Study effect and mechanism of ofloxacin and levofloxacin on development of Rana nigromaculata tadpoles based on the hypothalamus-pituitary-thyroid axis.

Excessive antibiotics transferred into the aquatic environment may affect the development of amphibians. Previous studies on the aquatic ecological risk of ofloxacin generally ignored its enantiomers. The purpose of this study was to compare the effects and mechanisms of ofloxacin (OFL) and levofloxacin (LEV) on the early development of Rana nigromaculata. After 28-day exposure at environmental levels, we found that LEV exerted more severe inhibitory effects on the development of tadpoles than OFL. According to the enrichment results of differentially expressed genes in the LEV and OFL treatments, LEV and OFL had different effects on the thyroid development of tadpoles. dio2 and trh were affected by the regulation of dexofloxacin instead of LEV. At the protein level, LEV was the main component that affected thyroid development-related protein, while dexofloxacin in OFL had little effect on thyroid development. Furthermore, molecular docking results further confirmed that LEV was a major component affecting thyroid development-related proteins, including DIO and TSH. In summary, OFL and LEV regulated the thyroid axis by differential binding to DIO and TSH proteins, thereby exerting differential effects on the thyroid development of tadpoles. Our research is of great significance for comprehensive assessment of chiral antibiotics aquatic ecological risk.

Removal of oxytetracycline and ofloxacin in wastewater by microalgae-bacteria symbiosis for bioenergy production.

The development of microalgae-bacteria symbiosis for treating wastewater is flourishing owing to its high biomass productivity and exceptional ability to purify contaminants. A nature-selected microalgae-bacteria symbiosis, mainly consisting of Dictyosphaerium and Pseudomonas, was used to treat oxytetracycline (OTC), ofloxacin (OFLX), and antibiotic-containing swine wastewater. Increased antibiotic concentration gradually reduced biomass productivity and intricately changed symbiosis composition, while 1 mg/L OTC accelerated the growth of symbiosis. The symbiosis biomass productivity reached 3.4-3.5 g/L (5.7-15.3 % protein, 18.4-39.3 % carbohydrate, and 2.1-3.9 % chlorophyll) when cultured in antibiotic-containing swine wastewater. The symbiosis displayed an excellent capacity to remove 76.3-83.4 % chemical oxygen demand, 53.5-62.4 % total ammonia nitrogen, 97.5-100.0 % total phosphorus, 96.3-100.0 % OTC, and 32.8-60.1 % OFLX in swine wastewater. The microbial community analysis revealed that the existence of OTC/OFLX increased the richness and evenness of microalgae but reduced bacteria species in microalgae-bacteria, and the toxicity of OFLX to bacteria was stronger than that of OTC.

Uptake and effects of a mixture of widely used therapeutic drugs in Eruca sativa L. and Zea mays L. plants.

Pharmaceutically active compounds (PACs) are continuously dispersed into the environment due to human and veterinary use, giving rise to their potential accumulation in edible plants. In this study, Eruca sativa L. and Zea mays L. were selected to determine the potential uptake and accumulation of eight different PACs (Salbutamol, Atenolol, Lincomycin, Cyclophosphamide, Carbamazepine, Bezafibrate, Ofloxacin and Ranitidine) designed for human use. To mimic environmental conditions, the plants were grown in pots and irrigated with water spiked with a mixture of PACs at concentrations found in Italian wastewaters and rivers. Moreover, 10× and 100× concentrations of these pharmaceuticals were also tested. The presence of the pharmaceuticals was tested in the edible parts of the plants, namely leaves for E. sativa and grains for Z. mays. Quantification was performed by liquid chromatography mass spectroscopy (LC/MS/MS). In the grains of 100× treated Z. mays, only atenolol, lincomycin and carbamazepine were above the limit of detection (LOD). At the same concentration in E. sativa plants the uptake of all PACs was >LOD. Lincomycin and oflaxacin were above the limit of quantitation in all conditions tested in E. sativa. The results suggest that uptake of some pharmaceuticals from the soil may indeed be a potential transport route to plants and that these environmental pollutants can reach different edible parts of the selected crops. Measurements of the concentrations of these pharmaceuticals in plant materials were used to model potential adult human exposure to these compounds. The results indicate that under the current experimental conditions, crops exposed to the selected pharmaceutical mixture would not have any negative effects on human health. Moreover, no significant differences in the growth of E. sativa or Z. mays plants irrigated with PAC-spiked vs. non-spiked water were observed.

Enantioselective recognition mechanism of ofloxacin via Cu(II)-modulated DNA.

The specific interactions of Cu(2+) with self-complementary DNA sequences involving d[G4C4(GC)2G4C4], d[(GC)10], and d[(AT)10], as well as the chiral recognition mechanism of ofloxacin enantiomers via the Cu(II)-modulated DNAs, were investigated using characterizations of circular dichroism, gel electrophoresis, FT-IR spectroscopy, UV melting measurement, electron paramagnetic resonance, and HPLC. The Cu(II)-coordinated GC-rich DNAs exhibit amplified enantioselectivity toward the S-enantiomer of ofloxacin. Especially in the case of d[G4C4(GC)2G4C4], ofloxacin enantiomers intercalate into the two adjacent guanine bases through the minor groove mediated by Cu(2+), which leads to a more favorable binding between S-ofloxacin and DNA. The highest ee value of ofloxacin enantiomers in the permeate after being adsorbed by the Cu(II)-DNA complex is obtained as 49.2% in the R-enantiomer at the [Cu(2+)]/[base] molar ratio of 0.25, while at the [Cu(2+)]/[base] molar ratio of 0.05 the highest ee value of ofloxacin enantiomers in the retentate reaches 26.3% in the S-enantiomer. This work illustrates a novel promising route to construct DNA-based chiral selectors toward certain drug enantiomers through the programmable enantioselective recognition on the basis of DNA chirality and the specific binding of transition metal ions.

Sensitivity and proportionality assessment of metabolites from microdose to high dose in rats using LC-MS/MS.

BACKGROUND: The objective of this study was to evaluate the sensitivity requirement for LC-MS/MS as an analytical tool to characterize metabolites in plasma and urine at microdoses in rats and to investigate proportionality of metabolite exposure from a microdose of 1.67 µg/kg to a high dose of 5000 µg/kg for atorvastatin, ofloxacin, omeprazole and tamoxifen. RESULTS: Only the glucuronide metabolite of ofloxacin, the hydroxylation metabolite of omeprazole and the hydration metabolite of tamoxifen were characterized in rat plasma at microdose by LC-MS/MS. The exposure of detected metabolites of omeprazole and tamoxifen appeared to increase in a nonproportional manner with increasing doses. Exposure of ortho- and para-hydroxyatorvastatin, but not atorvastatin and lactone, increased proportionally with increasing doses. CONCLUSION: LC-MS/MS has demonstrated its usefulness for detecting and characterizing the major metabolites in plasma and urine at microdosing levels in rats. The exposure of metabolites at microdose could not simply be used to predict their exposure at higher doses.

Antimutagenic activity and radical scavenging activity of water infusions and phenolics from ligustrum plants leaves.

Water infusions of Ligustrum delavayanum and Ligustrum vulgare leaves and eight phenolics isolated therefrom have been assayed in vitro on ofloxacin-induced genotoxicity in the unicellular flagellate Euglena gracilis. The tested compounds luteolin, quercetin, luteolin-7-glucoside, luteolin-7-rutinoside, quercetin-3-rutinoside, apigenin-7-rutinoside, tyrosol and esculetin inhibited the mutagenic activity of ofloxacin (43 microM) in E. gracilis. Water infusions from leaves of L. delavayanum and L. vulgare showed higher antimutagenic effect (p(t) < 0.001). The activity of these samples against ofloxacin (86 microM)-induced genotoxicity was lower, but statistically significant (p(t) < 0.05), excluding the water infusion of L. delavayanum leaves (p(t) < 0.01). Efficacy of quercetin, luteolin-7-rutinoside, apigenin-7-rutinoside was insignificant. The antimutagenic effect of most phenolics we studied could be clearly ascribed to their DPPH scavenging activity, substitution patterns and lipophilicity.

Levofloxacin-resistant invasive Streptococcus pneumoniae in the United States: evidence for clonal spread and the impact of conjugate pneumococcal vaccine.

The emergence of fluoroquinolone resistance in sterile-site isolates of Streptococcus pneumoniae is documented in this study characterizing all invasive levofloxacin-resistant (MIC, > or = 8 mg/liter) S. pneumoniae isolates (n = 50) obtained from the Centers for Disease Control and Prevention Active Bacterial Core Surveillance from 1998 to 2002. Resistance among all isolates increased from 0.1% in 1998 to 0.6% in 2001 (P = 0.008) but decreased to 0.4% in 2002, while resistance among vaccine serotypes continued to increase from 0.3% in 1998 to 1.0% in 2002, suggesting that fluoroquinolones continue to exert selective pressure on these vaccine serotypes. Only 22% of resistant isolates were not covered by the conjugate vaccine serogroups. Multilocus sequence typing revealed that 58% of resistant strains were related to five international clones identified by the Pneumococcal Molecular Epidemiology Network, with the Spain(23F)-1 clone being most frequent (16% of all isolates). Thirty-six percent of the isolates were coresistant to penicillin, 44% were coresistant to macrolides, and 28% were multiresistant to penicillin, macrolides, and fluoroquinolones. Fifty percent of the isolates were resistant to any three drug classes. Ninety-four percent of the isolates had multiple mutations in the quinolone resistance-determining regions of the gyrA, gyrB, parC, and parE genes. In 16% of the isolates, there was evidence of an active efflux mechanism. An unusual isolate was found that showed only a single parE mutation and for which the ciprofloxacin MIC was lower (2 mg/liter) than that of levofloxacin (8 mg/liter). Our results suggest that invasive pneumococcal isolates resistant to levofloxacin in the United States show considerable evidence of multiple resistance and of clonal spread.

Penetration of topically applied ciprofloxacin, norfloxacin and ofloxacin into the aqueous humor of the uninflamed human eye.

This study was undertaken to compare aqueous humor penetration of topical 0.3% ciprofloxacin, 0.3% norfloxacin and 0.3% ofloxacin in 63 patients undergoing cataract surgery. The patients were divided into two groups. Group 1 (long-term treatment) involved 30 patients undergoing cataract extraction who received either 0.3% ciprofloxacin, 0.3% norfloxacin or 0.3% ofloxacin topical drops. Each patient was preoperatively given a single drop per hour six times. At the time of surgery, 0.1 ml aqueous fluid was aspirated from the anterior chamber and immediately stored at -70 degrees C. Topically applied ciprofloxacin, ofloxacin and norfloxacin achieved mean aqueous humor levels of 2.80 +/- 1.07, 2.95 +/- 1.19 and 1.50 +/- 0.48 micrograms/ml respectively. There was no statistically significant difference in intraocular mean aqueous levels of ciprofloxacin versus ofloxacin. Topical ciprofloxacin and ofloxacin achieved mean aqueous humor levels significantly higher than norfloxacin (p < 0.001 and p < 0.0008 respectively). Group 2 (short-term treatment) involved 33 patients undergoing cataract extraction who received 0.3% ciprofloxacin, 0.3% ofloxacin and 0.3% norfloxacin topical drops. These patients were given one drop at 90 minutes and one drop 30 minutes preoperatively. At the time of surgery, 0.1 ml aqueous fluid was aspirated from the anterior chamber and immediately stored at -70 degrees C. Topically applied ciprofloxacin, ofloxacin and norfloxacin achieved mean aqueous humor levels of 1.11 +/- 0.50, 1.50 +/- 0.62 and 1.20 +/- 0.43 micrograms/ml respectively. There was no statistically significant difference in intraocular mean aqueous humor levels of ofloxacin versus norfloxacin and ciprofloxacin versus norfloxacin. Topical ofloxacin achieved a significantly higher mean aqueous humor level than ciprofloxacin (p < 0.03). All levels were above the minimum inhibitory concentrations of ciprofloxacin, ofloxacin and norfloxacin for most of the sensitive organisms.