347 research outputs found
Skin tear prevalence in an Australian acute care hospital : A 10-year analysis
Hospital-acquired skin tear prevalence is under-reported; thus, the aim of this study was to analyse skin tear point prevalence and characteristics in a tertiary acute care hospital in Queensland, Australia, over a 10-year period. All consenting adult inpatients received a full skin inspection and skin tear category, site, cause, treatment, and whether it was documented as hospital- or community-acquired were recorded. Eleven prevalence audits were analysed with a total sample of 3626 patients. An overall pooled prevalence of 8.9% (95% confidence interval [CI] 7.5-10.4) with an associated hospital-acquired pooled prevalence of 5.5% (95% CI 4.5-6.7) was found. In total, 616 skin tears were reported, of which 374 (60.7%) were hospital-acquired. Over a third of patients (38.7%) had multiple skin tears and most patients (84.8%) with at least one skin tear were aged ≥70 years. The largest proportion of skin tears (40.1%) was those with no skin flap. Of those documented, most were caused by falls or collisions, suggesting combined skin tear and falls prevention strategies may be effective. Over a decade, there was a downward trend in hospital-acquired skin tear, which is encouraging. Skin tear prevalence is recommended as a measure of care quality with an emphasis on good quality documentation
A Simple, Highly Efficient Method for Heterologous Expression in Mammalian Primary Neurons Using Cationic Lipid-mediated mRNA Transfection
Expression of heterologous proteins in adult mammalian neurons is a valuable technique for the study of neuronal function. The post-mitotic nature of mature neurons prevents effective DNA transfection using simple, cationic lipid-based methods. Adequate heterologous protein expression is often only achievable using complex techniques that, in many cases, are associated with substantial toxicity. Here, a simple method for high efficiency transfection of mammalian primary neurons using in vitro transcribed mRNA and the cationic lipid transfection reagent Lipofectamine™ 2000 is described. Optimal transfection conditions were established in adult mouse dissociated dorsal root ganglion (DRG) neurons using a 96-well based luciferase activity assay. Using these conditions, a transfection efficiency of 25% was achieved in DRG neurons transfected with EGFP mRNA. High transfection efficiencies were also obtained in dissociated rat superior cervical ganglion (SCG) neurons and mouse cortical and hippocampal cultures. Endogenous Ca2+ currents in EGFP mRNA-transfected SCG neurons were not significantly different from untransfected neurons, which suggested that this technique is well suited for heterologous expression in patch clamp recording experiments. Functional expression of a cannabinoid receptor (CB1R), a G protein inwardly rectifying K+ channel (GIRK4) and a dominant-negative G protein α-subunit mutant (GoA G203T) indicate that the levels of heterologous protein expression attainable using mRNA transfection are suitable for most functional protein studies. This study demonstrates that mRNA transfection is a straightforward and effective method for heterologous expression in neurons and is likely to have many applications in neuroscience research
Rapid Modification of Proteins Using a Rapamycin-Inducible Tobacco Etch Virus Protease System
The ability to disrupt the function of a specific protein on a rapid time scale provides a powerful tool for biomedical research. Specific proteases provide a potential method to selectively cleave a chosen protein, but rapid control of protease activity is difficult.A heterologous expression system for rapid target-directed proteolysis in mammalian cells was developed. The system consists of an inducible NIa protease from the tobacco etch virus (TEVp) and a chosen protein into which a TEVp substrate recognition sequence (TRS) has been inserted. Inducible activity was conferred to the TEVp using rapamycin-controlled TEVp fragment complementation. TEVp activity was assayed using a FRET-based reporter construct. TEVp expression was well tolerated by mammalian cells and complete cleavage of the substrate was possible. Cleavage at 37 degrees C proceeded exponentially with a time constant of approximately 150 minutes. Attempts to improve cleavage efficiency were hampered by substantial background activity, which was attributed to inherent affinity between the TEVp fragments. A second TEVp assay, based on changes in inactivation of a modified K(V)3.4 channel, showed that functional properties of a channel can be using altered using an inducible TEVp system. Similar levels of background activity and variability were observed in both electrophysiological and FRET assays.The results suggested that an optimum level of TEVp expression leading to sufficient inducible activity (with minimal background activity) exists but the variability in expression levels between cells makes the present system rather impractical for single cell experiments. The system is likely to be more suitable for experiments in which the cell-to-cell variability is less of an issue; for example, in experiments involving large populations of cells
Intranuclear Microinjection of DNA into Dissociated Adult Mammalian Neurons
Primary neuronal cell cultures are valuable tools to study protein function since they represent a more biologically relevant system compared to immortalized cell lines. However, the post-mitotic nature of primary neurons prevents effective heterologous protein expression using common procedures such as electroporation or chemically-mediated transfection. Thus, other techniques must be employed in order to effectively express proteins in these non-dividing cells
Soluble α2-macroglobulin receptor is increased in endotracheal aspirates from infants and children after cardiopulmonary bypass
ObjectiveCytokine dysregulation contributes to the systemic inflammatory response after cardiopulmonary bypass. Clearance of cytokine binding proteins may be important in the resolution of inflammation. Our aim was to determine whether the cytokine binding protein α2-macroglobulin and its soluble receptor were upregulated in endotracheal aspirates from infants and children undergoing cardiopulmonary bypass.MethodsSeventy tracheal aspirates were collected before and after cardiopulmonary bypass from 35 infants and children undergoing surgical correction of congenital heart defects. α2-Macroglobulin and the soluble α2-macroglobulin receptor were identified by Western blot. With the use of multi-analyte cytokine profiling, pro-inflammatory and anti-inflammatory cytokines were quantified, normalized to total protein, and expressed as ratios. Paired t tests and Wilcoxon signed-rank tests were performed between prebypass and postbypass samples. Correlations were examined among α2-macroglobulin, soluble α2-macroglobulin receptor, cytokine ratios, and the clinical variables of cardiopulmonary bypass, aortic crossclamp, and circulatory arrest times.Resultsα2-Macroglobulin increased by 50% (mean densitometry increase 82,683 ± 184,594, P = .012), and soluble α2-macroglobulin receptor increased by 17% (mean densitometry increase 506,148 ± 687,037, P = .0001) after cardiopulmonary bypass. The ratio of interleukin-8/interleukin-4 increased by 136% (P = .0001), and interleukin-8/interleukin-10 increased by 102% (P = .001). The increase in soluble α2-macroglobulin receptor was positively correlated with the ratios of interleukin-8/interleukin-4 and interleukin-8/interleukin-10. There were no statistically significant positive correlations between the increase in α2-macroglobulin or soluble α2-macroglobulin receptor and measured clinical variables.ConclusionsWe report for the first time the upregulation of α2-macroglobulin and soluble α2-macroglobulin receptor in tracheal aspirates after cardiopulmonary bypass in infants and children. Soluble α2-macroglobulin receptor correlates with increased α2-macroglobulin and a disproportionate increase in pro-inflammatory to anti-inflammatory cytokine ratios
A narrative review of the surgical management of Paget-Schroetter syndrome: case series and long-term follow-up
BACKGROUND AND OBJECTIVE: Paget-Schroetter syndrome (PSS) is an uncommon disorder which causes thrombosis of the subclavian vein (SV). This is due to compression of the SV by the surrounding anatomical structures. The optimal management of PSS remains subject to debate, with endovascular intervention and open surgical decompression being favoured current options. This review article evaluates both approaches to the management of PSS, while also presenting a case series with long-term follow-up of patients that underwent open surgical intervention for PSS. METHODS: The clinical outcomes of PSS patients undergoing different 4 surgical approaches to perform surgical decompression are included. A literature review, across publications from PubMed, Embase, and Web of Science, was conducted with specific criteria to facilitate evaluation of both open surgical and endovascular approaches to the management of PSS. KEY CONTENT AND FINDINGS: Evaluation of data from the included case series and available literature suggests that endovascular thrombolytic devices offer better clinical results, however, SV decompression is still required for successful resolution. CONCLUSIONS: An approach to PSS encompassing endovascular intervention followed by surgical anatomical decompression may provide optimal outcomes as both intrinsic lesions and extrinsic compression of the SV is treated. However, further prospective investigation into this field is warranted
The TRiC/CCT chaperone is implicated in Alzheimer's disease based on patient GWAS and an RNAi screen in Aβ-expressing Caenorhabditis elegans.
The human Aβ peptide causes progressive paralysis when expressed in the muscles of the nematode worm, C. elegans. We have exploited this model of Aβ toxicity by carrying out an RNAi screen to identify genes whose reduced expression modifies the severity of this locomotor phenotype. Our initial finding was that none of the human orthologues of these worm genes is identical with the genome-wide significant GWAS genes reported to date (the "white zone"); moreover there was no identity between worm screen hits and the longer list of GWAS genes which included those with borderline levels of significance (the "grey zone"). This indicates that Aβ toxicity should not be considered as equivalent to sporadic AD. To increase the sensitivity of our analysis, we then considered the physical interactors (+1 interactome) of the products of the genes in both the worm and the white+grey zone lists. When we consider these worm and GWAS gene lists we find that 4 of the 60 worm genes have a +1 interactome overlap that is larger than expected by chance. Two of these genes form a chaperonin complex, the third is closely associated with this complex and the fourth gene codes for actin, the major substrate of the same chaperonin
Antibacterial and antibiofilm potency of XF drugs, impact of photodynamic activation and synergy with antibiotics.
With increasing incidence of antimicrobial resistance, there is an urgent need for novel and effective antibacterials. Destiny Pharma plc have developed a series of porphyrin-based XF drugs, some with dual mechanisms of antibacterial action. An innate mechanism acts through binding to the outer bacterial membrane and a separate, light-activated, photodynamic (PD) mechanism, acts via the generation of reactive oxygen species. This study aimed to assess the innate and PD associated antibacterial activity of XF drugs against planktonic bacteria, their biofilms and combinational effects with conventional antibiotics. Minimum inhibitory concentrations (MICs) were determined for 3 XF drugs against 114 bacterial isolates. MICs for XF-73 and XF-70 were determined (± PD). DPD-207 was designed to not exhibit PD action due to its structure. XF-drugs (± PD) were further assessed for synergy with conventional antibiotics (using a checkerboard assay) and antibiofilm activity against susceptible strains. XF drugs were innately active against all tested Gram-positive isolates. PD action significantly increased bacterial susceptibility to XF-73 and XF-70 for all Gram-positive isolates. Generally, the XF drugs exhibited higher MICs against Gram-negative isolates, however PD significantly enhanced potency, particularly for XF-70. XF-73 and XF-70 exhibited synergy with ertapenem against a methicillin resistant Staphylococcus aureus (MRSA) strain (± PD) and XF-73 with polymyxin B (± PD) against Pseudomonas aeruginosa. No antagonism was seen between the XF drugs and any of the 5 antibiotics tested. The antibiofilm effect of XF drugs was also observed for all Staphylococcus isolates tested. Generally, PD did not enhance activity for other bacterial isolates tested with the exception of XF-73 against Acinetobacter baumannii biofilms. XF drugs exhibited significant antimicrobial activity against Gram-positive bacteria, with PD enhancement of bacterial susceptibility. Additionally, XF drugs displayed synergy with conventional antibiotics and demonstrated antibiofilm effects
A psychometric evaluation of emotional responses to horror music
This research explores and designs an effective experimental interface to evaluate people's emotional responses to horror music. We studied methodological approaches by using traditional psychometric techniques to measure emotional responses, including self-reporting, and galvanic skin response (GSR). GSR correlates with psychological arousal. It can help circumvent a problem in self-reporting where people are unwilling to report particular felt responses, or confuse perceived and felt responses. We also consider the influence of familiarity. Familiarity can induce learned emotional responses rather than listeners describing how it actually makes them feel. The research revealed different findings in self-reports and GSR data. Both measurements had an interaction between music and familiarity but show inconsistent results from the perspective of simple effects
- …