The venom composition of the parasitic wasp Chelonus inanitus resolved by combined expressed sequence tags analysis and proteomic approach

<p>Abstract</p> <p>Background</p> <p>Parasitic wasps constitute one of the largest group of venomous animals. Although some physiological effects of their venoms are well documented, relatively little is known at the molecular level on the protein composition of these secretions. To identify the majority of the venom proteins of the endoparasitoid wasp <it>Chelonus inanitus </it>(Hymenoptera: Braconidae), we have randomly sequenced 2111 expressed sequence tags (ESTs) from a cDNA library of venom gland. In parallel, proteins from pure venom were separated by gel electrophoresis and individually submitted to a nano-LC-MS/MS analysis allowing comparison of peptides and ESTs sequences.</p> <p>Results</p> <p>About 60% of sequenced ESTs encoded proteins whose presence in venom was attested by mass spectrometry. Most of the remaining ESTs corresponded to gene products likely involved in the transcriptional and translational machinery of venom gland cells. In addition, a small number of transcripts were found to encode proteins that share sequence similarity with well-known venom constituents of social hymenopteran species, such as hyaluronidase-like proteins and an Allergen-5 protein.</p> <p>An overall number of 29 venom proteins could be identified through the combination of ESTs sequencing and proteomic analyses. The most highly redundant set of ESTs encoded a protein that shared sequence similarity with a venom protein of unknown function potentially specific of the <it>Chelonus </it>lineage. Venom components specific to <it>C. inanitus </it>included a C-type lectin domain containing protein, a chemosensory protein-like protein, a protein related to yellow-e3 and ten new proteins which shared no significant sequence similarity with known sequences. In addition, several venom proteins potentially able to interact with chitin were also identified including a chitinase, an imaginal disc growth factor-like protein and two putative mucin-like peritrophins.</p> <p>Conclusions</p> <p>The use of the combined approaches has allowed to discriminate between cellular and truly venom proteins. The venom of <it>C. inanitus </it>appears as a mixture of conserved venom components and of potentially lineage-specific proteins. These new molecular data enrich our knowledge on parasitoid venoms and more generally, might contribute to a better understanding of the evolution and functional diversity of venom proteins within Hymenoptera.</p

Body Fluid Cytokine Levels in Mild Cognitive Impairment and Alzheimer’s Disease: a Comparative Overview

This article gives a comprehensive overview of cytokine and other inflammation associated protein levels in plasma, serum and cerebrospinal fluid (CSF) of patients with Alzheimer's disease (AD) and mild cognitive impairment (MCI). We reviewed 118 research articles published between 1989 and 2013 to compare the reported levels of 66 cytokines and other proteins related to regulation and signaling in inflammation in the blood or CSF obtained from MCI and AD patients. Several cytokines are evidently regulated in (neuro-) inflammatory processes associated with neurodegenerative disorders. Others do not display changes in the blood or CSF during disease progression. However, many reports on cytokine levels in MCI or AD are controversial or inconclusive, particularly those which provide data on frequently investigated cytokines like tumor necrosis factor alpha (TNF-α) or interleukin-6 (IL-6). The levels of several cytokines are possible indicators of neuroinflammation in AD. Some of them might increase steadily during disease progression or temporarily at the time of MCI to AD conversion. Furthermore, elevated body fluid cytokine levels may correlate with an increased risk of conversion from MCI to AD. Yet, research results are conflicting. To overcome interindividual variances and to obtain a more definite description of cytokine regulation and function in neurodegeneration, a high degree of methodical standardization and patients collective characterization, together with longitudinal sampling over years is essential

Cerebrospinal fluid markers of neuroinflammation in delirium:A role for interleukin-1β in delirium after hip fracture

AbstractObjectiveExaggerated central nervous system (CNS) inflammatory responses to peripheral stressors may be implicated in delirium. This study hypothesised that the IL-1β family is involved in delirium, predicting increased levels of interleukin-1β (IL-1β) and decreased IL-1 receptor antagonist (IL-1ra) in the cerebrospinal fluid (CSF) of elderly patients with acute hip fracture. We also hypothesised that Glial Fibrillary Acidic Protein (GFAP) and interferon-γ (IFN-γ) would be increased, and insulin-like growth factor 1 (IGF-1) would be decreased.MethodsParticipants with acute hip fracture aged >60 (N=43) were assessed for delirium before and 3–4 days after surgery. CSF samples were taken at induction of spinal anaesthesia. Enzyme-linked immunosorbent assays (ELISA) were used for protein concentrations.ResultsPrevalent delirium was diagnosed in eight patients and incident delirium in 17 patients. CSF IL-1β was higher in patients with incident delirium compared to never delirium (incident delirium 1.74pg/ml (1.02–1.74) vs. prevalent 0.84pg/ml (0.49–1.57) vs. never 0.66pg/ml (0–1.02), Kruskal–Wallis p=0.03). CSF:serum IL-1β ratios were higher in delirious than non-delirious patients. CSF IL-1ra was higher in prevalent delirium compared to incident delirium (prevalent delirium 70.75pg/ml (65.63–73.01) vs. incident 31.06pg/ml (28.12–35.15) vs. never 33.98pg/ml (28.71–43.28), Kruskal–Wallis p=0.04). GFAP was not increased in delirium. IFN-γ and IGF-1 were below the detection limit in CSF.ConclusionThis study provides novel evidence of CNS inflammation involving the IL-1β family in delirium and suggests a rise in CSF IL-1β early in delirium pathogenesis. Future larger CSF studies should examine the role of CNS inflammation in delirium and its sequelae

In Vitro Effect of Instrumentation Using Ultrasonication with and without Hydrogen Peroxide on the Removal of Biofilms and Spread of Viable Microorganisms in Aerosols.

PURPOSE To evaluate the use of hydrogen peroxide as an adjunct to ultrasonication (US) in biofilm removal and whether it can limit the spread of viable microorganisms in the aerosol. MATERIALS AND METHODS Multi-species biofilms were formed on dentin disks and titanium disks fixed on a plastic surface. After placing the specimens in a periodontal pocket model, an ultrasonic scaler was applied for 30 s, in part combined with 0.25% or 0.5% H2O2. After treatment, the remaining biofilm was analysed for bacterial counts (colony forming units [CFU]), biofilm quantity and metabolic activity. Further, the cytotoxic effect of hydrogen peroxide on periodontal ligament fibroblasts was assessed and the spread of bacteria in aerosol was quantified. RESULTS Ultrasonication reduced bacterial counts in biofilm, biofilm mass and metabolic activity on both dentin and titanium disks. Adjunctive use of 0.25% and 0.5% H2O2 more effectively reduced the viable bacteria in biofilm than ultrasonication alone; this was also found on both dentin and titanium. The different concentrations of H2O2 did not lead to corresponding differences in bacterial mass and metabolic activity. The spread of bacteria through aerosols was statistically significantly reduced when adjunctive H2O2 was used. However, a certain cytotoxic effect on periodontal ligament fibroblasts by H2O2 could not be ruled out. CONCLUSIONS Irrigating with H2O2 during periodontal instrumentation with an ultrasonic scaler increases the reduction of viable bacteria within biofilms. It might limit bacterial spreading via aerosols

Experimentelle Untersuchungen \ufcber die Degeneration der Prothorakaldr\ufcsen nach der Adulth\ue4utung bei der Schabe Nauphoeta cinerea

Volume: 77Start Page: 616End Page: 62