Role of MTHFR 677C>T and 1298A>C gene polymorphisms on renal toxicity caused by lead exposure in wastewater treatment plant workers.

Environmental and occupational lead (Pb) exposures continue to pose major public health problems. Wastewater treatment plant (WWTP) workers proved are exposing to high Pb concentrations in sludge departments. The aim of the work was to investigate the role of MTHFR C677T and MTHFR A1298C gene polymorphisms on alteration of oxidative stress and homocysteine levels in WWTP workers exposed to high Pb concentrations, and study its relations with renal functions. The study included 90 WWTP workers from Abu-Rawash WWTP. Homocysteine, creatinine, urea, malondialdehyde (MDA), and total antioxidant capacity (TAC) were measured. Polymorphisms of MTHFR C677CT and MTHFR A1298C genes were studied using PCR/RFLP. Urine Pb concentrations were also measured. About 32.2% of the workers were with detectable Pb levels. Pb, homocysteine, and MDA levels were significantly higher among workers carrying TT polymorphism compared to other MTHFR C677T gene polymorphisms, while TAC was significantly lower among them compared to other polymorphisms. The same results were found among workers carrying CC compared to other MTHFR A1298C gene polymorphisms. WWTP workers carrying MTHFR 677TT and MTHFR 1298CC are more susceptible to elevation of homocysteine and the urinary Pb compared to the workers with the other polymorphisms. Furthermore, those workers were found to have increase in urea and creatinine. Therefore, MTHFR C677T and MTHFR A1298C gene polymorphisms could be used for prediction of the susceptibility to the risk of kidney impairments among WWTP workers in the sludge departments caused by their exposure to high Pb in their workplace.

Carbamazepine, lamotrigine, levetiracetam and valproic acid in dried blood spots with liquid chromatography tandem mass spectrometry; method development and validation.

Monitoring of antiepileptic drugs in children with epilepsy require multiple visits at a clinic for blood collection. Dried blood spot sampling is an alternative way of collection, performed at home by self-collection and can save time and costs for patients and family members. The aim was to develop and validate an LC-MS/MS dried blood spot method for carbamazepine, lamotrigine, levetiracetam and valproic acid with the requirements of using standard equipment and material in a routine laboratory setting. Whatman-903 filter paper was utilized, and discs were punched into a 96 well plate with an automated puncher and barcode reading. Extraction with methanol/water solution including internal standards on an orbital shaker was followed by a vacuum centrifuge step and reconstitution in mobile phase. Bioanalytical validation was performed according to guidelines from European Medicines Agency and additional dried blood spot specific validation. Calibration curves of the four included drugs had R2 values ≥0.994. Therapeutic relevant concentrations were well within measuring ranges. Within and -between run precision had %CV:s of 2.9-10.5%. Accuracy (%bias) was between -16.5% (lower limit of quantification) to +7.4%. Blood spots in a volume range of 15-50µL with hematocrit in expected ranges for this patient group were within precision and accuracy limits. To test the method, concentrations from dried blood spot venous and capillary patient samples (n=50) were compared with plasma concentrations. Good correlations for all four drugs with R2 of >0.92 was shown. In summary, a fast method for dried blood spots based on a 96 well format was developed for four commonly prescribed antiepileptic drugs. This validated method with traceability in sample preparation by bar code reading makes it suitable for the clinical laboratory.