IFN-�� signaling, with the synergistic contribution of TNF-��, mediates cell specific microglial and astroglial activation in experimental models of Parkinson's disease

To through light on the mechanisms underlying the stimulation and persistence of glial cell activation in Parkinsonism, we investigate the function of IFN-γ and TNF-α in experimental models of Parkinson''s disease and analyze their relation with local glial cell activation. It was found that IFN-γ and TNF-α remained higher over the years in the serum and CNS of chronic Parkinsonian macaques than in untreated animals, accompanied by sustained glial activation (microglia and astroglia) in the substantia nigra pars compacta. Importantly, Parkinsonian monkeys showed persistent and increasing levels of IFN-γR signaling in both microglial and astroglial cells. In addition, experiments performed in IFN-γ and TNF-α KO mice treated with MPTP revealed that, even before dopaminergic cell death can be observed, the presence of IFN-γ and TNF-α is crucial for microglial and astroglial activation, and, together, they have an important synergistic role. Both cytokines were necessary for the full level of activation to be attained in both microglial and astroglial cells. These results demonstrate that IFN-γ signaling, together with the contribution of TNF-α, have a critical and cell-specific role in stimulating and maintaining glial cell activation in Parkinsonism.     

The tropical caprellid amphipod Paracaprella pusilla: a new alien crustacean in the Mediterranean Sea

Paracaprella pusilla Mayer (Fauna und Flora des Golfes von Neapel 17:1–55, 1890), originally described from Brazil, is one of the most abundant caprellid amphipod species in tropical and subtropical seas around the world. During a survey of caprellid amphipods from marinas along the Balearic Island (Western Mediterranean Sea) carried out between November 2011 and August 2012, we found two established populations of P. pusilla in Mallorca and Ibiza, respectively. So far, its occurrence in European waters was reported only from southwestern Spain in 2010. This record represents a northward range expansion of the species’ distribution, which is found for the first time in the Mediterranean. This is also the first record of the genus Paracaprella in the Mediterranean Sea. The most probable introduction vector was ship fouling. We also found the invasive caprellid Caprella scaura Templeton (Trans Entomol Soc Lond 1:185–198, 1836) in Mallorca and Menorca, which is recorded for the first time in the Balearic Islands, confirming its rapid expansion along the Mediterranean. When comparing reproductive traits between both alien species, we found that P. pusilla has a higher fecundity than C. scaura for the same female size. Taking into account this evidence, the species may be expected to appear in other Mediterranean and adjacent areas.     

Long-distance dispersal, low connectivity and molecular evidence of a new cryptic species in the obligate rafter Caprella andreae Mayer, 1890 (Crustacea: Amphipoda: Caprellidae)

The amphipod Caprella andreae Mayer, 1890 was recorded for the first time in Southern Iberian Peninsula (36°44′15″N, 3°59′38″W). This species is the only obligate rafter of the suborder Caprellidea and has been reported to attach not only to floating objects such as ropes or driftwoods but also to turtle carapaces. Mitochondrial and nuclear markers were used to examine dispersal capabilities and population genetic structure of C. andreae across seven localities in the Mediterranean and Atlantic Ocean collected from floating substrata with different dispersal patterns. The strong population differentiation with no haplotypes shared between populations suggests that C. andreae is quite faithful to the substratum on which it settles. In addition, the proportionally higher genetic diversity displayed in populations living on turtles as well as the presence of highly differentiated haplotypes in the same turtle population may be indicative that these populations survive longer, which could lead C. andreae to prefer turtles instead of floating objects to settle and disperse. Therefore, rafting on floating objects may be sporadic, and ocean currents would not be the most important factor shaping patterns of connectivity and population structure in this species. Furthermore, molecular phylogenetic analyses revealed the existence of a cryptic species whose estimates of genetic divergence are higher than those estimated between C. andreae and other congeneric species (e.g. Caprella dilatata and Caprella penantis). Discovery of cryptic species among widely distributed small marine invertebrates is quite common and, in this case, prompts for a more detailed phylogenetic analysis and taxonomic revision of genus Caprella. On the other hand, this study also means the first record of the gammarids Jassa cadetta and Elasmopus brasiliensis and the caprellid Caprella hirsuta on drifting objects.     

Modified hormonal balance in rooting-recalcitrant rac mutant tobacco shoots

ABSTRACT

The rooting-recalcitrant rac tobacco mutant has been multiplied in vitro via outgrowth of axillary buds in parallel to the D8 wild-type. The mutant shoots grew at a lower rate and did not root whatever the treatments, whereas the wild-type shoots rooted spontaneously during the culture cycle without auxin treatment. The mutant and wild-type shoots showed similar peroxidase variations along the culture cycle (21 days) but with higher levels of activity for the rac mutant: minimum peroxidase activity occurred at day 14 in whole shoots of both tobacco genotypes, but already at day 7 in the basal parts of the stems (where roots appear) of the wild-type tobacco, while it was delayed in the mutant. Free and conjugated auxin and polyamine levels were also determined in whole shoots and basal parts of the stems. The rac mutant was characterised by higher auxin and polyamine contents. A peak of auxins and polyamines appeared at day 14 in the whole shoots whatever the tobacco genotype. This peak was delayed in the basal parts of the rac stems compared to the wild-type ones. The mutant shoots contained higher levels of benzyladenine and isopentenyladenosine at the end of the culture cycle, whereas zeatin riboside was more abundant in wild-type shoots. In response to increasing concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA), only the wild-type shoots responded by an increase in growth rate followed by inhibition at high concentrations. The rac shoot responses were very low or nonexistent. Peroxidase activity was also measured in the basal parts of tobacco stems grown in the presence of IBA. Results suggest growth inhibition related to auxin accumulation, possibly combined with elevated putrescine content. Second, rooting induction seems to take place in both tobacco genotypes; however, the process of root formation is blocked in the mutant. The lack of initiation and expression phases of rooting in relation to auxin content in the mutant is discussed.     

Stress profiling of longevity mutants identifies Afg3 as a mitochondrial determinant of cytoplasmic mRNA translation and aging

Although environmental stress likely plays a significant role in promoting aging, the relationship remains poorly understood. To characterize this interaction in a more comprehensive manner, we examined the stress response profiles for 46 long‐lived yeast mutant strains across four different stress conditions (oxidative, ER, DNA damage, and thermal), grouping genes based on their associated stress response profiles. Unexpectedly, cells lacking the mitochondrial AAA protease gene AFG3 clustered strongly with long‐lived strains lacking cytosolic ribosomal proteins of the large subunit. Similar to these ribosomal protein mutants, afg3Δ cells show reduced cytoplasmic mRNA translation, enhanced resistance to tunicamycin that is independent of the ER unfolded protein response, and Sir2‐independent but Gcn4‐dependent lifespan extension. These data demonstrate an unexpected link between a mitochondrial protease, cytoplasmic mRNA translation, and aging.     

Molecular mechanisms underlying genotype‐dependent responses to dietary restriction

Dietary restriction (DR) increases lifespan and attenuates age‐related phenotypes in many organisms; however, the effect of DR on longevity of individuals in genetically heterogeneous populations is not well characterized. Here, we describe a large‐scale effort to define molecular mechanisms that underlie genotype‐specific responses to DR. The effect of DR on lifespan was determined for 166 single gene deletion strains in Saccharomyces cerevisiae. Resulting changes in mean lifespan ranged from a reduction of 79% to an increase of 103%. Vacuolar pH homeostasis, superoxide dismutase activity, and mitochondrial proteostasis were found to be strong determinants of the response to DR. Proteomic analysis of cells deficient in prohibitins revealed induction of a mitochondrial unfolded protein response (mtUPR), which has not previously been described in yeast. Mitochondrial proteotoxic stress in prohibitin mutants was suppressed by DR via reduced cytoplasmic mRNA translation. A similar relationship between prohibitins, the mtUPR, and longevity was also observed in Caenorhabditis elegans. These observations define conserved molecular processes that underlie genotype‐dependent effects of DR that may be important modulators of DR in higher organisms.     

In silico comparison of marine, terrestrial and synthetic compounds using ChemGPS-NP for navigating chemical space

Nature represents a vast source of chemical diversity, which is supposed to cover broader areas of chemical space than synthetically obtained substances typical of medicinal chemistry. With regard to drug discovery from nature, the terrestrial environment has been the most and longest studied source, while the investigation of compounds produced by marine organisms is still in its infancy. With the objective of demonstrating the enormous chemical diversity of nature, in particular that of the marine environment, we used the chemical space navigation tool ChemGPS-NP to compare sets of marine, terrestrial and synthetic compounds with respect to physico-chemical properties and their occupation of the biologically relevant chemical space. Despite considerable overlap, the three datasets clearly differ from each other by occupying and extending into different, specific, regions in chemical space. Synthetic compounds are e.g. comparably small, with some of them being highly flexible, while marine and terrestrial products are larger and characterised by higher and lower molecular flexibility, respectively, with increasing size. Moreover, the three datasets differ to some degree in polarity, aromaticity and heteroatom content. Taken together, ChemGPS-NP has been proven to be a useful tool for navigating large volumes of biologically relevant chemical space. In this study we demonstrated the chemical uniqueness and differences of large sets of natural products, with particular emphasis on marine substances. The hence de-veiled differences further underline the relevance of natural products, of both marine and terrester origin, for future drug discovery.     

The essential role of the phosphorylated pathway of serine biosynthesis in Arabidopsis

In plants, 3 different pathways of serine biosynthesis have been described: the Glycolate pathway, which is associated with photorespiration, and 2 non-photorespiratory pathways, the Glycerate and the Phosphorylated pathways. The Phosphorylated Pathway of Serine Biosynthesis (PPSB) has been known since the 1950s, but has been studied relatively little, probably because it was considered of minor significance as compared with the Glycolate pathway. In the associated study¹, we described for the first time in plants the in vivo functional characterization of the PPSB, by targeting the phosphoserine phosphatase (PSP1), the last enzyme of the pathway. Following a gain- and loss-of-function approach in Arabidopsis, we provided genetic and molecular evidence for the essential role of PSP1 for embryo and pollen development, and for proper root growth. A metabolomics study indicated that the PPSB affects glycolysis, the Krebs cycle, and the biosynthesis of several amino acids, which suggests that this pathway is an important link connecting metabolism and development. The mechanisms underlying the essential functions of PSP1 are discussed.     

Change Detection: Training and Transfer

Observers often fail to notice even dramatic changes to their environment, a phenomenon known as change blindness. If training could enhance change detection performance in general, then it might help to remedy some real-world consequences of change blindness (e.g. failing to detect hazards while driving). We examined whether adaptive training on a simple change detection task could improve the ability to detect changes in untrained tasks for young and older adults. Consistent with an effective training procedure, both young and older adults were better able to detect changes to trained objects following training. However, neither group showed differential improvement on untrained change detection tasks when compared to active control groups. Change detection training led to improvements on the trained task but did not generalize to other change detection tasks.     

Production and First-in-Man Use of T Cells Engineered to Express a HSVTK-CD34 Sort-Suicide Gene

Suicide gene modified donor T cells can improve immune reconstitution after allogeneic haematopoietic stem cell transplantation (SCT), but can be eliminated in the event of graft versus host disease (GVHD) through the administration of prodrug. Here we report the production and first-in-man use of mismatched donor T cells modified with a gamma-retroviral vector expressing a herpes simplex thymidine kinase (HSVTK):truncated CD34 (tCD34) suicide gene/magnetic selection marker protein. A stable packaging cell line was established to produce clinical grade vector pseudotyped with the Gibbon Ape Leukaemia Virus (GALV). T cells were transduced in a closed bag system following activation with anti-CD3/CD28 beads, and enriched on the basis of CD34 expression. Engineered cells were administered in two escalating doses to three children receiving T-depleted, CD34 stem cell selected, mismatched allogeneic grafts. All patients had pre-existing viral infections and received chemotherapy conditioning without serotherapy. In all three subjects cell therapy was tolerated without acute toxicity or the development of acute GVHD. Circulating gene modified T cells were detectable by flow cytometry and by molecular tracking in all three subjects. There was resolution of virus infections, concordant with detectable antigen-specific T cell responses and gene modified cells persisted for over 12 months. These findings highlight the suitability of tCD34 as a GMP compliant selection marker and demonstrate the feasibility, safety and immunological potential of HSVTK-tCD34 suicide gene modified donor T cells.

Trial Registration

ClinicalTrials.gov NCT01204502 <NCT01204502>