CONSORT Item Reporting Quality in the Top Ten Ranked Journals of Critical Care Medicine in 2011: A Retrospective Analysis

IntroductionReporting randomised controlled trials is a key element in order to disseminate research findings. The CONSORT statement was introduced to improve the reporting quality. We assessed the adherence to the CONSORT statement of randomised controlled trials published 2011 in the top ten ranked journals of critical care medicine (ISI Web of Knowledge 2011, Thomson Reuters, London UK).MethodsDesign. We performed a retrospective cross sectional data analysis. Setting. This study was executed at the University Hospital of RWTH, Aachen. Participants. We selected the following top ten listed journals according to ISI Web of Knowledge (Thomson Reuters, London, UK) critical care medicine ranking in the year 2011: American Journal of Respiratory and Critical Care Medicine, Critical Care Medicine, Intensive Care Medicine, CHEST, Critical Care, Journal of Neurotrauma, Resuscitation, Pediatric Critical Care Medicine, Shock and Minerva Anestesiologica. Main outcome measures. We screened the online table of contents of each included journal, to identify the randomised controlled trials. The adherence to the items of the CONSORT Checklist in each trial was evaluated. Additionally we correlated the citation frequency of the articles and the impact factor of the respective journal with the amount of reported items per trial.ResultsWe analysed 119 randomised controlled trials and found, 15 years after the implementation of the CONSORT statement, that a median of 61,1% of the checklist-items were reported. Only 55.5% of the articles were identified as randomised trials in their titles. The citation frequency of the trials correlated significantly (rs = 0,433; p<0,001 and r = 0,331; p<0,001) to the CONSORT statement adherence. The impact factor showed also a significant correlation to the CONSORT adherence (r = 0,386; p<0,001).ConclusionThe reporting quality of randomised controlled trials in the field of critical care medicine remains poor and needs considerable improvement.     

Secretome analysis of Anabaena sp. PCC 7120 and the involvement of the TolC‐homologue HgdD in protein secretion

Secretion of proteins is a central strategy of bacteria to influence and respond to their environment. Until now, there has been very few discoveries regarding the cyanobacterial secrotome or the secretion machineries involved. For a mutant of the outer membrane channel TolC‐homologue HgdD of Anabaena sp. PCC 7120, a filamentous and heterocyst‐forming cyanobacterium, an altered secretome profile was reported. To define the role of HgdD in protein secretion, we have developed a method to isolate extracellular proteins of Anabaena sp. PCC 7120 wild type and an hgdD loss‐of‐function mutant. We identified 51 proteins of which the majority is predicted to have an extracellular secretion signal, while few seem to be localized in the periplasmic space. Eight proteins were exclusively identified in the secretome of wild‐type cells, which coincides with the distribution of type I secretion signal. We selected three candidates and generated hemagglutinin‐tagged fusion proteins which could be exclusively detected in the extracellular protein fraction. However, these proteins are not secreted in the hgdD‐mutant background, where they are rapidly degraded. This confirms a direct function of HgdD in protein secretion and points to the existence of a quality control mechanism at least for proteins secreted in an HgdD‐dependent pathway.     

The preoperative activity of Th1 and Th17 cytokine axes in prediction of sepsis after radical cystectomy

The aim of the study was to correlate the preoperative activity of Th1 and Th17 cytokine axes with the development of sepsis after radical cystectomy. The study involved twenty patients with the infiltrative transitional cell carcinoma of the urinary bladder without previous radiotherapy/chemotherapy, who underwent open radical cystectomy with urinary diversion. Preoperative plasma concentrations of Th1 cytokines interleukin 12 (IL-12) and interferon gamma (IFN-γ), and Th17 cytokines IL-23 and IL-17, were measured using ELISA. Preoperative expression of mRNA for IL-12p35, IFN-γ, IL-23p19 and IL-17 was quantified by real-time RT-PCR using mRNA extracted from peripheral blood mononuclear cells. Eight patients developed postoperative sepsis, diagnosed within two weeks post-operation as systemic inflammatory response syndrome in the presence of local or systemic infection. The preoperative basal plasma concentrations of Th1 and Th17 cytokines were slightly above the detection limits, with a tendency toward lower concentrations in patients who developed sepsis, but the difference was not significant (p>0.05). The preoperative expression of mRNA encoding IL-12p35 and IL-17 was significantly lower in patients who developed sepsis (p=0.003 and p=0.028, respectively). The similar trend was observed for IL-23p19 and IFN-γ, but the differences did not reach the statistical significance (p=0.051 and p=0.172, respectively). These data suggest that determination of preoperative Th1 and Th17 cytokine mRNA levels might be useful in predicting sepsis development after radical cystectomy.     

Aminoguanidine and N-acetyl-cysteine supress oxidative and nitrosative stress in EAE rat brains

Experimental autoimmune encephalomyelitis (EAE) is a well-established animal model of human multiple sclerosis (MS). We have evaluated the role of oxidative and nitrosative stress, as the causal factors in the development of EAE, responsible for the damage of cardinal cellular components, such as lipids, proteins and nucleic acids, resulting in demyelination, axonal damage, and neuronal death. EAE was induced in female Sprague-Dawley rats, 3 months old (300±20 g), by immunization with myelin basic protein in combination with Complete Freund's adjuvant (CFA). The animals were divided into seven groups: control, EAE, CFA, EAE+aminoguanidine (AG), AG, EAE+N-acetyl-L-cysteine (NAC) and NAC. The animals were sacrificed 15 days after EAE induction, and the levels of nitrosative and oxidative stress were determined in 10% homogenate of the whole encephalitic mass. In EAE rats, brain NO production and MDA level were significantly increased (P<0.001) compared to the control values, whereas AG and NAC treatment decreased both parameters in EAE rats compared to EAE group (P<0.001). Glutathione (GSH) was reduced (P<0.001) in EAE rats in comparison with the control and CFA groups, but increased in EAE+AG and EAE+NAC group compared to the EAE group (P<0.01). Superoxide dismutase (SOD) activity was significantly decreased (P<0.001) in the EAE group compared to all other experimental groups. The clinical expression of EAE was significantly decreased (P<0.05) in the EAE groups treated with AG and NAC compared to EAE rats, during disease development. The obtained results prove an important role of oxidative and nitrosative stress in the pathogenesis of EAE, whereas AG and NAC protective effects offer new possibilities for a modified combined approach in MS therapy.     

Evaluation of the DNA damaging effects of amitraz on human lymphocytes in the Comet assay

Amitraz is formamidine pesticide widely used as insecticide and acaricide. In veterinary medicine, amitraz has important uses against ticks, mites and lice on animals. Also, amitraz is used in apiculture to control Varroa destructor. It this study, the alkaline Comet assay was used to evaluate DNA damaging effects of amitraz in human lymphocytes. Isolated human lymphocytes were incubated with varying concentrations of amitraz (0.035, 0.35, 3.5, 35 and 350 μg/mL). The Comet assay demonstrated that all concentrations of amitraz caused statistically significant increase in the level of DNA damage, thus indicating that amitraz possesses genotoxic potential. The concentration of amitraz that produced the highest DNA damage (3.5 μg/mL) was chosen for further analysis with the antioxidant catalase. The obtained results showed that co-treatment with antioxidant catalase (100 IU/mL or 500 IU/mL) significantly reduced the level of DNA damage, indicating the possible involvement of reactive oxygen species in DNA damaging effects of amitraz. Flow cytometric analysis revealed increase of the apoptotic index following treatment with amitraz. However, co-treatment with catalase reduced the apoptotic index, while treatment with catalase alone reduced the percentage of apoptotoc cells even in comparison with the negative control. Therefore, catalase had protective effects against ROS-mediated DNA damage and apoptosis.     

Monte Carlo investigation of electron specific energy distribution in a single cell model

Radiation and Environmental Biophysics - Knowledge of microdosimetric quantities of certain radionuclides is important in radio immune cancer therapies. Specific energy distribution of...     

Selective ultrasound-assisted extractions of lipophilic constituents from Betula alleghaniensis and B. papyrifera wood at low temperatures

Betula alleghaniensis and B. papyrifera are widely distributed in the province of Québec (Canada) and, since these trees are valuable exports for the local lumber industry, large amounts of their residual ligneous biomass are available for further exploitation. Betula species are well known for their significant concentrations of triterpenes, some of which were recently discovered to present promising bioactivity. The secondary transformation of birch biomass could therefore become important for many industries, particularly the pharmaceutical industry. In the present study, extracts from birch sawdust were obtained using an optimised ultrasound-assisted extraction in which the careful choice of temperature permitted a selective extraction of the targeted triterpenes. Moreover, compared with the classical Soxhlet method, higher extraction yields were obtained in a shorter time. The lipophilic extracts obtained using dichloromethane as a solvent were analysed by GC-MS and the major compounds identified as lupane-type cyclic triterpenoids accompanied by the non-cyclic triterpene squalene. Numerous aliphatic long-chain fatty acids were also found in the extracts together with phytosterols. Betulonic acid and squalene, the major extract constituents for both B. alleghaniensis and B. papyrifera, are both bioactive molecules.     

Efficacy of Imiquimod-Based Transcutaneous Immunization Using a Nano-Dispersed Emulsion Gel Formulation

Background

Transcutaneous immunization (TCI) approaches utilize skin associated lymphatic tissues to elicit specific immune responses. In this context, the imidazoquinoline derivative imiquimod formulated in Aldara applied onto intact skin together with a cytotoxic T lymphocyte (CTL) epitope induces potent CTL responses. However, the feasibility and efficacy of the commercial imiquimod formulation Aldara is limited by its physicochemical properties as well as its immunogenicity.

Methodology/Principal Findings

To overcome these obstacles, we developed an imiquimod-containing emulsion gel (IMI-Gel) and characterized it in comparison to Aldara for rheological properties and in vitro mouse skin permeation in a Franz diffusion cell system. Imiquimod was readily released from Aldara, while IMI-Gel showed markedly decreased drug release. Nevertheless, comparing vaccination potency of Aldara or IMI-Gel-based TCI in C57BL/6 mice against the model cytotoxic T-lymphocyte epitope SIINFEKL, we found that IMI-Gel was equally effective in terms of the frequency of peptide-specific T-cells and in vivo cytolytic activity. Importantly, transcutaneous delivery of IMI-Gel for vaccination was clearly superior to the subcutaneous or oral route of administration. Finally, IMI-Gel based TCI was at least equally effective compared to Aldara-based TCI in rejection of established SIINFEKL-expressing E.G7 tumors in a therapeutic setup indicated by enhanced tumor rejection and survival.

Conclusion/Significance

In summary, we developed a novel imiquimod formulation with feasible pharmaceutical properties and immunological efficacy that fosters the rational design of a next generation transcutaneous vaccination platform suitable for the treatment of cancer or persistent virus infections.     

Genetic diversity and relationships among species of the genus Thymus L. (section Serpyllum)

Amplified fragment length polymorphism (AFLP) markers were used to assess genetic relations of 32 Thymus populations belonging to seven species of genus Thymus, section Serpyllum, from Serbia. The collected species belong to three subsections: (1) Alternantes Klokov (Th. pulegioides L.), (2) Isolepides (Borbás) Halácsy (Th. glabrescens Willd., Th. marschallianus Willd., Th. pannonicus All.), and (3) Pseudomarginati (Braun ex Borbás) Jalas (Th. balcanus Borbás, Th. moesiacus Velen., Th. praecox Opiz). Analysis of molecular variance (AMOVA) revealed that most of the genetic diversity was attributable to differences among individuals within populations, while the remaining variability was almost equally distributed among species and among populations within species. The pairwise AMOVA analyses further showed that species differentiation was significant in all cases, except between Th. marschallianus and Th. pannonicus. All the individuals belonging to Th. pulegioides (subsection Alternantes) formed a well-supported clade, the most divergent from all other species in this study. This genetic differentiation of Th. pulegioides is supported by its morphological features because it is the only analyzed taxon having hairs only on the stem edges, and can easily be recognized by this character. According to our results, the subsection Isolepides is polyphyletic. Th. glabrescens is clearly separated from the other two species of this subsection (Th. marschallianus, Th. pannonicus). Although morphologically distinguishable by leaf indumentum, individuals belonging to Th. marschallianus and Th. pannonicus could not be differentiated at the molecular level. The species belonging to subsection Pseudomarginati formed a monophyletic clade, although not supported by high bootstrap value. The clade is further divided into three well-supported clades representing distinct species (Th. balcanus, Th. praecox and Th. moesiacus). In our study, the AFLP markers demonstrated that they could be suitable for studying complex genetic relationships, including frequent interspecies hybridization events, although strict reticulate evolutionary history could not be determined. All the population genetic parameters in the section Serpyllum suggest that the reproductive incompatibility between the species is very weak, which indicates a strong reticulating system, especially in the case when two or more species occur sympatrically.