Reactions and coping strategies in lay rescuers who have provided CPR to out-of-hospital cardiac arrest victims: A qualitative study

Objective: Cardiopulmonary resuscitation (CPR) provided by community citizens is of paramount importance for out-of-hospital cardiac arrest (OHCA) victims’ survival. Fortunately, CPR rates by community citizens seem to be rising. However, the experience of providing CPR is rarely investigated. The aim of this study was to explore reactions and coping strategies in lay rescuers who have provided CPR to OHCA victims. Methods, participants: This is a qualitative study of 20 lay rescuers who have provided CPR to 18 OHCA victims. We used a semistructured interview guide focusing on their experiences after providing CPR. Setting: The study was conducted in the Stavanger region of Norway, an area with very high bystander CPR rates. Results: Three themes emerged from the interview analysis: concern, uncertainty and coping strategies. Providing CPR had been emotionally challenging for all lay rescuers and, for some, had consequences in terms of family and work life. Several lay rescuers experienced persistent mental recurrences of the OHCA incident and had concerns about the outcome for the cardiac arrest victim. Unknown or fatal outcomes often caused feelings of guilt and were particularly difficult to handle. Several reported the need to be acknowledged for their CPR attempts. Health-educated lay rescuers seemed to be less affected than others. A common coping strategy was confiding in close relations, preferably the health educated. However, some required professional help to cope with the OHCA incident. Conclusions: Lay rescuers experience emotional and social challenges, and some struggle to cope in life after providing CPR in OHCA incidents. Experiencing a positive patient outcome and being a health-educated lay rescuer seem to mitigate concerns. Common coping strategies are attempts to reduce uncertainty towards patient outcome and own CPR quality. Further studies are needed to determine whether an organised professional follow-up can mitigate the concerns and uncertainty of lay rescuers.publishedVersio

Reactions and coping strategies in lay rescuers who have provided CPR to out-of-hospital cardiac arrest victims: A qualitative study

Objective Cardiopulmonary resuscitation (CPR) provided by community citizens is of paramount importance for out-of-hospital cardiac arrest (OHCA) victims' survival. Fortunately, CPR rates by community citizens seem to be rising. However, the experience of providing CPR is rarely investigated. The aim of this study was to explore reactions and coping strategies in lay rescuers who have provided CPR to OHCA victims. Methods, participants This is a qualitative study of 20 lay rescuers who have provided CPR to 18 OHCA victims. We used a semistructured interview guide focusing on their experiences after providing CPR. Setting The study was conducted in the Stavanger region of Norway, an area with very high bystander CPR rates. Results Three themes emerged from the interview analysis: concern, uncertainty and coping strategies. Providing CPR had been emotionally challenging for all lay rescuers and, for some, had consequences in terms of family and work life. Several lay rescuers experienced persistent mental recurrences of the OHCA incident and had concerns about the outcome for the cardiac arrest victim. Unknown or fatal outcomes often caused feelings of guilt and were particularly difficult to handle. Several reported the need to be acknowledged for their CPR attempts. Health-educated lay rescuers seemed to be less affected than others. A common coping strategy was confiding in close relations, preferably the health educated. However, some required professional help to cope with the OHCA incident. Conclusions Lay rescuers experience emotional and social challenges, and some struggle to cope in life after providing CPR in OHCA incidents. Experiencing a positive patient outcome and being a health-educated lay rescuer seem to mitigate concerns. Common coping strategies are attempts to reduce uncertainty towards patient outcome and own CPR quality. Further studies are needed to determine whether an organised professional follow-up can mitigate the concerns and uncertainty of lay rescuers.publishedVersio

Expression of endoglucanases in Pichia pastoris under control of the GAP promoter

BACKGROUND: Plant-derived biomass is a potential alternative to fossil feedstocks for a greener economy. Enzymatic saccharification of biomass has been studied extensively and endoglucanases have been found to be a prerequisite for quick initial liquefaction of biomass under industrial conditions. Pichia pastoris, widely used for heterologous protein expression, can be utilized for fungal endoglucanase production. The recently marketed PichiaPink™ expression system allows for rapid clone selection, and employs the methanol inducible AOX1 promoter to ensure high protein expression levels. However, methanol is toxic and poses a fire hazard, issues which become more significant at an industrial scale. It is possible to eliminate these risks and still maintain high productivity by switching to the constitutive GAP promoter. RESULTS: In the present study, a plasmid carrying the constitutive GAP promoter was created for PichiaPink™. We then studied expression of two endoglucanases, AfCel12A from Aspergillus fumigatus and TaCel5A from Thermoascus aurantiacus, regulated by either the AOX1 promoter or the GAP promoter. Initial experiments in tubes and small bioreactors showed that the levels of AfCel12A obtained with the constitutive promoter were similar or higher, compared to the AOX1 promoter, whereas the levels of TaCel5A were somewhat lower. After optimization of cultivation conditions using a 15-l bioreactor, the recombinant P. pastoris strains utilizing the GAP promoter produced ca. 3–5 g/l of total secreted protein, with CMCase activity equivalent to 1200 nkat/ml AfCel12A and 170 nkat/ml TaCel5A. CONCLUSIONS: We present a strategy for constitutive recombinant protein expression in the novel PichiaPink™ system. Both AfCel12A and TaCel5A were successfully expressed constitutively in P. pastoris under the GAP promoter. Reasonable protein levels were reached after optimizing cultivation conditions

Genome-wide analysis of signal peptide functionality in Lactobacillus plantarum WCFS1

<p>Abstract</p> <p>Background</p> <p><it>Lactobacillus plantarum </it>is a normal, potentially probiotic, inhabitant of the human gastrointestinal (GI) tract. The bacterium has great potential as food-grade cell factory and for <it>in situ </it>delivery of biomolecules. Since protein secretion is important both for probiotic activity and in biotechnological applications, we have carried out a genome-wide experimental study of signal peptide (SP) functionality.</p> <p>Results</p> <p>We have constructed a library of 76 Sec-type signal peptides from <it>L. plantarum </it>WCFS1 that were predicted to be cleaved by signal peptidase I. SP functionality was studied using staphylococcal nuclease (NucA) as a reporter protein. 82% of the SPs gave significant extracellular NucA activity. Levels of secreted NucA varied by a dramatic 1800-fold and this variation was shown not to be the result of different mRNA levels. For the best-performing SPs all produced NucA was detected in the culture supernatant, but the secretion efficiency decreased for the less well performing SPs. Sequence analyses of the SPs and their cognate proteins revealed four properties that correlated positively with SP performance for NucA: high hydrophobicity, the presence of a transmembrane helix predicted by TMHMM, the absence of an anchoring motif in the cognate protein, and the length of the H+C domain. Analysis of a subset of SPs with a lactobacillal amylase (AmyA) showed large variation in production levels and secretion efficiencies. Importantly, there was no correlation between SP performance with NucA and the performance with AmyA.</p> <p>Conclusion</p> <p>This is the first comprehensive experimental study showing that predicted SPs in the <it>L. plantarum </it>genome actually are capable of driving protein secretion. The results reveal considerable variation between the SPs that is at least in part dependent on the protein that is secreted. Several SPs stand out as promising candidates for efficient secretion of heterologous proteins in <it>L. plantarum</it>. The results for NucA provide some hints as to the sequence-based prediction of SP functionality, but the general conclusion is that such prediction is difficult. The vector library generated in this study is based on exchangeable cassettes and provides a powerful tool for rapid experimental screening of SPs.</p

Incidence and risk factors of intraoperative periprosthetic femoral fractures during primary total hip arthroplasty: 218,423 cases reported to the Norwegian Arthroplasty Register between 1987 and 2020

Background and purpose: Intraoperative periprosthetic femoral fractures (IPFFs) can occur during primary total hip arthroplasty (THA). We describe the incidence of IPFFs during THA in Norway and estimate potential risk factors that could be associated with IPFF Patients and methods — Data from the Norwegian Arthoplasty Register (1987–2020) was used: 2,268 IPFFs from 218,423 primary THAs in 172,598 patients. The following factors were analyzed: sex, age, diagnosis, previous operation on the same hip, surgical approach, and stem fixation technique. Association of these factors with IPFF risk was assessed using multivariable Poisson regression. Results: IPFF occurred during 2,268 operations with an incidence of 1.0% among all primary THAs. The risk of IPFF was associated with female sex (relative risk 1.8; 99% CI 1.5–2.1), age 80–90 years and age over 90 years (compared with age 60–70 years: 1.3; CI 1.0–1.6 and 2.6; CI 1.6–4.3, respectively), non-osteoarthritis diagnoses (2.2; CI 1.9–2.6), previous surgery to the same hip (1.8; CI 1.5–2.2), lateral approach (compared with the posterior approach: 1.5; CI 1.1–2.0), and cementless stem fixation (2.7; CI 2.0–3.6). Interpretation: Surgeons should be aware of the factors associated with an increased risk of IPFF: female sex, age above 80 years, non-osteoarthritis diagnoses, and previous surgery to the same hip. Cemented stem fixation and posterior approach should be favored in high-risk patients, such as elderly women.publishedVersio

Heterologous expression of Oenococcus oeni malolactic enzyme in Lactobacillus plantarum for improved malolactic fermentation

Lactobacillus plantarum is involved in a multitude of food related industrial fermentation processes including the malolactic fermentation (MLF) of wine. This work is the first report on a recombinant L. plantarum strain successfully conducting MLF. The malolactic enzyme (MLE) from Oenococcus oeni was cloned into the lactobacillal expression vector pSIP409 which is based on the sakacin P operon of Lactobacillus sakei and expressed in the host strain L. plantarum WCFS1. Both recombinant and wild-type L. plantarum strains were tested for MLF using a buffered malic acid solution in absence of glucose. Under the conditions with L-malic acid as the only energy source and in presence of Mn2+ and NAD+, the recombinant L. plantarum and the wild-type strain converted 85% (2.5 g/l) and 51% (1.5 g/l), respectively, of L-malic acid in 3.5 days. Furthermore, the recombinant L. plantarum cells converted in a modified wine 15% (0.4 g/l) of initial L-malic acid concentration in 2 days. In conclusion, recombinant L. plantarum cells expressing MLE accelerate the malolactic fermentation

Identification of proteins related to the stress response in Enterococcus faecalis V583 caused by bovine bile

<p>Abstract</p> <p>Background</p> <p><it>Enterococcus faecalis </it>is an opportunistic pathogen and one of the most important causes of hospital infections. Bile acids are a major stress factor bacteria have to cope with in order to colonize and survive in the gastro-intestinal tract. The aim of this study was to investigate the effects of bile acids on the intracellular proteome of <it>E. faecalis </it>V583.</p> <p>Results</p> <p>The proteomes of cells challenged with 1% bile were analyzed after 20 - 120 minutes exposure, using 2D gel electrophoresis and mass spectrometry. Among the approximately 500 observed proteins, 53 unique proteins were found to be regulated in response to bile and were identified with mass spectrometry. The identified proteins belonged to nine different functional classes, including fatty acid- and phospholipid-biosynthesis, energy metabolism, and transport and binding. Proteins involved in fatty acid and phospholipid biosynthesis pathways were clearly overrepresented among the identified proteins and all were down-regulated upon exposure to bile. The proteome data correlated reasonably well with data from previous transcriptome experiments done under the same conditions, but several differences were observed.</p> <p>Conclusion</p> <p>The results provide an overview of potentially important proteins that <it>E. faecalis </it>V583 needs to regulate in order to survive and adapt to a bile-rich environment, among which are several proteins involved in fatty acid and phospholipid biosynthesis pathways. In addition, this study reveals several hypothetical proteins, which are both abundant and clearly regulated and thus stand out as targets for future studies on bile stress.</p

Quantitative transcript analysis of the inducible expression system pSIP: comparison of the overexpression of Lactobacillus spp. β-galactosidases in Lactobacillus plantarum

<p>Abstract</p> <p>Background</p> <p>Two sets of overlapping genes, <it>lacLMReu </it>and <it>lacLMAci</it>, encoding heterodimeric β-galactosidases from <it>Lactobacillus reuteri </it>and <it>Lactobacillus acidophilus</it>, respectively, have previously been cloned and expressed using the pSIP vector system and <it>Lactobacillus plantarum </it>WCSF1 as host. Despite the high similarity between these <it>lacLM </it>genes and the use of identical cloning and expression strategies, strains harboring <it>lacLMReu </it>produced about twenty-fold more β-galactosidase than strains containing <it>lacLMAci</it>.</p> <p>Results</p> <p>In this study, the plasmid copy numbers (PCN) of expression vectors pEH9R (<it>lacLMReu</it>) and pEH9A (<it>lacLMAci</it>) as well as the transcription levels of both <it>lacLM </it>genes were compared using quantitative PCR methods. Analyses of parallel fermentations of <it>L. plantarum </it>harboring either pEH9R or pEH9A showed that the expression plasmids were present in similar copy numbers. However, transcript levels of <it>lacLM </it>from <it>L. reuteri </it>(pEH9R) were up to 18 times higher than those of <it>lacLM </it>from <it>L. acidophilus </it>(pEH9A). As a control, it was shown that the expression levels of regulatory genes involved in pheromone-induced promoter activation were similar in both strains.</p> <p>Conclusion</p> <p>The use of identical expression strategies for highly similar genes led to very different mRNA levels. The data indicate that this difference is primarily caused by translational effects that are likely to affect both mRNA synthesis rates and mRNA stability. These translational effects thus seem to be a dominant determinant for the success of gene expression efforts in lactobacilli.</p

Heterologous expression of a recombinant lactobacillal -galactosidase in Lactobacillus plantarum: effect of different parameters on the sakacin P-based expression system

Background: Two overlapping genes lacL and lacM (lacLM) encoding for heterodimeric -galactosidase from Lactobacillus reuteri were previously cloned and over-expressed in the food-grade host strain Lactobacillus plantarum WCFS1, using the inducible lactobacillal pSIP expression system. In this study, we analyzed different factors that affect the production of recombinant L. reuteri -galactosidase. Results: Various factors related to the cultivation, i.e. culture pH, growth temperature, glucose concentration, as well as the induction conditions, including cell concentration at induction point and inducer concentration, were tested. Under optimal fermentation conditions, the maximum -galactosidase levels obtained were 130 U/mg protein and 3540 U/ml of fermentation broth corresponding to the formation of approximately 200 mg of recombinant protein per litre of fermentation medium. As calculated from the specific activity of the purified enzyme (190 U/mg), -galactosidase yield amounted to roughly 70% of the total soluble intracellular protein of the host organism. It was observed that pH and substrate (glucose) concentration are the most prominent factors affecting the production of recombinant -galactosidase. Conclusions: The over-expression of recombinant L. reuteri -galactosidase in a food-grade host strain was optimized, which is of interest for applications of this enzyme in the food industry. The results provide more detailed insight into these lactobacillal expression systems and confirm the potential of the pSIP system for efficient, tightly controlled expression of enzymes and proteins in lactobacilli.(VLID)90704