Among the strategies targeting vector control, the exploitation of the endosymbiont Wolbachia to produce sterile males and/or invasive females with reduced vector competence seems to be promising. A new Aedes albopictus transinfection (ARwP-M) was generated by introducing wMel Wolbachia in the ARwP line which had been established previously by replacing wAlbA and wAlbB Wolbachia with the wPip strain. Various infection and fitness parameters were studied by comparing ARwP-M, ARwP and wild-type (S_ANG population) Ae. albopictus sharing the same genetic background. Moreover, the vector competence of ARwP-M related to chikungunya, dengue and zika viruses was evaluated in comparison with ARwP. ARwP-M showed a 100% rate of maternal inheritance of wMel and wPip Wolbachia. Survival, female fecundity and egg fertility did not show to differ between the three Ae. albopictus lines. Crosses between ARwP-M males and S_ANG females were fully unfertile regardless of male age while egg hatch in reverse crosses increased from 0 to about 17% with S_ANG males aging from 3 to 17 days. When competing with S_ANG males for S_ANG females, ARwP-M males induced a level of sterility significantly higher than that expected for an equal mating competitiveness (mean Fried index of 1.71 instead of 1). The overall Wolbachia density in ARwP-M females was about 15 fold higher than in ARwP, mostly due to the wMel infection. This feature corresponded to a strongly reduced vector competence for chikungunya and dengue viruses (in both cases, 5 and 0% rates of transmission at 14 and 21 days post infection) with respect to ARwP females. Results regarding Zika virus did not highlight significant differences between ARwP-M and ARwP. However, none of the tested ARwP-M females was capable at transmitting ZIKV. These findings are expected to promote the exploitation of Wolbachia to suppress the wild-type Ae. albopictus populations. Aedes albopictus is one of the major human disease vectors and, despite substantial control efforts, it is rapidly spreading worldwide and increasing its epidemiological role. Thus, innovative approaches to fight this mosquito are urgently needed. Among the available control strategies, the exploitation of the endosymbiotic bacterium Wolbachia seems to be promising. In nature, the infection by Wolbachia is generally not detrimental, instead, it causes a series of modifications in host physiology promoting the spread of the infection in uninfected populations. Herein, we report on the artificial transinfection of specific Wolbachia strains in Ae. albopictus to replace its native Wolbachia infection type. This manipulation aimed at exploiting the expected modifications in the reproductive biology and vector competence of the species to contribute to reduce its epidemiological role. Specifically, we found that the new double Wolbachia infection did not affect Ae. albopictus fitness. The males belonging to the manipulated line, ARwP-M, induced full egg infertility in the wild-type females they mate with and showed increased male mating competitiveness. Remarkably, the ARwP-M females demonstrated significantly reduced competence for chikungunya and dengue viruses while both tested Ae. albopictus lines showed a very low susceptibility for Zika virus. These findings may encourage the use of ARwP-M Ae. albopictus as a highly efficient and safe biocide to suppress the wild-type populations.

A better understanding of locally advanced cervical cancer (LACC) is mandatory for further improving the rates of disease control, since a significant proportion of patients still fail to respond or undergo relapse after concurrent chemoradiation treatment (CRT), and survival for these patients has generally remained poor. To identify specific markers of CRT response, we compared pretreatment biopsies from LACC patients with pathological complete response (sensitive) with those from patients showing macroscopic residual tumor (resistant) after neoadjuvant CRT, using a proteomic approach integrated with gene expression profiling. The study of the underpinning mechanisms of chemoradiation response was carried out through in vitro models of cervical cancer. We identified annexin A2 (ANXA2), N-myc downstream regulated gene 1 (NDRG1) and signal transducer and activator of transcription 1 (STAT1) as biomarkers of LACC patients’ responsiveness to CRT. The dataset collected through qPCR on these genes was used as training dataset to implement a Random Forest algorithm able to predict the response of new patients to this treatment. Mechanistic investigations demonstrated the key role of the identified genes in the balance between death and survival of tumor cells. Our results define a predictive gene signature that can help in cervical cancer patient stratification, thus providing a useful tool towards more personalized treatment modalities.

Plant cultivation on spacecraft or planetary outposts is a promising and actual perspective both for food and bioactive molecules production. To this aim, plant response to ionizing radiations, as an important component of space radiation, must be assessed through on-ground experiments due to the potentially fatal effects on living systems. Hereby, we investigated the effects of X-rays and γ-rays exposure on tomato “hairy root” cultures (HRCs), which represent a solid platform for the production of pharmaceutically relevant molecules, including metabolites and recombinant proteins. In a space application perspective, we used an HRC system previously fortified through the accumulation of anthocyanins, which are known for their anti-oxidant properties. Roots were independently exposed to different photon radiations, namely X-rays (250 kV) and γ-rays (Co⁶⁰, 1.25 MeV), both at the absorbed dose levels of 0.5, 5, and 10 Gy. Molecular changes induced in the proteome of HRCs were investigated by a comparative approach based on two-dimensional difference in-gel electrophoresis (2D-DIGE) technology, which allowed to highlight dynamic processes activated by these environmental stresses. Results revealed a comparable response to both photon treatments. In particular, the presence of differentially represented proteins were observed only when roots were exposed to 5 or 10 Gy of X-rays or γ-rays, while no variations were appreciated at 0.5 Gy of both radiations, when compared with unexposed control. Differentially represented proteins were identified by mass spectrometry procedures and their functional interactions were analyzed, revealing variations in the activation of stress response integrated mechanisms as well as in carbon/energy and protein metabolism. Specific results from above-mentioned procedures were validated by immunoblotting. Finally, a morphometric analysis verified the absence of significant alterations in the development of HRCs, allowing to ascribe the observed variations of protein expression to processes of acclimation to ionizing radiations. Overall results contribute to a meaningful risk evaluation for biological systems exposed to extra-terrestrial environments, in the perspective of manned interplanetary missions planned for the near future.

Background: Releasing considerable numbers of radiation-sterilized males is a promising strategy to suppress mosquito vectors. However, releases may also include small percentages of biting females, which translate to non-negligible numbers when releases are large. Currently, the effects of irradiation on host-seeking and host-biting behaviors have not been exhaustively investigated. Information is also lacking regarding the effects of sterilizing treatment on the endosymbiotic bacteriumWolbachia, which is known to affect the vector competence of infected mosquitos.Methods: To ascertain the effects of irradiation on females, the pupae of twoAedes albopictusstrains, differing in their natural or artificialWolbachiainfection type, andAedes aegypti—which is not infected byWolbachia—were treated with various doses of X-rays and monitored for key fitness parameters and biting behavior over a period of 2 weeks. The effect of radiation onWolbachiawas investigated by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH) analysis.Results: PartialAedes albopictusfemale sterility was achieved at 28 Gy, but the number of weekly bites more than doubled compared to that of the controls. Radiation doses of 35 and 45 Gy completely inhibited progeny production but did not significantly affect the survival or flight ability ofAe. albopictusfemales and caused a tripling of the number of bites per female per week (compared to untreated controls). These results were also confirmed inAe. aegyptiafter treatment at 50 Gy.Wolbachiadensity decreased significantly in 45-Gy-irradiated females, with the greatest decreases in the early irradiation group (26 ± 2-h-old pupae).Wolbachiadensity also decreased as adults aged. This trend was confirmed in ovaries but not in extra-ovarian tissues. FISH analysis showed a strongly reducedWolbachia-specific fluorescence in the ovaries of 13 ± 1-day-old females.Conclusions: These results suggest that, under sterile insect technique (SIT) programs, the vector capacity of a target population could increase with the frequency of the irradiated females co-released with the sterile males due to an increased biting rate. In the context of successful suppression, the related safety issues are expected to be generally negligible, but they should be conservatively evaluated when large-scale programs relying on imperfect sexing and high overflooding release ratios are run for long periods in areas endemic for arboviral diseases. Also, the effects of irradiation on the vector competence deserve further investigation.Graphical