Using FTA® cards to store avian blood samples for genetic studies. Their application in sex determination

FTA® cards were used for long‐term storage of avian blood samples. Blood DNA was extracted by a simple method and used in PCR for sex identification of adult and nestling Great Grey Shrikes Lanius excubitor.     

The CR3 motif of Rrp44p is important for interaction with the core exosome and exosome function

The 10-subunit RNA exosome is involved in a large number of diverse RNA processing and degradation events in eukaryotes. These reactions are carried out by the single catalytic subunit, Rrp44p/Dis3p, which is composed of three parts that are conserved throughout eukaryotes. The exosome is named for the 3′ to 5′ exoribonuclease activity provided by a large C-terminal region of the Rrp44p subunit that resembles other exoribonucleases. Rrp44p also contains an endoribonuclease domain. Finally, the very N-terminus of Rrp44p contains three Cys residues (CR3 motif) that are conserved in many eukaryotes but have no known function. These three conserved Cys residues cluster with a previously unrecognized conserved His residue in what resembles a metal-ion-binding site. Genetic and biochemical data show that this CR3 motif affects both endo- and exonuclease activity in vivo and both the nuclear and cytoplasmic exosome, as well as the ability of Rrp44p to associate with the other exosome subunits. These data provide the first direct evidence that the exosome-Rrp44p interaction is functionally important and also provides a molecular explanation for the functional defects when the conserved Cys residues are mutated.     

A methodological approach to assess the small mammal community diversity in the temperate rainforest of Patagonia

Assessing small mammal diversity is a common procedure, which usually employs widespread standard techniques, for gathering information for a wide range of studies. Traditional methods, however, may be biased against capturing arboreal marsupials, such as Dromiciops gliroides, an endemic marsupial currently considered a rare species in the Patagonian temperate rainforest due to the low abundances reported previously. I tested a new capturing methodology to assess the small mammal diversity of an old-growth forest in Patagonia, based on a randomized and balanced design, which incorporated a combination of different trap types, bait types, and placement heights. The proposed methodology included four trap types (two for live-capturing: wire-mesh and Sherman traps, and two sign-recording traps for tracks and hair), two types of bait (banana and rolled oats), and two trap placements (ground level and 1.5–2.5 m above the ground). Trap type, bait type, and height of placement all had significantly different effects on capturing and detecting rodents or marsupials; environmental variables at the trap location also affected the ability to detect rodents and marsupials. Traditional methods used for sampling small mammals performed well for rodents but are not effective for capturing marsupials and vice versa, showing species-specific sampling protocols. There is no single combination of trap-bait-height capable to assess the entire small mammal community, but the combination of the most effective protocol for rodents and the most effective protocol for marsupials guarantee better results.     

Liver Accumulation of Plasmodium chabaudi-Infected Red Blood Cells and Modulation of Regulatory T Cell and Dendritic Cell Responses

It is postulated that accumulation of malaria-infected Red Blood Cells (iRBCs) in the liver could be a parasitic escape mechanism against full destruction by the host immune system. Therefore, we evaluated the in vivo mechanism of this accumulation and its potential immunological consequences. A massive liver accumulation of P. c. chabaudi AS-iRBCs (Pc-iRBCs) was observed by intravital microscopy along with an over expression of ICAM-1 on day 7 of the infection, as measured by qRT-PCR. Phenotypic changes were also observed in regulatory T cells (Tregs) and dendritic cells (DCs) that were isolated from infected livers, which indicate a functional role for Tregs in the regulation of the liver inflammatory immune response. In fact, the suppressive function of liver-Tregs was in vitro tested, which demonstrated the capacity of these cells to suppress naive T cell activation to the same extent as that observed for spleen-Tregs. On the other hand, it is already known that CD4+ T cells isolated from spleens of protozoan parasite-infected mice are refractory to proliferate in vivo. In our experiments, we observed a similar lack of in vitro proliferative capacity in liver CD4+ T cells that were isolated on day 7 of infection. It is also known that nitric oxide and IL-10 are partially involved in acute phase immunosuppression; we found high expression levels of IL-10 and iNOS mRNA in day 7-infected livers, which indicates a possible role for these molecules in the observed immune suppression. Taken together, these results indicate that malaria parasite accumulation within the liver could be an escape mechanism to avoid sterile immunity sponsored by a tolerogenic environment.     

Biomass-size spectra in aquatic communities in shallow fluctuating Mediterranean salt marshes (Emporda wetlands, NE Spain)

The distribution of biomass among phytoplankton and free-livinginvertebrates was analysed in a shallow Mediterranean salt marshsubmitted to fluctuating water level. Among phytoplankton, biomassaccumulated in sizes dominated by mixotrophic species, indicatinga competitive advantage for these species, which also prey onother smaller primary producers. Among invertebrates, biomassaccumulated in the larger sizes, corresponding to species whichpartially exploit other nearby systems, such as the aerial environment(insects), or to those able to exploit particulated organicmatter in marsh-bed sediment (amphipods). Biomass distributionmodels developed for pelagic systems are discussed in relationto fluctuating temporary waters. The integrated spectrum approximated(r² = 0.96) a Pareto distribution with a slope of c = 1.38.Intense disturbances caused a decrease in r² and an increasein c. Under stable conditions, two different tendencies wereobserved, depending on the degree of eutrophy of the basins:higher values of c were measured in the more eutrophic basins,and lower values in the less eutrophic ones. We hypothesizethat highly irregular nutrient input could explain these differences.     

Potential long-acting contraceptive agents: esters of testosterone with alkoxy- and halogeno-substituted carboxylic acids

The chemical synthesis and physical data of several new esters of testosterone (17 beta-hydroxyandrost-4-en-3-one), which contain either a halogeno or an alkoxy substituent in the acid chain, are reported.