Conformational variability of the stationary phase survival protein E from Xylella fastidiosa revealed by X‐ray crystallography,small‐angle X‐ray scattering studies,and normal mode analysis

Xylella fastidiosa is a xylem‐limited bacterium that infects a wide variety of plants. Stationary phase survival protein E is classified as a nucleotidase, which is expressed when bacterial cells are in the stationary growth phase and subjected to environmental stresses. Here, we report four refined X‐ray structures of this protein from X. fastidiosa in four different crystal forms in the presence and/or absence of the substrate 3′‐AMP. In all chains, the conserved loop verified in family members assumes a closed conformation in either condition. Therefore, the enzymatic mechanism for the target protein might be different of its homologs. Two crystal forms exhibit two monomers whereas the other two show four monomers in the asymmetric unit. While the biological unit has been characterized as a tetramer, differences of their sizes and symmetry are remarkable. Four conformers identified by Small‐Angle X‐ray Scattering (SAXS) in a ligand‐free solution are related to the low frequency normal modes of the crystallographic structures associated with rigid body‐like protomer arrangements responsible for the longitudinal and symmetric adjustments between tetramers. When the substrate is present in solution, only two conformers are selected. The most prominent conformer for each case is associated to a normal mode able to elongate the protein by moving apart two dimers. To our knowledge, this work was the first investigation based on the normal modes that analyzed the quaternary structure variability for an enzyme of the SurE family followed by crystallography and SAXS validation. The combined results raise new directions to study allosteric features of XfSurE protein.     

Gemcitabine anti-proliferative activity significantly enhanced upon conjugation with cell-penetrating peptides

Gemcitabine proven efficiency against a wide range of solid tumors and undergoes deamination to its inactive uridine metabolite, which underlies its low bioavailability, and tumour resistance was also associated with nucleoside transporter alterations. Hence, we have conjugated gemcitabine to cell-penetrating peptides (CPP), in an effort to both mask its aniline moiety and facilitate its delivery into cancer cells. Two CPP-drug conjugates have been synthesized and studied regarding both the time-dependent kinetics of gemcitabine release and their anti-proliferative activity on three different human cancer cell lines. Results obtained reveal a dramatic increase in the anti-proliferative activity of gemcitabine in vitro, upon conjugation with the CPPs. As such, CPP-gemcitabine conjugates emerge as promising leads for cancer therapy.     

Devoted fathers or selfish lovers? Conflict between mating effort and parental care in a harem‐defending arachnid

When there is a temporal trade‐off between mating effort and parental care, theoretical models predict that intense sexual selection on males leads to reduced paternal care. Thus, high‐quality males should invest more in mating effort because they have higher chances of acquiring mates, whereas low‐quality males should bias their investment towards parental care. Once paternal care has evolved, offspring value should also influence males’ decisions to invest in offspring attendance. Here, we performed a manipulation under field conditions to investigate the factors that influence male allocation in either mating effort or parental care. We predicted that facultative paternal care in the harem‐holding harvestman Serracutisoma proximum would be negatively influenced by male attractiveness and positively influenced by offspring value. We found that attractive males were less likely to engage in egg attendance and that the higher the perceived paternity, the higher the caring frequency. Finally, egg mortality was not related to caring frequency by males, but predation pressure was much lower than that recorded in previous studies with the same population. Thus, the benefits of facultative male care may be conditional to temporal variation in the intensity of egg predation. In conclusion, males adjust their investment in either territory defence or egg attendance according to their recent mating history and perceived paternity. Our findings suggest that exclusive paternal care can evolve from facultative paternal care only if the trade‐off between mating effort and parental care is circumvented.     

Incipient speciation driven by hypertrophied lips in Midas cichlid fishes?

Sympatric speciation has been debated in evolutionary biology for decades. Although it has gained in acceptance recently, still only a handful of empirical examples are seen as valid (e.g. crater lake cichlids). In this study, we disentangle the role of hypertrophied lips in the repeated adaptive radiations of Nicaraguan crater lake cichlid fish. We assessed the role of disruptive selection and assortative mating during the early stages of divergence and found a functional trade‐off in feeding behaviour between thick‐ and thin‐lipped ecotypes, suggesting that this trait is a target of disruptive selection. Thick‐lipped fish perform better on nonevasive prey at the cost of a poorer performance on evasive prey. Using enclosures in the wild, we found that thick‐lipped fish perform significantly better in rocky than in sandy habitats. We found almost no mixed pairs during two breeding seasons and hence significant assortative mating. Genetic differentiation between ecotypes seems to be related to the time since colonization, being subtle in L. Masaya (1600 generations ago) and absent in the younger L. Apoyeque (<600 generations ago). Genome‐wide differentiation between ecotypes was higher in the old source lakes than in the young crater lakes. Our results suggest that hypertrophied lips might be promoting incipient sympatric speciation through divergent selection (ecological divergence in feeding performance) and nonrandom mating (assortative mating) in the young Nicaraguan crater lakes. Nonetheless, further manipulative experiments are needed in order to confirm the role of hypertrophied lips as the main cue for assortative mating.     

Changes in the realized niche of the invasive succulent CAM plant Furcraea foetida

Furcraea foetida (Asparagaceae) is a native plant of Central America and northern South America but there is no information about its country of origin. The species was introduced into Brazil and is now considered invasive, particularly in coastal ecosystems. To date, nothing is known about the environmental factors that constrain its distribution and there is only inconclusive information about its location of origin. We used reciprocal distribution models (RDM) to assess invasion risk of F. foetida across Brazil and to identify source regions in its native range. We also tested the niche conservatism hypothesis using Principal Components Analyses and statistical tests of niche equivalency and similarity between its native and invaded ranges. For RDM analysis, we built two models using maximum entropy, one using records in the native range to predict the invaded distribution (forward‐Ecological Niche Model or forward‐ENM) and one using records in the invaded range to predict the native distribution (reverse‐ENM). Forward‐ENM indicated invasion risk in the Cerrado region and the innermost region of the Atlantic Forest, however, failed to predict the current occurrence in southern Brazil. Reverse‐ENM supported an existing hypothesis that F. foetida originated in the Orinoco river basin, Amazon basin and Caribbean islands. Prediction errors in the RDM and multivariate analysis indicated that the species expanded its realized niche in Brazil. The niche similarity test further suggested that the niche differences are because of differences in habitat availability between the two ranges, not because of evolutionary changes. We hypothesize that physiological pre‐adaptation (especially, the crassulacean acid metabolism), human‐driven propagule pressure and high competitive ability are the main factors determining the current spatial distribution of the species in Brazil. Our study highlights the need to include F. foetida in plant invasion monitoring programs, especially in priority conservation areas where the species has still not been introduced.     

The multidrug resistance transporters CgTpo1_1 and CgTpo1_2 play a role in virulence and biofilm formation in the human pathogen Candida glabrata

The mechanisms of persistence and virulence associated with Candida glabrata infections are poorly understood, limiting the ability to fight this fungal pathogen. In this study, the multidrug resistance transporters CgTpo1_1 and CgTpo1_2 are shown to play a role in C. glabrata virulence. The survival of the infection model Galleria mellonella, infected with C. glabrata, was found to increase upon the deletion of either CgTPO1_1 or CgTPO1_2. The underlying mechanisms were further explored. In the case of CgTpo1_1, this phenotype was found to be consistent with the observation that it confers resistance to antimicrobial peptides (AMP), such as the human AMP histatin‐5. The deletion of CgTPO1_2, on the other hand, was found to limit the survival of C. glabrata cells when exposed to phagocytosis and impair biofilm formation. Interestingly, CgTPO1_2 expression was found to be up‐regulated during biofilm formation, but and its deletion leads to a decreased expression of adhesin‐encoding genes during biofilm formation, which is consistent with a role in biofilm formation. CgTPO1_2 expression was further seen to decrease plasma membrane potential and affect ergosterol and fatty acid content. Altogether, CgTpo1_1 and CgTpo1_2 appear to play an important role in the virulence of C. glabrata infections, being at the cross‐road between multidrug resistance and pathogenesis.     

Snakebites in “Invisible Populations”: A cross-sectional survey in riverine populations in the remote western Brazilian Amazon

In the Brazilian Amazon, long distances, low healthcare coverage, common use of ineffective or deleterious self-care practices, and resistance to seeking medical assistance have an impact on access to antivenom treatment. This study aimed to estimate snakebite underreporting, and analyze barriers that prevent victims from obtaining healthcare in communities located in 15 municipalities on the banks of the Solimões, Juruá and Purus Rivers, in the remote Western Brazilian Amazon. Information on the participants’ demographics, previous snakebites, access to healthcare, time taken to reach medical assistance, use of self-care practices, and the reason for not accessing healthcare were collected through semi-structured interviews. In the case of deaths, information was collected by interviewing parents, relatives or acquaintances. A total of 172 participants who reported having suffered snakebites during their lifetime were interviewed. A total of 73 different treatment procedures was reported by 65.1% of the participants. Participants living in different river basins share few self-care procedures that use traditional medicine, and 91 (52.9%) participants reported that they had access to healthcare. Living in communities along the Juruá River [OR = 12.6 (95% CI = 3.2–49.7; p<0.001)] and the use of traditional medicine [OR = 11.6 (95% CI = 3.4–39.8; p<0.001)] were variables that were independently associated to the lack of access to healthcare. The main reasons for not accessing healthcare were the pprioritization of traditional treatments (70.4%), and the failure to recognize the situation as being potentially severe (50.6%). Four deaths from complications arising from the snakebite were reported, and three of these were from communities on the banks of the Juruá River. Only one of these received medical assistance. We found an unexpectedly high underreporting of snakebite cases and associated deaths. Snakebite victims utilized three main different healing systems: 1) self-care using miscellaneous techniques; 2) official medical healthcare generally combined with traditional practices; and 3) self-care using traditional practices combined with Western medicines. To mitigate snakebite burden in the Brazilian Amazon, an innovative intervention that would optimize timely delivery of care, including antivenom distribution among existing community healthcare centers, is needed.     

Microsatellite loci in the cactophilic species Drosophila antonietae (Diptera; Drosophilidae)

Morphometric and isoenzymatic data have showed little variation among Drosophila antonietae populations. This suggests historic gene flow, due to the distribution of these populations, which are associated with cacti along rivers, and natural selection on these markers. Microsatellite loci have high variability and are ideal for detecting gene flow and analysing population structure. Here we describe the isolation of seven polymorphic microsatellite loci in D. antonietae.     

Microsatellite loci from the pink shrimp Farfantenaeus notialis (Crustacea,Decapoda)

Farfantepenaeus notialis is an important resource for fisheries in Cuba. For this reason and for a sustainable exploitation it is important to study their population structure and genetic variability. We report and characterize microsatellites as genetic markers from this species. Fifteen microsatellite polymerase chain reaction (PCR) primers were designed and tested in some individuals from different populations. Seven pair of primers showed reliable amplification products and five were polymorphic. The allele number ranged from 4 to 33, and the observed and expected heterozygosities were relatively high with values between 0.63 and 0.74 and 0.56 and 0.81, respectively. Departures from Hardy–Weinberg equilibrium were observed for all loci.     

Identification of microsatellite loci in apricot

The sequences of 21 primer pairs of microsatellite loci screened from a genomic library of apricot (Prunus armeniaca L.) are reported in this study. All the identified microsatellite loci were characterized in a set of 25 apricot cultivars and revealed to be polymorphic with 3–12 alleles per locus. These markers showed to be more informative than isozymes and restriction fragment length polymorphisms (RFLPs) reported in the literature.