The impact of short-term topical gatifloxacin and moxifloxacin on bacterial injection after hypodermic needle passage through human conjunctiva.

PURPOSE: To determine the bacterial contamination rate of a 27-gauge needle bore during conjunctival penetration in donor eye bank eyes and the effect of short-term use of topical 0.3% gatifloxacin and 0.5% moxifloxacin. METHODS: One hundred consecutive human donors had 10 conjunctival penetrations per 10 syringes per eye before antibiotic placement; this was repeated 15 min after antibiotic use. Samples were cultured by expressing 0.3 mL of saline through the needle. Positive cultures were speciated. RESULTS: There were 1,033 positive cultures (25.8% of all cultures); 568 (28.4%) pre-antibiotics, 249 (24.9%) after gatifloxacin (P=0.04, compared to the pre-antibiotic rate), and 216 (21.6%) after moxifloxacin (P<0.001). The most common organism was Staphylococcus epidermidis [334 positive cultures (8.4%)]. No antibiotic effect was seen on this or other organisms except S. aureus [4.6% pre-antibiotic, 2.8% after gatifloxacin (P=0.02), and 1.8% after moxifloxacin (P<0.001)] and other Staphylococcus species [5.3% pre-antibiotic, 3.6% after gatifloxacin (P=0.04), and 3.2% after moxifloxacin (P=0.01)]. CONCLUSIONS: Transconjunctival penetration often results in needle bore contamination; bacteria are included in an injected solution. Fifteen minutes of exposure to 2 topical antibiotics had a minimal effect on bacterial contamination and no significant effect on many common pathogens.

Ceramide mediates vascular dysfunction in diet-induced obesity by PP2A-mediated dephosphorylation of the eNOS-Akt complex.

Vascular dysfunction that accompanies obesity and insulin resistance may be mediated by lipid metabolites. We sought to determine if vascular ceramide leads to arterial dysfunction and to elucidate the underlying mechanisms. Pharmacological inhibition of de novo ceramide synthesis, using the Ser palmitoyl transferase inhibitor myriocin, and heterozygous deletion of dihydroceramide desaturase prevented vascular dysfunction and hypertension in mice after high-fat feeding. These findings were recapitulated in isolated arteries in vitro, confirming that ceramide impairs endothelium-dependent vasorelaxation in a tissue-autonomous manner. Studies in endothelial cells reveal that de novo ceramide biosynthesis induced protein phosphatase 2A (PP2A) association directly with the endothelial nitric oxide synthase (eNOS)/Akt/Hsp90 complex that was concurrent with decreased basal and agonist-stimulated eNOS phosphorylation. PP2A attenuates eNOS phosphorylation by preventing phosphorylation of the pool of Akt that colocalizes with eNOS and by dephosphorylating eNOS. Ceramide decreased the association between PP2A and the predominantly cytosolic inhibitor 2 of PP2A. We conclude that ceramide mediates obesity-related vascular dysfunction by a mechanism that involves PP2A-mediated disruption of the eNOS/Akt/Hsp90 signaling complex. These results provide important insight into a pathway that represents a novel target for reversing obesity-related vascular dysfunction.