicator fura-2 and the O2-sensitive dye dihydrorhodamine-123, which becomes fluorescent in its oxidized form, rhodamine-123. We describe a new double-dye technique whereby the kinetics of both [Ca2+]i levels and O2. production can be monitored simultaneously. This technique was developed in the dimethylsulfoxide-differentiated monocytic-like U-937 cell line (not equal to U-937), validated by comparison with single dye measurements and applied to human alveolar macrophages. The chemotactic peptide N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine induced in both cell types a similar transient elevation in [Ca2+]i, followed within seconds by a sustained increase in O2 production, which was however 4-fold weaker in not equal to U-937 cells. These results indicate that O2 production is an early event following the stimulation of human alveolar macrophages. This new double-dye technique may be relevant to other O2 ion-producing cells and could help to define more precisely the kinetics of the events leading to this biological response.

Volatile flavour compounds in heated blackberry juices.

Volatile compounds were isolated from the blackberry juices of two wild varieties and one cultivated variety by simultaneous distillations/extractions. All extracts were freed from acids, fractionated, analysed by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-infrared spectrometry (GC-IR) and sensorially assessed at the sniffing-port of the chromatograph. Alcohols, furans and aldehydes are the predominant chemical classes of all kinds of heated juices, the most abundant compound being 2-heptanol in the cultivated variety and furfural in the wild blackberry varieties. The addition of 50% (w/w) sugar during the heating treatment does not modify the odorous profile.

Pyrazines responsible for the potatolike odor produced by some serratia and cedecea strains.

The pyrazines responsible for the potatolike odor produced by some Serratia and Cedecea cultures were identified by gas-liquid chromatography (with flame ionization and thermoionic ionization detectors) and mass spectrometry. Alkylpyrazines were produced by the six strains studied irrespective of their odors. The major alkyl-methoxypyrazine produced by Cedecea davisae was 3-sec-butyl-2-methoxypyrazine, and that produced by odoriferous Serratia strains (S. rubidaea, S. odorifera, and S. ficaria) was 3-isopropyl-2-methoxy-5-methyl-pyrazine. Other pyrazines produced by some strains were 3-isopropyl-2-methoxypyrazine, 3-isobutyl-2-methoxypyrazine, 3-sec-butyl-2-methoxy-5-methylpyrazine, and 3-isobutyl-2-methoxy-6-methylpyrazine. Some of these pyrazines had not previously been found as natural products or to be produced by bacteria.