Reliability of AMH in Serum after Long-term Storage at -80°C and an Extended Thawing Episode

Background: Measurement of anti-Müllerian hormone (AMH) is a valuable clinical tool for evaluating ovarian function. The present study aims to evaluate the reliability of AMH measurements obtained from samples kept for long-term storage with or without intermittent thawing.Methods and findings: Serum samples from 35 young female cancer patients were prospectively collected and stored at -80°C from 2007 until 2010. In 2011, AMH was analyzed with the DSL assay. During storage, the samples were exposed to a freezer error in 2013 that resulted in their being thawed up to 11°C for a maximum of 21 days and then refrozen. In 2014, the same samples (new aliquots) were analyzed with the Ansh-AMH assay. To test the reliability of the results from 2014, we conducted a thawing experiment on serum samples from 10 randomly selected females and compared the Ansh-AMH results for samples stored in a freezer with aliquots from the same samples that were stored at 11°C for 1, 3, 7, 14, and 21 days, respectively. Average AMH levels were 1.6 times higher when assayed with the Ansh-AMH compared with the DSL-AMH, which is in line with reported agreement between these types of assays. The same difference between the assays was found in samples that differed two years in storage time. The Ansh-AMH levels from ten serum samples without long-term storage were not influenced by exposure to 11°C for up to 21 days.Conclusions: The results indicate that long-term storage at -80°C and episodes of thawing have little impact on AMH levels analyzed with current methods. These data are reassuring and enable longitudinal studies to be planned that will analyze all collected serum samples simultaneously

Severe gonadotoxic insult manifests early in young girls treated for Ewing sarcoma

We prospectively investigated anti-Müllerian hormone (AMH) as a measure of ovarian insult in young females during and after treatment for Wilms tumor (WT), osteosarcoma (OS), and Ewing sarcoma (ES). Twenty-one female childhood cancer patients, with a mean age of 7.9 years (range 0.6-17), entered the study. Levels of AMH, follicle-stimulating hormone (FSH), and luteinizing hormone were monitored at diagnosis and every 3 to 4 months during, and regularly for a mean of 2.6 years after treatment. A profound decline in AMH was seen in the majority of the 21 study patients 3 to 4 months after the beginning of treatment, the exception being patients with WT, of whom 60% showed no such decline. During the remaining treatment, all patients except those with WT not treated with whole abdominal radiotherapy or stem cell transplantation (SCT) had AMH below detection limit. After completion of treatment, patients with OS and WT (without whole abdominal radiotherapy and SCT) recovered in AMH and had FSH in the normal range. In contrast, ES patients showed no AMH recovery and highly fluctuating FSH in the first years of follow-up, except for the 2 youngest patients, who had a late, slow AMH recovery. In conclusion, young female ES patients already showed signs of severe ovarian dysfunction during the first years after cancer treatment similar to patients treated with SCT and abdominal radiotherapy, in contrast to females with WT and OS. Fertility counseling and information concerning fertility preservation procedures should be considered before starting to treat young females with ES

Premature ovarian failure after childhood cancer and risk of metabolic syndrome : A cross-sectional analysis

Objective: Female childhood cancer survivors (CCS) are at risk of several late effects, such as metabolic syndrome (MetS) and premature ovarian insufficiency (POI). The objective is to study if POI is associated with risk of MetS and increased cardiovascular risk in CSS. Design: A cross-sectional study with a median time since the cancer diagnosis of 25 (12-41) years. Patients and controls were recruited from the South Medical Region of Sweden. Methods: The study included 167 female CCS, median age 34 (19-57) years, diagnosed with childhood cancer at median age 8.4 (0.1-17.9) years together with 164 controls, matched for age, sex, ethnicity, residence, and smoking habits. All subjects were examined with fasting glucose, insulin, HbA1c, and lipid profile. Fat mass was calculated with dual-energy X-ray absorptiometry (DXA), and questionnaires for medication were obtained. Detailed information of cancer treatment was available. Results: POI was present in 13% (22/167) among CCS (hypothalamic/pituitary cause excluded) and in none among controls. MetS was present in 14% (24/167) among all CCS (P = 0.001), in 23% (5/22) of those with POI (P < 0.001), compared with 4% (6/164) among controls. OR for MetS in all CCS compared with controls was 4.4 (95% CI: 1.8, 11.1) (P = 0.002) and among CCS with POI the OR was 7.7 (CI: 2.1, 28.1) (P = 0.002). Conclusion: The prevalence of MetS was higher in females treated for childhood cancer compared with controls, and the presence of POI significantly increased the risk of developing MetS