The expression and activity of thioredoxin reductase 1 splice variants v1 and v2 regulate the expression of genes associated with differentiation and adhesion

The mammalian redox-active selenoprotein thioredoxin reductase (TrxR1) is a main player in redox homoeostasis. It transfers electrons from NADPH to a large variety of substrates, particularly to those containing redox-active cysteines. Previously, we reported that the classical form of cytosolic TrxR1 (TXNRD1_v1), when overexpressed in human embryonic kidney cells (HEK-293), prompted the cells to undergo differentiation [Nalvarte et al. (2004) J. Biol. Chem. 279, 54510–54517]. In the present study, we show that several genes associated with differentiation and adhesion are differentially expressed in HEK-293 cells stably overexpressing TXNRD1_v1 compared with cells expressing its splice variant TXNRD1_v2. Overexpression of these two splice forms resulted in distinctive effects on various aspects of cellular functions including gene regulation patterns, alteration of growth rate, migration and morphology and susceptibility to selenium-induced toxicity. Furthermore, differentiation of the neuroblastoma cell line SH-SY5Y induced by all-trans retinoic acid (ATRA) increased both TXNRD1_v1 and TXNRD1_v2 expressions along with several of the identified genes associated with differentiation and adhesion. Selenium supplementation in the SH-SY5Y cells also induced a differentiated morphology and changed expression of the adhesion protein fibronectin 1 and the differentiation marker cadherin 11, as well as different temporal expression of the studied TXNRD1 variants. These data suggest that both TXNRD1_v1 and TXNRD1_v2 have distinct roles in differentiation, possibly by altering the expression of the genes associated with differentiation, and further emphasize the importance in distinguishing each unique action of different TrxR1 splice forms, especially when studying the gene silencing or knockout of TrxR1.     

Mining citizen science data to explore stopover sites and spatiotemporal variation in migration patterns of the red-footed falcon

Citizen science data have already been used to effectively address questions regarding migration, a fundamental stage in the life history of birds. In this study, we use data from eBird and from 3 additional regional citizen science databases to describe the migration routes and timing of the red-footed falcon Falco vespertinus in the Mediterranean region across 8 years (2010–2017). We further examine the seasonal and yearly variation in migration patterns and explore sites used during the species migration. Our results suggest that the autumn passage is spatially less variable and temporally more consistent among years than in spring and that birds migrate faster in spring than in autumn. The species seems to be more prevalent along the Central Mediterranean during spring migration, probably as a result of the clockwise loop migration that red-footed falcons perform. There was a high variation in annual median migration dates for both seasons as well as in migration routes across years and seasons. Higher variation was exhibited in the longitudinal component thus indicating flexibility in migration routes. In addition, our results showed the species’ preference for lowlands covered with cropland and mosaics of cropland and natural vegetation as stopover sites during migration. Stopover areas predicted from our distribution modeling highlight the importance of the Mediterranean islands as stopover sites for sea-crossing raptors, such as the red-footed falcon. This study is the first to provide a broad-scale spatiotemporal perspective on the species migration across seasons, years and flyways and demonstrates how citizen science data can inform future monitoring and conservation strategies.     

Photosynthetic generation of heterologous terpenoids in cyanobacteria

The work aims to convert the secondary slow metabolism of the terpenoid biosynthetic pathway into a primary activity in cyanobacteria and to generate heterologous products using these photosynthetic microorganisms as cell factories. Case study is the production of the 10-carbon monoterpene β-phellandrene (PHL) in Synechocystis sp. PCC 6803 (Synechocystis). Barriers to this objective include the slow catalytic activity of the terpenoid metabolism enzymes that limit rates and yield of product synthesis and accumulation. “Fusion constructs as protein overexpression vectors” were applied in the overexpression of the geranyl diphosphate synthase (GPPS) and β-phellandrene synthase (PHLS) genes, causing accumulation of GPPS up to 4% and PHLS up to 10% of the total cellular protein. Such GPPS and PHLS protein overexpression compensated for their slow catalytic activity and enabled transformant Synechocystis to constitutively generate 24 mg of PHL per g biomass (2.4% PHL:biomass, w-w), a substantial improvement over earlier yields. The work showed that a systematic overexpression, at the protein level, of the terpenoid biosynthetic pathway genes is a promising approach to achieving high yields of prenyl product biosynthesis, on the way to exploiting the cellular terpenoid metabolism for commodity product generation.     

Diet overlap between small ruminants and the European hare in a Mediterranean shrubland

Seasonal diets of goats, sheep and European hares (Lepus europaeus) were examined using microhistological analysis of feces collected when these herbivores grazed together in a typical Mediterranean shrubland. Approximately half of the total diet content of goats was shrubs (mainly kermes oak, Quercus coccifera), while that of hares was grasses (mostly brush grass, Chrysopogon gryllus). Sheep had a more balanced diet consisting mainly of grasses, forbs, and shrubs. Dietary overlap between goats and sheep was high throughout the year. In contrast, there was very low dietary overlap between small ruminants and hares. Dietary diversity was high in spring and low in winter across all species, with sheep in general displaying higher dietary diversity across all seasons than goats and hares. Goats had intermediate and hares had low dietary diversity across all seasons. Communal grazing by small ruminants and hares ensures that there is a more uniform use of the available forage resources than if a single herbivore is left to graze an area.     

Sunbathing,a possible risk factor of murine typhus infection in Greece

BackgroundThere are few studies about the presence of murine typhus in Greece. Our objective was to conduct a large scale retrospective investigation to determine the clinical and epidemiological features of patients diagnosed with murine typhus in Greece.Methodology/Principal findingsFrom 2012 to 2019 serum samples from hospitalized patients and outpatients throughout Greece suspected for murine typhus infection were tested by immunofluorescence assay for Rickettsia typhi. Immunofluorescence positive samples obtained since 2016 were also tested by qPCR targeting R. typhi. Clinical and epidemiological data were retrospectively collected for the patients with confirmed murine typhus. Overall, we tested 5,365 different patients and, in total, 174 patients from all geographic regions of Greece were diagnosed with murine typhus. The most frequently reported sign or symptom was fever (89%), followed by headache (84%) and rash (81%). The classical triad of fever, headache, and rash was present in 72% of patients during their illness. Severe infections with complications including acute renal failure or septic shock were not recorded. The majority of cases (81%) occurred during May–October and peaked in June and September. Most of patients (81%) infected in Athens, recalled that their only activity the last weeks before symptoms onset was swimming on the beach and 59% of them also reported an insect bite while sunbathing.Conclusions/SignificanceOur results may reflect the reemergence of murine typhus in Greece and we highlight the importance of awareness of this difficult-to-recognize undifferentiated febrile illness.     

Heterologous synthesis of geranyllinalool,a diterpenol plant product,in the cyanobacterium Synechocystis

Cyanobacteria are industrially robust photosynthetic microorganisms that can be genetically programmed to synthesize commodity products for domestic and industrial consumption. In the present work, Synechocystis was endowed with the synthesis of the plant secondary metabolite geranyllinalool, a diterpene alcohol of commercial interest. Total average yields of 360 μg of geranyllinalool per gram of dry cell weight were obtained in the course of a 48-h cultivation period. Geranyllinalool was primarily sequestered inside the transformant cells, corresponding to 60–70% of the total heterologous product, instead of being entirely exuded, as the case is with shorter heterologous terpene hydrocarbons. Extraction of geranyllinalool necessitated disruption of the cells in order to release and isolate this chemical product. Moreover, geranyllinalool accumulation in the cells caused a mild inhibitory effect on cell fitness and biomass growth rate, such that the duplication time of Synechocystis transformants was 1.4-fold longer than that of the control. The remaining 30–40% of the geranyllinalool product was found to float on the surface of sealed transformant cultures, where it was siphoned off by applying a hydrophobic overlayer, with no need to disrupt the cells in this case. Concluding, the work extended efforts to heterologously produce terpene and terpenol products in cyanobacteria, and addressed possibilities and constrains inherent to this production system.

     

A Generalized Estimator of the Attributable Benefit of an Optimal Treatment Regime

Summary For many diseases where there are several treatment options often there is no consensus on the best treatment to give individual patients. In such cases, it may be necessary to define a strategy for treatment assignment; that is, an algorithm that dictates the treatment an individual should receive based on their measured characteristics. Such a strategy or algorithm is also referred to as a treatment regime. The optimal treatment regime is the strategy that would provide the most public health benefit by minimizing as many poor outcomes as possible. Using a measure that is a generalization of attributable risk (AR) and notions of potential outcomes, we derive an estimator for the proportion of events that could have been prevented had the optimal treatment regime been implemented. Traditional AR studies look at the added risk that can be attributed to exposure of some contaminant; here we will instead study the benefit that can be attributed to using the optimal treatment strategy. We will show how regression models can be used to estimate the optimal treatment strategy and the attributable benefit of that strategy. We also derive the large sample properties of this estimator. As a motivating example, we will apply our methods to an observational study of 3856 patients treated at the Duke University Medical Center with prior coronary artery bypass graft surgery and further heart‐related problems requiring a catheterization. The patients may be treated with either medical therapy alone or a combination of medical therapy and percutaneous coronary intervention without a general consensus on which is the best treatment for individual patients.     

Hotspots of endemic terrestrial invertebrates in southern Greece

In the present paper we identify biodiversity hotspots of endemic terrestrial invertebrates in southern Greece, using four different criteria and five taxonomic groups. The hotspots identification procedure was facilitated by the use of the MapInfoPro G.I.S. software. We mapped the known distribution of 424 endemic species on a digital map of southern Greece in 548 10 × 10 km² grids. The total number of registrations (records per grid) is 3826 in 465 grids. Registrations are unevenly distributed in the five sub-regions of the study area, a fact that results from both the unequal collecting effort applied in each sub-region and their different levels of endemism. We first identified the hotspots for each individual taxonomic group, and then for the total species pool. Finally, we analyzed the species saturation pattern in order to find the minimum number of grids needed for the inclusion of 85% of the total number of species. The most important hotspots are mostly at the higher mountains of the study area, especially on Taygetos and Idi Mts. We also give a brief evaluation of the performance of the four criteria used in the case of endemic species analyses.