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Ocrelizumab is a monoclonal anti-CD20 antibody approved for the treatment of multiple sclerosis (MS). The clinical value of therapeutic drug monitoring (TDM) for this antibody in treatment of MS is unknown, and an adequately specific and precise quantitation method for ocrelizumab in patient serum could facilitate investigation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quantitation methods have been shown to have higher analytic specificity and precision than enzyme-linked immunosorbent assays.To establish and validate an LC-MS/MS-based quantitation method for ocrelizumab.We present an LC-MS/MS-based quantitation method using immunocapture purification followed by trypsinization and analysis by a triple quadrupole mass analyzer obtaining results within the same day.We found that the ocrelizumab peptide GLEWVGAIYPGNGDTSYNQK (Q1/Q3 Quantifier ion: 723.68We have established a reliable quantitation method for serum ocrelizumab that can be applied in clinical studies, facilitating the evaluation of ocrelizumab TDM in MS.

Tumours of uncertain differentiation represent a markedly heterogeneous group of neoplasms that show no identifiable line of differentiation by currently available technologies, and do not resemble any specific normal cells or tissues. Continuing advances in molecular genetics have significantly aided to our understanding of these tumours, enabling refinements in classification and better insights into their biology, and furthering the development of ancillary immunohistochemical and molecular diagnostic tests and targeted therapies in the clinical setting. This review summarizes key genetic, morphologic and immunohistochemical features for a selection of cutaneous neoplasms of uncertain differentiation, with emphasis on new entities and developments in the diagnostic setting.

Priming with elicitors, including phytohormones, is known to induce physiological changes in plants that affect resource allocation and nutrient uptake strategies. However, effects of these changes on belowground microbial communities, particularly those involved in nitrogen (N) transformations, remain poorly characterized. Here, we tested the hypothesis that treating seeds with elicitors would affect N-transforming communities associated to roots and rhizosphere, influencing soil N availability and thus plant performance. We measured the effects of treating winter wheat seeds with cis-jasmone, methyl jasmonate or methyl salicylate on plant traits and on the genetic potential for ammonia oxidation by ammonia-oxidizing bacteria (AOB) and archaea (AOA), the initial inorganic N transformation process resulting in N loss. Elicitors reduced the genetic potential for ammonia oxidation by AOB in root-associated microbial communities and increased traits reflecting N uptake in winter wheat. Proposed mechanisms include increased exploitative competition for ammonium, limiting substrate availability for microorganisms, and induced biological nitrification inhibition capacity actively suppressing ammonia oxidizers. There were minor effects on the composition of the AOB and AOA communities, but an indicator species analysis showed that several AOB and a few AOA genotypes were indicative of elicitor treatments. Seed treatment with elicitors may represent an overlooked strategy to minimize nitrification to mitigate N losses from arable soils.

Background Primary leiomyosarcoma (LMS) of the gastrointestinal (GI) tract is rare. Limited literature exists regarding the clinical characteristics and outcome for patients with localised and metastatic disease. Methods A retrospective chart review was performed for patients greater than 18 years of age diagnosed with GI LMS at The Royal Marsden Hospital between 1 January 2000–1 May 2020. Descriptive statistics were performed. Patients were censored at data cut-off date of 27 June 2020. Results Forty-six patients with a median age at diagnosis of 54 years (range 25–85) were identified. Fifteen percent (n = 7) of patients previously received abdominal radiation for an unrelated cancer. All patients with localised disease (n = 36) had resection with oncological margins. For patients who underwent potentially curative surgery, median recurrence-free survival (mRFS) was 13 months (0.4–183 months), and half of these patients (n = 18) developed recurrent disease post resection (distant n = 16, local n = 2). Median overall survival (mOS) was 27 months for patients with distant recurrence. Twenty-one percent (n = 10) of patients presented with synchronous metastatic disease and their mOS was 19 months. Median progression-free survival (mPFS) for patients treated with conventional chemotherapy ranged from 2.0 to 8.0 months. Conclusion The risk of recurrence is significant, and recurrence-free survival was short even with complete oncologic resection. The relationship of prior abdominal radiotherapy to the development of GI LMS warrants further investigation. Outcomes with systemic therapy for metastatic disease were poor and there is a need for the development of more effective systemic therapies.

All multivariate extensions of the univariate theory of risk measurement run into the same fundamental problem of the absence, in dimension d > 1 , of a canonical ordering of R d . Based on measure transportation ideas, several attempts have been made recently in the statistical literature to overcome that conceptual difficulty. In Hallin (2017), the concepts of center-outward distribution and quantile functions are developed as generalizations of the classical univariate concepts of distribution and quantile functions, along with their empirical versions. The center-outward distribution function F ± is a cyclically monotone mapping from R d to the open unit ball B d , while its empirical counterpart F ± ( n ) is obtained as a cyclically monotone mapping from the sample to a regular grid over B d ; in dimension d = 1 , F ± reduces to 2 F − 1 . Based on the concept of Moreau envelope, a smooth interpolation F ˜ ± of F ± has been proposed in del Barrio et al. (2018). Here, we suggest to adapt the definition of the empirical F ± ( n ) so as to relax the presence of ties, which is impractical in the context of risk measurement and propose a class of smooth approximations F n , ξ ( ξ a smoothness index) of F ± ( n ) as an alternative to the F ˜ ± interpolation. Associated with the concepts of center-outward distribution and quantile functions and the associated convex potentials, we construct measures of risk of the maximum correlation type and their estimators based on F ± ( n ) and F n , ξ . We also discuss the use of the volumes of the resulting empirical quantile regions. Some simulations and applications to case studies illustrate the value of the approach.

Agricultural soils are a main source of nitrous oxide (N2O), a potent greenhouse gas and the dominant ozone -depleting substance emitted to the atmosphere. The only known sink of N2O in soil is the microbial reduction of N2O to N2. Carbon (C) availability is a key factor in determining microbial community composition in soil. However, its role in shaping the structure of N2O reducing communities in soil is unexplored. In this study, a microcosm experiment was set up in which two arable soils with contrasting edaphic properties were incubated anaerobically for 83 days with four different C substrates: glucose, acetate, hydroxyethylcellulose (HEC) and mixture of the three. We show that the effect of C addition on the abundance and diversity of clade I and clade II nosZ genes, encoding different variants of the N2O reductase, varies across the different C substrates differently in contrasting soil types, yet still plays an important role in selecting specific taxa of N2O reducers under deni-trifying conditions. We observed an increase of betaproteobacterial clade I and II N2O reducing species with addition of HEC, whereas alphaproteobacterial clade I species and clade II species within other Proteobacteria and Bacteriodetes were associated with glucose and acetate. These results show that specific C-substrates select for certain lineages of nitrous oxide reducers and influence patterns of niche partitioning within clades of N2O reducers, whereas other soil factors drive differences between clade I and II nosZ communities.

Background Initial grading of retroperitoneal leiomyosarcoma (LMS) is performed by core biopsy (CB) however, discrepancy between grade of tumour at initial CB and surgical excision is recognised, raising concerns about the accuracy of CB for directing neoadjuvant therapy. The histological grading system used for staging LMS consists of 3 components: tumour differentiation, mitotic index and proportion of necrosis. We postulate that assessment of necrosis by histopathology alone is inadequate, resulting in under-grading of LMS. We propose and assess a combined grading system that incorporates CT scan findings into pre-surgical grading. Methods Retrospective, blinded review of CT, CB histology and final surgical histology of patients with retroperitoneal LMS was undertaken. A modified grading system, CTH-Grade, was derived by replacing the CB necrosis score with a CT-derived necrosis score. The sensitivity and specificity of CTH-Grade, the standard histopathology scoring, H-grade were compared. Inter-observer variability in assessment of CT necrosis was also assessed. Results 53 patients fulfilled criteria for inclusion. CT was more sensitive at detection of necrosis than CB histology alone with sensitivity of 100% vs 53%. The use of CTHGrade resulted in increased detection of high-grade tumours with CTH-grade having sensitivities of 80% and 35% for Grade 2 and 3 tumours respectively vs 53% and 15% with H-Grade. Assessment of reader agreement demonstrated Kappa scores of 0.8. Conclusion Histology from CB under-grades LMS due to undersampling of tumour necrosis. CT is more sensitive in assessing necrosis and its incorporation into a modified CT-histopathology grading system (CTH-Grade) improves accuracy of grading with significant implications for patient management.

Studies of acute gastroenteritis (AGE) are hampered by the lack of routine diagnostic methods with good sensitivity and specificity. Molecular methods are increasingly used for clinical purposes, but the clinical significance of a positive result remains a challenge. In this study we aimed to compare results of routine diagnostic methods and molecular methods in symptomatic children and asymptomatic controls.Patients presenting to the pediatric emergency departments of two university hospitals in Brussels with AGE were recruited prospectively from May 2015 to October 2016; asymptomatic controls were recruited from the same hospitals. Stool analyses were performed for all participants for common pathogenic bacteria (culture), virus (immunochromatography) and parasites (microscopy). Stools were also analysed with the Luminex Gastrointestinal Pathogen Panel, a multiplex-PCR for common enteropathogens.Stools from 178 patients and 165 controls were analysed. An enteropathogen was detected in 62.4% (111/178) of cases when combining the two methods (56.2% (100/178) by Luminex, 42.7% (76/178) with routine methods) and 29.1% (48/165) of controls (24.2% (40/165) by Luminex and 10.3% (17/165) by routine methods). Some pathogens were detected more often with Luminex than with routine methods, such as Salmonella (16.3% (29/178) with Luminex and 3.9% (7/178) with routine method, p 0.05), whereas others identified by culture methods, such as Campylobacter, Shigella, Yersinia, were missed by Luminex.Molecular tools seem attractive methods, providing high positivity and a rapid turn-around time for the diagnosis of AGE. However, high rates of positivity in both cases and controls highlight the difficulty in interpreting results. Pathogens missed by Luminex but detected by culture methods raise more questions about the true clinical interest of the technique for our patients.

Constructed floating wetlands (CFWs) have been tested in different climatic regions and aquatic habitat types for nitrogen (N) removal from surface water, but there is limited knowledge about their applicability for N removal in cold climate regions. Most CFWs studies are conducted at the micro- or mesocosm scale, while the application of CFWs at in situ is rare. Moreover, most CFWs studies have focused on plant N accumulation without considering macrophyte root-associated denitrification as a possible N removal pathway. Here, we study the N removal potential of CFWs through N accumulation by macrophytes and potential denitrification activity (PDA) associated with plants. At a mining area in the sub-arctic region of Sweden receiving N-rich mine effluents, we tested the concept of CFWs and evaluated the performance of six native, emerging macrophyte species planted in CFWs. The CFWs were deployed in two types of systems: in situ in the recipient lake, subjected to ambient N concentrations, and CFWs placed in water-side “eco-tanks”, subjected to higher N concentrations. We showed that macrophyte establishment in CFWs is feasible under cold climatic conditions, both in situ and eco-tanks. The standing biomass of macrophytes, bulk N accumulation in plant biomass and PDA in mesocosms were 0.54–2.25 kg m−2, 7.56–24.75 mg N m−2 d−1 and 31.82–2250.77 mg N2O-N m−2 d−1, respectively. In the recipient, the variation was larger and the values were higher (standing biomass, 0.37–6.74 kg m−2; bulk N accumulation, 8.09–106.93 mg N m−2 d−1; PDA, 11.89–8446.15 mg N m−2 d−1). Macrophyte root-associated denitrification was the main N removal pathway in the CFWs. Given the demonstrated applicability of CFWs and the high denitrification rates that can be obtained, future studies should focus on designing CFWs to enhance denitrification as this process leads to permanent removal of N from the water phase.

Management of chronic pain is restricted by the lack of effective tools. This study evaluated the efficacies of sinomenine combined gabapentin or ligustrazine hydrochloride in treating peripheral and central chronic neuropathic pain. The study was conducted in mice with photochemically induced sciatic nerve injury, and in rats with photochemically induced spinal cord injury. For assessing the effectiveness of combined therapy, sinomenine, gabapentin or ligustrazine hydrochloride was injected intraperitoneally (i.p.), and pain behavioral tests were performed. At sub-effective dosages, pre-administration of sinomenine (for 60 min) plus gabapentin or ligustrazine hydrochloride, generated significant anti-allodynic effects in mice or rats with peripheral or central neuropathic pain. However, these effects were abolished when gabapentin or ligustrazine hydrochloride was pre-administered, and then sinomenine was given 60 min later. The combined efficacies of sinomenine and gabapentin or ligustrazine hydrochloride, cannot be blocked or reversed by the naloxone, suggesting the underlying mechanism is not mediated by opioid receptors. Moreover, following repeated treatments, sinomenine and gabapentin combination increased the baseline mechanical threshold, while generating prolonged analgesia, without introducing notable side effects. Sinomenine can enhance the efficacy of gabapentin or ligustrazine hydrochloride in rodent models of peripheral or central neuropathic pain, without introducing tolerance or other notable side effects. Findings of current study suggest that combing sinomenine and gabapentin or ligustrazine hydrochloride could be highly beneficial in neuropathic pain therapies.

Explosives used in mining operations release reactive nitrogen (N) that discharge into surrounding waters. Existing pond systems at mine sites could be used for N removal through denitrification and we investigated capacity in tailings and clarification pond sediments at an iron-ore mine site. Despite differences in microbial community structure in the two ponds, the potential denitrification rates were similar, although carbon limited. Therefore, a microcosm experiment in which we amended sediment from the clarification pond with acetate, cellulose or green algae as possible carbon sources was conducted during 10 weeks under denitrifying conditions. Algae and acetate treatments showed efficient nitrate removal and increased potential denitrification rates, whereas cellulose was not different from the control. Denitrifiers were overall more abundant than bacteria performing dissimilatory nitrate reduction to ammonium (DNRA) or anaerobic ammonium oxidation, although DNRA bacteria increased in the algae treatment and this coincided with accumulation of ammonium. The algae addition also caused higher emissions of methane (CH4) and nitrous oxide (N2O). The bacterial community in this treatment had a large proportion of Bacteroidia, sulfate reducing taxa and bacteria known as fermenters. Functional gene abundances indicated an imbalance between organisms that produce N2O in relation to those that can reduce it, with the algae treatment showing the lowest relative capacity for N2O reduction. These findings show that pond sediments have the potential to contribute to mitigating nitrate levels in water from mining industry, but it is important to consider the type of carbon supply as it affects the community composition, which in turn can lead to unwanted processes and increased greenhouse gas emissions.

This study investigates the impact of beach sediment supply on dune volume evolution through data analysis and model simulations of the Kennemer dunes in the Netherlands. A cross-shore sediment transport model (the CS-model) is applied with local time-averaged longshore sediment transport gradients derived from bathymetric and topographic observations. The model is used to study the relative importance of different transport processes on dune volume evolution, assuming that aeolian transport from the beach to the dune is supply-limited. The wave-driven longshore transport gradients are found to explain a large part of the observed variation in the dune evolution within the study area. In accreting parts of the coast, dunes are growing due to sediment supply from longshore transport, whereas in eroding parts, dune growth depends on supply from artificial nourishments. Seasonal constructions on the beach and vegetation removal from the dunes partly impede dune growth along the considered stretch of coast. The model performance is satisfactory, being able to reproduce a considerable part of the large variation in the alongshore dune response observed in the study area. Overall, the results are a promising contribution to the capability of simulating decadal-scale dune evolution, which is important for long-term flood risk assessments and safe designs of nature-based solutions.