Characterization of DNA concentration in urine and dried blood samples to detect the c.577 deletion within the EPO gene.

The EPO gene variant, c.577del (VAR-EPO), was discovered in the Chinese population in 2021. The mutated protein is naturally present in urine from individuals heterozygous for the variant. Electrophoresis methods currently applied in anti-doping laboratories produce a pattern in samples from individuals carrying VAR-EPO that cannot be unambiguously distinguished from individuals who received recombinant EPO doses. Consequently, the analysis of blood samples is obligatory to facilitate interpretation of suspicious findings from urine samples. However, this complicates the process and delays the reporting. Objective of this study was to develop EPO c.577del detection in urine and dried blood samples (DBS) in order to facilitate and accelerate EPO results management. Moreover, estimation of the success rate of sequencing regarding concentration of DNA in urine and DBS was evaluated. Conclusive results regarding Sanger sequencing were obtained for all samples with DNA concentrations above 0.024 ng/µL DNA in 80% of urines samples from volunteers. The potential success of DNA sequencing rate in athletes' urines was investigated. A total of 191 urine samples were considered. DNA concentration exceeding 0.024 ng/µL was detected in 85% of the samples. Interestingly, in-competition samples had a significantly higher DNA concentration than out-of-competition male urine samples (0.330 vs. 0.084 ng/µL). Moreover, conclusive EPO sequences were obtained for 100% of DBS (cellulose and polymer matrices). In conclusion, method for detection of EPO gene variant was developed in urine and DBS. Characterization of DNA concentration was performed in order to evaluate the probability of success of sequencing EPO gene in anti-doping field.

Mortality and functional consequences after revision osteosynthesis for peritrochanteric fractures treated by intramedullary nail: A retrospective study of 312 patients.

INTRODUCTION: Osteosynthesis of peritrochanteric fractures by intramedullary nail is associated with a 3 to 15% rate of complications requiring revision surgery. The objective of our study was to compare mortality and autonomy between patients who required revision surgery after osteosynthesis, and patients without revision surgery. HYPOTHESES: Patients who underwent revision surgery after osteosynthesis of a peritrochanteric fracture by intramedullary nail did not exhibit excess mortality or loss of autonomy compared to those who did not undergo revision surgery. METHODS: Between January 2017 and October 2019, 312 consecutive patients were operated on for a peritrochanteric fracture by intramedullary nail. Among them, 23 patients (7.4%) required revision surgery (change, nail removal, total hip arthroplasty). Mortality at 90 days and at one year was evaluated and compared between the group of "revision" patients and "no revision" patients. The level of autonomy was assessed by the Parker and Palmer score, one year postoperatively and compared between the 2 groups. RESULTS: The patients requiring revision surgery were younger: 73 years old vs. 86 years old (p<0.011). The overall mortality was 7.25% at 90 days after surgery and 15% at one year. Mortality was zero at 90 days, and at 1 year, postoperatively for patients requiring revision surgery. There was no significant difference for the mortality at 90 days (p=0.39) between the 2 groups, however at 1 year, it was significantly less for the revision patients (0% vs. 17%, p=0.032). There was no significant difference for the Parker score between the "revision" group; 5 (4-9) and the "no revision" group; 4 (3-7), at one year postoperatively (p=0.24). CONCLUSION: This study did not show any excess mortality, nor loss of autonomy at 1 year postoperatively, for patients who required osteosynthesis for a trochanteric fracture, and who presented with a complication requiring revision surgery. LEVEL OF EVIDENCE: III, comparative retrospective study.

Eliminating Quantum Phase Slips in Superconducting Nanowires.

In systems with reduced dimensions, quantum fluctuations have a strong influence on the electronic conduction, even at very low temperatures. In superconductors, this is especially interesting, since the coherent state of the superconducting electrons strongly interacts with these fluctuations and therefore is a sensitive tool to study them. In this paper, we report on comprehensive measurements of superconducting nanowires in the quantum phase slip regime. Using an intrinsic electromigration process, we have developed a method to lower the nanowire's resistance in situ and therefore eliminate quantum phase slips in small consecutive steps. We observe critical (Coulomb) blockade voltages and superconducting critical currents, in good agreement with theoretical models. Between these two regimes, we find a continuous transition displaying a nonlinear metallic-like behavior. The reported intrinsic electromigration technique is not limited to low temperatures, as we find a similar change in resistance that spans over 3 orders of magnitude also at room temperature. Aside from superconducting quantum circuits, such a technique to reduce the resistance may also have applications in modern electronic circuits.

Current-Resistance Effects Inducing Nonlinear Fluctuation Mechanisms in Granular Aluminum Oxide Nanowires.

The unusual superconducting properties of granular aluminum oxide have been recently investigated for application in quantum circuits. However, the intrinsic irregular structure of this material requires a good understanding of the transport mechanisms and, in particular, the effect of disorder, especially when patterned at the nanoscale level. In view of these aspects, electric transport and voltage fluctuations have been investigated on thin-film based granular aluminum oxide nanowires, in the normal state and at temperatures between 8 and 300 K. The nonlinear resistivity and two-level tunneling fluctuators have been observed. Regarding the nature of the noise processes, the experimental findings give a clear indication in favor of a dynamic random resistor network model, rather than the possible existence of a local ordering of magnetic origin. The identification of the charge carrier fluctuations in the normal state of granular aluminum oxide nanowires is very useful for improving the fabrication process and, therefore, reducing the possible sources of decoherence in the superconducting state, where quantum technologies that are based on these nanostructures should work.

Incidence of peri-operative paediatric cardiac arrest and the influence of a specialised paediatric anaesthesia team: Retrospective cohort study.

BACKGROUND: Peri-operative critical events are still a major problem in paediatric anaesthesia care. Access to more experienced healthcare teams might reduce the adverse event rate and improve outcomes. OBJECTIVE: The current study analysed incidences of peri-operative paediatric cardiac arrest before and after implementation of a specialised paediatric anaesthesia team and training programme. DESIGN: Retrospective cohort study with before-and-after analysis. SETTING: Department of Anaesthesiology and Intensive Care Medicine, University Hospital of Cologne, Germany. PATIENTS: A total of 36 243 paediatric anaesthetics (0 to 18 years) were administered between 2008 and 2016. INTERVENTION: Implementation of a specialised paediatric anaesthesia team and training programme occurred in 2014 This included hands-on supervised training in all fields of paediatric anaesthesia, double staffing for critical paediatric cases and a 24/7 emergency team. A logistic regression analysis with risk factors (age, ASA physical status, emergency) was used to evaluate the impact of implementation of the specialised paediatric anaesthesia team. MAIN OUTCOME MEASURES: Incidences of peri-operative paediatric cardiac arrest and anaesthesia-attributable cardiac arrest before and after the intervention. RESULTS: Twelve of 25 paediatric cardiac arrests were classified as anaesthesia-attributable. The incidence of overall peri-operative paediatric cardiac arrest was 8.1/10 000 (95% CI 5.2 to 12.7) in the period 2008 to 2013 and decreased to 4.6/10 000 (95% CI 2.1 to 10.2) in 2014 to 2016. Likewise, the incidence of anaesthesia-attributable cardiac arrest was lower after 2013 [1.6/10 000 (95% CI 0.3 to 5.7) vs. 4.3/10 000 (95% CI 2.3 to 7.9)]. Using logistic regression, children anaesthetised after 2013 had nearly a 70% lower probability of anaesthesia-attributable cardiac arrest (odds ratio 0.306, 95% CI 0.067 to 1.397; P = 0.1263). For anaesthesia-attributable cardiac arrest, young age was the most contributory risk factor, whereas in overall paediatric cardiac arrest, ASA physical statuses 3 to 5 played a more important role. CONCLUSION: In this study on incidences of peri-operative paediatric cardiac arrest from a European tertiary care university hospital, implementation of a specialised paediatric anaesthesia team and training programme was associated with lower incidences of peri-operative paediatric cardiac arrest and a reduced probability of anaesthesia-attributable cardiac arrest.

Patellofemoral design enhancements reduce long-term complications of postero-stabilized total knee arthroplasty.

PURPOSE: Few studies investigated whether trochlear and patellar design enhancements improve long-term outcomes of total knee arthroplasty (TKA). This study aimed to compare the long-term survival and complication rates of two consecutive generations of the same TKA system with identical tibiofemoral geometry, but different patellofemoral designs. METHODS: The authors retrieved the records of 93 patients (104 knees) operated with the HLS II system and 116 patients (122 knees) operated with HLS Evolution system. Patients were evaluated preoperatively and at a minimum of 10 years noting all complications. Kaplan-Meier (KM) survival was compared for two endpoints: (1) revision of all components and (2) revision of any component. RESULTS: From the HLS II series, the incidence of revision of all components was 6.4%, and of any component was 9.8%. From the HLS Evolution series, the incidence of revision of all components was 4.1%, and of any component was 5.1%. Comparing the survival at equivalent follow-up of 14 years, considering revision of all components, the HLS II had higher survival than the HLS Evolution (98.9% vs 95.9%), while considering revision of any component, the HLS II had lower survival than the HLS Evolution (93.0% vs 94.9%). The differences in survival of the two implants were not significant, neither at equivalent follow-up of 14 years (n.s.), nor at maximum follow-up of each cohort (n.s.). The complication rate was higher for the HLS II series compared to the HLS Evolution (28% vs 12%, p = 0.009), but patellofemoral complications were not more frequent (8% vs 6%, n.s.). CONCLUSIONS: Though the differences in survival of the two implants were not significant, conflicting findings are observed due to partial revisions for patellar fractures (5 in the HLS II series and 1 in the HLS Evolution series) which could be related to patellofemoral design enhancements. This study highlights the importance of patello-femoral geometry, which is often overlooked in TKA. LEVEL OF EVIDENCE: Retrospective comparative study, Level III.

Arthroscopic All-Inside Repair of Medial Meniscus Grade 2 Horizontal Cleavage Tear Using Additional Posteromedial Portal.

Management of intrasubstance horizontal cleavage meniscal lesions of microtraumatic origin remains poorly defined in young patients. For grade 2 lesions resistant to conservative measures, the standard technique is debridement of the intrasubstance tear and open suture repair via a posteromedial approach. The objective of this Technical Note is to propose an arthroscopic alternative to this open technique, using an arthroscopic additional posteromedial portal. This technique facilitates an approach to the lesion via its peripheral portion without creating an iatrogenic lesion of the free edge of the meniscus, which is located in the white zone and thus exhibits limited vascularity.

Incidence and risk factors of anaesthesia-related perioperative cardiac arrest: A 6-year observational study from a tertiary care university hospital.

BACKGROUND: In recent decades, the incidences of anaesthesia-related perioperative mortality and adverse outcomes have decreased drastically. However, to date, data on perioperative cardiac arrest and risk factors of perioperative cardiac arrest from European countries are scarce. OBJECTIVES: To determine the incidences of perioperative cardiac arrest and rates of anaesthesia-related and anaesthesia-contributory cardiac arrest. Identification of pre-existing risk factors leading to perioperative cardiac arrest. DESIGN: Retrospective cohort study. SETTING: Department of Anaesthesiology and Intensive Care Medicine, University Hospital of Cologne, Germany. INTERVENTIONS: Perioperative critical incident reports between 2007 and 2012 were screened, and reports on cardiac arrest within 24 h postoperatively were identified. Cardiac arrests were classified as 'anaesthesia-related', 'anaesthesia-contributory' or 'anaesthesia-unrelated' by two reviewers independently. Univariate and multi-variate logistic regression analysis was used to identify risk factors associated with perioperative cardiac arrest. RESULTS: Analysis of 318 critical incidents from 169 500 anaesthetics revealed 99 perioperative cardiac arrests. This is an overall incidence of perioperative cardiac arrest of 5.8/10 000 anaesthetics [95% confidence interval (CI), 4.7 to 7.0]. The rate of anaesthesia-related cardiac arrest was 0.7/10 000 (95% CI, 0.3 to 1.1), and the rate of anaesthesia-contributory cardiac arrest was 1.7/10 000 (95% CI, 1.1 to 2.3). Most cardiac arrests related to anaesthesia were due to respiratory events. From the multi-variate analysis, American Society of Anesthesiologists physical status grade at least 3 [P = 0.007, odds ratio (OR) 2.59 (95% CI, 1.29 to 5.19)], emergency surgery [P < 0.001, OR 4.00 (95% CI, 2.15 to 7.54)] and pre-existing cardiomyopathy [P < 0.001, OR 17.48 (95% CI, 6.18 to 51.51)] emerged as predictors of cardiac arrest. CONCLUSION: These first available European data on perioperative cardiac arrest from a large unselected cohort indicate that the overall perioperative incidence of cardiac arrest at our institution was slightly lower than published in the literature, whereas rates of anaesthesia-related and anaesthesia-contributory cardiac arrest were comparable. Most cardiac arrests related to anaesthesia were due to respiratory events. American Society of Anesthesiologists physical status grade at least 3, emergency surgery and pre-existing cardiomyopathy appear to be relevant risk factors for cardiac arrest.

Classification and Surgical Repair of Ramp Lesions of the Medial Meniscus.

Ramp lesions of the medial meniscus are commonly associated with anterior cruciate ligament ruptures and consist of longitudinal peripheral tears of the posterior horn of the medial meniscus. Given the frequency of partial-thickness tears, they can be difficult to diagnose arthroscopically from the anterior compartment. We describe a classification of the different types of ramp lesions depending on both tear pattern (partial- or full-thickness tear) and associated meniscotibial ligament disruption. An original technique of arthroscopic suture placement through a single posteromedial portal with a 25° curved suture hook device is described.

A Fluoroscopy-Free Technique for Percutaneous Screw Positioning During Arthroscopic Treatment of Depression Tibial Plateau Fractures.

This article aims to describe a simple and reliable technique that helps in positioning the cannulated percutaneous screws during fixation of depression-type tibial plateau fractures. After fracture reduction under arthroscopic control, an outside-in anterior cruciate ligament femoral guide is introduced through the tibial cortical metaphyseal window and positioned under endoscopic control just underneath the elevated fragment. When proper height is achieved, a guide pin is drilled from lateral to medial through the sleeve, 1 to 2 cm distal to the articular surface of the depressed fragment. The cannulated screw can then be introduced under endoscopic control, without fluoroscopic assistance, just under the previously elevated joint surface. This technique ensures optimal placement of the cannulated screw in the middle of the bony tunnel to obtain optimal subchondral bone support during fixation of the depressed tibial plateau fracture.

Repair of Meniscal Ramp Lesions Through a Posteromedial Portal During Anterior Cruciate Ligament Reconstruction: Outcome Study With a Minimum 2-Year Follow-up.

PURPOSE: To evaluate the results of arthroscopic all-inside suture repair of medial meniscal ramp lesions through a posteromedial portal during anterior cruciate ligament (ACL) reconstruction. METHODS: All patients who underwent a suture of the posterior segment of the medial meniscus using a suture hook device through a posteromedial portal during ACL reconstruction with minimum 2 year-follow-up were included in the study. Repair was performed for longitudinal tears within the rim of less than 3 mm (capsulomeniscal junction or red-red zone) or 3 to 5 mm (red-white zone) of an unstable torn meniscus. Patients were assessed pre- and postoperatively with IKDC score and Tegner activity scale. Instrumented knee testing was performed with the Rolimeter arthrometer. Complications including reoperation for failed meniscal repair were also recorded. RESULTS: One hundred thirty-two patients met the inclusion criteria. The mean follow-up time was 27 months (range, 24 to 29 months). The average subjective IKDC rose from 63.8 ± 13.5 (range, 27 to 92) preoperatively to 85.7 ± 12 (range, 43 to 100) at last follow-up (P < .0001). The Rolimeter test decreased from a side-to-side difference in anterior knee laxity of 7 mm (range, 5 to 14 mm) to a mean value of 0.4 mm (range, -3 to 5 mm) at last follow-up (P < .0001). The Tegner activity scale at the last follow-up (6.9 ± 1.72) was slightly lower than that before surgery (7.2 ± 1.92; P = .0017). Nine patients (6.8%) had failure of the meniscal repair. In 5 cases, recurrent tears were related to a newly formed tear located anterior to the initial tear. CONCLUSIONS: Our results show that arthroscopic meniscal repair of ramp lesions during ACL reconstruction through a posteromedial portal provided a high rate of meniscus healing at the level of the tear and appeared to be safe and effective in this group of patients. LEVEL OF EVIDENCE: Level IV, therapeutic study, case series (no control group).

Evaluation of longitudinal steroid profiles from male football players in UEFA competitions between 2008 and 2013.

Testosterone and related compounds are the most recurrent doping substances. The steroid profile, consisting of the quantification of testosterone and its metabolites, has been described as the most significant biomarker to detect doping with pseudo-endogenous anabolic steroids. The steroidal module of the Athlete Biological Passport (ABP) was launched by the World Anti-Doping Agency (WADA) in 2014. To assess the value of introducing the module to its anti-doping programme, the Union of European Football Associations (UEFA) decided to analyze retrospectively the steroid profile data of 4195 urine samples, collected from 879 male football players and analyzed in 12 WADA-accredited laboratories between 2008 and mid-2013. This study focused on the evaluation of T/E ratios. The coefficient of variation (CV) and the adaptive model were the two statistical models used to study the longitudinal follow-up. A CV of 46% was determined to be the maximal natural intra-individual variation of the T/E when the sequence consisted of single data points analyzed in different laboratories. The adaptive model showed some profiles with an atypical T/E sequence and also enabled an estimate of the prevalence of external factors impacting the T/E sequences. Despite the limitations of this retrospective study, it clearly showed that the longitudinal and individual follow-up of the T/E biomarker of the players is a good tool for target testing in football. UEFA has therefore decided to implement the steroidal module of the ABP from the start of the next European football season in September 2015. Copyright © 2015 John Wiley & Sons, Ltd.

Antidoping programme and biological monitoring before and during the 2014 FIFA World Cup Brazil.

BACKGROUND: The FIFA has implemented an important antidoping programme for the 2014 FIFA World Cup. AIM: To perform the analyses before and during the World Cup with biological monitoring of blood and urine samples. METHODS: All qualified players from the 32 teams participating in the World Cup were tested out-of-competition. During the World Cup, 2-8 players per match were tested. Over 1000 samples were collected in total and analysed in the WADA accredited Laboratory of Lausanne. RESULTS: The quality of the analyses was at the required level as described in the WADA technical documents. The urinary steroid profiles of the players were stable and consistent with previously published papers on football players. During the competition, amphetamine was detected in a sample collected on a player who had a therapeutic use exemption for attention deficit hyperactivity disorder. The blood passport data showed no significant difference in haemoglobin values between out-of-competition and postmatch samples. CONCLUSIONS: Logistical issues linked to biological samples collection, and the overseas shipment during the World Cup did not impair the quality of the analyses, especially when used as the biological passport of football players.

Untargeted profiling of urinary steroid metabolites after testosterone ingestion: opening new perspectives for antidoping testing.

AIM: Antidoping procedures are expected to greatly benefit from untargeted metabolomic approaches through the discovery of new biomarkers of prohibited substances abuse. RESULTS: Endogenous steroid metabolites were monitored in urine samples from a controlled elimination study of testosterone undecanoate after ingestion. A platform coupling ultra-high pressure LC with high-resolution quadrupole TOF MS was used and high between-subject metabolic variability was successfully handled using a multiblock data analysis strategy. Links between specific subsets of metabolites and influential genetic polymorphisms of the UGT2B17 enzyme were highlighted. CONCLUSION: This exploratory metabolomic strategy constitutes a first step toward a better understanding of the underlying patterns driving the high interindividual variability of steroid metabolism. Promising biomarkers were selected for further targeted study.

Circulating microRNAs as long-term biomarkers for the detection of erythropoiesis-stimulating agent abuse.

MicroRNAs (miRNAs) are small, non-protein coding transcripts involved in many cellular and physiological mechanisms. Recently, a new class of miRNA called 'circulating miRNAs' was found in cell-free body fluids such as plasma and urine. Circulating miRNAs have been shown to be very stable, specific, and sensitive biomarkers. In this paper, we investigate whether circulating miRNAs can serve as biomarkers for erythropoiesis-stimulating agent abuse. To this end, we analyzed miRNA levels in plasma by miRNA microarrays and quantitative real-time polymerase chain reaction (PCR). Plasma samples are derived from a clinical study with healthy subjects injected with erythropoiesis-stimulating agent (C.E.R.A.). Based on microarray results, we observed a significant difference in the levels of miRNAs in plasma after C.E.R.A. injection. We demonstrated that a specific miRNA, miR-144, exhibit a high increase that lasts 27 days after C.E.R.A. stimulation. Considering the fact that miR-144 is an essential erythropoiesis agent in different organisms, these findings suggest the possibility of using miR-144 as a sensitive and informative biomarker to detect C.E.R.A. abuse.

Use of forensic investigations in anti-doping.

The fight against doping is mainly focused on direct detection, using analytical methods for the detection of doping agents in biological samples. However, the World Anti-Doping Code also defines doping as possession, administration or attempted administration of prohibited substances or methods, trafficking or attempted trafficking in any prohibited substance or methods. As these issues correspond to criminal investigation, a forensic approach can help assessing potential violation of these rules. In the context of a rowing competition, genetic analyses were conducted on biological samples collected in infusion apparatus, bags and tubing in order to obtain DNA profiles. As no database of athletes' DNA profiles was available, the use of information from the location detection as well as contextual information were key to determine a population of suspected athletes and to obtain reference DNA profiles for comparison. Analysis of samples from infusion systems provided 8 different DNA profiles. The comparison between these profiles and 8 reference profiles from suspected athletes could not be distinguished. This case-study is one of the first where a forensic approach was applied for anti-doping purposes. Based on this investigation, the International Rowing Federation authorities decided to ban not only the incriminated athletes, but also the coaches and officials for 2 years.

Extraskeletal osteosarcoma has a favourable prognosis when treated like conventional osteosarcoma.

PURPOSE: The aims of this analysis were to investigate the clinical features of extraskeletal osteosarcoma (ESOS) and examine the outcome after multi-modal therapy. METHODS: The co-operative osteosarcoma study-group database was searched for patients with extraskeletal osteosarcoma. Eligible patients were included in a retrospective analysis of patient, tumour and treatment related variables and outcome. As for conventional osteosarcoma, scheduled treatment included surgery and multi-agent chemotherapy. RESULTS: Seventeen eligible patients were identified with a median age of 44 years (range, 3-65 years). The thigh was the commonest tumour site. Two patients had a history of previous malignancies and two had primary metastases. Median follow-up was 3.2 years (range: 0.6-7.4 years) and at last follow-up, 11 patients were alive in complete remission, 3 patients were alive with disease and 3 patients had died of their disease. Three-year overall actuarial and event-free survival rates were 77% and 56%, respectively. Patients with macroscopically complete surgical remission had an improved overall survival (P = 0.0004). CONCLUSIONS: The patients in this retrospective study had a surprisingly good survival rate. This may be due to the combination of multi-agent chemotherapy with surgery, and we recommend this approach in the treatment of ESOS.

Delaying surgery with chemotherapy for osteosarcoma of the extremities.

Osteosarcoma is the most frequent primary cancer of bone. When treated by surgery alone, it is almost invariably followed by metastatic dissemination and death. This dismal prognosis can be improved dramatically by including chemotherapy in an interdisciplinary regimen. Today, two-thirds of patients with localised extremity primaries can achieve long-term survival with such intensive multimodal therapy. This article provides a summary overview of current pharmacotherapy in osteosarcoma of the extremities, focussing on the approach of preoperative 'neoadjuvant' chemotherapy and thus, the potential benefits and pitfalls of delaying surgery. Prospective, multi-institutional trials are essential in guaranteeing that as many patients as possible can benefit from modern, efficacious interdisciplinary therapeutic regimens and that further progress can be made.

Acute lymphoblastic leukemia and Klinefelter syndrome in children: two cases and review of the literature.

The occurrence of mediastinal germ cell tumor and breast cancer have been repeatedly reported in men with Klinefelter syndrome (KS) but this association is debated controversially for patients with hematologic malignancies. The authors describe 2 tall adolescents in whom diagnostic workup for acute lymphoblastic leukemia (ALL) revealed 47,XXY and 47,XXY/48,XXXYkaryotype, respectively. Among 4195 registered male patients in the ALL-BFM study group since 1983, no further patients with ALL and KS were identified. Given the lack of epidemiological data, this retrospective analysis illustrates the association of previously described cases of hematologic malignancies with KS. In contrast to other chromosomal aberrations, the incidence of ALL does not seem to be increased in pediatric patients with KS.