Specific versus Non-Specific Immune Responses in an Invertebrate Species Evidenced by a Comparative de novo Sequencing Study

Our present understanding of the functioning and evolutionary history of invertebrate innate immunity derives mostly from studies on a few model species belonging to ecdysozoa. In particular, the characterization of signaling pathways dedicated to specific responses towards fungi and Gram-positive or Gram-negative bacteria in Drosophila melanogaster challenged our original view of a non-specific immunity in invertebrates. However, much remains to be elucidated from lophotrochozoan species. To investigate the global specificity of the immune response in the fresh-water snail Biomphalaria glabrata, we used massive Illumina sequencing of 5′-end cDNAs to compare expression profiles after challenge by Gram-positive or Gram-negative bacteria or after a yeast challenge. 5′-end cDNA sequencing of the libraries yielded over 12 millions high quality reads. To link these short reads to expressed genes, we prepared a reference transcriptomic database through automatic assembly and annotation of the 758,510 redundant sequences (ESTs, mRNAs) of B. glabrata available in public databases. Computational analysis of Illumina reads followed by multivariate analyses allowed identification of 1685 candidate transcripts differentially expressed after an immune challenge, with a two fold ratio between transcripts showing a challenge-specific expression versus a lower or non-specific differential expression. Differential expression has been validated using quantitative PCR for a subset of randomly selected candidates. Predicted functions of annotated candidates (approx. 700 unisequences) belonged to a large extend to similar functional categories or protein types. This work significantly expands upon previous gene discovery and expression studies on B. glabrata and suggests that responses to various pathogens may involve similar immune processes or signaling pathways but different genes belonging to multigenic families. These results raise the question of the importance of gene duplication and acquisition of paralog functional diversity in the evolution of specific invertebrate immune responses

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Purification and characterization of a novel calcium binding protein from the extrapallial fluid of mollusc Mytilus edulis

In the bivalve mollusc Mytilus edulis shell thickening occurs from the extrapallial (EP) fluid wherein secreted shell matrix macromolecules are thought to self-assemble into a framework that regulates the growth of CaCO3 crystals, which eventually constitute ∼95% of the mature shell. Herein is the initial report on the purification and characterization of a novel EP fluid glycoprotein, which is likely a building block of the shell-soluble organic matrix. This primary EP fluid protein comprises 56% of the total protein in the fluid and is shown to be a dimer of 28,340 Da monomers estimated to be 14.3% by weight carbohydrate. The protein is acidic (pI = 4.43) and rich in histidine content (11.14%) as well as in Asx and Glx residues (25.15% total). The N terminus exhibits an unusual repeat sequence of histidine and aspartate residues that occur in pairs: NPVDDHHDDHHDAPIVEHHD∼. Ultracentrifugation and polyacrylamide gel electrophoresis demonstrate that the protein binds calcium and in so doing assembles into a series of higher order protomers, which appear to have extended structures. Circular dichroism shows that the protein-calcium binding/protomer formation is coupled to a significant rearrangement in the protein\u27s secondary structure in which there is a major reduction in β-sheet with an associated increase in α-helical content of the protein. A model for shell organic matrix self-assembly is proposed

Patterns of non-syndromic permanent tooth agenesis in a large orthodontic population

OBJECTIVE: The aim of this study is to explore patterns of non-syndromic permanent tooth agenesis in a large orthodontic patient group. DESIGN: A record review was performed in various orthodontic clinics to identify white patients with non-syndromic permanent tooth agenesis, excluding 3rd molars. Four hundred and fourteen subjects fulfilled the inclusion criteria. RESULTS: In the 414 subjects with tooth agenesis, approximately 70% presented 1-2 missing teeth. Symmetric agenesis patterns were often observed in the sample (by jaw, by side, or crossed quadrants), with prevalence approaching 30% for cases with contralateral tooth agenesis within a jaw. In cases with 1 or 2 missing teeth, from the total number of potential tooth agenesis patterns in the sample, a certain part was evident, limiting the variation to 27.8% (44/158). In the entire sample, both in the maxilla and the mandible a certain incisor/premolar agenesis phenotype was observed in 59.0% of cases in isolated form. CONCLUSIONS: Although there was variation in the tooth agenesis patterns, our findings suggest the involvement of particular genetic, epigenetic, and/or environmental factors in the formation of the entire dentition, which often lead to specific tooth agenesis phenotypes in cases where this process is disrupted. The present study provides a comprehensive categorization of orthodontic cases with tooth agenesis and can assist in planning future epidemiological and genetic studies

In Vitro Activity of Caffeic Acid Phenethyl Ester against Different Oral Microorganisms

This was an in vitro study that aimed to evaluate the antimicrobial effect of the propolis extract caffeic acid phenethyl ester (CAPE) on four different oral microorganisms. Seven different concentrations of CAPE (0.2, 0.5, 1, 1.5, 2, 3, and 4 mg/mL) for use against Staphylococcus aureus, Streptococcus mutans, Streptococcus oralis, and Streptococcus salivarius were determined using minimum inhibition concentration (MIC), minimum bactericidal concentration (MBC), broth microdilution, and well diffusion tests over 1, 3, 6, 12, and 24 h, while NaF at 0.05 percent was used as a positive control. Staphylococcus aureus was most affected by CAPE’s inhibitory effect on bacterial growth, whereas S. mutans was the least affected. S. mutans and S. oralis had similar CAPE MIC and MBC values of 1 mg/mL and 1.5 mg/mL, respectively. The most resistant bacteria to CAPE were S. salivarius and S. aureus, with MIC and MBC values of 3 mg/mL and 4 mg/mL, respectively. S. oralis, followed by S. salivarius, S. mutans, and S. aureus, had the highest viable count following exposure to CAPE’s MBC values, while S. aureus had the lowest. The current results of the inhibitory effect of CAPE on bacterial growth are promising, and the values of both CAPE MBC and MIC against the related four cariogenic bacterial organisms are significant. CAPE can be employed as an adjunct dental hygiene substance for maintaining good oral hygiene, and has a potential therapeutic effect in the field of oral health care

ESI and MALDI LC/MS-MS approaches for large scale protein, identification and quantification : Are they equivalent?

An approach for large scale protein identification and quantification was described. The peptide mixtures from the strong cation exchange (SCX) fractionated protein digests were separated and analyzed by HPLC MS/MS. The human fibrobalst nuclei were purified from stimulated and non-stimulated cells. The quantification results with MALDI and ESI LC/MS were shown to be identical within experimental error