Safety Assessment of Compounds after In Vitro Metabolic Conversion Using Zebrafish Eleuthero Embryos.

Zebrafish-based platforms have recently emerged as a useful tool for toxicity testing as they combine the advantages of in vitro and in vivo methodologies. Nevertheless, the capacity to metabolically convert xenobiotics by zebrafish eleuthero embryos is supposedly low. To circumvent this concern, a comprehensive methodology was developed wherein test compounds (i.e., parathion, malathion and chloramphenicol) were first exposed in vitro to rat liver microsomes (RLM) for 1 h at 37 °C. After adding methanol, the mixture was ultrasonicated, placed for 2 h at -20 °C, centrifuged and the supernatant evaporated. The pellet was resuspended in water for the quantification of the metabolic conversion and the detection of the presence of metabolites using ultra high performance liquid chromatography-Ultraviolet-Mass (UHPLC-UV-MS). Next, three days post fertilization (dpf) zebrafish eleuthero embryos were exposed to the metabolic mix diluted in Danieau's medium for 48 h at 28 °C, followed by a stereomicroscopic examination of the adverse effects induced, if any. The novelty of our method relies in the possibility to quantify the rate of the in vitro metabolism of the parent compound and to co-incubate three dpf larvae and the diluted metabolic mix for 48 h without inducing major toxic effects. The results for parathion show an improved predictivity of the toxic potential of the compound.

The health risk for seafood consumers under future ocean acidification (OA) scenarios: OA alters bioaccumulation of three pollutants in an edible bivalve species through affecting the in vivo metabolism.

The current knowledge about the effect of pCO2-driven ocean acidification on the bioaccumulation of pollutants in marine species is still scarce, as only limited types of pollutants have been investigated. Therefore, to obtain a better understanding of the effect of ocean acidification on the process of bioaccumulation and subsequent food safety, the accumulation of benzo[a]pyrene (B[a]P), chloramphenicol (CAP), and nitrofurazone (NFZ) in an edible bivalve species, Tegillarca granosa, under present and near-future ocean acidification scenarios was investigated in the present study. The health risks associated with consuming contaminated blood clams were also assessed using target hazard quotient (THQ), lifetime cancer risk (CR), or margin of exposure (MoE). To explain the alterations in bioaccumulation of these pollutants, the expressions of genes encoding corresponding key metabolic proteins were analyzed as well. The results obtained showed that ocean acidification exerted a significant effect on the accumulation of B[a]P, NFZ, and CAP in the clams. After four-week exposure to B[a]P, NFZ, or CAP contaminated seawater acidified with CO2 at pH 7.8 and 7.4, significantly greater amounts of B[a]P and lower amounts of NFZ and CAP were accumulated in the clams compared to that in the control. Although no non-carcinogenic risk of consuming B[a]P-contaminated blood clams was detected using the THQ values obtained, the CR values obtained indicated a high life-time risk in all groups. In addition, according to the MoE values obtained, the health risks in terms of consuming NFZ- and CAP-contaminated clams were significantly reduced under ocean acidification scenarios but still cannot be ignored, especially for children. The gene expression results showed that the ability of clams to eliminate B[a]P may be significantly constrained, whereas the ability to eliminate NFZ and CAP may be enhanced under ocean acidification scenarios, indicating that the changes in the accumulation of these pollutants may be due to the altered in vivo metabolism.

Phospholipid vesicle-based permeation assay and EpiSkin® in assessment of drug therapies destined for skin administration.

Cost-effective and efficient methods for permeability screening are crucial during early development of drugs, drug formulations, and cosmeceuticals. Alternatives to animal experiments are impelled for both economical and ethical reasons. The aim of this study was to determine the ability of the phospholipid vesicle-based permeation assay (PVPA) to assess the effect of different formulations on drug permeability and thus establish its utility in formulation development. Three model drugs were tested in solutions and as liposomal formulations. The permeability results for the PVPA models were compared with the results for the reconstructed human skin model, EpiSkin(®). The drugs were ranked based on their estimated penetration potentials, and the results were in accordance with what was expected considering the physicochemical properties of the drugs. PVPAs (E-80, ceramide, cholesterol, cholesteryl sulfate, and palmitic acid) was able to distinguish between drug solutions and liposomal formulations; however, EpiSkin(®) detected only small differences between the drugs in solution and formulations. In contrast with EpiSkin(®), which is limited by a 3-day testing window, PVPA barriers can be stored frozen for up to 2 weeks or even up to 16 months, depending on their compositions. The PVPA models are thus more cost effective and efficient than the EpiSkin(®) model for permeability screening during early drug development.

Relationships between chemical oxygen demand (COD) components and toxicity in a sequential anaerobic baffled reactor/aerobic completely stirred reactor system treating Kemicetine.

In this study the interactions between toxicity removals and Kemicetine, COD removals, intermediate products of Kemicetine and COD components (CODs originating from slowly degradable organics, readily degradable organics, inert microbial products and from the inert compounds) were investigated in a sequential anaerobic baffled reactor (ABR)/aerobic completely stirred tank reactor (CSTR) system with a real pharmaceutical wastewater. The total COD and Kemicetine removal efficiencies were 98% and 100%, respectively, in the sequential ABR/CSTR systems. 2-Amino-1 (p-nitrophenil)-1,3 propanediol, l-p-amino phenyl, p-amino phenol and phenol were detected in the ABR as the main readily degradable inter-metabolites. In the anaerobic ABR reactor, the Kemicetin was converted to corresponding inter-metabolites and a substantial part of the COD was removed. In the aerobic CSTR reactor the inter-metabolites produced in the anaerobic reactor were completely removed and the COD remaining from the anerobic reactor was biodegraded. It was found that the COD originating from the readily degradable organics did not limit the anaerobic degradation process, while the CODs originating from the slowly degradable organics and from the inert microbial products significantly decreased the anaerobic ABR reactor performance. The acute toxicity test results indicated that the toxicity decreased from the influent to the effluent of the aerobic CSTR reactor. The ANOVA test statistics showed that there was a strong linear correlation between acute toxicity, CODs originating from the slowly degradable organics and inert microbial products. A weak correlation between acute toxicity and CODs originating from the inert compounds was detected.

Start-up strategies of UASB reactor for treatment of pharmaceutical wastewater.

Two start-up strategies of upflow anaerobic sludge blanket (UASB) reactor for treatment of pharmaceutical wastewater were investigated. The results showed that both of them were workable. Compared with the strategy that started up the reactor directly using chloromycetin wastewater, the strategy that started up the reactor first using mixed wastewater and then using chloromycetin wastewater could save time by 23%. When the latter strategy was adopted the development of sludge activity fluctuated more largely and its final activity was lower, but the sludge grew faster in the course of start-up.

Assessment of pancreatic ductal penetration of antibiotics.

Pancreatic ductal penetration of antibiotics is not uniform. Antibiotic therapy for pancreatic and pancreatic related infections, theoretically, is enhanced by drugs that reach the ductal system. The pancreatic ductal penetrance of Cefamandol (1 gm), Amikacin (7.5 mg/kg), and Chloramphenicol (1 gm) given as a single intravenous dose prior to endoscopic retrograde cholangiopancreatography was studied in ten patients. Serum and pancreatic juice were collected simultaneously, frozen, and later assayed for antibiotic concentration. Each antibiotic achieved its expected therapeutic serum level. In contrast, pancreatic ductal levels of Cefamandol and Amikacin were subtherapeutic, whereas Chloramphenicol levels were therapeutic. Further studies are needed to identify other antibiotics with good pancreatic ductal penetrance.

Pharmacokinetic assessment of immunosuppressive activity of antibiotics in human plasma by a modification of the mixed lymphocyte reaction.

Antibiotics may impair the development and expression of specific or nonspecific immune responses. Prophylactic administration of antibacterial antibiotics is widely used in ICUs. We studied the immunosuppressive activities of cefotaxime, chloramphenicol, gentamicin, metronidazole, and rifamycin as a function of time after the administration of these drugs to ICU patients, finding that the last 4 drugs had an immunosuppressive activity detectable up to 8 h by a mixed lymphocyte reaction. When these antimicrobial agents were added to normal pooled plasma in concentrations similar to those obtained in vivo, a similar degree of inhibition was observed.

Bioavailability and generic prescribing.

Although oral drug bioinequivalence has been attributed to a number of causes (excipients, dosage form, variation in dissolution time, and aging) less is known about bioavailability problems of topical medications in ophthalmology. Factors that can alter drug absorption from solutions (pH, partition coefficient, container impurities, contact time, etc.) are noted, and cases in which bioavailability problems should be considered as causes of therapeutic failure are discussed. Various attitudes representing pharmaceutical companies, the federal government, pharmacists, consumers and physicians toward the related problems of bioinequivalence and generic prescribing are examined. Techniques for in vivo and in vitro drug testing and for establishing uniform conditions of drug manufacture and storage can contribute to identification and minimization of bioavailability problems. A rational program based on a combination of such techniques could, ultimately, lead to establishment of the terms "generic equivalency" and "therapeutic equivalency" as synonymous.