Tick prevalence and species diversity on Aldabran giant tortoises (Dipsochelys dussumieri) in relation to host range and host size in a restored ecosystem, Kenya

Tick species density and diversity on Aldabran tortoises was investigated in relation to the habitat range and size of each tortoise. Identification of tick infestation patterns forms an important aspect of effective tick control. Ten Aldabran tortoises were de‐ticked and monitored over the course of 2 months. Tick species found were Amblyomma sparsum, Amblyomma nuttalli, Amblyomma hebraeum and Boophilus decoloratus, with the most prevalent species found being A. sparsum. Tick loads varied considerably from 20 to 214 ticks per tortoise, with most ticks collected from the head/neck region. Tortoises ranging outside Haller Park had higher tick loads (70–214) compared with tortoises ranging within Haller Park (20–99). Tick load was not correlated with tortoise size. Results indicate that tick loads are related to the habitat range of the tortoises and may indirectly also be related to food preference and host food availability. Implications of the findings and appropriate tick control measures are discussed.     

Peptidyl-prolyl isomerase 1 (Pin1) preserves the phosphorylation state of tissue factor and prolongs its release within microvesicles

The exposure and release of TF is regulated by post-translational modifications of its cytoplasmic domain. Here, the potential of Pin1 to interact with the cytoplasmic domain of TF, and the outcome on TF function was examined. MDA-MB-231 and transfected-primary endothelial cells were incubated with either Pin1 deactivator Juglone, or its control Plumbagin, as well as transfected with Pin1-specific or control siRNA. TF release into microvesicles following activation, and also phosphorylation and ubiquitination states of cellular-TF were then assessed. Furthermore, the ability of Pin1 to bind wild-type and mutant forms of overexpressed TF-tGFP was investigated by co-immunoprecipitation. Additionally, the ability of recombinant or cellular Pin1 to bind to peptides of the C-terminus of TF, synthesised in different phosphorylation states was examined by binding assays and spectroscopically. Finally, the influence of recombinant Pin1 on the ubiquitination and dephosphorylation of the TF-peptides was examined. Pre-incubation of Pin1 with Juglone but not Plumbagin, reduced TF release as microvesicles and was also achievable following transfection with Pin1-siRNA. This was concurrent with early ubiquitination and dephosphorylation of cellular TF at Ser253. Pin1 co-immunoprecipitated with overexpressed wild-type TF-tGFP but not Ser258 → Ala or Pro259 → Ala substituted mutants. Pin1 did interact with Ser258-phosphorylated and double-phosphorylated TF-peptides, with the former having higher affinity. Finally, recombinant Pin1 was capable of interfering with the ubiquitination and dephosphorylation of TF-derived peptides. In conclusion, Pin1 is a fast-acting enzyme which may be utilised by cells to protect the phosphorylation state of TF in activated cells prolonging TF activity and release, and therefore ensuring adequate haemostasis.     

Bacterial endophytes from rice cut grass (<Emphasis Type="Italic">Leersia oryzoides</Emphasis> L.) increase growth,promote root gravitropic response,stimulate root hair formation,and protect rice seedlings from disease

Background and Aims

Leersia oryzoides, a wild relative of rice (Oryza sativa), may carry potential seed-borne bacterial endophytes which could be used to enhance growth of rice. We hypothesized that seed-associated bacteria from L. oryzoides would be compatible with rice and promote seedling growth, development, and survival.

Methods

We isolated bacteria from seed of L. oryzoides and checked compatibility with rice as well as Bermuda grass seeds for seedling growth promotion. Internal colonisation of bacteria into root cells was observed by ROS staining and microscopic observation. Growth promoting bacteria were evaluated for IAA production, phosphate solubilization and antifungal activities.

Results

Overall, ten bacteria were found to be growth promoting in rice seedlings with effects including restoration of root gravitropic response, increased root and shoot growth, and stimulation of root hair formation. All bacteria were identified by 16S rDNA sequencing. Six bacteria were found to become intracellular in root parenchyma and root hairs in rice and in Bermuda grass seedlings. Six bacteria were able to produce IAA in LB broth with highest (47.06 ± 1.99 μg ml^?1) by LTE3 (Pantoea hericii). Nine isolates solubilized phosphate and inhibited at least one soil borne fungal pathogen.

Conclusions

Seed bacteria of L. oryzoides are compatible with rice. Many of these bacteria become intracellular, induce root gravitropic response, increase root and shoot growth, and stimulate root hair formation in both rice and Bermuda grass seedlings. Presence of bacteria protects seedlings from soil pathogens during seedling establishment. This research suggests that bioprospecting microbes on near relatives of rice and other crop plants may be a viable strategy to obtain microbes to improve cultivation of crops.

     

University Sports Rivalries Provide Insights on Coalitional Psychology

Sports are an excellent venue for demonstrating evolutionary principles to audiences not familiar with academic research. Team sports and sports fandom feature dynamics of in-group loyalty and intergroup competition, influenced by our evolved coalitional psychology. We predicted that reactions to expressions signaling mutual team/group allegiance would vary as a function of the territorial context. Reactions should become more prevalent, positive, and enthusiastic as one moves from the home territory to a contested area, and from a contested area to a rival’s territory during active rival engagement. We also predicted that men would be more responsive than women based on sex differences in evolved coalitional psychology. The research team visited public places immediately prior to 2016–2017 collegiate football and basketball games. A male research confederate wore a sweatshirt displaying the logo of one of the competing university teams and vocalized the team’s most popular slogan when he saw a fan displaying similar logos. Observers followed 5 m behind, recording reactions (N?=?597) and response positivity/enthusiasm. Reaction tone was most positive in the rival territory, least positive in the home territory, and intermediate in the periphery and contested territory. Rates of “no reaction” were lowest in the rival territory but were highest in the periphery. Men had higher reaction rates and more positive and enthusiastic reaction tones than women. Reactions generally followed predictions based on expected signal value. This project provides evidence that coalitional psychology influences dynamics related to university sports team rivalries and that context matters for expressions of alliance.     

One-step affinity purification of fusion proteins with optimal monodispersity and biological activity: application to aggregation-prone HPV E6 proteins

Background

Bacterial expression and purification of recombinant proteins under homogeneous active form is often challenging. Fusion to highly soluble carrier proteins such as Maltose Binding Protein (MBP) often improves their folding and solubility, but self-association may still occur. For instance, HPV E6 oncoproteins, when produced as MBP-E6 fusions, are expressed as mixtures of biologically inactive oligomers and active monomers. While a protocol was previously developed to isolate MBP-E6 monomers for structural studies, it allows the purification of only one MBP-E6 construct at the time. Here, we explored a parallelizable strategy more adapted for biophysical assays aiming at comparing different E6 proteins.

Results

In this study, we took advantage of the distinct size and diffusion properties of MBP-E6 monomers and oligomers to separate these two species using a rapid batch preparation protocol on affinity resins. We optimized resin reticulation, contact time and elution method in order to maximize the proportion of monomeric MBP-E6 in the final sample. Analytical size-exclusion chromatography was used to quantify the different protein species after purification. Thus, we developed a rapid, single-step protocol for the parallel purification of highly monomeric MBP-E6 samples. MBP-fused HPV16 E6 samples obtained by this approach were validated by testing the binding to their prototypical peptide targets (the LXXLL motif from ubiquitine ligase E6AP) by BIAcore-SPR assay.

Conclusions

We have designed a rapid single-step batch affinity purification approach to isolate biologically active monomers of MBP-fused E6 proteins. This protocol should be generalizable to isolate the monomer (or the minimal biologically active oligomer) of other proteins prone to self-association.

     

Assessment of an ultra-sensitive IFNγ immunoassay prototype for latent tuberculosis diagnosis

Worldwide there are about 1.7 billion individuals with latent tuberculosis infection (LTBI) and only 5% to 15% will develop active tuberculosis (TB). It is recommended to treat only those most at risk of developing active TB to avoid problems of drug resistance. LTBI diagnosis involves reviewing the individual’s medical history, physical examination, and biological tests. Interferon gamma release assays (IGRA) can yield “undeterminate” or “uncertain” results, which makes clinical management decisions difficult. We assessed an ultra-sensitive immunoassay prototype based on single molecule array (SiMoA) technology to evaluate its overall performance, and in particular, its performance for indeterminate and uncertain positive or negative samples, as classified by the results from the current ELISA technique used for IFNγ quantification. We analyzed samples from hospitalized or consulting patients and healthcare workers from three hospitals in Paris, previously classified as negative (n = 30), positive (n = 35), uncertain negative (n = 25), uncertain positive (n = 31), or indeterminate (n = 30). We observed that with the SiMoA assay 83.3% of the indeterminate samples became interpretable and could be classified as negative, whereas 74% of uncertain positive samples were classified as positive. Most uncertain negative samples (72%) were reclassified as uncertain positive (68%) or positive (4%). The results suggest that the ultra-sensitive SiMoA IFNγ assay could represent a useful tool for the identification of true positive and negative samples among those giving indeterminate or uncertain results with the TB IGRA assay currently used.     

Stable isotope composition and parasitic infections of harbor seal young‐of‐the‐year used as prey‐based diet indicators

The stable isotope composition (δ¹³C and δ¹⁵N values) of harbor seals (Phoca vitulina) is influenced by their diet. Young‐of‐the‐year during lactation and postweaning fast are expected be enriched in ¹⁵N compared to foraging seals. We studied the temporal variation of stable isotope composition of young‐of‐the‐year and adults to determine from which point in time the young‐of‐the‐year tissues (i.e., muscles and vibrissae) are influenced by independent foraging only. These results were compared with the development of trophically transmitted parasitic infections. The δ¹⁵N values in young‐of‐the‐year muscles decreased from June (20.3‰ ± 0.5‰) to October (18.5‰ ± 0.4‰), while those of foraging seals were all year long below 19.2‰. This decrease coincides with the increase of parasitic infections in young‐of‐the‐year, reflecting a shift to fish diet. Together these results suggest that the muscles of the young‐of‐the‐year older than 5–6 mo reflect independent foraging and that they can therefore be used in community diet studies. The nursing signal in vibrissae was unclear, as the δ¹⁵N values of young‐of‐the‐year were stable over time, whereas those of adults varied seasonally. However, δ¹⁵N values of nursing pups were significantly higher than those of adults in May and June, maybe due to their reliance on milk.     

Evaluation and optimization of a vibrotactile signal in an autonomous driving context

An important issue in autonomous vehicle development is the transition phase between autonomous and manual driving. Here, we focused on a vibrotactile signal that warns the driver 1 min before they must take over. Our goal was to design a signal detectable by, and satisfactory for, all drivers. After listing parameters defining a vibrotactile signal, we performed a design‐of‐experiments study to select 16 signals for evaluation by 80 participants of different ages, body mass indexes, and genders. In a car on the road, autonomous driving conditions were simulated with the participant in the front passenger seat, who performed a task unrelated to driving. Whenever the participant detected a signal, they gave a satisfaction score. Modeling enabled us to evaluate the effects of the signal parameters and participant characteristics on satisfaction and detection. Moreover, we obtained specifications for the design of a tactile signal according to our two criteria.

Practical applications

To the best of our knowledge, this is the first study to focus on the sensory aspect of a tactile signal in a car environment. Both functional (i.e., signal detection) and hedonic aspects are important. We investigated the influence of gender, age, and body mass index of the participants on signal detection and satisfaction score, and optimized the satisfaction score under detection constraints. We use a 3‐point method to reach those two goals:

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Sequence diversity of five <Emphasis Type="Italic">Medicago sativa</Emphasis> genes involved in agronomic traits to set up allele mining in breeding

Lucerne (Medicago sativa) is an autotetraploid forage legume, whose breeding mainly relies on phenotypic recurrent selection to create synthetic populations. Allele mining could be a strategy to take advantage of the large genetic diversity of the species. This paper reports the polymorphism of five genes impacting the phenotype for selected traits: CAD1 and CCoaOMT (digestibility), CONSTANS-like (forage yield), NHX1 (salt tolerance), and WXP1 (drought tolerance). Complete genes were sequenced for 387 genotypes of 43 cultivated accessions and 20 genotypes of wild accessions. Wild versus cultivated polymorphism were compared, adaptive evolution was evaluated by comparing M. sativa and Medicago truncatula sequences and variants of the cultivated pool were characterized. We showed that the wild pool was more variable than the cultivated pool, with 36.6% and 8.4% of the variants that were specific to the wild pool and the cultivated pool, respectively. This result confirmed a bottleneck effect during domestication and selection. We also found that CAD1, CCoaOMT, and NHX1 were under a strong purifying selection and contained few non-synonymous variants (8, 5, and 8, respectively), while CONSTANS-like and WXP1 were under a less pronounced purifying selection and were more polymorphic (45 and 91 non-synonymous variants, respectively). This result suggests that adaptive evolution could be an indicator of expected polymorphism. The application of allele mining strategy for plant breeding in autotetraploid species is discussed.