Search

Abstract Background Adhesive capsulitis (AC) is a disease of the glenohumeral joint that is characterized by pain and both passive and active global stiffness with a slow and insidious onset. The disease can occur spontaneously (primary AC) or it can be secondary to other comorbidities, surgery, or trauma, such as fracture or dislocation. Multiple treatment approaches have been suggested: intra-articular steroid injection, physical therapy, manipulation under total anesthesia, and arthroscopic or open surgery. Shoulder manipulation under anesthesia is usually proposed to patients that suffer from severe AC and have already undergone several nonoperative treatments without benefit. Different techniques have been proposed. This study presents our manipulation technique and the clinical results we achieved after shoulder mobilization under brachial plexus block in patients with phase III primary AC. Materials and methods A retrospective cohort study was performed on a sample of 110 patients with phase III AC who were treated with this manipulation and followed up for 1 year. Patients underwent two assessments—before the procedure (T0) and 4 months after it (T1)—based on the Numerical Rating Scale, Simple Shoulder Test, and joint range of motion to assess shoulder pain, function, and joint articulation, respectively. Furthermore, the patients had to express their degree of satisfaction with the procedure and the results achieved. Results Positive and statistically significant results were recorded in terms of pain reduction (ΔNPRS = − 5.4; p

Metatranscriptomic methods involving RNA sequencing (RNA-seq) are powerful tools for evaluating the gene expression profiles of transcriptionally-active microbes that inhabit the tissues of animals. Bivalve molluscs, like all invertebrates, are holobionts and sites of interactions between host animals and both prokaryotic and eukaryotic symbionts. The present communication describes the metatranscriptomic profile of the resident microorganisms that inhabit the gut of blue mussels, Mytilus edulis, under standard laboratory conditions. Each of the eight mussels described herein were housed in isolated microcosms with routine husbandry for 14 days before their gut tissues were sampled and subjected to RNA-seq. Subsequent mRNA reads that aligned to the mussel genome were removed, and the non-host reads were annotated for function and pathway analyses. Under laboratory conditions, the resident gut microbiota expressed genes associated mostly with aerobic energy processes, with other notable contributions from metabolism and protein processing genes. At the pathway level, the most abundant complete pathways expressed in the resident gut microbial communities were related to aerobic cellular respiration, nucleotide biosynthesis, and catabolism. These data represent novel, baseline microbial gene expression information from the gut of mussels, which are crucial for future research examining the mussel holobiont and bivalve microbial ecology. Public retrieval and secondary analyses of these metatranscriptomic profiles are highly encouraged.

The 2010 Deepwater Horizon (DwH) Oil spill released an enormous volume of oil into the Gulf of Mexico (GoM), prompting the widespread use of chemical dispersants like Corexit® EC9500A. The ecological consequences of this treatment, especially when combined with natural factors such as sunlight, remain unexplored in the context of marine bacterial communities’ dynamics. To address this knowledge gap, our study employed a unique metaproteomic approach, investigating the combined effects of sunlight, crude Macondo surrogate oil, and Corexit on GoM microbiome across different mesocosms. Exposure to oil and/or Corexit caused a marked change in community composition, with a decrease in taxonomic diversity relative to controls in only 24 hours. Hydrocarbon (HC) degraders, particularly those more tolerant to Corexit and phototoxic properties of crude oil and/or Corexit, proliferated at the expense of more sensitive taxa. Solar radiation exacerbated these effects in most taxa. We demonstrated that sunlight increased the dispersant’s toxicity, impacting on community structure and functioning. These functional changes were primarily directed by oxidative stress with upregulated proteins and enzymes involved in protein turnover, general stress response, DNA replication and repair, chromosome condensation, and cell division. These factors were more abundant in chemically treated conditions, especially in the presence of Corexit compared to controls. Oil treatment significantly enhanced the relative abundance of Alteromonas, an oil-degrading Gammaproteobacteria. In combined oil-Corexit treatments, the majority of identified protein functions were assigned to Alteromonas, with strongly expressed proteins involved in membrane transport, motility, carbon and amino acid metabolism and cellular defense mechanisms. Marinomonas, one of the most active genera in dark conditions, was absent from the light treatment. Numerous metabolic pathways and HC-degrading genes provided insights into bacterial community adaptation to oil spills. Key enzymes of the glyoxylate bypass, enriched in contaminant-containing treatments, were predominantly associated with Rhodobacterales and Alteromonadales. Several proteins related to outer membrane transport, photosynthesis, and nutrient metabolisms were characterized, allowing predictions of the various treatments on biogeochemical cycles. The study also presents novel perspectives for future oil spill clean-up processes.

The nuclear safety assessment involving large transient simulations is forcing the community to develop methods for coupling thermal-hydraulics and neutronic codes and three-dimensional (3D) Computational Fluid Dynamics (CFD) codes. In this paper a set of dynamic boundary conditions are implemented in OpenFOAMⓇ in order to apply zero-dimensional (0D) approaches coupling with 3D thermal-hydraulic simulation in a single framework. This boundary conditions are applied to model pipelines, tanks, pumps, and heat exchangers. On a first stage, four tests are perform in order to assess the implementations. The results are compared with experimental data, full 3D CFD, and system code simulations, finding a general good agreement. The semi-implicit implementation nature of these boundary conditions has shown robustness and accuracy for large time steps. Finally, an application case, consisting of a simplified open pool with a cooling external circuit is solved to remark the capability of the tool to simulate thermal hydraulic systems commonly found in nuclear installations.

Abstract The multiscale method called Pseudo-Direct Numerical Simulation (P-DNS) is presented as a Reduced Order Model (ROM) aiming to solve problems obtaining similar accuracy to a solution with many degrees of freedom (DOF). The theoretical basis of P-DNS is other than any standard ROM. However, from a methodological point of view, P-DNS shares the idea of an offline computation, as ROM does, providing the set of coefficients, as a database or table, needed to solve the main problem. This work highlights the advantages and disadvantages of both methodologies. In particular, the drawback of the standard ROM concerning problems where space and time are not separated variables is discussed. The so-called Idelsohn’s benchmark is possibly the most elemental test that can be proposed to point out this drawback. This one-dimensional heat transfer problem with a moving heat source shows that, unlike ROMs, P-DNS can solve it by reducing the number of degrees of freedom as much as needed.

The increased potential for contamination of seawater by crude oils requires studies of bacterial biodegradation potential, but little is known of the differential negative impacts of oils on bacterial growth. No two wells generate chemically identical oils; and importantly, solar exposure of crude oil may differentially affect the bacterial response. Elucidating the role that sunlight plays on the potential toxicity of spilled crude oils is imperative to understanding how oil spills might affect microbes in the tropical and subtropical waters of Florida. This study examined light exposure of six different crude oils, and subsequent microbial responses to altered oils. Marine bacterioplankton heterotrophic activities were measured via3H-leucine incorporation after the addition of oils’ water accommodated fractions (WAFs) that were created under varied solar conditions. Inhibition of production increased with higher concentrations of WAFs, but dose-response trends varied among the oils. Increased solar exposure during WAF preparation generally led to more inhibition, but trends varied among oils. WAFs were also prepared under different parts of the solar spectrum. Solar-irradiated WAFs resulted in significant but variable acute toxicity vs. dark counterparts. Solar-induced toxicity was primarily a result of visible and not ultraviolet light exposure. Results indicate responses to oil spills are highly dependent on the source of the oil and solar conditions at the time and location of the spill. The data presented here demonstrate the importance of photochemical changes and oil source in modulating microbial activity and bioremediation potential.

This paper evaluates the performance of a low-cost technology in rural Benin (West Africa) that locally produces chlorine (sodium hypochlorite) and then injects it into the community water tanks. Thirty-one water tanks were selected in cooperation with local authorities to receive electro-chlorinator devices (WATA™). Water samples were tested at two points: before chlorination and at the most distant terminal after chlorination. Residual chlorine control tests and microbiology control tests were performed. Water samples that tested positive for the presence of microorganisms were analyzed at a laboratory when possible. The water provided to the community was not always chlorinated, and over half of the tanks lacked access to chlorine powder. Among the sites using the technology, 30% (9/31) of them had structural problems that prevented the existing system from performing adequately. Furthermore, 60% of the water samples collected before chlorination were positive for microbiological contamination. All samples collected from systems where proper chlorination was taking place tested negative for microbiological contamination. However, the water from six tanks presenting structural problems continued to be distributed to the population despite contamination. The average residual chlorine level analyzed at the most distant terminal fountain was 0.29±0.2 mg/L, which is within the country's reference level 0.1–0.8 mg/L. The installation of water-chlorinator devices (WATA™) produces water without microbiological contamination and with chlorine levels within the WHO's recommended values. However, the success of the technology depends on the pre-existing structure of the water tank. HIGHLIGHTS Chlorine powder shortage was experienced by 52% of the 31 tanks studied.; 60% of water samples collected before chlorination had microbiological contamination.; After installation, the average residual chlorine level was 0.29±0.2 mg/L.; Infrastructural problems were present in 30% (9/31) of sites.;

In deep ocean hypersaline basins, the combination of high salinity, unusual ionic composition and anoxic conditions represents significant challenges for microbial life. We used geochemical porewater characterization and DNA sequencing based taxonomic surveys to enable environmental and microbial characterization of anoxic hypersaline sediments and brines in the Orca Basin, the largest brine basin in the Gulf of Mexico. Full-length bacterial 16S rRNA gene clone libraries from hypersaline sediments and the overlying brine were dominated by the uncultured halophilic KB1 lineage, Deltaproteobacteria related to cultured sulfate-reducing halophilic genera, and specific lineages of heterotrophic Bacteroidetes. Archaeal clones were dominated by members of the halophilic methanogen genus Methanohalophilus, and the ammonia-oxidizing Marine Group I (MG-I) within the Thaumarchaeota. Illumina sequencing revealed higher phylum- and subphylum-level complexity, especially in lower-salinity sediments from the Orca Basin slope. Illumina and clone library surveys consistently detected MG-I Thaumarchaeota and halotolerant Deltaproteobacteria in the hypersaline anoxic sediments, but relative abundances of the KB1 lineage differed between the two sequencing methods. The stable isotopic composition of dissolved inorganic carbon and methane in porewater, and sulfate concentrations decreasing downcore indicated methanogenesis and sulfate reduction in the anoxic sediments. While anaerobic microbial processes likely occur at low rates near their maximal salinity thresholds in Orca Basin, long-term accumulation of reaction products leads to high methane concentrations and reducing conditions within the Orca Basin brine and sediments.

The present paper describes and validates an efficient CFD implementation to replicate the working fluid-dynamics of a real four-stroke internal combustion engine. To do this, experimental data obtained on a single-cylinder engine are used to validate the proposed computational approach. The engine domain is divided into regions according to each moving zone, and these are coupled using a pseudo-supermesh interface presented in a previous work by the authors. In this work, the original pseudo-supermesh strategy is enhanced by introducing the dual-boundary concept to model the valve opening/closing events to increase the accuracy and simplicity of the simulation procedure. The results produced by the proposed software tool show a good correlation to the experimental measurements of the complete engine cycle. Macroscopic quantities of the in-cylinder flow are accurately replicated as well as the instantaneous evolution of the in-cylinder and intake manifold pressure. Furthermore, the present work shows that the computational efficiency and scalability of the enhanced pseudo-supermesh approach are preserved even when applied to more complex real problems. In this sense, this work contributes to a new engineering tool promoting the enhanced pseudo-supermeshes as an effective tool for the design, development, and optimization of internal combustion engines.

Recently, the interest in improving the content of bioactive compounds for enhancing the nutritional value of cereal-based products has largely increased, and several strategies, both genetic and agronomic, were proposed. Here, we report the preliminary results of the effect of two sowing periods (winter and spring) on the accumulation of bioactive compounds (i.e., anthocyanins, carotenoids, minerals) and grain morphology in three pigmented cereal species (barley, bread and durum wheat). The results showed that with delayed sowing time, a significant reduction in yield was observed in all species, less in barley (50.4%) than in wheat (70.3% and 66.5% for bread and durum, respectively). On the contrary, the carotenoids accumulation was favored (increased by 12.2%, 15.7% and 27.8% for barley, bread and durum wheat, respectively), as well as those of anthocyanins although strongly dependent on the genotype (i.e., considering only the purple genotypes, the anthocyanins increase by 4.6%, 35.4% and 72.8% in barley, bread and durum wheat, respectively). For minerals compounds, our study highlighted a strong environmental influence even when the pigmented genotypes accumulated more minerals than commercial ones, while the morphometric seed traits were less influenced by this agronomic practice. Therefore, by exploiting the genetic variability for the accumulation of anthocyanins and carotenoids in the grain and by optimizing the sowing time, it could be possible to increase the nutritional value of the final cereal-based products.

Atrial fibrillation (AF) and coronary artery disease requiring percutaneous coronary intervention (PCI) and stenting often coexist in older patients. This poses the difficult problem of concurrent anticoagulant and double antiplatelet therapy (triple therapy). Current treatment guidelines do recommend triple therapy, especially in the course of acute coronary syndrome (ACS), with limitations due to an excessive risk of bleeding associated with this therapeutic regimen. This review summarizes randomized clinical trials and observational studies that compared triple therapy with a variety of different therapeutic options. Although the available evidence is not completely satisfactory and other studies are urgently needed, alternative regimens to triple therapy in AF patients undergoing PCI and stenting are promising, at least in terms of safety.

Deep-sea hypersaline anoxic basins (DHABs) and other hypersaline environments contain abundant and diverse microbial life that has adapted to these extreme conditions. The bacterial Candidate Division KB1 represents one of several uncultured groups that has been consistently observed in hypersaline microbial diversity studies. Here we report the phylogeography of KB1, its phylogenetic relationships to Candidate Division OP1 Bacteria, and its potential metabolic and osmotic stress adaptations based on a partial single cell amplified genome (SAG) of KB1 from Orca Basin, the largest hypersaline seafloor brine basin in the Gulf of Mexico. Our results are consistent with the hypothesis – previously developed based on 14C incorporation experiments with mixed-species enrichments from Mediterranean seafloor brines - that KB1 has adapted its proteins to elevated intracellular salinity, but at the same time KB1 apparently imports glycine betaine; this compatible solute is potentially not limited to osmoregulation but could also serve as a carbon and energy source.

The Ostracoda (Crustacea; Class Ostracoda) is a diverse, frequently abundant, and ecologically important component of the marine zooplankton assemblage. There are more than 200 described species of marine planktonic ostracods, many of which (especially conspecific species) can be identified only by microscopic examination and dissection of fragile morphological characters. Given the complexity of species identification and increasing lack of expert taxonomists, DNA barcodes (short DNA sequences for species discrimination and identification) are particularly useful and necessary. Results are reported from analysis of 210 specimens of 78 species of marine planktonic ostracods, including two novel species, and 51 species for which barcodes have not been previously published. Specimens were collected during 2006 to 2008 from the Atlantic, Indian, and Southern Oceans, Greenland Sea and Gulf of Alaska. Samples were collected from surface to 5,000 m using various collection devices. DNA sequence variation was analyzed for a 598 base-pair region of the mitochondrial cytochrome oxidase subunit I (COI) gene. Kimura-2-Parameter (K2P) genetic distances within described species (mean = 0.010 ± 0.017 SD) were significantly smaller than between species (0.260 + 0.080), excluding eight taxa hypothesized to comprise cryptic species due to morphological variation (especially different size forms) and/or collection from different geographic regions. These taxa showed similar K2P distance values within (0.014 + 0.026) and between (0.221 ± 0.068) species. All K2P distances > 0.1 resulted from comparisons between identified or cryptic species, with no overlap between intra- and interspecific genetic distances. A Neighbor Joining tree resolved nearly all described species analyzed, with multiple sequences forming monophyletic clusters with high bootstrap values (typically 99%). Based on taxonomically and geographically extensive sampling and analysis (albeit with small sample sizes), the COI barcode region was shown to be a valuable character for discrimination, recognition, identification, and discovery of species of marine planktonic ostracods.

Oligodendroglioma poses a biological conundrum for malignant adult human gliomas: it is a tumor type that is universally incurable for patients, and yet, only a few of the human tumors have been established as cell populations in vitro or as intracranial xenografts in vivo. Their survival, thus, may emerge only within a specific environmental context. To determine the fate of human oligodendroglioma in an experimental model, we studied the development of an anaplastic tumor after intracranial implantation into enhanced green fluorescent protein (eGFP) positive NOD/SCID mice. Remarkably after nearly nine months, the tumor not only engrafted, but it also retained classic histological and genetic features of human oligodendroglioma, in particular cells with a clear cytoplasm, showing an infiltrative growth pattern, and harboring mutations of IDH1 (R132H) and of the tumor suppressor genes, FUBP1 and CIC. The xenografts were highly invasive, exhibiting a distinct migration and growth pattern around neurons, especially in the hippocampus, and following white matter tracts of the corpus callosum with tumor cells accumulating around established vasculature. Although tumors exhibited a high growth fraction in vivo, neither cells from the original patient tumor nor the xenograft exhibited significant growth in vitro over a six-month period. This glioma xenograft is the first to display a pure oligodendroglioma histology and expression of R132H. The unexpected property, that the cells fail to grow in vitro even after passage through the mouse, allows us to uniquely investigate the relationship of this oligodendroglioma with the in vivo microenvironment.

Oligodendroglial tumors form a distinct subgroup of gliomas, characterized by a better response to treatment and prolonged overall survival. Most oligodendrogliomas and also some oligoastrocytomas are characterized by a unique and typical unbalanced translocation, der(1,19), resulting in a 1p/19q co-deletion. Candidate tumor suppressor genes targeted by these losses, CIC on 19q13.2 and FUBP1 on 1p31.1, were only recently discovered. We analyzed 17 oligodendrogliomas and oligoastrocytomas by applying a comprehensive approach consisting of RNA expression analysis, DNA sequencing of CIC, FUBP1, IDH1/2, and array CGH. We confirmed three different genetic subtypes in our samples: i) the "oligodendroglial" subtype with 1p/19q co-deletion in twelve out of 17 tumors; ii) the "astrocytic" subtype in three tumors; iii) the "other" subtype in two tumors. All twelve tumors with the 1p/19q co-deletion carried the most common IDH1 R132H mutation. In seven of these tumors, we found protein-disrupting point mutations in the remaining allele of CIC, four of which are novel. One of these tumors also had a deleterious mutation in FUBP1. Only by integrating RNA expression and array CGH data, were we able to discover an exon-spanning homozygous microdeletion within the remaining allele of CIC in an additional tumor with 1p/19q co-deletion. Therefore we propose that the mutation rate might be underestimated when looking at sequence variants alone. In conclusion, the high frequency and the spectrum of CIC mutations in our 1p/19q-codeleted tumor cohort support the hypothesis that CIC acts as a tumor suppressor in these tumors, whereas FUBP1 might play only a minor role.

Two of the signature genetic events that occur in human gliomas, EGFR amplification and IDH mutation, are poorly represented in experimental models in vitro. EGFR amplification, for example, occurs in 40 to 50% of GBM, and yet, EGFR amplification is rarely preserved in cell cultures derived from human tumors. To analyze the fate of EGFR amplified and IDH mutated cells in culture, we followed the development over time of cultures derived from human xenografts in nude rats enriched for tumor cells with EGFR amplification and of cultures derived from patient samples with IDH mutations, in serum monolayer and spheroid suspension culture, under serum and serum free conditions. We observed under serum monolayer conditions, that nestin positive or nestin and SMA double positive rat stromal cells outgrew EGFR amplified tumor cells, while serum spheroid cultures preserved tumor cells with EGFR amplification. Serum free suspension culture exhibited a more variable cell composition in that the resultant cell populations were either predominantly nestin/SOX2 co-expressing rat stromal cells or human tumor cells, or a mixture of both. The selection for nestin/SMA positive stromal cells under serum monolayer conditions was also consistently observed in human oligodendrogliomas and oligoastrocytomas with IDH mutations. Our results highlight for the first time that serum monolayer conditions can select for stromal cells instead of tumor cells in certain brain tumor subtypes. This result has an important impact on the establishment of new tumor cell cultures from brain tumors and raises the question of the proper conditions for the growth of the tumor cell populations of interest.

We measured potential nitrate removal and denitrification rates in hydrothermally altered sediments inhabited by Beggiatoa mats and adjacent brown oil stained sediments from the Guaymas Basin, Gulf of California. Sediments with Beggiatoa maintained slightly higher rates of potential denitrification than did brown sediments at 31.2 (±12.1) versus 21.9 (±1.4) μM N d-1, respectively. In contrast, the nitrate removal rates in brown sediments were higher than those observed in mat-hosting sediments at 418 ± 145 versus 174 ± 74 μM N d-1, respectively. Additional experiments were conducted to assess the responses of denitrifying communities to environmental factors (i.e. nitrate, sulfide, and dissolved organic carbon (DOC) concentration). The denitrifying community had a high affinity for nitrate (Km = 137 ± 91 μΜ ΝΟ3-), in comparison to other environmental communities of denitrifiers, and were capable of high maximum rates of denitrification (vmax = 1164 ± 153 μM N d-1). The presence of sulfide resulted in significantly lower denitrification rates. Microorganisms with the potential to perform denitrification were assessed in these sediments using the bacterial16S rRNA gene and nitrous oxide reductase (nosZ) functional gene libraries. The bacterial 16S rRNA gene clone library was dominated by Epsilonproteobacteria (38%), some of which (e.g. Sulfurimonas sp.) have a potential for sulfide dependent denitrification. The nosZ clone library did not contain clones similar to pure culture denitrifiers; these clones were most closely associated with environmental clones.