The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi.

Carbohydrate active enzymes (CAZymes) are vital for the lignocellulose-based biorefinery. The development of hypersecreting fungal protein production hosts is therefore a major aim for both academia and industry. However, despite advances in our understanding of their regulation, the number of promising candidate genes for targeted strain engineering remains limited. Here, we resequenced the genome of the classical hypersecreting Neurospora crassa mutant exo-1 and identified the causative point of mutation to reside in the F-box protein-encoding gene, NCU09899. The corresponding deletion strain displayed amylase and invertase activities exceeding those of the carbon catabolite derepressed strain Δcre-1, while glucose repression was still mostly functional in Δexo-1 Surprisingly, RNA sequencing revealed that while plant cell wall degradation genes are broadly misexpressed in Δexo-1, only a small fraction of CAZyme genes and sugar transporters are up-regulated, indicating that EXO-1 affects specific regulatory factors. Aiming to elucidate the underlying mechanism of enzyme hypersecretion, we found the high secretion of amylases and invertase in Δexo-1 to be completely dependent on the transcriptional regulator COL-26. Furthermore, misregulation of COL-26, CRE-1, and cellular carbon and nitrogen metabolism was confirmed by proteomics. Finally, we successfully transferred the hypersecretion trait of the exo-1 disruption by reverse engineering into the industrially deployed fungus Myceliophthora thermophila using CRISPR-Cas9. Our identification of an important F-box protein demonstrates the strength of classical mutants combined with next-generation sequencing to uncover unanticipated candidates for engineering. These data contribute to a more complete understanding of CAZyme regulation and will facilitate targeted engineering of hypersecretion in further organisms of interest.

Child abuse and neglect: A comparative study between dental and medical students' competencies in Germany.

INTRODUCTION: Despite children's right to a non-violent upbringing, they become daily victims of violence. Physical violence concerns mostly the head-especially the orofacial area. Therefore, dentists and paediatricians should be able to suspect possible abuse and to initiate child protection. This study aimed to record and compare the training situation and competencies of dental and medical students at Jena University Hospital regarding child abuse and neglect (CAN). MATERIALS AND METHODS: Using a three-part questionnaire about CAN, 123 medical and 77 dental students were surveyed anonymously after completing all courses on the topic. The question sets included as follows: (1) educational experiences, training content and satisfaction; (2) current knowledge regarding diagnostic, management and legal concerns; (3) self-evaluation, needs of further education and wishes. Reliability of the questionnaire was tested with kappa-statistics and assessed as good. RESULTS: Dental and medical students' overall satisfaction with CAN-related training is low. Although medical students had more knowledge on CAN, all participants showed large deficits. Better diagnostic than management skills were reported in both groups. Only 1.3% dental and 16.3% medical students felt adequately prepared to deal with CAN. 7% of all study participants stated that they can report CAN without any help. 87.0% of medical and 79.2% of dental students expressed a desire for further education. To improve their knowledge, both courses prefer seminars, followed by expert talks, lectures and simulation-based training (SkillsLab). CONCLUSION: Dental and medical students are inadequately prepared to suspect possible abuse and to deal with possible signs of CAN. Mandatory interdisciplinary courses and lectures addressing CAN are recommendable for both medical and dental curricula.

Converting to reverse shoulder arthroplasty from primary anatomic shoulder arthroplasty and fracture hemiarthroplasty: a radiographic and clinical outcome analysis at 8-years.

INTRODUCTION: Revision shoulder arthroplasty can be challenging. One of the main considerations for surgeons is the type of implant that was placed in the initial surgery. Anatomic shoulder arthroplasty (ASA) is used for cases of osteoarthritis as well as for fractures of the humeral head. Hemiarthroplasty can be used for complex proximal humerus fractures. The purpose of this study is to determine whether there is a difference in clinical and radiographic outcomes between patients that failed primary fracture hemiarthroplasty (FHA), or ASA for osteoarthritis and then required reoperation with a conversion to reverse shoulder arthroplasty (RSA). METHODS: Patients with failed anatomic shoulder replacement, who had undergone conversion to RSA, were enrolled after a mean follow-up of 107 (85-157) months. Two different groups, one with failed ASA implanted for osteoarthritis and one with failed FHA, were created. At follow-up patients were assessed with standard radiographs and clinical outcome scores. RESULTS: Twenty-nine patients (f = 17, m = 12; 51%) suffered from a failed ASA (Group A), while the remaining 28 patients (f = 21, m = 74; 49%) had been revised due to a failed FHA (Group B). Patients of Group B had a poorer Constant score (Group A: 60 vs. Group B: 46; p = 0.02). Abduction (Group A: 115° vs. Group B: 89°; p = 0.02) was worse after conversion of a failed FHA to RSA in comparison to conversions of failed ASA. The mean bone loss of the lateral metaphysis was higher in patients with failed FHA (Group A: 5 mm vs. Group B: 20 mm; p = 0.0). CONCLUSION: The initial indication for anatomic shoulder arthroplasty influences the clinical and radiological outcome after conversion to RSA. Conversion of failed FHA to RSA is related to an increased metaphyseal bone loss, decreased range of motion and poorer clinical outcomes when compared to conversions of failed ASA implanted for osteoarthritis. LEVEL OF EVIDENCE: III Retrospective Cohort Comparison Study.

ROAR ED-SANE: A Retrospective Observational Assessment Review of an Emergency Department Sexual Assault Nurse Examiner Program's Adherence to Centers for Disease Control and Prevention Guidelines for Sexually Transmitted Infections.

BACKGROUND: Sexual assault survivors are at increased risk for sexually transmitted infections. Sexual Assault Nurse Examiner programs guide sexually transmitted infection treatment, monitoring, and follow-up scheduling according to guidelines by the Centers for Disease Control and Prevention (CDC). Reported low rates of provider adherence to CDC treatment guidelines and patient adherence to follow-up necessitate a review of medication prescribing and follow-up scheduling practices, especially at smaller community hospitals in the United States. METHODS: A retrospective medical record review was conducted to assess adherence rates to CDC guidelines for prescribing practices, scheduling, and follow-up of sexual assault survivors. We included pediatric and adult patients presenting to the emergency department (ED) and participating in the ED Sexual Assault Nurse Examiner program at a rural, community-based teaching hospital in La Crosse, WI, from January 2018 to December 2021. Descriptive statistics were used to evaluate results. RESULTS: Analysis included 103 patients. Prescribing adherence to CDC guidelines was >80% for all except human immunodeficiency virus (53.4%), trichomoniasis (68.1%), and hepatitis B (69%). Of the 38 patients who had a follow-up scheduled during their ED encounter, 78.9% attended their scheduled follow-up and 94.7% of those appointments were scheduled within the CDC-recommended time frame, leading to an overall adherence of 40%. CONCLUSIONS: Adherence rates were high for most prescribing practices, and attendance of scheduled follow-up was higher than expected. Opportunities to improved adherence to CDC guidelines were identified in prescribing for 3 disease states (human immunodeficiency virus, trichomoniasis, and hepatitis B) and in scheduling of follow-up.

Comparison of the Soluble fms-Like Tyrosine Kinase 1/Placental Growth Factor Ratio Alone versus a Multi-Marker Regression Model for the Prediction of Preeclampsia-Related Adverse Outcomes after 34 Weeks of Gestation.

INTRODUCTION: The objective of this retrospective study was to compare the predictive performance of the soluble fms-like tyrosine kinase 1 (sFlt-1)/placental growth factor (PlGF) ratio alone or in a multi-marker regression model for preeclampsia-related maternal and/or fetal adverse outcomes in women >34 weeks of gestation. METHODS: We analyzed the data collected from 655 women with suspected preeclampsia. Adverse outcomes were predicted by multivariable and univariable logistic regression models. The outcome of patients was evaluated within 14 days after presentation with signs and symptoms of preeclampsia or diagnosed preeclampsia. RESULTS: The full model integrating available, standard clinical information and the sFlt-1/PlGF ratio had the best predictive performance for adverse outcomes with an AUC of 72.6%, which corresponds to a sensitivity of 73.3% and specificity of 66.0%. The positive predictive value of the full model was 51.4%, and the negative predictive value was 83.5%. 24.5% of patients, who did not experience adverse outcomes but were classified as high risk by sFlt-1/PlGF ratio (≥38), were correctly classified by the regression model. The sFlt-1/PlGF ratio alone had a significantly lower AUC of 65.6%. CONCLUSIONS: Integrating angiogenic biomarkers in a regression model improved the prediction of preeclampsia-related adverse outcomes in women at risk after 34 weeks of gestation.

Improved Detection of Child Maltreatment with Routine Screening in a Tertiary Care Pediatric Hospital.

OBJECTIVES: To evaluate the impact of layering routine child abuse screening on top of a preexisting electronic health record-embedded Child Abuse Clinical Decision Support System (CA-CDSS) in a pediatric emergency department. STUDY DESIGN: The Pittsburgh Child Abuse Screening Tool (P-CAST) was performed in all children aged <13 years and in nonverbal children aged ≥13 years who presented to a pediatric tertiary care center over a 6-month period. The P-CAST was layered on top of a preexisting CA-CDSS that included passive triggers, alerts, and abuse-specific order sets. RESULTS: Of the 28 797 screens performed, 1.8% were positive in children aged <13 years and 1.6% were positive in nonverbal children aged ≥13 years. One-half of the children with a positive P-CAST also triggered the CA-CDSS; the other one-half triggered only because of the P-CAST. Nineteen percent of the patients with a positive P-CAST were reported to Child Protective Services (CPS). There was no relationship between race and the odds of a positive P-CAST or between race and the likelihood of a report being made to CPS. CONCLUSIONS: Active routine child abuse screening improves identification of suspected child maltreatment in a children's hospital above and beyond what is identified with a CA-CDSS, which depends on passive triggers. The lack of a relationship between race and a positive P-CAST or a report to CPS suggest that systematic child abuse screening may mitigate well-recognized racial disparities in identifying and reporting suspected child maltreatment.

Emission, Transport, and Deposition of visible Plastics in an Estuary and the Baltic Sea-a Monitoring and Modeling Approach.

AIM: was to assess whether a comprehensive approach linking existing knowledge with monitoring and modeling can provide an improved insight into coastal and marine plastics pollution. We focused on large micro- and mesoplastic (1-25 mm) and selected macroplastic items. Emission calculations, samplings in the Warnow river and estuary (water body and bottom sediments) and a flood accumulation zone monitoring served as basis for model simulations on transport and behavior in the entire Baltic Sea. Considered were the most important pathways, sewage overflow and stormwater. The coastline monitoring together with calculations allowed estimating plastics emissions for Rostock city and the Warnow catchment. Average concentrations at the Warnow river mouth were 0.016 particles/m³ and in the estuary 0.14 particles/m³ (300 µm net). The estuary and nearby Baltic Sea beaches were hot-spots for plastic accumulation with 6-31 particles/m². With increasing distance from the estuary, the concentrations dropped to 0.3 particles/m². This spatial pattern, the plastic pollution gradients and the observed annual accumulation values were consistent with the model results. Indicator items for sewer overflow and stormwater emissions exist, but were only found at low numbers in the environment. The considered visible plastics alone can hardly serve as indicator for microplastic pollution (<1 mm). The use of up-scaled emission data as input for Baltic Sea model simulations provided information on large scale emission, transport and deposition patterns of visible plastics. The results underline the importance of plastic retention in rivers and estuaries.

Y-Site Compatibility of Intravenous Levetiracetam With Commonly Used Critical Care Medications.

Purpose: Levetiracetam is an antiepileptic medication commonly used in critical care areas for seizure treatment or prophylaxis. Compatibility data of levetiracetam with other critical care medications are limited, which can make administration challenging. This study aims to assess the physical Y-site compatibility of intravenous levetiracetam with some other commonly used critical care medications. Methods: Y-site administration was simulated by independently mixing levetiracetam with each of 11 selected medications in a 4-dram, colorless, screw-cap, glass vial, at a 1:1 ratio. Clinically used concentrations of each medication were compounded in 0.9% sodium chloride following United States Pharmacopeia chapter 797 standards. Physical compatibility was observed and assessed at 0, 15, and 30 minutes after mixing. Medication mixtures were considered physically incompatible if there was visual evidence of color change, gas evolution, haze, or particulate formation, pH change >10%, or if they had an absorbance value >0.010 A. Results: No evidence of physical incompatibility was observed during simulated Y-site testing with cisatracurium 1 mg/mL, dexmedetomidine 4 µg/mL, fosphenytoin 15 mg PE/mL, norepinephrine 16 mg/mL, norepinephrine 32 mg/mL, norepinephrine 64 mg/mL, piperacillin-tazobactam 33.75 mg/mL, propofol 10 mg/mL, vancomycin 5 mg/mL, or vasopressin 1 unit/mL when tested in 0.9% sodium chloride. Levetiracetam was incompatible with piperacillin-tazobactam 45 mg/mL. Conclusion: Levetiracetam 5 mg/mL in 0.9% sodium chloride was found to be physically compatible for 30 minutes with 10 of the 11 medications tested during simulated Y-site administration.

Psychiatric and Psychosomatic Consultation-Liaison Services in General Hospitals: A Systematic Review and Meta-Analysis of Effects on Symptoms of Depression and Anxiety.

BACKGROUND: Psychiatric and psychosomatic consultation-liaison services (CL) are important providers of diagnosis and treatment for hospital patients with mental comorbidities and psychological burdens. OBJECTIVE: To perform a systematic review and meta-analysis investigating the effects of CL on depression and anxiety. METHODS: Following PRISMA guidelines, a systematic literature search was conducted until 2017. Included were published randomized controlled trials using CL interventions with adults in general hospitals, treatment as usual as control groups, and depression and/or anxiety as outcomes. Risk of bias was assessed using the Cochrane Risk of Bias Tool. Level of integration was assessed using the Standard Framework for Levels of Integrated Healthcare. Meta-analyses were performed using random effects models and meta-regression for moderator effects. RESULTS: We included 38 studies (9,994 patients). Risk of bias was high in 17, unclear in 15, and low in 6 studies. Studies were grouped by type of intervention: brief interventions tailored to the patients (8), interventions based on specific treatment manuals (19), and integrated, collaborative care (11). Studies showed small to medium effects on depression and anxiety. Meta-analyses for depression yielded a small effect (d = -0.19, 95% CI: -0.30 to -0.09) in manual studies and a small effect (d = -0.33, 95% CI: -0.53 to -0.13) in integrated, collaborative care studies, the latter using mostly active control groups with the possibility of traditional consultation. CONCLUSIONS: CL can provide a helpful first treatment for symptoms of depression and anxiety. Given that especially depressive symptoms in medically ill patients are long-lasting, the results underline the benefit of integrative approaches that respect the complexity of the illness.

The Mediated Amplification of a Crisis: Communicating the A/H1N1 Pandemic in Press Releases and Press Coverage in Europe.

In the aftermath of the A/H1N1 pandemic, health authorities were criticized for failures in crisis communication efforts, and the media were accused of amplifying the pandemic. Considering these criticisms, A/H1N1 provides a suitable case for examining risk amplification processes that may occur in the transfer of information from press releases to print news media during a health crisis. We integrated the social amplification of risk framework with theories of news decisions (news values, framing) in an attempt to contribute to existing research both theoretically and empirically. We conducted a quantitative content analysis of press releases disseminated by health and governmental authorities, as well as the quality and tabloid press in 10 European countries between March 2009 and March 2011. Altogether 243 press releases, 1,243 quality press articles, and 834 tabloid press articles were coded. Consistent with research on news values and framing the results suggest that quality and tabloid papers alike amplified A/H1N1 risks by emphasizing conflict and damage, presenting information in a more dramatized way, and using risk-amplifying frames to a greater extent and risk-attenuating frames to a lesser extent than press releases. To some extent, the quality and tabloid press differed in how risk information was presented. While tabloid press articles seemed to follow the leading quality press with regards to content and framing of health crisis coverage, they exhibited a stronger emphasis on drama and emotion in the way they presented information.

Inpatient and Day Hospital Treatment of Patients with Depression and Job-related Burnout.

OBJECTIVE: Burnout is a process of physical and emotional exhaustion that often results in clinical depression. Detailed descriptions and evaluations of specialized psychosomatic treatment are rare. This pilot study investigates the feasibility of inpatient and day hospital treatment of patients with burnout syndrome. Additionally, we present results of an initial, noncontrolled, pre-post-evaluation of changes in symptoms and individual work-related risk factors for burnout. METHODS: Sixty-four consecutive patients with burnout syndrome were assessed before and after specialized multimodal treatment using a clinical symptom checklist (ICD-10 Symptom Rating) and burnout-specific instruments (Maslach Burnout Inventory, Occupational Stress & Coping Inventory). RESULTS: Patients' average age was 45 (range 23 to 61), 70% were currently employed, 24% in managerial positions or self-employed, and 89% diagnosed with an affective disorder. The average length of time off work due to illness in the past year was 13 weeks. Treatment lasted five weeks on average. After treatment, depression (p < 0.001; effect size d = 0.79), emotional exhaustion (p = 0.001; d = 0.41), and the subjective significance of work (p = 0.001; d = 0.36) decreased, while emotional distancing (p < 0.001; d = 0.56), balance and mental stability (p < 0.001; d = 0.38) and life satisfaction (p < 0.001; d = 0.37) increased. Clinical significance was determined using the Reliable Change Index. CONCLUSIONS: The treatment program described here is well accepted by patients with severe burnout. It contributes to positive changes in symptoms and work-related risk factors. Controlled studies are necessary to establish treatment efficacy.

The sFlt-1/PlGF-ratio and the risk of preeclampsia-related adverse outcomes in subsequent pregnancies with signs and symptoms of a preeclampsia.

OBJECTIVES: This study characterizes the outcome of two subsequent pregnancies with suspected preeclampsia (PE). We investigated the diagnostic accuracy of clinical signs, Doppler examinations, and the soluble fms-like tyrosine kinase-1 (sFlt-1) to placental growth factor (PlGF)-ratio to predict PE-related adverse outcomes (AO). The sFlt-1/PlGF-ratio of the first pregnancy was compared to the outcome of the subsequent pregnancy. STUDY DESIGN: A total of 1928 patients at risk for preeclampsia were screened, of them 1117 were eligible for inclusion. Of these, 84 women presented with suspected PE in two subsequent pregnancies. OUTCOME MEASURES: Diagnostic accuracy of clinical markers was assessed. Associations between the sFlt-1/PlGF-ratio in the first and the odds of an AO in the subsequent pregnancy were investigated with logistic regression. RESULTS: The prevalence of AOs decreased from 27.4 % in the first to 17.9 % in the second pregnancy. Comparison of the accuracy of the different clinical markers for an AO showed a high specificity for an sFlt-1/PlGF-ratio at the cut-off of ≥ 85 in both pregnancies (81.3 %, 95 % CI 63.6-92.8 vs 92.6 %,95 % CI 83.7-97.6), but a lower sensitivity in the second pregnancy (92.9 %, 95 % CI 66.1-99.8 vs 33.3%, 95 % CI 11.8-61.6). An elevated sFlt-1/PlGF-ratio in the first did not increase the odds of an AO in the subsequent pregnancy. CONCLUSIONS: The prevalence of AOs decreases in subsequent pregnancies. Our finding that the sFlt-1/PlGF-ratio of the first was not related to the outcome of the subsequent pregnancy suggests that angiogenic markers are only a within-pregnancy short-term tool to assess AOs.

Implementation of a screening, brief intervention, and referral to treatment program using the electronic medical record in a pediatric trauma center.

The purpose of this project was to develop and implement a consistent process for (1) screening adolescents by history for alcohol and substance abuse and (2) providing a motivational interview for change and appropriate referrals as needed. In the 18 months since we implemented the program, 534 patients were eligible for screening. Of these, 442 actually underwent screening and of these, 32 screened positive, thus receiving a brief intervention by social work and referral for further treatment. Use of the electronic medical record was key to the implementation and sustainability of this project.

Effects of Messaging and Communication Strategy Training on Interaction in Teams With Deaf and Hearing College Students.

The study investigated effects of computer-based messaging and training in communication strategies on interactions of deaf and typically hearing (TH) teammates in completing decision-making tasks without interpreter support. Fifteen teams, two deaf and two TH college students each, completed three decision-making tasks, one without messaging, one with messaging, and one with messaging and training in communication strategies. Each interaction was coded for (a) communication method, (b) to whom the interaction was directed, and (c) the function of the interaction. Without messaging, teams used speech, sign, or paper and pencil; with messaging, they predominantly used this technology to communicate with each other. Without messaging, teammates directed communications to members of the same hearing status; with messaging, they directed communications to the whole team. Teammates made fewer communication repairs with messaging than without. In focus groups conducted after the decision-making tasks, participants noted messaging's limitations and benefits.

Correlation between placental weight and angiogenic markers sFlt-1 and PlGF in women with preeclampsia and fetal growth restriction.

OBJECTIVES: The angiogenic factors sFlt-1 (soluble fms-like tyrosine kinase) and PlGF (Placental Growth Factor) play a key role in the pathophysiology, prediction and diagnosis of preeclampsia-associated pregnancy disorders. However, the correlation between maternal serum levels and the placental weight, especially in hypertensive pregnancy disorders is still unclear. STUDY DESIGN: Retrospectively, we analyzed data from a real-world cohort of patients with preeclampsia (PE), intrauterine growth restriction (IUGR), PE + IUGR and controls giving birth within 14 days from inclusion. Herein, correlational analyses were calculated between placental weight, maternal serum levels of sFlt-1, PlGF and the respective sFlt-1/PlGF-ratios. MAIN OUTCOME MEASURES AND RESULTS: This study included n = 328 patients (n = 134 with PE, n = 40 with IUGR and n = 25 showed PE + IUGR) and n = 129 controls. The gestational age-adjusted placental weight was significantly decreased in patients with PE ± IUGR, but not in PE alone, when comparing to controls. Correlation between PlGF and the placental weight was significantly positive and increasing with severity of disease (controls 0.134, p = 0.131, PE 0.419, p < 0.01, IUGR 0.517, p < 0.01, PE + IUGR r = 0.723, p < 0.01). Furthermore, an inverse correlation between the sFlt-1/PlGF-ratio and the placental weight was found. The sFlt-1/PlGF-ratio per gram placental weight was highest in patients with PE + IUGR and lowest in controls (0.6 (IQR 0.4-1.8) vs. 0.05 (IQR 0.02-0.15)). CONCLUSION: A correlation between the serum levels of sFlt-1 and PlGF and the placental weight is present in PE-associated pregnancy disorders. This mirrors the model of an angiogenic continuum in the placenta where the serum sFlt-1 to PlGF ratio increases with severity of the disease.

Validation of a CE-IVD, urine exosomal RNA expression assay for risk assessment of prostate cancer prior to biopsy.

Improved risk stratification of patients suspected of prostate cancer prior to biopsy continues to be an unmet clinical need. ExoDx Prostate (IntelliScore) "EPI" is a non-invasive urine test utilizing RNA from exosomes to provide a risk score that correlates with the likelihood of finding high grade prostate cancer at biopsy. Here, we present the results from a prospective clinical validation study of EPI-CE, a CE-marked in-vitro diagnostic (IVD) assay, specifically developed for use in European clinical laboratories. The study (NCT04720599) enrolled patients with ≥ 50 years, PSA 2-10 ng/mL, prior to MRI, who were scheduled for initial biopsy. First catch urine samples were collected from participants without prior digital rectal examination or prostate massage. Exosomal RNA was isolated and expression levels of three biomarkers ERG, PCA3 and SPDEF were analyzed according to the EPI-CE Instructions For Use. In the study cohort of N = 109 patients, EPI-CE was validated to have a Negative Predictive Value of 89%, a Sensitivity of 92% and a superior performance to two commonly used multiparametric risk calculators (PCPT and ERSPC) in both Receiver Operating Characteristics with a higher Area Under the Curve for EPI-CE 0.67 (95% CI 0.56-0.77) versus PCPT 0.59 (95% CI 0.47-0.71) and ERSPC 0.60 (95% CI 0.49-0.72) and higher Net Benefits analysis across a wide range of risk acceptance levels. This is the first clinical study reporting on the performance of EPI-CE. We demonstrate that EPI-CE provides information beyond standard clinical parameters and provides a better risk assessment prior to MRI, of patients suspected of prostate cancer, than the commonly used multiparametric risk calculators.

The Manufacturing of 3D Printed models for the Neurotraumatological Education of Military Surgeons.

INTRODUCTION: When deployed abroad, military surgeons frequently have to deal with casualties involving head trauma. The emergency treatments, as well as craniotomies, are often performed by non-neurosurgeons qualified with basic neurotraumatological skills. Previous neurotrauma courses for education of non-neurosurgeons in Germany teach surgical emergency skills but do not include the training of skills needed to successfully utilize imaging in surgical planning, which is of importance for the safety and success of the treatment. To overcome these limitations, 3D printed models of neurotrauma cases were fabricated for application in the training of non-neurosurgeons. MATERIALS AND METHODS: Five models of actual neurotrauma cases from our neurosurgical department were segmented from CT scans and 3D printed using multi-part fused deposition modeling. Model quality was assessed with respect to the representation of pre-defined anatomical landmarks. The models were then fixed to a wooden mount with a central light source and covered by a latex mask for skin simulation. Surgical planning by means of craniometric measurements on the basis of available CT scans of the corresponding patients was then applied to the model. RESULTS: The 3D printed models precisely represented the cranium, the lesion, and anatomical landmarks, which are taken into consideration during surgical planning. Surface covering with washable latex masks ensured sufficient masking of the now non-noticeable lesion within the semi-translucent skull. Surgical planning was performed using washable marker drawings. When lighted, the otherwise non-visible lesion within the semi-translucent 3D printed craniums became visible and facilitated immediate success control for the course participants. CONCLUSION: The presented method provided a way to fabricate precise 3D models of neurotrauma cases, which are suitable to teach the application of medical imaging in surgical planning. For further benefit analysis, the application of the presented education tool needs to be investigated within a neurotrauma course.

Extracellular vesicles from plasma have higher tumour RNA fraction than platelets.

In addition to Circulating Tumour Cells (CTCs), cell-free DNA (cfDNA) and Extracellular Vesicles (EVs), the notion of "Tumour-Educated Platelets" (TEP) has recently emerged as a potential source of tumour-derived biomarkers accessible through blood liquid biopsies. Here we sought to confirm the suitability of the platelet blood fraction for biomarker detection in comparison to their corresponding EV fraction. As publications have claimed that tumour RNA and other tumour-derived material are transferred from tumour cells to the platelets and that tumour-derived transcripts can be detected in platelets, we chose to focus on RNA carrying a mutation as being of bona fide tumour origin. After informed consent, we collected prospective blood samples from a cohort of 12 melanoma patients with tissue-confirmed BRAF V600E mutation. Each blood specimen was processed immediately post collection applying two published standard protocols in parallel selecting for EVs and platelets, respectively. The RNA of each fraction was analysed by a highly sensitive ARMS RT-qPCR enabling the quantification of the mutant allele fraction (%MAF) of BRAF V600E down to 0.01%. In a direct comparative analysis, the EV fraction contained detectable BRAF V600E in 10 out of 12 patients, whereas none of the patient platelet fractions resulted in a mutant allele signal. The platelet fraction of all 12 patients contained high amounts of wild-type BRAF signal, but no mutation signal above background was detectable in any of the samples. Our observations suggest that the phenomenon of tumour RNA transfer to platelets occurs below detection limit since even a very sensitive qPCR assay did not allow for a reliable detection of BRAF V600E in the platelet fraction. In contrast, EV fractions derived from the same patients allowed for detection of BRAF V600E in 10 of 12 blood specimens.

Use of extracellular vesicles from lymphatic drainage as surrogate markers of melanoma progression and BRAF V600E mutation.

Liquid biopsies from cancer patients have the potential to improve diagnosis and prognosis. The assessment of surrogate markers of tumor progression in circulating extracellular vesicles could be a powerful non-invasive approach in this setting. We have characterized extracellular vesicles purified from the lymphatic drainage also known as exudative seroma (ES) of stage III melanoma patients obtained after lymphadenectomy. Proteomic analysis showed that seroma-derived exosomes are enriched in proteins resembling melanoma progression. In addition, we found that the BRAFV600E mutation can be detected in ES-derived extracellular vesicles and its detection correlated with patients at risk of relapse.