Antimutagenicity of ursolic acid and oleanolic acid against doxorubicin-induced clastogenesis in Balb/c mice

Ursolic acid (UA) and oleanolic acid (OA) are triterpenoid compounds found in food, medicinal herbs and various other plants in free form or bound to glycosides. Both substances are known for their antimicrobial, hepatoprotective, anti-inflammatory, antiallergic, antiviral and cytotoxic activities. In the present study, we evaluated the antimutagenic potential of UA and OA using the micronucleus test in peripheral blood and bone marrow of Balb/c mice. The animals were divided into 10 treatment groups: mice treated with UA (80 mg/kg b.w.); OA (80 mg/kg b.w.); a mixture of UA and OA (80 mg/kg b.w.); the antineoplastic agent doxorubicin (DXR, 90 mg/kg b.w.); DMSO and DXR; UA and DXR; OA and DXR; UA, OA and DXR, and negative and solvent controls. UA, OA and a mixture of UA and OA were administered to the animals by gavage, followed by the intraperitoneal injection of DXR. The results showed a significant reduction in micronucleus frequency in the groups concomitantly treated with the triterpenoid compounds and DXR compared to that treated with DXR alone. The present results demonstrate the antimutagenic activity of UA and OA under the experimental conditions used in this study.     

Heat shock triggers MAPK activation and MKP-1 induction in Leydig testicular cells

Testicular function is highly dependent on temperature control. In Leydig testicular cells, the signaling pathway activated by heat stress is poorly defined. Here we describe the molecular events triggered by heat shock (HS, 10 min, 45 degrees C) in MA-10 cells. HS produced a rapid and transient activation of ERK1/2 and JNK kinases, and also increased MAP kinase phosphatase-1 (MKP-1) protein and mRNA levels. The effect of HS on MKP-1 messenger reached significance at 15 min, peaked (3.5-fold) at 60 min, and was partially dependent on ERK1/2 activity. The temporal profiles of MKP-1 protein levels and MAPKs phospho-dephosphorylation suggest that MKP-1 induction could contribute to ERK1/2 and JNK inactivation after HS. In summary, this study indicates that the response to heat stress in Leydig cells includes the activation of MAPKs related to cell survival (ERK1/2) and death (JNK), and the induction of a MAPK activity inhibitory loop.     

Isolation and characterization of circulating tumor cells using a novel workflow combining the CellSearch® system and the CellCelector™

Circulating tumor cells (CTC) are rare cells which have left the primary tumor to enter the blood stream. Although only a small CTC subgroup is capable of extravasating, the presence of CTCs is associated with an increased risk of metastasis and a shorter overall survival. Understanding the heterogeneous CTC biology will optimize treatment decisions and will thereby improve patient outcome. For this, robust workflows for detection and isolation of CTCs are urgently required. Here, we present a workflow to characterize CTCs by combining the advantages of both the CellSearch^® and the CellCelector? micromanipulation system. CTCs were isolated from CellSearch^® cartridges using the CellCelector? system and were deposited into PCR tubes for subsequent molecular analysis (whole genome amplification (WGA) and massive parallel multigene sequencing). By a CellCelector? screen we reidentified 97% of CellSearch^® SKBR‐3 cells. Furthermore, we isolated 97% of CellSearch^®‐proven patient CTCs using the CellCelector? system. Therein, we found an almost perfect correlation of R² = 0.98 (Spearman's rho correlation, n = 20, p < 0.00001) between the CellSearch^® CTC count (n = 271) and the CellCelector? detected CTCs (n = 252). Isolated CTCs were analyzed by WGA and massive parallel multigene sequencing. In total, single nucleotide polymorphisms (SNPs) could be detected in 50 genes in seven CTCs, 12 MCF‐7, and 3 T47D cells, respectively. Taken together, CTC quantification via the CellCelector? system ensures a comprehensive detection of CTCs preidentified by the CellSearch^® system. Moreover, the isolation of CTCs after CellSearch^® using the CellCelector? system guarantees for CTC enrichment without any contaminants enabling subsequent high throughput genomic analyses on single cell level. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:125–132, 2017