bluuebunny/crossref_metadata_2025 huggingface dataset:
Escape box and puzzle design as educational methods for engagement and satisfaction of medical student learners in emergency medicine: survey study
Authors: Cantwell Christina , Saadat Soheil , Sakaria Sangeeta , Wiechmann Warren , Sudario Gabriel
Published: July 2022

Background

Gamification in medical education has gained popularity over the past several years. We describe a virtual escape box in emergency medicine clerkship didactics to teach chest pain and abdominal pain and compare this instructional method to a traditional flipped classroom format.

Methods

A virtual escape box was designed at our institution and incorporated into the mandatory two-week emergency medicine clerkship. The game consisted of a PDF with four cases containing puzzles to unlock a final clue. Likert scale surveys were administered to assess participants’ perceptions of the escape box format; of clerkship didactics as a whole; and of the clerkship overall. These responses were compared to the prior year’s evaluations on flipped classroom didactics and clerkship.

Results

One hundred thirty-four learners participated in the escape box and completed the survey. Eighty-six percent strongly agreed with feeling more engaged with the escape box, 84% strongly agreed with learning something new, 81% strongly agreed with finding the escape box to be satisfying, 78% strongly agreed with being able to apply knowledge gained, and 74% strongly agreed with wanting more escape boxes incorporated into medical education. The escape box showed a higher average score (3.6 ± 0.63) compared to chest pain (3.5 ± 0.67) and abdominal pain (3.2 ± 0.77) flipped classroom sessions (p = 0.0491) for the category of “lecturer explaining content clearly and at the proper level of complexity.” For the category of “lecturer provided effective instructional materials,” the escape box showed higher scores (3.6 ± 0.63) compared to flipped classroom for chest pain (3.4 ± 0.77) and abdominal pain (3.1 ± 0.80) (p < 0.001).

Conclusions

Escape boxes are adaptable to a virtual format and can teach abstract concepts such as teamwork and communication in addition to traditional lecture content. Ratings of didactics were higher for the escape box compared to the flipped classroom, while ratings of overall clerkship experience were not found to change significantly.
Undergraduate perceptions on transitioning into E-learning for continuation of higher education during the COVID pandemic in a developing country: a cross-sectional study from Sri Lanka
Authors: Subashini Nirma , Udayanga Lahiru , De Silva L. H. N. , Edirisinghe J. C. , Nafla M. N.
Published: July 2022

Background

The higher education was significantly influenced by the COVID pandemic within many developing countries, including Sri Lanka, necessitating to adopt more E-Learning opportunities. Therefore, this study was conducted to characterize the perceptions of Sri Lankan undergraduates to accept E-Learning in higher education, during the COVID crisis.

Method

A total of 657 randomly selected undergraduates of three state universities in Sri Lanka, were recruited as the study population. Information on Socio-demographic factors, Knowledge, Attitudes and Practices on E-Learning methods were acquired using an interviewer administered questionnaire. A Probit regression model was fitted to identify the driving factors for the willingness of undergraduates to engage in E-Learning.

Results

Around, 80.2% of the respondents were females. Majority were residing in semi-urban areas (44.7%), followed by rural areas (39.9%). More than two third of the respondents were familiar with E-Learning and different Learning Management Systems (LMS) that facilitate E-Learning (68.9%). Majority of the respondents (73.7%) were using different E-Learning platforms, mostly 2 to 3 days per week (25.7%). Only around one third (36.4%) had received any formal training in using LMS or other E-Learning platforms. Smart phones (77.8%) were found to be the most preferred device used for E-Learning activities, followed by computers and laptops (21.3%). Meanwhile, LMS/MOODLE (45.4%), WhatsApp/Viber (33.0%) and Zoom (32.7%), were the most commonly used E-Learning platforms. The average acceptance of E-Learning methods was found to be 70.7% among Sri Lankan undergraduates during the COVID epidemic. Based on the Probit regression, nature of the residing locality (P=0.009), family income (P=0.048), academic field (P<0.001) and year (P=0.028), knowledge on Information and Communications Technology [ICT] (P=0.012), and previous training experiences on E-Learning (P<0.001) were found as significant drivers that influence the acceptance of E-Learning practices of the Sri Lankan undergraduates.

Conclusion

Adopting E-learning into higher education sector could be recognized as a viable solution to facilitate the higher education during a crisis like COVID. However, relevant authorities in Sri Lanka should take immediate actions to empower the physical resources for E-Learning, improve the basic telecommunication infrastructure and conduct appropriate training programmes to promote E-Learning among Sri Lankan undergraduates.
Full-scale simulations to improve disaster preparedness in hospital pharmacies
Authors: Schumacher Laurence , Senhaji Salim , Gartner Birgit Andrea , Carrez Laurent , Dupuis Arnaud , Bonnabry Pascal , Widmer Nicolas
Published: July 2022

Purpose

Assess whether full-scale simulation exercises improved hospital pharmacies’ disaster preparedness.

Methods

Swiss hospital pharmacies performed successive full-scale simulation exercises at least four months apart. An interprofessional team created two scenarios, each representing credible regional-scale disasters involving approximately fifty casualties (a major road accident and a terrorist attack). Four exercise assessors used appraisal forms to evaluate participants’ actions and responses during the simulation (rating them using five-point Likert scales).

Results

Four hospital pharmacies performed two full-scale simulation exercises each. Differences between exercises one and two were observed. On average, the four hospitals accomplished 69% ± 6% of the actions expected of them during exercise one. The mean rate of expected actions accomplished increased to 84% ± 7% (p < 0.005) during exercise two. Moreover, the average quality of actions improved from 3.0/5 to 3.6/5 (p = 0.01), and the time required to gather a crisis management team drastically decreased between simulations (from 23 to 5 min). The main challenges were communication (reformulation) and crisis management. Simulation exercise number one resulted in three hospital pharmacies creating disaster action plans and the fourth improving its already existing plan.

Conclusion

This study highlighted the value of carrying out full-scale disaster simulations for hospital pharmacies as they improved overall institutional preparedness and increased staff awareness. The number of expected actions accomplished increased significantly. In the future, large-scale studies and concept dissemination are warranted.
Emergency departments in the United States treating high proportions of patients with ambulatory care sensitive conditions: a retrospective cross-sectional analysis
Authors: Hsuan Charleen , Zebrowski Alexis , Lin Michelle P. , Buckler David G. , Carr Brendan G.
Published: July 2022

Background

One in nine emergency department (ED) visits by Medicare beneficiaries are for ambulatory care sensitive conditions (ACSCs). This study aimed to examine the association between ACSC ED visits to hospitals with the highest proportion of ACSC visits (“high ACSC hospitals) and safety-net status.

Methods

This was a cross-sectional study of ED visits by Medicare fee-for-service beneficiaries ≥ 65 years using 2013–14 claims data, Area Health Resources File data, and County Health Rankings. Logistic regression estimated the association between an ACSC ED visit to high ACSC hospitals, accounting for individual, hospital, and community factors, including whether the visit was to a safety-net hospital. Safety net status was measured by Disproportionate Share Hospital (DSH) index patient percentage; public hospital status; and proportion of dual-eligible beneficiaries. Hospital-level correlation was calculated between ACSC visits, DSH index, and dual-eligible patients. We stratified by type of ACSC visit: acute or chronic.

Results

Among 5,192,729 ACSC ED visits, the odds of visiting a high ACSC hospital were higher for patients who were Black (1.37), dual-eligible (1.18), and with the highest comorbidity burden (1.26, p < 0.001 for all). ACSC visits had increased odds of being to high ACSC hospitals if the hospitals were high DSH (1.43), served the highest proportion of dual-eligible beneficiaries (2.23), and were for-profit (relative to non-profit) (1.38), and lower odds were associated with public hospitals (0.64) (p < 0.001 for all). This relationship was similar for visits to high chronic ACSC hospitals (high DSH: 1.59, high dual-eligibility: 2.60, for-profit: 1.41, public: 0.63, all p < 0.001) and to a lesser extent, high acute ACSC hospitals (high DSH: 1.02; high dual-eligibility: 1.48, for-profit: 1.17, public: 0.94, p < 0.001). The proportion of ACSC visits at all hospitals was weakly correlated with DSH proportion (0.2) and the proportion of dual-eligible patients (0.29), and this relationship was also seen for both chronic and acute ACSC visits, though stronger for the chronic ACSC visits.

Conclusion

Visits to hospitals with a high proportion of acute ACSC ED visits may be less likely to be to hospitals classified as safety net hospitals than those with a high proportion of chronic ACSC visits.
Initial presentation for acute low back pain: is early physical therapy associated with healthcare utilization and spending? A retrospective review of a National Database
Authors: Marrache Majd , Prasad Niyathi , Margalit Adam , Nayar Suresh K. , Best Matthew J. , Fritz Julie M. , Skolasky Richard L.
Published: July 2022

Background

Early initiation of physical therapy (PT) has been associated with lower healthcare costs and utilization; however, these studies have been limited to single institutions or healthcare systems. Our goal was to assess healthcare utilization and spending among patients who present for the first time with low back pain (LBP), according to whether they received early physical therapy (PT), using a large, nationwide sample; and geographic variation in rates of early PT and 30-day LBP-related spending.

Methods

Using the Truven MarketScan database, we identified nearly 980,000 US adults ages 18–64 years who initially presented with acute LBP from 2010 through 2014 and did not have nonmusculoskeletal causes of LBP. Approximately 110,000 patients (11%) received early PT (≤2 weeks after presentation). We compared healthcare utilization and spending at 30 days and 1 year after presentation between patients who received early PT and those who did not. Alpha = 0.05.

Results

At 30 days, early PT was associated with lower odds of chiropractor visits (odds ratio [OR] = 0.41, 95% confidence interval [CI] = 0.40–0.42), pain specialist visits (OR = 0.49, 95% CI = 0.47–0.51), emergency department visits (OR = 0.51, 95% CI = 0.49–0.54), advanced imaging (OR = 0.57, 95% CI = 0.56–0.58), orthopaedist visits (OR = 0.67, 95% CI = 0.66–0.69), and epidural steroid injections (OR = 0.68, 95% CI = 0.65–0.70). At 1 year, early PT was associated with less healthcare utilization. At 30 days, patients with early PT had lower mean LBP-related spending ($1180 ± $1500) compared with those without early PT ($1250 ± $2560) (P < 0.001). At 1 year, LBP-related spending was significantly less among patients who did not receive early PT ($2510 ± $3826) versus those who did ($2588 ± $3704). Early PT rates (range, 4–25%; P < 0.001) and 30-day LBP-related spending differed by state (range, $421 to −$410; P < 0.001).

Conclusion

Early PT for acute LBP was associated with less 30-day and 1-year healthcare utilization and less 30-day LBP-related spending. Early PT rates and 30-day spending differed by US state.

Level of evidence

IV
Evaluation of new and old biomarkers in dogs with degenerative mitral valve disease
Authors: Klein Stephanie , Nolte Ingo , Granados-Soler José Luis , Lietz Philipp , Sehn Maximiliane , Raue Jonathan Friedemann , Rohn Karl , Packeiser Eva-Maria , Bach Jan-Peter
Published: July 2022

Background

Dogs with degenerative mitral valve disease are commonly presented to small animal clinicians. Diagnosis, clinical staging, and therapeutic design are based on a combination of clinical examination, radiography, and echocardiography. To support diagnosis and clinical monitoring, a multi-marker-based approach would be conceivable. The aim of this study was to investigate the suitability of Galectin-3 and interleukin-1 receptor-like 1 protein (ST2) in dogs with degenerative mitral valve disease in accordance with N-terminal-prohormone-B-type natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI). For this purpose, serum concentrations of Galectin-3 and ST2 of 64 dogs with different stages of mitral valve disease and 21 dogs without cardiac disease were analyzed at the first examination and six months later. Echocardiography, blood cell count and clinical chemistry were performed and established biomarkers NT-proBNP and cTnI were measured additionally. Differences in the biomarker concentrations between all groups at both timepoints and the change in biomarker concentrations from first to second evaluation was investigated. Furthermore, correlations of each biomarker, between biomarkers and echocardiographic measurements, were calculated. Finally, the receiver-operating characteristic curve and the area under the curve analysis were performed to differentiate between disease stages and controls.

Results

Serum concentrations of Galectin-3 and ST2 were not statistically different between canine patients in the respective stages of mitral valve disease or in comparison to dogs in the control group at any timepoint. A significant increase in ST2 concentrations from the baseline to the follow-up examination was observed in dogs classified as stage B1 and the control group. The concentrations of NT-proBNP and cTnI in stage C dogs were significantly increased in comparison to the other groups.

Conclusions

In this study, no relation between Galectin-3 and ST2 levels to the presence or stage of mitral valve disease could be detected. Nevertheless, considering the increase in ST2 concentrations from the first to second measurement, its value on monitoring disease progress could be feasible. In agreement with previous studies, NT-proBNP and cTnI have once more proven their utility in assessing disease severity. The approach of examining new cardiac biomarkers in dogs is still worth pursuing.
APEX1 regulates alternative splicing of key tumorigenesis genes in non-small-cell lung cancer
Authors: Peng Li , Liu Yuwei , Chen Jing , Cheng Mengxin , Wu Ying , Chen Min , Zhong Ya , Shen Dan , Chen Ling , Ye Xujun
Published: July 2022

Background

Aberrant alternative splicing (AS) contributes to tumor progression. Previous studies have shown that apurinic-apyrimidinic endonuclease-1 (APEX1) is involved in tumor progression. It is unknown whether APEX1 functions in tumor progression by regulation of AS. It is also unknown whether APEX1 can regulate non-small-cell lung cancer (NSCLC) proliferation and apoptosis. We analyzed APEX1 expression levels in 517 lung NSCLC samples from the TCGA (Cancer Genome Atlas) database. The impact of APEX1 over expression on A549 cell proliferation and apoptosis was detected by the methyl thiazolyl tetrazolium assay and by flow cytometry. The transcriptome of A549 cells with and without APEX1 over expression was determined by Illumina sequencing, followed by analysis of AS. RT-qPCR validated expression of APEX1-related genes in A549 cells. We have successfully applied RNA-seq technology to demonstrate APEX1 regulation of AS.

Results

APEX1 expression was shown to be upregulated in NSCLC samples and to reduce cell proliferation and induce apoptosis of A549 cells. In addition, APEX1 regulated AS of key tumorigenesis genes involved in cancer proliferation and apoptosis within MAPK and Wnt signaling pathways. Each of these pathways are involved in lung cancer progression. Furthermore, validated AS events regulated by APEX1 were in key tumorigenesis genes; AXIN1 (axis inhibition protein 1), GCNT2 (N-acetyl glucosaminyl transferase 2), and SMAD3 (SMAD Family Member 3). These genes encode signaling pathway transcription regulatory factors.

Conclusions

We found that increased expression of APEX1 was an independent prognostic factor related to NSCLC progression. Therefore, APEX1 regulation of AS may serve as a molecular marker or therapeutic target for NSCLC treatment.
Association between lipid profiles and viral respiratory infections in human sputum samples
Authors: Humes Sara T. , Iovine Nicole , Prins Cindy , Garrett Timothy J. , Lednicky John A. , Coker Eric S. , Sabo-Attwood Tara
Published: July 2022

Background

Respiratory infections such as influenza account for significant global mortality each year. Generating lipid profiles is a novel and emerging research approach that may provide new insights regarding the development and progression of priority respiratory infections. We hypothesized that select clusters of lipids in human sputum would be associated with specific viral infections (Influenza (H1N1, H3N2) or Rhinovirus).

Methods

Lipid identification and semi-quantitation was determined with liquid chromatography and high-resolution mass spectrometry in induced sputum from individuals with confirmed respiratory infections (influenza (H1N1, H3N2) or rhinovirus). Clusters of lipid species and associations between lipid profiles and the type of respiratory viral agent was determined using Bayesian profile regression and multinomial logistic regression.

Results

More than 600 lipid compounds were identified across the sputum samples with the most abundant lipid classes identified as triglycerides (TG), phosphatidylethanolamines (PE), phosphatidylcholines (PC), Sphingomyelins (SM), ether-PC, and ether-PE. A total of 12 lipid species were significantly different when stratified by infection type and included acylcarnitine (AcCar) (10:1, 16:1, 18:2), diacylglycerols (DG) (16:0_18:0, 18:0_18:0), Lysophosphatidylcholine (LPC) (12:0, 20:5), PE (18:0_18:0), and TG (14:1_16:0_18:2, 15:0_17:0_19:0, 16:0_17:0_18:0, 19:0_19:0_19:0). Cluster analysis yielded three clusters of lipid profiles that were driven by just 10 lipid species (TGs and DGs). Cluster 1 had the highest levels of each lipid species and the highest prevalence of influenza A H3 infection (56%, n = 5) whereas cluster 3 had lower levels of each lipid species and the highest prevalence of rhinovirus (60%; n = 6). Using cluster 3 as the reference group, the crude odds of influenza A H3 infection compared to rhinovirus in cluster 1 was significantly (p = 0.047) higher (OR = 15.00 [95% CI: 1.03, 218.29]). After adjustment for confounders (smoking status and pulmonary comorbidities), the odds ratio (OR) became only marginally significant (p = 0.099), but the magnitude of the effect estimate was similar (OR = 16.00 [0.59, 433.03]).

Conclusions

In this study, human sputum lipid profiles were shown to be associated with distinct types of viral infection. Better understanding the relationship between respiratory infections of global importance and lipids contributes to advancing knowledge of pathogenesis of infections including identifying populations with increased susceptibility and developing effective therapeutics and biomarkers of health status.
Sequence, structure, and function of the Dps DNA-binding protein from Deinococcus wulumuqiensis R12
Authors: Chen Yao , Yang Zhihan , Zhou Xue , Jin Mengmeng , Dai Zijie , Ming Dengming , Zhang Zhidong , Zhu Liying , Jiang Ling
Published: July 2022

Deinococcus wulumuqiensisR12, which was isolated from arid irradiated soil in Xinjiang province of China, belongs to a genus that is well-known for its extreme resistance to ionizing radiation and oxidative stress. The DNA-binding protein Dps has been studied for its great contribution to oxidative resistance. To explore the role of Dps inD. wulumuqiensisR12, the Dps sequence and homology-modeled structure were analyzed. In addition, thedpsgene was knocked out and proteomics was used to verify the functions of Dps inD. wulumuqiensisR12. Docking data and DNA binding experiments in vitro showed that the R12 Dps protein has a better DNA binding ability than the Dps1 protein fromD. radioduransR1. When thedpsgene was deleted inD. wulumuqiensisR12, its resistance to H2O2and UV rays was greatly reduced, and the cell envelope was destroyed by H2O2treatment. Additionally, the qRT-PCR and proteomics data suggested that when thedpsgene was deleted, the catalase gene was significantly down-regulated. The proteomics data indicated that the metabolism, transport and oxidation–reduction processes ofD. wulumuqiensisR12 were down-regulated after the deletion of thedpsgene. Overall, the data conformed that Dps protein plays an important role inD. wulumuqiensisR12.
Flagella disruption in Bacillus subtilis increases amylase production yield
Authors: Fehler Annaleigh Ohrt , Kallehauge Thomas Beuchert , Geissler Adrian Sven , González-Tortuero Enrique , Seemann Stefan Ernst , Gorodkin Jan , Vinther Jeppe
Published: July 2022

Background

Bacillus subtilis is a Gram-positive bacterium used as a cell factory for protein production. Over the last decades, the continued optimization of production strains has increased yields of enzymes, such as amylases, and made commercial applications feasible. However, current yields are still significantly lower than the theoretically possible yield based on the available carbon sources. In its natural environment, B. subtilis can respond to unfavorable growth conditions by differentiating into motile cells that use flagella to swim towards available nutrients.

Results

In this study, we analyze existing transcriptome data from a B. subtilis α-amylase production strain at different time points during a 5-day fermentation. We observe that genes of the fla/che operon, essential for flagella assembly and motility, are differentially expressed over time. To investigate whether expression of the flagella operon affects yield, we performed CRISPR-dCas9 based knockdown of the fla/che operon with sgRNA target against the genes flgE, fliR, and flhG, respectively. The knockdown resulted in inhibition of mobility and a striking 2–threefold increase in α-amylase production yield. Moreover, replacing flgE (required for flagella hook assembly) with an erythromycin resistance gene followed by a transcription terminator increased α-amylase yield by about 30%. Transcript levels of the α-amylase were unaltered in the CRISPR-dCas9 knockdowns as well as the flgE deletion strain, but all manipulations disrupted the ability of cells to swim on agar.

Conclusions

We demonstrate that the disruption of flagella in a B. subtilis α-amylase production strain, either by CRISPR-dCas9-based knockdown of the operon or by replacing flgE with an erythromycin resistance gene followed by a transcription terminator, increases the production of α-amylase in small-scale fermentation.
Impact of COVID-19 among people who use drugs: A qualitative study with harm reduction workers and people who use drugs
Authors: Conway Fiona N. , Samora Jake , Brinkley Katlyn , Jeong Haelim , Clinton Nina , Claborn Kasey R.
Published: July 2022

Background

Fatal drug overdoses in the USA hit historical records during the COVID-19 pandemic. Throughout the pandemic, people who used drugs had greater odds of contracting COVID-19, increased drug use due to COVID-related stress, and heightened levels of anxiety and depression. This qualitative study examined the specific ways the pandemic negatively impacted people who use drugs.

Methods

Qualitative interviews with 24 people who use drugs and 20 substance use harm reduction workers were conducted. Data from the qualitative interviews were analyzed using applied thematic analysis to identify emergent themes based on the a priori research goals.

Results

Thematic analysis identified several common experiences during the pandemic among people who use drugs. These included mental distress due to financial strain and social isolation; increased drug use; increased risky drug-seeking and use behaviors due to changes in the drug markets; and reduced access to harm reduction, treatment, and recovery support services.

Conclusions

Our study highlighted critical systemic failures that contributed to the rise in overdose deaths during the COVID-19 pandemic. Addressing these challenges through policy reform and improved funding models will ensure the sustainability of harm reduction services and increase access to substance use treatment among highly vulnerable people who use drugs.
Spermidine reduces neuroinflammation and soluble amyloid beta in an Alzheimer’s disease mouse model
Authors: Freitag Kiara , Sterczyk Nele , Wendlinger Sarah , Obermayer Benedikt , Schulz Julia , Farztdinov Vadim , Mülleder Michael , Ralser Markus , Houtman Judith , Fleck Lara , Braeuning Caroline , Sansevrino Roberto , Hoffmann Christian , Milovanovic Dragomir , Sigrist Stephan J. , Conrad Thomas , Beule Dieter , Heppner Frank L. , Jendrach Marina
Published: July 2022

Background

Deposition of amyloid beta (Aβ) and hyperphosphorylated tau along with glial cell-mediated neuroinflammation are prominent pathogenic hallmarks of Alzheimer’s disease (AD). In recent years, impairment of autophagy has been identified as another important feature contributing to AD progression. Therefore, the potential of the autophagy activator spermidine, a small body-endogenous polyamine often used as dietary supplement, was assessed on Aβ pathology and glial cell-mediated neuroinflammation.

Results

Oral treatment of the amyloid prone AD-like APPPS1 mice with spermidine reduced neurotoxic soluble Aβ and decreased AD-associated neuroinflammation. Mechanistically, single nuclei sequencing revealed AD-associated microglia to be the main target of spermidine. This microglia population was characterized by increased AXL levels and expression of genes implicated in cell migration and phagocytosis. A subsequent proteome analysis of isolated microglia confirmed the anti-inflammatory and cytoskeletal effects of spermidine in APPPS1 mice. In primary microglia and astrocytes, spermidine-induced autophagy subsequently affected TLR3- and TLR4-mediated inflammatory processes, phagocytosis of Aβ and motility. Interestingly, spermidine regulated the neuroinflammatory response of microglia beyond transcriptional control by interfering with the assembly of the inflammasome.

Conclusions

Our data highlight that the autophagy activator spermidine holds the potential to enhance Aβ degradation and to counteract glia-mediated neuroinflammation in AD pathology.
Protocol: rhPCR and SNaPshot assays to distinguish Plasmodiophora brassicae pathotype clusters
Authors: Tso Heather H. , Galindo-González Leonardo , Locke Troy , Strelkov Stephen E.
Published: July 2022

Background

Clubroot of canola (Brassica napus), caused by the soilborne pathogen Plasmodiophora brassicae, has become a serious threat to canola production in Canada. The deployment of clubroot-resistant (CR) cultivars is the most commonly used management strategy; however, the widespread cultivation of CR canola has resulted in the emergence of new pathotypes of P. brassicae capable of overcoming resistance. Several host differential sets have been reported for pathotype identification, but such testing is time-consuming, labor-intensive, and based on phenotypic classifications. The development of rapid and objective methods that allow for efficient, cost-effective and convenient pathotyping would enable testing of a much larger number of samples in shorter times. The aim of this study was to develop two pathotyping assays, an RNase H2-dependent PCR (rhPCR) assay and a SNaPshot assay, which could quickly differentiate P. brassicae pathotypes.

Results

Both assays clearly distinguished between pathotype clusters in a collection of 38 single-spore isolates of P. brassicae. Additional isolates pathotyped from clubbed roots and samples from blind testing also were correctly clustered. The rhPCR assay generated clearly differentiating electrophoretic bands without non-specific amplification. The SNaPshot assay was able to detect down to a 10% relative allelic proportion in a 10:90 template mixture with both single-spore isolates and field isolates when evaluated in a relative abundance test.

Conclusions

This study describes the development of two rapid and sensitive technologies for P. brassicae pathotyping. The high-throughput potential and accuracy of both assays makes them promising as SNP-based pathotype identification tools for clubroot diagnostics. rhPCR is a highly sensitive approach that can be optimized into a quantitative assay, while the main advantages of SNaPshot are its ability to multiplex samples and alleles in a single reaction and the detection of up to four allelic variants per target site.
Delayed laparoscopic peritoneal washout in non-operative management of blunt abdominal trauma: a scoping review
Authors: Chu Megan , How Nathan , Laviolette Alysha , Bilic Monika , Tang Jennifer , Khalid Maham , Bos Cecily , Rice Timothy J. , Engels Paul T.
Published: July 2022

Objectives

Non-operative management (NOM) of blunt abdominal trauma has become increasingly common in hemodynamically stable patients. There are known complications of NOM from undrained intra-abdominal fluid accumulations including hemorrhage and peritonitis that require delayed operation. Thus, delayed operation can be considered as part of the overall management plan, instead of failure, of NOM. The aim of this scoping review is to establish key concepts regarding delayed laparoscopic peritoneal washout (DLPW) following NOM of blunt abdominal trauma patients.

Methods

MEDLINE, EMBASE, CENTRAL, and gray literature were systematically searched. Studies were included if they investigated or reported on the use of delayed laparoscopy involving peritoneal washout following NOM of blunt abdominal trauma patients. Bibliographies of included studies were manually reviewed to identify additional articles for inclusion.

Results

From 910 citations, 28 studies met inclusion criteria. This included seven case reports, eleven case series or observational cohort studies, six review articles, two management guidelines, one textbook chapter, and one randomized clinical trial. For those reported, medium grade liver injuries proved most common (95.2%). Indications for DLPW were primarily clinical features and changes in imaging findings, highlighting the importance of close observation. Authors reported clinical improvement after DLPW regarding symptomatology, vital signs, and biochemistry. A relatively high transfusion demand was reported with a mean of four units of packed red blood cells pre-operatively. Length of stay and post-operative complications were consistent with previously reported experiences with blunt abdominal injuries.

Conclusions

DLPW is beneficial in blunt abdominal trauma patients following NOM with improvement in symptoms, SIRS features, and a possible reduction in hospital length of stay. This study is limited by low-quality evidence and skewing of data toward isolated hepatic injuries. Future prospective cohort study comparing NOM with and without DLPW is required.
Response to PEEP in COVID-19 ARDS patients with and without extracorporeal membrane oxygenation. A multicenter case–control computed tomography study
Authors: Richard Jean-Christophe , Sigaud Florian , Gaillet Maxime , Orkisz Maciej , Bayat Sam , Roux Emmanuel , Ahaouari Touria , Davila Eduardo , Boussel Loic , Ferretti Gilbert , Yonis Hodane , Mezidi Mehdi , Danjou William , Bazzani Alwin , Dhelft Francois , Folliet Laure , Girard Mehdi , Pozzi Matteo , Terzi Nicolas , Bitker Laurent
Published: July 2022

Background

PEEP selection in severe COVID-19 patients under extracorporeal membrane oxygenation (ECMO) is challenging as no study has assessed the alveolar recruitability in this setting. The aim of the study was to compare lung recruitability and the impact of PEEP on lung aeration in moderate and severe ARDS patients with or without ECMO, using computed tomography (CT).

Methods

We conducted a two-center prospective observational case–control study in adult COVID-19-related patients who had an indication for CT within 72 h of ARDS onset in non-ECMO patients or within 72  h after ECMO onset. Ninety-nine patients were included, of whom 24 had severe ARDS under ECMO, 59 severe ARDS without ECMO and 16 moderate ARDS.

Results

Non-inflated lung at PEEP 5 cmH2O was significantly greater in ECMO than in non-ECMO patients. Recruitment induced by increasing PEEP from 5 to 15 cmH2O was not significantly different between ECMO and non-ECMO patients, while PEEP-induced hyperinflation was significantly lower in the ECMO group and virtually nonexistent. The median [IQR] fraction of recruitable lung mass between PEEP 5 and 15 cmH2O was 6 [4–10]%. Total superimposed pressure at PEEP 5 cmH2O was significantly higher in ECMO patients and amounted to 12 [11–13] cmH2O. The hyperinflation-to-recruitment ratio (i.e., a trade-off index of the adverse effects and benefits of PEEP) was significantly lower in ECMO patients and was lower than one in 23 (96%) ECMO patients, 41 (69%) severe non-ECMO patients and 8 (50%) moderate ARDS patients. Compliance of the aerated lung at PEEP 5 cmH2O corrected for PEEP-induced recruitment (CBABY LUNG) was significantly lower in ECMO patients than in non-ECMO patients and was linearly related to the logarithm of the hyperinflation-to-recruitment ratio.

Conclusions

Lung recruitability of COVID-19 pneumonia is not significantly different between ECMO and non-ECMO patients, with substantial interindividual variations. The balance between hyperinflation and recruitment induced by PEEP increase from 5 to 15 cmH2O appears favorable in virtually all ECMO patients, while this PEEP level is required to counteract compressive forces leading to lung collapse. CBABY LUNGis significantly lower in ECMO patients, independently of lung recruitability.
Proteomic response of Turicibacter bilis MMM721 to chicken bile and its bile acids
Authors: Maki Joel J. , Lippolis John D. , Looft Torey
Published: July 2022

Objective

Bile and its individual components, mainly bile acids, are important for digestion and drive bacterial community dynamics in the upper gastrointestinal tract of chickens. However, specific responses to bile acids have been characterized in only a few commensal bacteria, and it is unclear how other members of the microbiota respond to biliary stress. Here, we used label-free LC–MS/MS to assess the proteomic response of a common inhabitant of the chicken small intestine,Turicibacter bilisMMM721, to 24 h of growth in anaerobic growth media supplemented with 0.1% whole chicken bile, 0.1% taurochenodeoxycholic acid (TCDCA), or 0.1% taurocholic acid (TCA).

Results

Seventy, 46, and 10 differentially expressed proteins were identified inTuricibacter bilisMMM721 cultured with supplements of chicken bile, TCDCA, and TCA, respectively, when compared to unsupplemented controls. Many differentially expressed proteins were predicted to be involved in ribosomal processes, post-translational modifications and chaperones, and modifications to the cell surface. Ultimately, theT. bilisMMM721 response to whole bile and bile acids is complex and may relate to adaptations for small intestine colonization, with numerous proteins from a variety of functional categories being impacted.
Coexistence of cutaneous endometriosis and ovarian endometrioma: a case report
Authors: Mohaghegh Fatemeh , Hatami Parvaneh , Rajabi Parvin , Aryanian Zeinab
Published: July 2022

Background

Umbilical endometriosis is a rare entity accounting for 0.5–4% of cases with endometriosis.

Case presentation

Here we report a rare case of umbilical endometriosis with concurrent ovarian endometriomas in a 37-year old primiparous Iranian woman.

Conclusion

This interesting coexistence reflects the importance of thorough gynecological assessment in patients with cutaneous endometriosis to enable appropriate management.
High-throughput muscle fiber typing from RNA sequencing data
Authors: Oskolkov Nikolay , Santel Malgorzata , Parikh Hemang M. , Ekström Ola , Camp Gray J. , Miyamoto-Mikami Eri , Ström Kristoffer , Mir Bilal Ahmad , Kryvokhyzha Dmytro , Lehtovirta Mikko , Kobayashi Hiroyuki , Kakigi Ryo , Naito Hisashi , Eriksson Karl-Fredrik , Nystedt Björn , Fuku Noriyuki , Treutlein Barbara , Pääbo Svante , Hansson Ola
Published: July 2022

Background

Skeletal muscle fiber type distribution has implications for human health, muscle function, and performance. This knowledge has been gathered using labor-intensive and costly methodology that limited these studies. Here, we present a method based on muscle tissue RNA sequencing data (totRNAseq) to estimate the distribution of skeletal muscle fiber types from frozen human samples, allowing for a larger number of individuals to be tested.

Methods

By using single-nuclei RNA sequencing (snRNAseq) data as a reference, cluster expression signatures were produced by averaging gene expression of cluster gene markers and then applying these to totRNAseq data and inferring muscle fiber nuclei type via linear matrix decomposition. This estimate was then compared with fiber type distribution measured by ATPase staining or myosin heavy chain protein isoform distribution of 62 muscle samples in two independent cohorts (n = 39 and 22).

Results

The correlation between the sequencing-based method and the other two were rATPas = 0.44 [0.13–0.67], [95% CI], and rmyosin = 0.83 [0.61–0.93], with p = 5.70 × 10–3 and 2.00 × 10–6, respectively. The deconvolution inference of fiber type composition was accurate even for very low totRNAseq sequencing depths, i.e., down to an average of ~ 10,000 paired-end reads.

Conclusions

This new method (https://github.com/OlaHanssonLab/PredictFiberType) consequently allows for measurement of fiber type distribution of a larger number of samples using totRNAseq in a cost and labor-efficient way. It is now feasible to study the association between fiber type distribution and e.g. health outcomes in large well-powered studies.
Ten actions to achieve gender equity among intensivists: the French Society of Intensive Care (FICS) model
Authors: Hamzaoui Olfa , Boissier Florence , Salmon Gandonnière Charlotte , Aubron Cécile , Bodet-Contentin Laetitia , Fartoukh Muriel Sarah , Faure Mélanie , Jourdain Mercedes , Le Marec Julien , Tamion Fabienne , Terzi Nicolas , Hauw-Berlemont Caroline , Aissaoui Nadia ,   FEMMIR Group for the SRLF Trial Group
Published: July 2022

In our recent survey, we aimed to collect information on perceived inequity as well as professional and personal fulfillment among women intensivists in France. For the 371 respondents out of the 732 persons who received the survey, the findings were unequivocal: for one-third of the respondents, being a woman was considered as an obstacle to careers or academic advancement, and for two thirds, pregnancy was viewed as a barrier to their career advancement. Gender discrimination had been experienced by 55% of the respondents. In 2019, to promote and achieve gender equity in the French Intensive Care Society (FICS), ten actions were initiated and are detailed in the present manuscript together with supporting data: (1) creation of a working group: the FEMMIR group; (2) promotion of mentorship; (3) implementation of concrete sponsorship; (4) transparency and public reporting of gender ratios in editorial boards; (5) workshops dedicated to unconscious gender bias; (6) workshops dedicated to improved women assertiveness; (7) role models; (8) creation of educational/information programs for young intensivists; (9) development of research on gender inequity and, as a perspective; and (10) development of a wide-ranging program. This review is aimed at providing a toolbox of organizational best practices designed to achieve gender equity. It is particularly important to share promising practical action engaged in our FEMMIR group with other concerned professionals around the world.
CDK6 is upregulated and may be a potential therapeutic target in enzalutamide-resistant castration-resistant prostate cancer
Authors: Chen Xi , Wu Yechen , Wang Xinan , Xu Chengdang , Wang Licheng , Jian Jingang , Wu Denglong , Wu Gang
Published: July 2022

Background

Androgen deprivation therapy (ADT) is still the first-line treatment of prostate cancer (PCa). However, after a certain period of therapy, primary PCa inevitably progresses into castration-resistant PCa (CRPC). Enzalutamide (Enz) is an androgen receptor (AR) signal inhibitor which can delay the progression of CRPC and increase survival of patients with metastatic CRPC. However, the mechanisms involved in enzalutamide-resistant (EnzR) CRPC are still controversial. In the study, we used bioinformatic methods to find potential genes that correlated with the occurrence of EnzR CRPC.

Methods

We collected RNA sequencing data of the EnzR CRPC cell line LNCaP (EnzR LNCaP) from GSE44905, GSE78201, and GSE150807. We found the hub genes from the three datasets. Then we tested the expression of the hub genes in different databases and the potential drugs that can affect the hub genes. Finally, we verified the hub gene expression and drug function.

Results

From GSE44905, GSE78201 and GSE150807, we found 45 differentially expressed genes (DEGs) between LNCaP and EnzR LNCaP. Ten hub genes were found in the protein–protein interaction (PPI) network. The expression of hub gene and survival analysis were analyzed by different databases. We found that cyclin-dependent kinase 6 (CDK6) was highly expressed in both the EnzR LNCaP cell and PCa patients. Ten potential small molecules could suppress CDK6 expression as per “CLUE COMMAND” findings. Finally, we found the expression of CDK6 increased in both PCa patients’ samples, CRPC and EnzR PCa cell lines. Three potential CDK6 inhibitors, namely apigenin, chrysin and fisetin, can decrease cell proliferation.

Conclusions

The study proved that the abnormal overexpression of CDK6 may be a reason behind EnzR CRPC occurrence and suppression CDK6 expression may help treat EnzR CRPC.
A phase Ib dose-escalation study of troriluzole (BHV-4157), an oral glutamatergic signaling modulator, in combination with nivolumab in patients with advanced solid tumors
Authors: Silk Ann W. , Saraiya Biren , Groisberg Roman , Chan Nancy , Spencer Kristen , Girda Eugenia , Shih Weichung , Palmeri Marisa , Saunders Tracie , Berman Robert M. , Coric Vlad , Chen Suzie , Zloza Andrew , Vieth Joshua , Mehnert Janice M. , Malhotra Jyoti
Published: July 2022

Background

Glutamate signaling activates MAPK and PI3K/AKT pathways in tumor cells. Treatment with riluzole, a glutamate release inhibitor, has been previously shown to be safe in melanoma patients and produced biologic effects, but did not lead to radiographic responses, possibly due to poor pharmacokinetic properties. Therefore, we conducted a phase Ib trial to determine the safety and tolerability of the combination of the riluzole prodrug troriluzole (BHV-4157, trigriluzole) and the PD-1 antibody nivolumab in patients with advanced solid tumors.

Methods

Patients with advanced or refractory solid tumors and measurable disease per RECIST 1.1 were treated with increasing doses of troriluzole using a semi-Bayesian modified toxicity probability interval dose escalation procedure. Troriluzole monotherapy was orally self-administered for a 14-day lead-in period followed by continuation of troriluzole in combination with nivolumab 240 mg IV every 2 weeks. Endpoints included safety, pharmacokinetics (PK) and efficacy.

Results

We enrolled 14 patients with advanced solid tumors (melanoma = 3, NSCLC = 3, renal cell carcinoma = 2, bladder/urothelial = 2, ovarian cancer = 1, adenoid cystic carcinoma = 1, pleural mesothelial = 1, head and neck cancer = 1). Eleven patients had cancer progression on prior therapy with PD-1 or PD-L1 agent. Patients received troriluzole total daily doses from 140 to 560 mg (divided). The most common treatment-related adverse events (TRAE) occurring in ≥ 5 patients (> 35%) were transaminitis and increased lipase. DLT (dose-limiting toxicity) occurred in 3 patients: (1) grade 3 anorexia, (2) grade 3 fatigue and, (3) grade 3 atrial fibrillation. Six patients were treated at the MTD (maximum tolerated dose). No subjects discontinued treatment due to AEs. One response occurred (7%), which was a partial response in a subject who had PD-1 refractory disease. The 6-month PFS rate was 21%. PK data showed that the prodrug troriluzole was efficiently cleaved into riluzole by 2-h post-dosing in all dose cohorts tested.

Conclusion

The combination of troriluzole and nivolumab was safe and well-tolerated. The MTD of troriluzole was determined to be 420 mg total daily dose. The observed antitumor activity, primarily disease stabilization, is of interest in patients with PD-1 resistant tumors.

Trial Registration ClinicalTrials.gov Identifier NCT03229278.
Evaluation of l-arginine supplement on the growth rate, biofilm formation, and antibiotic susceptibility in Streptococcus mutans
Authors: Vaziriamjad Samaneh , Solgi Mobina , Kamarehei Farideh , Nouri Fatemeh , Taheri Mohammad
Published: July 2022

Introduction

Bacteria associated with dental caries have a high ability to produce organic acids from dietary carbohydrates during growth and metabolism under acidic conditions. In contrast, many symbiotic bacteria produce ammonia through the arginine deiminase (ADS) system, which modulates the pH of the oral cavity. l-Arginine metabolism by ADS is a significant inhibitor in the progression of tooth decay. This study aimed to investigate the effect of l-arginine on growth, biofilm formation, and antibiotic susceptibility in Streptococcus mutans.

Methods

In this study, the effect of l-arginine in different concentrations on the growth rate, antibiotic susceptibility, and inhibition of biofilm formation in S. mutans was investigated.

Results

The bacterial exponential growth rate was enhanced by 100 μM l-arginine (P > 0.05). The growth inhibition zone diameter of CAZ, CTR, AMP, and AMC-Clav antibiotics was reduced after 24 h of exposure in the presence of various concentrations of l-arginine specifically at 100 μM. l-Arginine also enhanced biofilm development at 5 and 10 μM concentrations, but reduced it at 50 and 100 μM concentrations.

Conclusion

According to the results of the present study, optimization of l-arginine concentration and its use as an adjunctive therapy or in combination with mouthwash or varnish is recommended to prevent oral caries.