Genome-wide mapping reveals single-origin chromosome replication in Leishmania, a eukaryotic microbe

Background DNA replication initiates on defined genome sites, termed origins. Origin usage appears to follow common rules in the eukaryotic organisms examined to date: all chromosomes are replicated from multiple origins, which display variations in firing efficiency and are selected from a larger pool of potential origins. To ask if these features of DNA replication are true of all eukaryotes, we describe genome-wide origin mapping in the parasite Leishmania. Results Origin mapping in Leishmania suggests a striking divergence in origin usage relative to characterized eukaryotes, since each chromosome appears to be replicated from a single origin. By comparing two species of Leishmania, we find evidence that such origin singularity is maintained in the face of chromosome fusion or fission events during evolution. Mapping Leishmania origins suggests that all origins fire with equal efficiency, and that the genomic sites occupied by origins differ from related non-origins sites. Finally, we provide evidence that origin location in Leishmania displays striking conservation with Trypanosoma brucei, despite the latter parasite replicating its chromosomes from multiple, variable strength origins. Conclusions The demonstration of chromosome replication for a single origin in Leishmania, a microbial eukaryote, has implications for the evolution of origin multiplicity and associated controls, and may explain the pervasive aneuploidy that characterizes Leishmania chromosome architecture

Socio‐cultural impacts of peer‐to‐peer accommodation on host communities

Peer-to-peer accommodation has recently emerged as a central focus of research in tourism and hospitality. However, research on socio-cultural impacts of peer-to-peer accommodation is fragmented. This study reviews the relevant literature on the socio-cultural impacts of peer-to-peer accommodation on host communities and proposes an integrative framework and a working definition for goal socio-cultural impacts of peer-to-peer accommodation. This literature review contributes to knowledge of how this business paradigm affects and frames host communities and integrates the factors that should be explored by researchers, policymakers, and organizations to manage the impacts of peer-to-peer accommodation on host communities.info:eu-repo/semantics/publishedVersio

Diverged composition and regulation of the Trypanosoma brucei origin recognition complex that mediates DNA replication initiation

Initiation of DNA replication depends upon recognition of genomic sites, termed origins, by AAA+ ATPases. In prokaryotes a single factor binds each origin, whereas in eukaryotes this role is played by a six-protein origin recognition complex (ORC). Why eukaryotes evolved a multisubunit initiator, and the roles of each component, remains unclear. In Trypanosoma brucei, an ancient unicellular eukaryote, only one ORC-related initiator, TbORC1/CDC6, has been identified by sequence homology. Here we show that three TbORC1/CDC6-interacting factors also act in T. brucei nuclear DNA replication and demonstrate that TbORC1/CDC6 interacts in a high molecular complex in which a diverged Orc4 homologue and one replicative helicase subunit can also be found. Analysing the subcellular localization of four TbORC1/CDC6-interacting factors during the cell cycle reveals that one factor, TbORC1B, is not a static constituent of ORC but displays S-phase restricted nuclear localization and expression, suggesting it positively regulates replication. This work shows that ORC architecture and regulation are diverged features of DNA replication initiation in T. brucei, providing new insight into this key stage of eukaryotic genome copying

Genome-scale RNA interference profiling of <i>Trypanosoma brucei</i> cell cycle progression defects

Trypanosomatids, which include major pathogens of humans and livestock, are flagellated protozoa for which cell cycle controls and the underlying mechanisms are not completely understood. Here, we describe a genome-wide RNA-interference library screen for cell cycle defects in Trypanosoma brucei. We induced massive parallel knockdown, sorted the perturbed population using high-throughput flow cytometry, deep-sequenced RNAi-targets from each stage and digitally reconstructed cell cycle profiles at a genomic scale; also enabling data visualisation using an online tool (https://tryp-cycle.pages.dev/). Analysis of several hundred genes that impact cell cycle progression reveals &gt;100 flagellar component knockdowns linked to genome endoreduplication, evidence for metabolic control of the G1-S transition, surface antigen regulatory mRNA-binding protein knockdowns linked to G2M accumulation, and a putative nucleoredoxin required for both mitochondrial genome segregation and for mitosis. The outputs provide comprehensive functional genomic evidence for the known and novel machineries, pathways and regulators that coordinate trypanosome cell cycle progression

Read, write, adapt:Challenges and opportunities during kinetoplastid genome replication

The genomes of all organisms are read throughout their growth and development, generating new copies during cell division and encoding the cellular activities dictated by the genome’s content. However, genomes are not invariant information stores but are purposefully altered in minor and major ways, adapting cellular behaviour and driving evolution. Kinetoplastids are eukaryotic microbes that display a wide range of such read–write genome activities, in many cases affecting critical aspects of their biology, such as host adaptation. Here we discuss the range of read–write genome changes found in two well-studied kinetoplastid parasites, Trypanosoma brucei and Leishmania, focusing on recent work that suggests such adaptive genome variation is linked to novel strategies the parasites use to replicate their unconventional genomes

Functional cell microcarriers: a new platform for cell separation and expansion

Publicado em "Journal of Tissue Engineering and Regenerative Medicine", vol. 7, supp. 1 (2013)The success of many stem cell applications in the biomedical field is highly dependent on the development of separation techniques for isolation and purification of cells with a very high yield and purity. Despite all the achievements made in the field over the past several years, new systems for effective cell separation are still needed. Previous work from our group demonstrated that functional chitosan films grafted with antibodies promote selective cell adhesion. 1 Herein we developed chitosan microparticles able to capture a specific cell types based in the concept of antibody coating for cell sorting. Our goal was to create new biomaterial surfaces capable of recruit a specific cell population within a mixture, reducing cell manipulation and time-consuming allowing at the same time cell expansion. Such system acts as a microcarrier for cell expansion of a specific cell target. Microcarrier culture system offers the advantage of providing a larger surface area for the growth of anchorage-dependent cells in a suspension culture system. Chitosan was chosen due to the excellent biocompatibility, gel forming properties, chemistry surface and low cell adhesion. This allows the modification with specific biochemical cues, for a controllable cell attachment. Here we develop functional biotinylated microparticles, such system allows tailoring microparticles to a variety of functional biomolecules. Here we tested the immobilization of antibodies to target specific cell types, CD31 for endothelial cells and CD90 for adipose stem cells. Primarily designed for an application in tissue engineering, two main challenges are accomplished with the herein presented microparticles: separation and scale-up expansion of specific cell type. The herein developed polymeric microparticles can also be used for directly deliver cells in vivo to repair and regenerate tissues

A multifactorial approach to untangle graphene oxide (GO) nanosheets effects on plants: plant growth-promoting bacteria inoculation, bacterial survival, and drought

Drought is a limiting factor for agricultural productivity. Climate change threatens to expand the areas of the globe subjected to drought, as well as to increase the severity and duration of water shortage. Plant growth-promoting bacteria (PGPB) are widely studied and applied as biostimulants to increase plant production and to enhance tolerance to abiotic and biotic constraints. Besides PGPB, studies on the potential of nanoparticles to be used as biostimulants are also thriving. However, many studies report toxicity of tested nanoparticles in bacteria and plants in laboratory conditions, but few studies have reported effects of nanoparticles towards bacterial cells and communities in the soil. The combined application of nanoparticles and PGPB as biostimulant formulations are poorly explored and it is important to unravel the potentialities of their combined application as a way to potentiate food production. In this study, Rhizobium sp. E20-8 and graphene oxide (GO) nanosheets were applied on container-grown maize seedlings in watered and drought conditions. Bacterial survival, seedling growth (dry weight), and biochemical endpoints (photosynthetic pigments, soluble and insoluble carbohydrates, proline, lipid peroxidation, protein, electron transport system, and superoxide dismutase) were evaluated. Results showed that the simultaneous exposure to GO and Rhizobium sp. E20-8 was able to alleviate the stress induced by drought on maize seedlings through osmotic and antioxidant protection by GO and mitigation of GO effects on the plant's biochemistry by Rhizobium sp. E20-8. These results constitute a new lead on the development of biostimulant formulations to improve plant performance and increase food production in water-limited conditions.publishe

Functional chitosan microcarriers for selective cell attachment and expansion

The success of many stem cell applications in the biomedical field is highly dependent on the development of reliable techniques either for isolation or selection of specific cell populations with a very high yield and purity.1 In this work we propose the use of chitosan microparticles (μPs) to capture a specific cell type based in the concept of antibody-antigen binding. Our goal was to create new biomaterials capable of selecting within a heterotypic cell suspension, a specific sub-population, and supporting subsequent cell expansion. Such system simultaneously allows the selection and acts as a microcarrier for a specific target, thus reducing cell manipulation and time-consumption

Geochemical fingerprinting of Roman pottery production from Manta Rota kilns (Southern Portugal)

The Roman site of Manta Rota, located in Algarve region, Portugal, is known from the XIXth century when Estacio da Veiga recovered a few pottery fragments and identified the foundations of Roman structures. References in early XXth century refer to a pottery kiln and to the remains of both amphorae and lamps. In 1992, emergency excavations directed by Cristina Garcia excavated a structure identified as a kiln and recovered abundant amphorae fragments of a locally produced Dressel 14 type. There is evidence of production of tiles, common ware and amphora. The study of the remains collected (terra sigillata and amphorae) in the excavations shows that Roman occupation of the site covers a period from the middle of the 1st century until the 5th century AD. The major production of the kiln area was the Dressel 14 fish sauce amphora centered in the middle/second half of the 1st century but the production of Almagro 51c is also attested. A preliminary macroscopic analysis of both the amphorae and the common ware show identical features. The aim of this paper is to present chemical characterization of the production of Manta Rota and to compare the results with different workshops in the Algarve region as Quinta do Lago, near Faro or S. Bartolomeu de Castro Marim, in the proximity of the studied area, in order to establish geochemical fingerprints of Manta Rota ceramic production center. Preliminary results point to a fine geochemical differentiation of the Manta Rota ceramics in comparison with the other two roman production centers of Algarve region

The effect of chitosan on the in vitro biological performance of chitosan-poly(butylene succinate) blends

Chitosan blends with synthetic biodegradable polymers have been proposed for various biomedical applications due to their versatile mechanical properties and easier processing. However, details regarding the main surface characteristics that may benefit from the blending of these two types of materials are still missing. Hence, this work aims at investigating the surface properties of chitosan-based blends, illustrating the way these properties determine the material-proteins interactions and ultimately the behavior of osteoblast-like cells. The surface characteristics of modified and nonmodified blends were assessed using complimentary techniques such as optical microscopy, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR-ATR), X-ray photoelectron spectroscopy (XPS), contact angle measurements and surface energy calculations. The adsorption of human serum albumin (HSA) and human plasma fibronectin (HFN) onto the different surfaces was quantified by association of an indirect method with a colorimetric assay. It was found that the presence of chitosan on the surface promoted the adsorption of proteins. Moreover, a preferential adsorption of albumin over fibronectin was registered. The in vitro biological performance of the studied materials was further investigated by a direct contact assay with an osteoblastic-like cell line (SaOs-2). A synergistic effect of the two components of the blend was observed. While the synthetic polyester promoted the adhesion of SaOs-2, the presence of chitosan significantly enhanced the osteoblastic activity of these cells. This work further confirmed the interest in designing polymeric blends with natural polymers as a successful strategy to enhance the biological performance of a biomaterial