Halofuginone and other febrifugine derivatives inhibit prolyl-tRNA synthetase

Febrifugine, the bioactive constituent of one of the 50 fundamental herbs of traditional Chinese medicine, has been characterized for its therapeutic activity, though its molecular target has remained unknown. Febrifugine derivatives have been used to treat malaria, cancer, fibrosis and inflammatory disease. We recently demonstrated that halofuginone (HF), a widely studied derivative of febrifugine, inhibits the development of T(H)17-driven autoimmunity in a mouse model of multiple sclerosis by activating the amino acid response (AAR) pathway. Here we show that HF binds glutamyl-prolyl-tRNA synthetase (EPRS), inhibiting prolyl-tRNA synthetase activity; this inhibition is reversed by the addition of exogenous proline or EPRS. We further show that inhibition of EPRS underlies the broad bioactivities of this family of natural product derivatives. This work both explains the molecular mechanism of a promising family of therapeutics and highlights the AAR pathway as an important drug target for promoting inflammatory resolution.     

Comparative analysis of phenolic profiles of ovipositional fluid of <Emphasis Type="Italic">Rhinusa pilosa</Emphasis> (Mecinini,Curculionidae) and its host plant <Emphasis Type="Italic">Linaria vulgaris</Emphasis> (Plantaginaceae)

Rhinusa pilosa (Gyllenhal) is a highly specific weevil that induces stem galls on the common toadflax Linaria vulgaris Mill. females oviposit the eggs near the apex of a growing shoot. The act of oviposition is accompanied by secretion of an ovipositional fluid, which is considered to be cecidogen, directly involved in gall induction. The remains of cecidogenic fluid were collected from the surface of the oviposition point on the stem. We performed a comparative analysis of the phenolics extracted from cecidogen, the stem and galls of L. vulgaris and adult and larva of R. pilosa by HPLC-DAD. One compound with A _max at 273, 332 nm (R _t 30.65 min) was exclusively found in the methanol extract of cecidogen. To further characterize the cecidogen and stem phenolic profiles, we used UHPLC coupled with an OrbiTrap mass analyzer. Among 49 phenolic compounds extracted from both the ovipositional fluid and the plant, protocatechuic acid and two phenolic glycosides were exclusively found in cecidogen: diosmetin-O-acetylrutinoside and an unidentified compound. The unknown compound produced an MS² base peak at 387 and 327 and 267 m/z base peaks at MS³ and MS⁴ fragmentation, respectively, and had the molecular formula C₃₂H₃₁O₁₈. The plausible role of phenolic compounds in the induction of gall formation on L. vulgaris is discussed.     

Loss of Y Chromosome in Peripheral Blood of Colorectal and Prostate Cancer Patients

Background

Although age-related loss of chromosome Y (LOY) in normal hematopoietic cells is a well-known phenomenon, the phenotypic consequences of LOY have been elusive. However, LOY has been found in association with smoking, shorter survival and higher risk of cancer. It was suggested that LOY in blood cells could become a predictive biomarker of male carcinogenesis.

Aims, Methods & Findings

To investigate the association of LOY in blood cells with the risk for development of colorectal (CC) and prostate cancers (PC), we have analyzed DNA samples from peripheral blood of 101 CC male patients (mean age 60.5±11.9 yrs), 70 PC patients (mean age 68.8±8.0 yrs) and 93 healthy control males (mean age 65.8±16.6 yrs). The methodology included co-amplification of homologous sequences on chromosome Y and other chromosomes using multiplex quantitative fluorescent (QF) PCR followed by automatic detection and analysis on ABI 3500 Genetic Analyzer. The mean Y/X ratio was significantly lower in the whole group of cancer patients (0.907±0.12; p = 1.17x10^-9) in comparison to the controls (1.015±0.15), as well as in CC (0.884±0.15; p = 3.76x10^-9) and PC patients (0.941±0.06; p = 0.00012), when analyzed separately. Multivariate logistic regression analysis adjusting for LOY and age showed that LOY is a more significant predictor of cancer presence than age, and that age probably does not contribute to the increased number of subjects with detectable LOY in cancer patients cohort.

Conclusion

In conclusion, our results support the recent findings of association of LOY in blood cells with carcinogenesis in males.     

Cross‐continental migratory connectivity and spatiotemporal migratory patterns in the great reed warbler

Migratory connectivity describes to which degree different breeding populations have distinct (non‐overlapping) non‐breeding sites. Uncovering the level of migratory connectivity is crucial for effective conservation actions and for understanding of the evolution of local adaptations and migratory routes. Here we investigate migration patterns in a passerine bird, the great reed warbler Acrocephalus arundinaceus, over its wide Western Palearctic breeding range using geolocators from Spain, Sweden, Czech Republic, Bulgaria and Turkey. We found moderate migratory connectivity: a highly significant spatial structure in the connections between breeding and sub‐Saharan non‐breeding grounds, but at the same time a partial overlap between individual populations, particularly along the Gulf of Guinea where the majority of birds from the Spanish, Swedish and Czech populations spent their non‐breeding period. The post‐breeding migration routes were similar in direction and rather parallel for the five populations. Birds from Turkey showed the most distinctive migratory routes and sub‐Saharan non‐breeding range, with a post‐breeding migration to east Africa and, together with birds from Bulgaria, a previously unknown pre‐breeding migration over the Arabian Peninsula indicating counter‐clockwise loop migration. The distances between breeding and sub‐Saharan non‐breeding sites, as well as between first and final sub‐Saharan non‐breeding sites, differed among populations. However, the total speed of migration did not differ significantly between populations; neither during post‐breeding migration in autumn, nor pre‐breeding migration in spring. There was also no significant relationship between the total speed of migration and distance between breeding and non‐breeding sites (neither post‐ nor pre‐breeding) and, surprisingly, the total speed of migration generally did not differ significantly between post‐breeding and pre‐breeding migration. Future challenges include understanding whether non‐breeding environmental conditions may have influenced the differences in migratory patterns that we observed between populations, and to which extent non‐breeding habitat fluctuations and loss may affect population sizes of migrants.     

Modulation of urokinase plasminogen activator system by poly(ADP-ribose)polymerase-1 inhibition

The urokinase plasminogen activator (uPA) system is a complex regulator of extracellular proteolysis which is involved in various physiological and pathological processes. The major components of this system are the serine protease uPA, two inhibitors PAI-1 and PAI-2, and the receptor uPAR. It has been previously shown by several groups that the uPA system has an important role in cancer progression and therefore its possible prognostic and therapeutic value has been evaluated. The aim of this study is to tackle the role of poly(ADP-ribosyl)ation in the induction of uPA activity in a glioblastoma cell line, A1235. This cell line is sensitive to alkylation damage and is a model for drug treatment. The components of the uPA system and the level of DNA damage were analyzed after alkylation agent treatment in combination with poly(ADP-ribose)polymerase-1 (PARP-1) inhibition. Here we show that the increase in uPA activity results from the net balance change between uPA and its inhibitor at mRNA level. Further, PARP-1 inhibition exerts its influence on uPA activity through DNA damage increase. Involvement of several signaling pathways, as well as cell specific regulation influencing the uPA system are discussed.