Quantitative monitoring of single nucleotide mutations by allele-specific quantitative PCR can be used for the assessment of minimal residual disease in patients with hematological malignancies throughout their clinical course

BackgroundMonitoring of minimal residual disease (MRD) in patients with hematological malignancies is important for evaluating the patients\u27 therapeutic response and risk of relapse. Single nucleotide mutations associated with leukemogenesis can be considered as applicable MRD markers.MethodsWe developed an allele-specific quantitative polymerase chain reaction (AS-qPCR) for FLT3 2503G > T, KIT 2446G > T, and KIT 2447A > T and compared the change in the expression levels of the FLT3 or KIT mutations assessed by AS-qPCR to those of the RUNX1–RUNX1T1 fusion gene and WT1 by conventional quantitative PCR.ResultsThe AS-qPCR using primers including template-mismatched nucleotide or template-mismatched nucleotide plus locked nucleic acid substituted nucleotide provided higher selectivity for mutant nucleotides. The change in the expression levels of the FLT3 or KIT mutations at the time of relapse and just after hematopoietic stem cell transplantation correlated well with that of the RUNX1–RUNX1T1 fusion gene and WT1. Moreover, during complete remission, only AS-qPCR could detect low-level expression of residual mutations.ConclusionsThe AS-qPCR for analyzing single nucleotide mutations contributes to the monitoring of MRD in patients without recurrent fusion gene throughout the clinical course and thus broadens the spectrum of patients in whom MRD can be monitored

Evaluation of the Relationship between Current Internal 137Cs Exposure in Residents and Soil Contamination West of Chernobyl in Northern Ukraine

After the Chernobyl Nuclear Power Plant accident, the residents living around the Chernobyl were revealed to have been internally exposed to 137Cs through the intake of contaminated local foods. To evaluate the current situation of internal 137Cs exposure and the relationship between the 137Cs soil contamination and internal exposure in residents, we investigated the 137Cs body burden in residents who were living in 10 selected cities from the northern part of the Zhitomir region, Ukraine, and collected soil samples from three family farms and wild forests of each city to measured 137Cs concentrations. The total number of study participants was 36,862, of which 68.9%of them were female. After 2010, the annual effective doses were less than 0.1 mSv in over 90% of the residents. The 137Cs body burden was significantly higher in autumn than other seasons (p < 0.001) and in residents living in more contaminated areas (p < 0.001). We also found a significant correlation between the proportion of residents in each city with an estimated annual exposure dose exceeding 0.1 mSv and 137Cs concentration of soil samples from family farms (r = 0.828, p = 0.003). In conclusion, more than 25 years after the Chernobyl accident, the internal exposure doses to residents living in contaminated areas of northern Ukraine is limited but still related to 137Cs soil contamination. Furthermore, the consumption of local foods is considered to be the cause of internal exposure

Missing western half of the Pacific Plate: Geochemical nature of the Izanagi-Pacific Ridge interaction with a stationary boundary between the Indian and Pacific mantles

The source mantle of the basaltic ocean crust on the western half of the Pacific Plate was examined using Pb–Nd–Hf isotopes. The results showed that the subducted Izanagi–Pacific Ridge (IPR) formed from both Pacific (180–∼80 Ma) and Indian (∼80–70 Ma) mantles. The western Pacific Plate becomes younger westward and is thought to have formed from the IPR. The ridge was subducted along the Kurile–Japan–Nankai–Ryukyu (KJNR) Trench at 60–55 Ma and leading edge of the Pacific Plate is currently stagnated in the mantle transition zone. Conversely, the entire eastern half of the Pacific Plate, formed from isotopically distinct Pacific mantle along the East Pacific Rise and the Juan de Fuca Ridge, largely remains on the seafloor. The subducted IPR is inaccessible; therefore, questions regarding which mantle might be responsible for the formation of the western half of the Pacific Plate remain controversial. Knowing the source of the IPR basalts provides insight into the Indian–Pacific mantle boundary before the Cenozoic. Isotopic compositions of the basalts from borehole cores (165–130 Ma) in the western Pacific show that the surface oceanic crust is of Pacific mantle origin. However, the accreted ocean floor basalts (∼80–70 Ma) in the accretionary prism along the KJNR Trench have Indian mantle signatures. This indicates the younger western Pacific Plate of IPR origin formed partly from Indian mantle and that the Indian–Pacific mantle boundary has been stationary in the western Pacific at least since the Cretaceous

Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension

OBJECTIVE: To evaluate whether eculizumab helps patients with anti-acetylcholine receptor-positive (AChR+) refractory generalized myasthenia gravis (gMG) achieve the Myasthenia Gravis Foundation of America (MGFA) post-intervention status of minimal manifestations (MM), we assessed patients' status throughout REGAIN (Safety and Efficacy of Eculizumab in AChR+ Refractory Generalized Myasthenia Gravis) and its open-label extension. METHODS: Patients who completed the REGAIN randomized controlled trial and continued into the open-label extension were included in this tertiary endpoint analysis. Patients were assessed for the MGFA post-intervention status of improved, unchanged, worse, MM, and pharmacologic remission at defined time points during REGAIN and through week 130 of the open-label study. RESULTS: A total of 117 patients completed REGAIN and continued into the open-label study (eculizumab/eculizumab: 56; placebo/eculizumab: 61). At week 26 of REGAIN, more eculizumab-treated patients than placebo-treated patients achieved a status of improved (60.7% vs 41.7%) or MM (25.0% vs 13.3%; common OR: 2.3; 95% CI: 1.1-4.5). After 130 weeks of eculizumab treatment, 88.0% of patients achieved improved status and 57.3% of patients achieved MM status. The safety profile of eculizumab was consistent with its known profile and no new safety signals were detected. CONCLUSION: Eculizumab led to rapid and sustained achievement of MM in patients with AChR+ refractory gMG. These findings support the use of eculizumab in this previously difficult-to-treat patient population. CLINICALTRIALSGOV IDENTIFIER: REGAIN, NCT01997229; REGAIN open-label extension, NCT02301624. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that, after 26 weeks of eculizumab treatment, 25.0% of adults with AChR+ refractory gMG achieved MM, compared with 13.3% who received placebo

Minimal Symptom Expression' in Patients With Acetylcholine Receptor Antibody-Positive Refractory Generalized Myasthenia Gravis Treated With Eculizumab

The efficacy and tolerability of eculizumab were assessed in REGAIN, a 26-week, phase 3, randomized, double-blind, placebo-controlled study in anti-acetylcholine receptor antibody-positive (AChR+) refractory generalized myasthenia gravis (gMG), and its open-label extension

Zoledronic Acid-Loaded β-TCP Inhibits Tumor Proliferation and Osteoclast Activation: Development of a Functional Bone Substitute for an Efficient Osteosarcoma Treatment

Osteosarcoma has a poor survival rate due to relapse and metastasis. Zoledronic acid (ZOL), an anti-resorptive and anti-tumor agent, is used for treating osteosarcoma. Delivery of ZOL to the target region is difficult due to its high binding affinity to bone minerals. This study developed a novel treatment for osteosarcoma by delivering ZOL to the target region locally and sustainably. In this study, we fabricated a novel bone substitute by loading ZOL on β-tricalcium phosphate (β-TCP). The ZOL-loaded β-TCP (ZOL/β-TCP) would be expected to express the inhibitory effects via both bound-ZOL (bound to β-TCP) and free-ZOL (release from ZOL/β-TCP). To explore the ability to release ZOL from the ZOL/β-TCP, the amount of released ZOL was measured. The released profile indicates that a small amount of ZOL was released, and most of it remained on the β-TCP. Our data showed that ZOL/β-TCP could successfully express the effects of ZOL via both bound-ZOL and free-ZOL. In addition, we examined the biological effects of bound/free-ZOL using osteosarcoma and osteoclasts (target cells). The results showed that two states of ZOL (bound/free) inhibit target cell activities. As a result, ZOL/β-TCP is a promising candidate for application as a novel bone substitute