Lead and its compounds – Determination of δ‐aminolevulinic acid in urine by HPLC with fluorescence detection : Biomonitoring Methods, 2018

The working group “Analyses in Biological Materials” of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area verified the presented biomonitoring method. The method described hereinafter permits the determination of δ‐aminolevulinic acid (ALA) in urine as a biological marker of effect to assess exposure to lead. An indicator of such exposure is the increased urinary excretion of ALA caused by lead‐induced inhibition of the enzyme δ‐aminolevulinic acid dehydratase. The determination of ALA in urine is based on a condensation reaction of ALA with formaldehyde and acetylacetone yielding a pyrrolizine derivate, which can be sensitively detected using fluorescence detection. 6‐Amino‐5‐oxohexanoic acid is used as an internal standard. Calibration standards are prepared in pooled urine and processed in the same way as the samples to be analysed. The method was extensively validated and the reliability data were confirmed by two independent laboratories, which have established and cross‐checked the whole procedure

Blei und seine Verbindungen – Bestimmung von δ‐Aminolävulinsäure in Urin mittels HPLC‐Fluoreszenzdetektion : Biomonitoring Methods in German language, 2018

The working group “Analyses in Biological Materials” of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area verified the presented biomonitoring method. The method described hereinafter permits the determination of δ‐aminolevulinic acid (ALA) in urine as a biological marker of effect to assess exposure to lead. An indicator of such exposure is the increased urinary excretion of ALA caused by lead‐induced inhibition of the enzyme δ‐aminolevulinic acid dehydratase. The determination of ALA in urine is based on a condensation reaction of ALA with formaldehyde and acetylacetone yielding a pyrrolizine derivate, which can be sensitively detected using fluorescence detection. 6‐Amino‐5‐oxohexanoic acid is used as an internal standard. Calibration standards are prepared in pooled urine and processed in the same way as the samples to be analysed. The method was extensively validated and the reliability data were confirmed by two independent laboratories, which have established and cross‐checked the whole procedure

Deep sequencing in conjunction with expression and functional analyses reveals activation of FGFR1 in ewing sarcoma

PURPOSE: A low mutation rate seems to be a general feature of pediatric cancers, in particular in oncofusion gene-driven tumors. Genetically, Ewing sarcoma is defined by balanced chromosomal EWS/ETS translocations, which give rise to oncogenic chimeric proteins (EWS-ETS). Other contributing somatic mutations involved in disease development have only been observed at low frequency. EXPERIMENTAL DESIGN: Tumor samples of 116 Ewing sarcoma patients were analyzed here. Whole-genome sequencing was performed on two patients with normal, primary, and relapsed tissue. Whole-exome sequencing was performed on 50 Ewing sarcoma and 22 matched normal tissues. A discovery dataset of 14 of these tumor/normal pairs identified 232 somatic mutations. Recurrent nonsynonymous mutations were validated in the 36 remaining exomes. Transcriptome analysis was performed in a subset of 14 of 50 Ewing sarcomas and DNA copy number gain and expression of FGFR1 in 63 of 116 Ewing sarcomas. RESULTS: Relapsed tumors consistently showed a 2- to 3-fold increased number of mutations. We identified several recurrently mutated genes at low frequency (ANKRD30A, CCDC19, KIAA0319, KIAA1522, LAMB4, SLFN11, STAG2, TP53, UNC80, ZNF98). An oncogenic fibroblast growth factor receptor 1 (FGFR1) mutation (N546K) was detected, and the FGFR1 locus frequently showed copy number gain (31.7%) in primary tumors. Furthermore, high-level FGFR1 expression was noted as a characteristic feature of Ewing sarcoma. RNA interference of FGFR1 expression in Ewing sarcoma lines blocked proliferation and completely suppressed xenograft tumor growth. FGFR1 tyrosine kinase inhibitor (TKI) therapy in a patient with Ewing sarcoma relapse significantly reduced 18-FDG-PET activity. CONCLUSIONS: FGFR1 may constitute a promising target for novel therapeutic approaches in Ewing sarcoma

Methotrexate, doxorubicin, and cisplatin (MAP) plus maintenance pegylated interferon alfa-2b versus MAP alone in patients with resectable high-grade osteosarcoma and good histologic response to preoperative MAP : first results of the EURAMOS-1 good respons randomized controlled trial

Purpose EURAMOS-1, an international randomized controlled trial, investigated maintenance therapy with pegylated interferon alfa-2b (IFN-α-2b) in patients whose osteosarcoma showed good histologic response (good response) to induction chemotherapy. Patients and Methods At diagnosis, patients age ≤ 40 years with resectable high-grade osteosarcoma were registered. Eligibility after surgery for good response random assignment included ≥ two cycles of preoperative MAP (methotrexate, doxorubicin, and cisplatin), macroscopically complete surgery of primary tumor, < 10% viable tumor, and no disease progression. These patients were randomly assigned to four additional cycles MAP with or without IFN-α-2b (0.5 to 1.0 μg/kg per week subcutaneously, after chemotherapy until 2 years postregistration). Outcome measures were event-free survival (EFS; primary) and overall survival and toxicity (secondary). Results Good response was reported in 1,041 of 2,260 registered patients; 716 consented to random assignment (MAP, n = 359; MAP plus IFN-α-2b, n = 357), with baseline characteristics balanced by arm. A total of 271 of 357 started IFN-α-2b; 105 stopped early, and 38 continued to receive treatment at data freeze. Refusal and toxicity were the main reasons for never starting IFN-α-2b and for stopping prematurely, respectively. Median IFN-α-2b duration, if started, was 67 weeks. A total of 133 of 268 patients who started IFN-α-2b and provided toxicity information reported grade ≥ 3 toxicity during IFN-α-2b treatment. With median follow-up of 44 months, 3-year EFS for all 716 randomly assigned patients was 76% (95% CI, 72% to 79%); 174 EFS events were reported (MAP, n = 93; MAP plus IFN-α-2b, n = 81). Hazard ratio was 0.83 (95% CI, 0.61 to 1.12; P = .214) from an adjusted Cox model. Conclusion At the preplanned analysis time, MAP plus IFN-α-2b was not statistically different from MAP alone. A considerable proportion of patients never started IFN-α-2b or stopped prematurely. Long-term follow-up for events and survival continues