Analysis of erythromycin and oleandomycin residues in food by high-performance liquid chromatography with fluorometric detection.

A reliable and sensitive procedure is presented for the analysis of erythromycin (ERY) and oleandomycin (OLE) in food of animal origin, such as meat, liver, kidney, raw milk and egg. The method is based on a solid-phase extraction clean-up with a cation exchange cartridge, a 9-fluoromethylchloroformate (FMOC) precolumn derivatization and a separation by HPLC with fluorometric detection. The selectivity is satisfactory enough to control ERY and OLE residues as not many interfering peaks are observed for various food matrices. The macrolides recoveries of the total procedure were low, although >50%. However, addition of an internal standard (roxithromycin) corrected for recovery to give satisfactory quantitative results for repeatability, linearity, detection and quantification limits and mainly accuracy.

Assessment of the contaminant concentration variability among Lake Geneva Arctic char using stable isotopic composition (delta15N and delta13C).

Measurements of organochlorine [polychlorinated biphenyls (PCBs) and dichloro-diphenyl-dichloroethylene (DDE)] and Hg concentrations and nitrogen and carbon stable isotopic compositions (delta15N and delta13C) were performed on 63 Arctic char (Salvelinus alpinus) from Lake Geneva. Fish exhibited a high interindividual variablity in pollutant concentrations. Since the accumulation of such persistent contaminants is obtained from food, the co-occurrence of dietary differentiation leading to the contaminant interindividual variability was suspected. delta15N and delta13C were used for assessing trophic position and food source differences among Arctic char. The low ranges of delta15N and delta13C could not explain the interindividual variability in pollutant concentrations. The lack of relation between delta15N and contaminant concentration did not suggest a trophic level biomagnification of PCB, DDE, and Hg. Lake Geneva spatial variability in pollutants may be an important factor of variability within the Arctic char population. The bioaccumulation pattern occurring for Hg was not apparent for PCB and DDE. Organochlorines are hydrophobic contaminants, and their bioaccumulation pattern may be masked by seasonal variations in fish lipid content.

Determination of streptomycin residues in food by solid-phase extraction and liquid chromatography with post-column derivatization and fluorometric detection.

A reliable and sensitive procedure is presented for the analysis of streptomycin (STP) in food of animal origin, like meat, milk and honey. The method is based on a separation by ion-pair liquid chromatography with beta-naphthoquinone-4-sulfonate (NQS) postderivatization and fluorescence detection. The clean-up of the extract is done by solid-phase extraction, firstly with a cation-exchange cartridge and secondly with an octadecyl cartridge. The selectivity is very good and not many interfering peaks are observed for various food matrices. The streptomycin recovery of the total procedure is superior to 80%. The procedure is quantitatively characterized and repeatability, linearity, detection and quantification limits are very satisfactory. A special focus is given to STP residues in honeys and a survey on 64 commercial honeys is presented. For honey analysis, the HPLC method is compared with an immunoassay test (ELISA), and the possibility of using this test for screening with and without solid-phase extraction clean-up is also discussed.

Expression of the insulin-like growth factor I receptor C terminus as a myristylated protein leads to induction of apoptosis in tumor cells.

The insulin-like growth factor I receptor (IGF-IR) has been shown to mediate mitogenesis and suppression of apoptosis. Certain mutations in the COOH terminus of the receptor abrogate the antiapoptotic activity but not the mitogenic activity. However, truncation of the receptor by deletion of the COOH-terminal 108 amino acids enhances suppression of apoptosis by the IGF-IR, which suggests that the COOH terminus has a negative regulatory role. To investigate this further, a series of mammalian expression vectors were generated that encoded either the COOH terminus of the receptor or the COOH terminus plus the kinase domain. In some cases, the first 16 amino acids of SRC were included at the NH2 terminus to provide a site for myristylation. In transient transfection assays, the membrane-targeted COOH-terminal construct, MyCF, was found to induce apoptosis in MCF-7 breast carcinoma cells and C6 glioblastoma cells, whereas the COOH-terminal construct without the myristylation signal, CF, was poorly cytotoxic. MyKCF, which encodes the kinase domain as well as the COOH terminus, had intermediate cytotoxicity. The cytotoxicity of MyCF was diminished by point mutations that were previously shown to abrogate suppression of apoptosis in the context of the full-length receptor. MCF-7 cells stably expressing the CF or the MyCF proteins exhibited decreased clonogenicity in soft agar and increased sensitivity to UV irradiation. These results indicate that expression of the IGF-IR COOH terminus promotes apoptosis of tumor cells, possibly by interfering with signals necessary for cell survival.

Anti-B4-blocked ricin synergizes with doxorubicin and etoposide on multidrug-resistant and drug-sensitive tumors.

Anti-B4-blocked ricin (anti-B4-bR) is an immunotoxin directed against CD19-positive cells that is currently being tested in several B-cell leukemia/lymphoma clinical trials. To explore the possibility of using anti-B4-bR in combination with chemotherapy protocols, we investigated the in vitro and in vivo cytotoxic effects of combining it with doxorubicin or etoposide using the lymphoma cell line Namalwa and a P-glycoprotein-expressing cell line, Namalwa/mdr-1, obtained by retroviral infection of Namalwa cells with the mdr-1 gene. Namalwa/mdr-1 cells were slightly more sensitive to anti-B4-bR than Namalwa cells; IC37 values were approximately 4 pmol/L and 8 pmol/L, respectively. When anti-B4-bR was combined simultaneously with doxorubicin or etoposide, additive to supra-additive killing of Namalwa and Namalwa/mdr-1 cells was observed. In xenografts of Namalwa/mdr-1 cells in severe combined immunodeficiency (SCID) mice, doxorubicin and etoposide at their maximum tolerated doses (3 mg/kg x 3 or 15 mg/kg x 3) showed no therapeutic effect. However, treatment with 5 daily bolus injections of anti-B4-bR (50 micrograms/kg) followed by treatment with doxorubicin or etoposide significantly increased the life span of the mice by 129% and 115%, respectively. After treatment with anti-B4-bR, the Namalwa/mdr-1 population expressed lower levels of P-glycoprotein, and this decrease may account for the synergistic action of the drug combinations. These results suggest that anti-B4-bR could be used to good effect in combination with current treatment regimens and further hint at a promising role for this immunotoxin in treatment of disease at the minimal residual disease stage, where cells may be resistant to chemotherapy.

Pesticide residues in food in Switzerland.

Every year, official Swiss laboratories are involved in the analysis of about 8000 food commodities in the context of pesticide residue surveys. Between 90 to 100 different active ingredients are found normally in total foodstuffs analysed and more than 6% of the investigated samples have a contamination level exceeding the federal tolerance limits. No real improvement in what concerns the quality of food with respect to pesticide contamination has been observed these last years.

Standardization of a simple method for the estimation of haemoglobin A2.

An improved method for the estimation of haemoglobin A2, at the same time precise, simple and cheap, is proposed. Haemoglobin A is separated from haemoglobin A2 by electrophoresis on Cellogel in discontinuous buffer at alkaline pH. The strips of cellulose acetate containing the haemoglobin fractions are completely dissolved in 80% acetic acid. The percentage of haemoglobin A2 present in each sample is calculated from the values for the spectrophotometric absorbance at 396 nm. The average percentage of haemoglobin A2 (+/- standard deviation) determined by this method was 2.31 +/- 0.37 in 51 normal subjects, and 4.64 +/- 0.53 in 29 subjects with heterozygous beta-thalassaemia.