Bacterial and Fungal Endophytic Microbiomes of Salicornia europaea

We examined Salicornia europaea, a nonmycorrhizal halophyte associated with specific and unique endophytic bacteria and fungi. The microbial community structure was analyzed at two sites differing in salinization history (anthropogenic and naturally saline site), in contrasting seasons (spring and fall) and in two plant organs (shoots and roots) via 16S rRNA and internal transcribed spacer amplicon sequencing. We observed distinct communities at the two sites, and in shoots and roots, while the season was of no importance. The bacterial community was less diverse in shoot libraries than in roots, regardless of the site and season, whereas no significant differences were observed for the fungal community. Proteobacteria and Bacteroidetes dominated bacterial assemblages, and Ascomycetes were the most frequent fungi. A root core microbiome operational taxonomic unit belonging to the genus Marinimicrobium was identified. We detected a significant influence of the Salicornia bacterial community on the fungal one by means of cocorrespondence analysis. In addition, pathways and potential functions of the bacterial community in Salicornia europaea were inferred and discussed. We can conclude that bacterial and fungal microbiomes of S. europaea are determined by the origin of salinity at the sites. Bacterial communities seemed to influence fungal ones, but not the other way around, which takes us closer to understanding of interactions between the two microbial groups. In addition, the plant organs of the halophyte filter the microbial community composition

Transcriptome profiling and environmental linkage to salinity across Salicornia europaea vegetation

Background: Salicornia europaea, a succulent obligatory halophyte is the most salt-tolerant plant species in the world. It survives salt concentrations of more than 1 M. Therefore, it is a suitable model plant to identify genes involved in salt tolerance mechanisms that can be used for the improvement of crops. The changes in a plant’s gene expression in response to abiotic stresses may depend on factors like soil conditions at the site, seasonality, etc. To date, experiments were performed to study the gene expression of S. europaea only under controlled conditions. Conversely, the present study investigates the transcriptome and physicochemical parameters of S. europaea shoots and roots from two different types of saline ecosystems growing under natural conditions. Results: The level of soil salinity was higher at the naturally saline site than at the anthropogenic saline site. The parameters such as ECe, Na+, Cl−, Ca+, SO4 2− and HCO3− of the soils and plant organs significantly varied according to sites and seasons. We found that Na+ mainly accumulated in shoots, whereas K+ and Ca2+ levels were higher in roots throughout the growing period. Moreover, changes in S. europaea gene expression were more prominent in seasons, than sites and plant organs. The 30 differentially expressed genes included enzymes for synthesis of Sadenosyl methionine, CP47 of light-harvesting complex II, photosystem I proteins, Hsp70 gene, ATP-dependent Clp proteases, ribulose bisphosphate carboxylase/oxygenase (Rubisco), phenylalanine ammonia-lyase (PAL), cytochrome c oxidase (COX) and ATP synthase. Conclusion: The comparisons made based on two seasons, plant organs and two different sites suggest the importance of seasonal variations in gene expression of S. europaea. We identify the genes that may play an important role in acclimation to season-dependent changes of salinity. The genes were involved in processes such as osmotic adjustment, energy metabolism and photosynthesis

Heme Oxygenase Isoforms Differ in Their Subcellular Trafficking during Hypoxia and Are Differentially Modulated by Cytochrome P450 Reductase

Heme oxygenase (HO) degrades heme in concert with NADPH cytochrome P450 reductase (CPR) which donates electrons to the reaction. Earlier studies reveal the importance of the hydrophobic carboxy-terminus of HO-1 for anchorage to the endoplasmic reticulum (ER) which facilitates the interaction with CPR. In addition, HO-1 has been shown to undergo regulated intramembrane proteolysis of the carboxy-terminus during hypoxia and subsequent translocation to the nucleus. Translocated nuclear HO-1 was demonstrated to alter binding of transcription factors and to alter gene expression. Little is known about the homologous membrane anchor of the HO-2 isoform. The current work is the first systematic analysis in a eukaryotic system that demonstrates the crucial role of the membrane anchor of HO-2 for localization at the endoplasmic reticulum, oligomerization and interaction with CPR. We show that although the carboxy-terminal deletion mutant of HO-2 is found in the nucleus, translocation of HO-2 to the nucleus does not occur under conditions of hypoxia. Thus, we demonstrate that proteolytic regulation and nuclear translocation under hypoxic conditions is specific for HO-1. In addition we show for the first time that CPR prevents this translocation and promotes oligomerization of HO-1. Based on these findings, CPR may modulate gene expression via the amount of nuclear HO-1. This is of particular relevance as CPR is a highly polymorphic gene and deficiency syndromes of CPR have been described in humans

Managing patients on extracorporeal membrane oxygenation support during the COVID-19 pandemic – a proposal for a nursing standard operating procedure

Background Extracorporeal membrane oxygenation (ECMO) is effective in a selected critically ill patient population with promising results in refractory hypoxemia related to the novel coronavirus disease (COVID-19). However, it requires specialized clinicians and resources in advanced technology. Moreover, the COVID-19 remains an ongoing global emergency, and there is no evidence-based practice in preparedness. This article proposes an innovative and optimized nursing care protocol, the Standard Operating Procedure (SOP), that regulates safety and efficiency in using personal protective equipment (PPE) during ECMO-relevant procedures while providing ECMO therapy for patients with COVID-19. Methods After performing a narrative literature search, we developed a high-fidelity translational simulation scenario. It included practicing appropriate donning and doffing PPE during work organization, ECMO-related procedures, and routine daily nursing care and management of ECMO over nine hours. In addition, we held supplementary constructive debrief meetings to consult international expert in the field. Results A proposal for nursing standardized operating procedures was created, divided into categories. They included work organization, workload references, competences, infrastructural conditions, cannulation equipment, daily routine nursing care, and procedures during ECMO. Conclusions High-fidelity medical simulation can play an important role in staff training, improvement in previously gained proficiency, and development of optimal SOP for nursing care and management during ECMO in patients with COVID-19. Optimal SOPs may further guide multidisciplinary teams, including intensive care units and interventional departments

Characterization of the structure and biochemical activity of lipid-protein nanodiscs prepared using the amphiphilic polymer DIBMA

Białka błonowe odgrywają ważną rolę w komórkach, ponieważ biorą udział między innymi w produkcji energii, transdukcji sygnału i utrzymaniu homeostazy komórkowej. W komórkach ludzkich często stanowią punkt uchwytu dla farmaceutyków. Z racji tego, że środowisko błonowe ma kluczowe znaczenie dla utrzymania struktury i funkcji białek błonowych, badania in vitro często stanowią wyzwanie. Dlatego konieczne jest opracowanie nowych narzędzi do symulacji ich natywnego środowiska. Na przestrzeni ostatnich dwóch dekad wielką nadzieją w dziedzinie badań strukturalnych okazały się być nanodyski. Nanodyski są modelami błon biologicznych, samoskładającymi strukturami, w których skład wchodzi dwuwarstwa lipidowa oraz stabilizujące ją cząsteczki polimeru amfifilowego np. DIBMA. Celem niniejszej pracy była optymalizacja tworzenia nanodysków z błon komórek eukariotycznych wspomaganych polimerem DIBMA uwzględniając zmienne środowisko jonowe, w którego skład wchodzą jednowartościowe jony K+, oraz jony dwuwartościowe Ca2+ i Mg2+. Nanodyski opisano pod względem homogenności oraz rozmiaru, wykorzystując przy tym metody dynamicznego rozpraszania światła (DLS) oraz sączenie molekularne. Udowodniono, że zastosowanie polimeru DIBMA do solubilizacji błon komórkowych umożliwia uformowanie nanodysków zawierających białka błonowe oraz podbłonowe. Dowiedziono, że modyfikacja składu lipidowego polegającego na usunięciu wybranych lipidów z błony komórkowej linii komórek HEK293 przy pomocy związków z rodziny β-cyklodekstryn powoduje powstawanie nanodysków o bardziej homogennym charakterze.Membrane proteins play an important role in cell physiology, including energy production, signal transduction and cell homeostasis. Additionally, they constitute a major group of drug targets for pharmaceuticals. As the membrane environment is crucial for the maintenance of the structure and function of membrane proteins, it is often a challenge to study them in vitro, and thus, it is necessary to develop new tools to simulate their natural setting. Over the last two decades, nanodiscs have been gaining increasing popularity, which are self-assembling structures comprising a lipid bilayer stabilized by amphiphilic polymers (DIBMA).The aim of this study was to optimize the production of nanodiscs from eukaryotic cell membranes aided by DIBMA polymer, taking account of various ionic conditions including monovalent K+ ions and divalent Ca2+ and Mg2+ ions. The nanodiscs were characterized in terms of size and homogeneity, employing dynamic light scattering and size-exclusion chromatography. It was proven that using DIBMA to solubilize the membranes enables the formation of nanodiscs containing membrane proteins, including submembrane proteins. Moreover, it was demonstrated that modifying the lipid composition of HEK293 cell membranes by removing certain lipids with β-cyclodextrins allows for the formation of more homogenous nanodiscs

Properties and functions of miropin in infections associated with periodontitis

Miropina jest serpiną prokariotyczną produkowaną przez bakterie Tannerella forsythia. Główną funkcją tego białka jest hamowanie szerokiej gamy proteaz serynowych i cysteinowych. Jest to ważne, gdyż środowisko, w którym bytuje T. forsythia, charakteryzuje się wysoką aktywnością proteolityczną. Ponadto ekspresja miropiny może stanowić czynnik odpowiedzialny za rozwój różnych chorób takich jak choroba Alzheimera, zapalenie płuc, reumatoidalne zapalenie stawów, miażdżyca.Miropin is a bacterial serpin produced by Tannerella forsythia. The main function of this protein is inhibition of a wide range of proteases. This is important because the environment in which T. forsythia lives has high proteolytic activity. In addition, the expression of miropin may be a factor responsible for the development of various diseases such as Alzheimer's disease, pneumonia, rheumatoid arthritis, atherosclerosis

MicroRNA as a Potential Therapeutic Molecule in Cancer

Small noncoding RNAs, as post-translational regulators of many target genes, are not only markers of neoplastic disease initiation and progression, but also markers of response to anticancer therapy. Hundreds of miRNAs have been identified as biomarkers of drug resistance, and many have demonstrated the potential to sensitize cancer cells to therapy. Their properties of modulating the response of cells to therapy have made them a promising target for overcoming drug resistance. Several methods have been developed for the delivery of miRNAs to cancer cells, including introducing synthetic miRNA mimics, DNA plasmids containing miRNAs, and small molecules that epigenetically alter endogenous miRNA expression. The results of studies in animal models and preclinical studies for solid cancers and hematological malignancies have confirmed the effectiveness of treatment protocols using microRNA. Nevertheless, the use of miRNAs in anticancer therapy is not without limitations, including the development of a stable nanoconstruct, delivery method choices, and biodistribution. The aim of this review was to summarize the role of miRNAs in cancer treatment and to present new therapeutic concepts for these molecules. Supporting anticancer therapy with microRNA molecules has been verified in numerous clinical trials, which shows great potential in the treatment of cancer

Role of Ascorbate in the Regulation of the Arabidopsis thaliana Root Growth by Phosphate Availability

Arabidopsis root system responds to phosphorus (P) deficiency by decreasing primary root elongation and developing abundant lateral roots. Feeding plants with ascorbic acid (ASC) stimulated primary root elongation in seedlings grown under limiting P concentration. However, at high P, ASC inhibited root growth. Seedlings of ascorbate-deficient mutant (vtc1) formed short roots irrespective of P availability. P-starved plants accumulated less ascorbate in primary root tips than those grown under high P. ASC-treatment stimulated cell divisions in root tips of seedlings grown at low P. At high P concentrations ASC decreased the number of mitotic cells in the root tips. The lateral root density in seedlings grown under P deficiency was decreased by ASC treatments. At high P, this parameter was not affected by ASC-supplementation. vtc1 mutant exhibited increased lateral root formation on either, P-deficient or P-sufficient medium. Irrespective of P availability, high ASC concentrations reduced density and growth of root hairs. These results suggest that ascorbate may participate in the regulation of primary root elongation at different phosphate availability via its effect on mitotic activity in the root tips