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Striatal cholinergic interneurons (CINs) use acetylcholine (ACh) and glutamate (Glut) to regulate the striatal network since they express vesicular transporters for ACh (VAChT) and Glut (VGLUT3). However, whether ACh and Glut are released simultaneously and/or independently from cholinergic varicosities is an open question. The answer to that question requires the multichannel detection of vesicular transporters at the level of single synaptic vesicle (SV). Here, we used super-resolution STimulated Emission Depletion microscopy (STED) to characterize and quantify the distribution of VAChT and VGLUT3 in CINs SVs. Nearest-neighbor distances analysis between VAChT and VGLUT3-immunofluorescent spots revealed that 34% of CINs SVs contain both VAChT and VGLUT3. In addition, 40% of SVs expressed only VAChT while 26% of SVs contain only VGLUT3. These results suggest that SVs from CINs have the potential to store simultaneously or independently ACh and/or Glut. Overall, these morphological findings support the notion that CINs varicosities can signal with either ACh or Glut or both with an unexpected level of complexity.

Free-living amoebae (FLA) are widely distributed protozoa in nature, known to cause severe eye infections and central nervous system disorders. There is growing attention to the potential role that these protozoa could act as reservoirs of pathogenic bacteria and, consequently, to the possibility that, the persistence and spread of the latter may be facilitated, by exploiting internalization into amoebae. Shiga toxin-producing strains of Escherichia coli (STEC) are zoonotic agents capable of causing serious diseases, such as hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). Cattle represent the main natural reservoir of STEC, which are frequently found also in other domestic and wild ruminants, often without causing any evident symptoms of disease. The aspects related to the ecology of STEC strains in animal reservoirs and the environment are poorly known, including the persistence of these microorganisms within niches unfavorable to survival, such as soils or waters. In this study we investigated the interaction between STEC strains of serotype O157: H7 with different virulence gene profiles, and a genus of a wild free-living amoeba, Acanthamoeba sp. Our results confirm the ability of STEC strains to survive up to 20 days within a wild Acanthamoeba sp., in a quiescent state persisting in a non-cultivable form, until they reactivate following some stimulus of an unknown nature. Furthermore, our findings show that during their internalization, the E. coli O157 kept the set of the main virulence genes intact, preserving their pathogenetic potential. These observations suggest that the internalization in free-living amoebae may represent a means for STEC to resist in environments with non-permissive growth conditions. Moreover, by staying within the protozoa, STEC could escape their detection in the vehicles of infections and resist to the treatments used for the disinfection of the livestock environment.

The detection of the ionic products of low-rate fusion reactions, and in particular of the 11B(p,α)2α, is one of the recognized main problems in experiments where these reactions are initiated by tailored interaction of intense and high-energy lasers with matter. A thorough description of this important issue, with a critical comparison of the diagnostic opportunities, is indeed so far. In this work, we describe the common diagnostic methodologies used for the detection of the alpha particles generated by the 11B(p,α)2α reaction and, for each, we outline advantages and limitations, with considerations that can also be applied to other low-rate fusion reactions. We show here that, in general, the univocal characterization of the α products coming from this reaction can be achieved by the simultaneous use of several diagnostic tools placed in close proximity.

The human penis is a main portal of entry for numerous pathogens, and vaccines able to control resulting infections locally are highly desirable. However, in contrast to the gastrointestinal or vaginal mucosa, the penile immune system and mechanisms inducing a penile immune response remain elusive. In this descriptive study, using multiparametric flow cytometry and immunohistochemistry, we characterized mucosal immune cells such as B, T, and natural killer (NK) cells from the urethra, fossa, and glans of human adult penile tissues. We show that memory B lymphocytes and CD138+ plasma cells are detected in all penile compartments. CD4+ and CD8+ T lymphocytes reside in the epithelium and lamina propria of the penile regions and have mostly a resting memory phenotype. All penile regions contain CD56dim NK cells surface expressing the natural cytotoxicity receptor NKp44 and the antibody-dependent cell cytotoxicity receptor CD16. These cells are also able to spontaneously secrete pro- and anti-inflammatory cytokines, such as IL-17 and IL-22. Finally, CCR10 is the main homing receptor detected in these penile cells although, together with CCR3, CCR6, and CCR9, their expression level differs between penile compartments. Unlike antigen-presenting cells which type differ between penile regions as we reported earlier, urethral, fossa, and glans content in immune B, T, and NK cells is comparable. However, median values per each analysis suggest that the glans, containing higher number and more activated NK cells together with higher number of terminally differentiate effector CD8+ T cells, is a superior effector site than the urethra and the fossa. Thus, the human penis is an immunologically active tissue containing the cellular machinery required to induce and produce a specific and effective response against mucosal pathogens. It can therefore be considered as a classic mucosal effector site, a feature that must be taken into account for the elaboration of efficient strategies, including vaccines, against sexually transmitted infections.

Speciation is thought to depend on general ecological segregation rather than on strict allopatry in species with planetary-scale foraging ranges, such as Pterodroma petrels. Separation in both breeding islands and foraging grounds are generally considered to be the conditions for prezygotic isolation. However, it is unclear how both can be maintained in a context of ongoing distributional range shifts due to global change, with range contractions and expansions caused by changing land use in breeding territories and oceanographic conditions impacting productivity areas. In this context, we expect that wherever the fragile balance of allopatric segregation breaks down, gene flow may be re-established across the borders of what is currently considered separated species. In these “melting pot” areas, fine-scale ecological differentiation is likely the only force opposing panmixia in otherwise highly similar organisms. In this study, we focus on six Polynesian Pterodroma species currently found on Motu Nui, a protected islet offshore of Rapa Nui (Easter Island), four of which have likely extended their breeding range to that island recently. To understand the fine-scale interaction between these closely-related species in their new contact zone, we used an integrative approach combining ecological and genetic methods, including morphometric, bioacoustics, stable isotope niche reconstruction, breeding site characterisation, phenology traits, and both nuclear and mitochondrial DNA markers. We found that significant amounts of gene flow occur across what is currently considered species boundaries, sometimes questioning the relevance of the accepted species classification. Interestingly, our results indicate that multivariate ecological distance between individuals in a group of closely related species (P. alba, P. atrata, P. heraldica, and P. neglecta) is a relevant predictor of gene flow intensity, while more distant species such as P. ultima and P. nigripennis seem to maintain reproductive isolation. Thus, the case of Pterodroma petrels supports the idea that incomplete allopatric speciation processes may be halted or even reversed on a secondary contact zone. Beyond their direct taxonomical relevance, our results underline the importance of considering fine ecological structures for biodiversity mapping and conservation policies.

BackgroundType 2 diabetes (T2D) and obesity induce left ventricular (LV) dysfunction. The underlying pathophysiological mechanisms remain unclear, but myocardial triglyceride content (MTGC) could be involved.ObjectivesThis study aimed to determine which clinical and biological factors are associated with increased MTGC and to establish whether MTGC is associated with early changes in LV function.MethodsA retrospective study was conducted using five previous prospective cohorts, leading to 338 subjects studied, including 208 well-phenotyped healthy volunteers and 130 subjects living with T2D and/or obesity. All the subjects underwent proton magnetic resonance spectroscopy and feature tracking cardiac magnetic resonance imaging to measure myocardial strain.ResultsMTGC content increased with age, body mass index (BMI), waist circumference, T2D, obesity, hypertension, and dyslipidemia, but the only independent correlate found in multivariate analysis was BMI (p=0.01; R²=0.20). MTGC was correlated to LV diastolic dysfunction, notably with the global peak early diastolic circumferential strain rate (r=-0.17, p=0.003), the global peak late diastolic circumferential strain rate (r=0.40, p