Analysis of P-gp genes relative expression associated to ivermectin resistance in Haemonchus contortus larval stages from in vitro cultures (L3 and xL3) and from gerbils (Meriones unguiculatus) (L4) as models of study.

The aim of the study was to compare the relative gene expression of Haemonchus contortus P-glycoprotein genes (Hco-pgp) between fourth (L4), infective (L3), and transitory infective (xL3) larval stages as laboratory models to study ivermectin (IVM) resistance. The H. contortus resistant to IVM (IVMr) and susceptible to IVM (IVMs) strains were used to develop xL3in vitro culture and to infect Meriones unguiculatus (gerbils) to collect L4 stages. Morphometric differences were evaluated from 25 individuals of H. contortus from each strain. Relative gene expression from xL3 and L4 was determined between comparison of IVMr stages and from IVMr vs IVMs stages. Seven Hco-pgp genes (1, 2, 3, 4, 9, 10, and 16) were analysed by RT-qPCR using L3 stage as control group, per strain, and GAPDH and ß-tubulin as constitutive genes. Morphological changes were confirmed between xL3 and L4 developing oral shape, oesophagus, and intestinal tube. In addition, the body length and width showed statistical differences (p < 0.05). The Hco-pgp1, 2, 3, and 4 genes (p < 0.05) were upregulated from 7.1- to 463.82-fold changes between IVMr stages, and Hco-pgp9 (13.12-fold) and Hco-pgp10 (13.56-fold) genes showed differences between L4 and xL3, respectively. The comparative study between IVMr vs IVMs strains associated to xL3 and L4 displayed significant upregulation for most of the Hco-pgp genes among 4.89-188.71 fold-change. In conclusion, these results suggest the use of H. contortus xL3 and L4 as suitable laboratory models to study IVMr associated with Hco-pgp genes to contribute to the understanding of anthelmintic resistance.

Treatment outcome of atypical EGFR mutations in the German National Network Genomic Medicine Lung Cancer (nNGM).

BACKGROUND: Atypical EGFR mutations occur in 10%-30% of non-small-cell lung cancer (NSCLC) patients with EGFR mutations and their sensitivity to classical epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKI) is highly heterogeneous. Patients harboring one group of uncommon, recurrent EGFR mutations (G719X, S768I, L861Q) respond to EGFR-TKI. Exon 20 insertions are mostly insensitive to EGFR-TKI but display sensitivity to exon 20 inhibitors. Clinical outcome data of patients with very rare point and compound mutations upon systemic treatments are still sparse to date. PATIENTS AND METHODS: In this retrospective, multicenter study of the national Network Genomic Medicine (nNGM) in Germany, 856 NSCLC cases with atypical EGFR mutations including co-occurring mutations were reported from 12 centers. Clinical follow-up data after treatment with different EGFR-TKIs, chemotherapy and immune checkpoint inhibitors were available from 260 patients. Response to treatment was analyzed in three major groups: (i) uncommon mutations (G719X, S7681, L861Q and combinations), (ii) exon 20 insertions and (iii) very rare EGFR mutations (very rare single point mutations, compound mutations, exon 18 deletions, exon 19 insertions). RESULTS: Our study comprises the largest thus far reported real-world cohort of very rare EGFR single point and compound mutations treated with different systemic treatments. We validated higher efficacy of EGFR-TKI in comparison to chemotherapy in group 1 (uncommon), while most exon 20 insertions (group 2) were not EGFR-TKI responsive. In addition, we found TKI sensitivity of very rare point mutations (group 3) and of complex EGFR mutations containing exon 19 deletions or L858R mutations independent of the combination partner. Notably, treatment responses in group 3 (very rare) were highly heterogeneous. Co-occurring TP53 mutations exerted a non-significant trend for a detrimental effect on outcome in EGFR-TKI-treated patients in groups 2 and 3 but not in group 1. CONCLUSIONS: Based on our findings, we propose a novel nNGM classification of atypical EGFR mutations.

Genomic analysis and proteomic response of the chromium-resistant and phenanthrene-degrading strain Streptomyces sp. MC1.

AIM: Chemically disparate toxic organic and/or inorganic molecules produced by anthropogenic activities often hinder the bioremediation process. This research was conducted to understand the capacity of Streptomyces sp. MC1 to remove chemically disparate toxics such as Cr(VI) or phenanthrene. METHODS AND RESULTS: Genomic, metabolic modeling and proteomic approaches were used in this study. Our results demonstrated that Streptomyces sp. MC1 has the genetic determinants to remove Cr(VI) or degrade phenanthrene. Proteomics showed that these genetic determinants were expressed. Metabolic versatility of the strain was confirmed by two metabolic models in complex and minimal media. Interestingly, our results also suggested a connection between the degradation of phenanthrene and synthesis of specialized metabolites. CONCLUSIONS: Streptomyces sp. MC1 has the genetic and physiological potential to remove Cr(VI) or degrade phenanthrene SIGNIFICANCE AND IMPACT OF STUDY: The probability of a microorganism to survive in the presence of different contaminants depends on its genetic potential and the ability to express it. The genetic and proteomic profiles obtained for Streptomyces sp. MC1 can be recommended as model and predict if other Streptomyces strains can be used in bioremediation processes. Our work also hypothesized that intermediates of the phenanthrene degradation serve as precursors for the specialized metabolism.

Opioid use prior to admission for chemotherapy induced febrile neutropenia is associated with increased documented infection, sepsis, and death.

PURPOSE: Cancer patients on chemotherapy are at risk for developing febrile neutropenia and infections. Opioids have been associated with immune suppression and risk of infection. We aimed to investigate opioid use associated with infections and death among cancer patients admitted with febrile neutropenia. METHODS: A total of 481 patients admitted for chemotherapy-induced febrile neutropenia were reviewed. There were 274 patients with opioid prescriptions (OP) within 10 days of hospitalization and 207 patients without opioid prescriptions (NOP) for >1 year of hospitalization. The primary outcomes were microbiologically and clinically documented infection as defined by the International Immunocompromised Host Society (IHS), sepsis by clinician, systemic inflammatory response syndrome (SIRS) criteria, and sequential organ failure assessment (SOFA) score. RESULTS: Documented infection occurred in 192 (70%) of patients with opioids compared to 99 (48%) with non-opioids, p < 0.001. Similar results were observed in sepsis by SOFA score with 173 (63%) opioids versus 92 (44%) non-opioids, p < 0.001, and sepsis by SIRS with 225 (82%) and 115 (56%) respectively, p < 0.001. Multivariable analysis showed opioid use has an increased adjusted odds of documented infection by 7.13 fold (95% CI 3.97-12.78), Sepsis by SOFA by 2.39 fold (95% CI 1.33-4.29), and Sepsis by SIRS by 1.87 fold (95% CI 1.13-3.10). Multivariable analysis for death/hospice showed that opioids had 2.30 fold (95% CI 1.16-4.57) increase in adjusted odds of death/hospice within 30 days of discharge. CONCLUSION: The data supports that patients with prior opioid use is associated with increased odds for infection, sepsis and death than non-opioid users admitted with febrile neutropenia.

Evaluation of the intraoperative human papillomavirus test as a marker of early cure at 12 months after electrosurgical excision procedure in women with cervical high-grade squamous intraepithelial lesion: a prospective cohort study.

OBJECTIVE: To evaluate if the intraoperative human papillomavirus (IOP-HPV) test has the same prognostic value as the HPV test performed at 6 months after treatment of high-grade squamous intraepithelial lesion (HSIL) to predict treatment failure. DESIGN: Prospective cohort study. SETTING: Barcelona, Spain. POPULATION: A cohort of 216 women diagnosed with HSIL and treated with loop electrosurgical excision procedure (LEEP). METHODS: After LEEP, an HPV test was performed using the Hybrid Capture 2 system. If this was positive, genotyping was performed with the CLART HPV2 technique. The IOP-HPV test was compared with HPV test at 6 months and with surgical margins. MAIN OUTCOME MEASURE: Treatment failure. RESULTS: Recurrence rate of HSIL was 6%. There was a strong association between a positive IOP-HPV test, a positive 6-month HPV test, positive HPV 16 genotype, positive surgical margins and HSIL recurrence. Sensitivity, specificity, and positive and negative predictive values of the IOP-HPV test were 85.7, 80.8,24.0 and 98.8% and of the HPV test at 6 months were 76.9, 75.8, 17.2 and 98.0%. CONCLUSION: Intraoperative HPV test accurately predicts treatment failure in women with cervical intraepithelial neoplasia grade 2/3. This new approach may allow early identification of patients with recurrent disease, which will not delay the treatment. Genotyping could be useful in detecting high-risk patients. TWEETABLE ABSTRACT: IOP-HPV test accurately predicts treatment failure in women with CIN 2/3.

Therapeutic patient education: the Avène-Les-Bains experience.

Originally developed to improve the management of serious chronic diseases such as diabetes mellitus, obesity, cardiovascular diseases, asthma and as aftercare following oncological treatments, therapeutic patient education (TPE) has been extended to other situations, including chronic dermatoses such as atopic dermatitis, eczema and psoriasis. In this article, we provide a brief overview of the development, implementation and evaluation of TPE at the Avène-Les-Bains Hydrotherapy Center, France.

Benefits of Avène thermal hydrotherapy in chronic skin diseases and dermatological conditions: an overview.

The beneficial effects of Avène Thermal Spring Water (TSW), a low mineral content spring water, on chronic skin diseases have been recognized for more than two centuries. This article provides a brief overview of efficacy and tolerance data for Avène TSW from clinical studies conducted at the Avène Hydrotherapy Center in patients with chronic inflammatory skin diseases or temporary skin injuries. Avène TSW hydrotherapy is effective as adjuvant management for chronic skin diseases and dermatological conditions, relieving subjective and physical symptoms with excellent tolerance.

Cure thermale et séquelles de brûlures: Thermal cures and sequelae of burns.

The sequelae of burns often are complicated by inflammatory and hypertrophic scars. Thermal cures can make a noticeable difference in improving these situations, which can be associated with considerable aesthetic damage and altered function. In this setting the benefits of the anti-inflammatory and antipruritic properties of thermal waters are combined with a high-pressure shower technique - the filiform shower - designed to soften and shape the infiltrated and hypertrophied skin areas. It is important to integrate the thermal cure at an early stage in the management of the condition, always considering it as an adjunct to the other therapeutic options available for the management of post-burn scarring. © 2020 Elsevier Masson SAS. All rights reserved.

La cure thermale en dermatologie, mode d'emploi: Use of thermal cures in dermatology.

Thermalism is one of the oldest medical disciplines with a history of thousands of years and whose benefits are well documented. The therapy is based on a program of daily care for a period of three weeks in a thermal station. Hydrotherapy is particularly suitable in dermatology since the thermal water will be in direct contact with the skin lesions. The main indications are eczema, psoriasis, chronic pruritus, and wound healing disorders with new indications emerging, such as for cancer treatment-related side effects and follow-up care after cancer treatment. Thermalism encompasses much more than hydrotherapy, acquiring over the past few years a distinct place in therapeutic education. The therapy, which for a long time has been seen as an alternative to pharmacological treatments, should be positioned as an adjunct treatment for chronic dermatoses, aiming at reducing in the mid and long term the severity of the disease and drug dependence. © 2020 Elsevier Masson SAS. Tous droits réservés.

Toxicités dermatologiques après prise en charge d'un cancer du sein : intérêt d'une cure thermale en soins oncologiques de support: Long lasting cutaneous adverse events after breast cancer and evaluation of hydrotherapy as supportive care.

Dermatological toxicities (affecting the skin, mucous membranes, nails or hair) are frequently associated with cancer treatments. They can represent a real burden for patients, with physical, social and psychological repercussions. These dermatological adverse events can also persist long after the treatment has ended, especially after treatment with cytotoxic chemotherapeutic agents such as taxanes. There is a clear need for the development of suitable supportive care measures to help manage these toxicities. The place of a hydrotherapy treatment in this context remains to be clarified. This article summarizes the main data available on the quality of life, and more specifically the dermatological quality of life, of patients for whom hydrotherapy was proposed after breast cancer. © 2020 Elsevier Masson SAS. All rights reserved.

Efficacy of a global supportive skin care programme with hydrotherapy after non-metastatic breast cancer treatment: A randomised, controlled study.

This study investigated the efficacy of post-treatment hydrotherapy as supportive care for management of persistent/long-lasting dermatologic adverse events (dAEs) induced in breast cancer survivors by adjuvant therapy, and its impact on quality of life (QoL). Patients in complete remission after standardised (neo)adjuvant chemotherapy, surgery and radiotherapy combination treatment for infiltrating HR+/HER2-breast carcinoma were enrolled in this randomised, multicentre controlled study 1-5 weeks after completing radiotherapy. The control group (CG, n = 33) received best supportive care and the treatment group (HG, n = 35) received 3-weeks of specific hydrotherapy. The primary criterion was change in QoL (QLQ-BR23) after hydrotherapy. Clinical grading of dAEs, cancer-related QoL (QLQ-C30), dermatologic QoL (DLQI) and general psychological well-being (PGWBI) were assessed. Significant dAEs were found at inclusion in both groups (n = 261). Most items showed significantly greater improvement in the HG versus CG group: QLQ-BR23 (breast [p = .0001] and arm symptoms [p = .0015], systemic therapy side effects [p = .0044], body image [p = .0139]), some dAE grading, DLQI (p = .0002) and PGWBI (p = .0028). Xerosis (88% of patients at inclusion) completely healed in all HG patients. Specific hydrotherapy is an effective supportive care for highly prevalent and long-lasting dAEs occurring after early breast cancer treatment, including chemotherapy, and leads to improved QoL and dermatologic toxicities.

Stent-based delivery of adeno-associated viral vectors with sustained vascular transduction and iNOS-mediated inhibition of in-stent restenosis.

In-stent restenosis remains an important clinical problem in the era of drug eluting stents. Development of clinical gene therapy protocols for the prevention and treatment of in-stent restenosis is hampered by the lack of adequate local delivery systems. Herein we describe a novel stent-based gene delivery platform capable of providing local arterial gene transfer with adeno-associated viral (AAV) vectors. This system exploits the natural affinity of protein G (PrG) to bind to the Fc region of mammalian IgG, making PrG a universal adaptor for surface immobilization of vector-capturing antibodies (Ab). Our results: 1) demonstrate the feasibility of reversible immobilization of AAV2 vectors using vector tethering by AAV2-specific Ab appended to the stent surface through covalently attached PrG, 2) show sustained release kinetics of PrG/Ab-immobilized AAV2 vector particles into simulated physiological medium in vitro and site-specific transduction of cultured cells, 3) provide evidence of long-term (12 weeks) arterial expression of luciferase with PrG/Ab-tethered AAV2Luc, and 4) show anti-proliferative activity and anti-restenotic efficacy of stent-immobilized AAV2iNOS in the rat carotid artery model of stent angioplasty.

Cytokine and antioxidant gene profiles from peripheral blood mononuclear cells of Pelibuey lambs after Haemonchus contortus infection.

The expression profiles of cytokines and antioxidant genes were determined from an experimental infection with H. contortus in Pelibuey lambs. The infection was followed for 34 days (d) to determine the number of eggs per gram (epg) and the packed cell volume (PCV). Differential white cell counts and expression profile estimations of IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, FCεR1A, GPX and SOD1 were determined at 0 hour, 4 hours, 2 days and 14 days post-infection (PI) in infected and control groups. Comparison of the fold change between 0 and 4-hours, 4-hours and 2-days and 2- and 14-days periods was performed. Significant differences (P<.05) between epg (>2000) and PCV (>30%) were determined after 21 days and were also observed with regard to monocyte and lymphocyte cells after 2 and 7 days PI. At 0 hour and 14 days PI, the GPX and IL-2 genes showed a 0.37- and 0.49-fold decrease in expression, respectively. In contrast, upregulation was observed at 4 hours of IL-8 (2.58) and FCεR1A (2.71), at 2 days for IL-4 (2.14) and IL-8 (4.02) and at 14 days for IL-2 (0.41), IL-10 (2.35) and FCεR1A (2.28). The comparison between the intervals of infection showed high expression values against H. contortus infection in Pelibuey sheep after the 2nd period of PI involving a dichotomy T cells.

Eau thermale d'Avène et dermatite atopique: Avène's thermal water and atopic dermatitis.

Atopic dermatitis (AD) is the most frequent disease treated at the Avène hydrotherapy center. Children represent a large part of the population due to the high prevalence of AD in early childhood. Avène thermal spring water (ATSW) has been known for its therapeutic effects since the middle of the 18th century. It has been greatly studied over the last decades, with a comprehensive fundamental, pharmaco-clinical and clinical approach. Cohort studies using the Scoring Atopic Dermatitis (SCORAD) clinical score and the Dermatology Life Quality Index (DLQI) or the Children's Dermatology Life Quality Index [CDLQI]) quality of life scores, allowed to confirm the clinical results obtained from the previous studies. These results were corroborated by clinical trials conducted in atopic patients outside the Avène hydrotherapy center, allowing to demonstrate the specific effect of the ATSW. Pharmacological and pharmaco-clinical studies evidenced several effects that could explain the healing effect of ATSW: effect on histamine release, anti-inflammatory effects on standardized models, immuno-modulation of some cytokines involved in DA physiopathology (interferon [INF], interleukin 2 and 4 [IL-2, IL-4]), improvement of keratinocyte differentiation, effect on the skin microbioma by promoting the development of a diversified non-pathogenic flora. In addition, an original microorganism, Aquaphilus dolomiae, never described in another medium, has very recently been identified in the ATSW. Aquaphilus dolomiae is responsible for significant pharmacological activities on inflammation, pruritus and enhancement of innate immunity. This set of works confirms the medical significance of the hydrotherapy which should be considered as a complementary care in the sometimes difficult management of AD.

Rotavirus activates dendritic cells derived from umbilical cord blood monocytes.

Rotavirus is the most common cause of acute infectious diarrhea in human neonates and infants. However, the studies aimed at dissecting the anti-virus immune response have been mainly performed in adults. Dendritic cells (DCs) play a crucial role in innate and acquired immune responses. Therefore, it is very important to determine the response of neonatal and infant DCs to rotavirus and to compare it to the response of adult DCs. Thus, we determined the response of monocyte-derived DCs from umbilical cord blood (UCB) and adult peripheral blood (PB) to rotavirus in vitro. It was found that the rotavirus and its genome, composed of segmented doubled stranded RNA (dsRNA), induced the activation of neonatal DCs, as these cells up-regulated the levels of CD40, CD86, MHC II, TLR-3 and TLR-4, the production of cytokines IL-6, IL-12/23p40, IL-10, TGF-ß (but not of IL-12p70), and the message for TNF-α and IFN-ß. This activation enabled the neonatal DCs to induce a strong proliferation of allogeneic CD4+ T cells and the production of IFN-γ. Moreover, neonatal DCs could be infected by rotavirus and sustain its replication. Neonatal DCs had a similar response as adult DCs towards rotavirus and its genome. However, adult DCs had a biased pro-inflammatory response compared to neonatal DCs, which showed a biased regulatory profile, as they produced higher levels of IL-10 and TGF-ß, and were less efficient in inducing a Th1 type response. So it can be concluded that rotavirus and its genome can induce the activation of neonatal DCs in spite of their tolerogenic bias.

Prevalence of and risk factors for pulmonary abnormalities in HIV-infected patients treated with antiretroviral therapy.

BACKGROUND: Pulmonary abnormalities are often present in patients infected with the human immunodeficiency virus (HIV). OBJECTIVES: The aim of the study was to determine the prevalence and characteristics of, and risk factors for, pulmonary abnormalities in HIV-positive patients. METHODS: A total of 275 HIV-positive patients [mean (± standard deviation) age 48.5 ± 6.6 years] were included in the study, of whom 95.6% had been receiving highly active antiretroviral therapy (HAART) for a mean (± standard deviation) duration of 11.9 ± 5.4 years. The median (interquartile range) CD4 lymphocyte count was 541 (392-813) cells/µL, and 92% of the patients had an undetectable viral load. We determined: (1) spirometry, static lung volumes, lung diffusing capacity, pulmonary gas exchange and exercise tolerance, and (2) the amount of emphysema via a computed tomography (CT) scan. RESULTS: Chronic cough and expectoration (47%) and breathlessness during exercise (33.9%) were commonly reported. Airflow limitation (AL) was present in 17.2%, low pulmonary diffusing capacity in 52.2% and emphysema in 10.5-37.7% of patients, depending on the method used for quantification. Most of these abnormalities had not been diagnosed or treated previously. Smoking exposure and previous tuberculosis were the main risk factors for AL, whereas smoking exposure and several variables related to HIV infection appeared to contribute to the risk of emphysema and low diffusing capacity. CONCLUSIONS: Despite HAART, pulmonary structural and functional abnormalities are frequent in HIV-positive patients. They are probably attributable to both environmental (smoking and tuberculosis) and HIV-related factors. Most of these abnormalities remain unnoticed and untreated. Given the relatively young age of these patients, these results anticipate a significant health problem in the next few years as, thanks to the efficacy of HAART, patients survive longer and experience the effects of aging.

Challenges of hepatitis C treatment in Native Americans in two North Dakota medical facilities.

INTRODUCTION: The prevalence of chronic liver disease (CLD) in the Aboriginal North American population is disproportionately higher than that of the non-indigenous population. Hepatitis C virus (HCV) is the second leading cause of CLD in American Indians or Alaska Natives (AIANs). This study described the experience of two teaching community medical centers in North Dakota in treating HCV infection among AIANs and compared treatment outcomes to a cohort of Caucasian patients. METHODS: The retrospective study described the characteristics and proportion of AIAN patients with HCV who received treatment. Documented reasons for not receiving treatment were analyzed. For those AIAN patients treated for HCV infection, responses to treatment, including rapid, early and sustained virological responses (SVRs), were compared with those of Caucasians. RESULTS: Only 22 (18%) of 124 AIANs with HCV infection received treatment. Common reasons for not receiving treatment include lack of access to specialists, concomitant or decompensated liver disease, alcohol and drug abuse and cost. There were no significant differences in the baseline characteristics and key predictors of SVR in AIANs compared to Caucasian controls. CONCLUSIONS: Most AIAN patients with HCV infection do not receive treatment despite comparable treatment response rates to Caucasians. Further population-based studies, addressing access to specialized hepatitis C treatment and public health concerns are warranted, as it is crucial to treat chronic HCV infection to decrease the burden of disease in the AIAN community.

Prognostic significance of thymidylate synthase polymorphisms in rectal cancer patients treated with neoadjuvant chemoradiotherapy.

AIM: There is a lack of prognostic factors of preoperative chemoradiation for locally advanced rectal cancer. Thymidylate synthase (TS) is the most important target of 5-fluorouracil; three main genetic polymorphisms of TS have been described. We analysed the prognostic value of these in patients with locally advanced rectal cancer treated with fluoropyrimidine-based chemoradiation. METHOD: Ninety-nine patients treated between November 2001 and March 2009 were included. All were treated by radiotherapy (5040 cGy) and concomitant fluoropyrimidine-based chemotherapy. Three polymorphisms were analysed: (i) a double (2R) or triple (3R) repeat of a 28 base pair (bp) tandem sequence upstream of the ATG codon initiation site in the 5'-terminal regulatory region, (ii) a functional G > C single nucleotide polymorphism present in the second repeat of the 3R alleles and (iii) a 6 bp deletion at nucleotide 1494 in the 3'-untranslated region. DNA was extracted from paraffin-embedded core biopsies taken from the tumour and the genotype was analysed using polymerase chain reaction restriction fragment length polymorphism. RESULTS: The 6 bp polymorphism was significantly associated with disease-free survival (+ 6 bp/+ 6 bp vs-6 bp/-6 bp, P = 0.032 logistic regression). No differences were found in disease-free survival according to the other polymorphisms studied. No relationship was observed between the different TS genotypes and pathological regression. CONCLUSION: The study suggests that the TS 6 bp polymorphism may be a predictor of disease-free survival in patients with locally advanced rectal cancer treated with fluoropyrimidine-based chemoradiation.

Real-world EGFR testing practices for non-small-cell lung cancer by thoracic pathology laboratories across Europe.

BACKGROUND: Testing for epidermal growth factor receptor (EGFR) mutations is an essential recommendation in guidelines for metastatic non-squamous non-small-cell lung cancer, and is considered mandatory in European countries. However, in practice, challenges are often faced when carrying out routine biomarker testing, including access to testing, inadequate tissue samples and long turnaround times (TATs). MATERIALS AND METHODS: To evaluate the real-world EGFR testing practices of European pathology laboratories, an online survey was set up and validated by the Pulmonary Pathology Working Group of the European Society of Pathology and distributed to 64 expert testing laboratories. The retrospective survey focussed on laboratory organisation and daily EGFR testing practice of pathologists and molecular biologists between 2018 and 2021. RESULTS: TATs varied greatly both between and within countries. These discrepancies may be partly due to reflex testing practices, as 20.8% of laboratories carried out EGFR testing only at the request of the clinician. Many laboratories across Europe still favour single-test sequencing as a primary method of EGFR mutation identification; 32.7% indicated that they only used targeted techniques and 45.1% used single-gene testing followed by next-generation sequencing (NGS), depending on the case. Reported testing rates were consistent over time with no significant decrease in the number of EGFR tests carried out in 2020, despite the increased pressure faced by testing facilities during the COVID-19 pandemic. ISO 15189 accreditation was reported by 42.0% of molecular biology laboratories for single-test sequencing, and by 42.3% for NGS. 92.5% of laboratories indicated they regularly participate in an external quality assessment scheme. CONCLUSIONS: These results highlight the strong heterogeneity of EGFR testing that still occurs within thoracic pathology and molecular biology laboratories across Europe. Even among expert testing facilities there is variability in testing capabilities, TAT, reflex testing practice and laboratory accreditation, stressing the need to harmonise reimbursement technologies and decision-making algorithms in Europe.