Search

Despite immunotherapy's promise in cancer treatment, patient responses vary substantially because of the individual nature of the immune system and the lack of reliable biomarkers. To address this issue, we developed a precision ex vivo platform that integrates patient-specific tumor and immune cells to study the mechanisms of antitumor immune response, predict immunotherapy outcomes, and identify effective treatments. This platform revealed unique single-cell immune response mechanisms and sensitivities to standard-of-care immunotherapies. Furthermore, we were able to identify a synergistic combination of anti-programmed cell death protein 1 (anti-PD-1) together with a Casitas B lineage lymphoma-b inhibitor that overcame anti-PD-1 resistance in selected patient samples. Activation of the interferon-γ-stimulated cytokines predicted combination efficacy, while immunosuppressive cytokines were associated with poor response. Our findings underscore the platform's potential in tailoring immunotherapies and advancing drug development, offering avenues for personalized cancer treatment.

The mRNA vaccines have proven to be very effective in preventing severe disease and death from SARS-CoV-2 in the general population. However, in patients with chronic kidney disease (CKD) in dialysis or with kidney transplants (KT) the vaccine responses vary, with severe breakthrough infections as a consequence. In this intervention study we investigated the magnitude and quality of the responses to mRNA vaccination administered prior to kidney replacement therapy (KRT). Twenty patients with CKD G4/5 and nine healthy controls were followed for 12 months after receiving two doses of BNT162b2 four weeks apart and a booster dose after 3-6 months. Induction of anti-Spike and anti-RBD IgG in plasma followed the same kinetics in CKD patients and controls, with a trend towards higher titers in controls. In accordance, there was no differences in the establishment of Spike-specific memory B-cells between groups. In contrast, the CKD patients showed lower levels of anti-Spike IgG in saliva and Spike-specific CD8 + T-cells in blood, possibly influencing the capacity of viral clearance which can contribute to an elevated risk of severe breakthrough infections. In conclusion, we found that CKD patients, despite having a reduced mucosal and cytotoxic immunity to BNT162b2, demonstrated a serological response in plasma similar to healthy controls. This suggests that immunization prior to RRT is efficient and motivated. (EudraCT-nr 2021-000988-68)., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Anja Rosdahl reports financial support was provided by Orebro Research Council. Fredrika Hellgren reports travel was provided by the Karolinska Institute Foundation for Virus Research, the Karolinska Institute Travle Grant, The Swedisch Society for Immunology and the Keystone Symposia. Karin Lore reports financial support was provided by 10.13039/501100004359Swedish Research Council and 10.13039/501100002794Swedish Cancer Society. Anja Rosdahl and Helena Hervius Askling reports a relationship with National Swedish Infectious Diseases Medical Doctors Vaccine Group that includes: board membership. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Gene regulatory elements, such as enhancers, greatly influence cell identity by tuning the transcriptional activity of specific cell types. Dynamics of enhancer landscape during early human Th17 cell differentiation remains incompletely understood. Leveraging ATAC-seq-based profiling of chromatin accessibility and comprehensive analysis of key histone marks, we identified a repertoire of enhancers that potentially exert control over the fate specification of Th17 cells. We found 23 SNPs associated with autoimmune diseases within Th17-enhancers that precisely overlapped with the binding sites of transcription factors actively engaged in T-cell functions. Among the Th17-specific enhancers, we identified an enhancer in the intron of RORA and demonstrated that this enhancer positively regulates RORA transcription. Moreover, CRISPR-Cas9-mediated deletion of a transcription factor binding site-rich region within the identified RORA enhancer confirmed its role in regulating RORA transcription. These findings provide insights into the potential mechanism by which the RORA enhancer orchestrates Th17 differentiation., Competing Interests: Declaration of competing interest Alexander Marson is a co-founder of Arsenal Biosciences, Spotlight Therapeutics, and Survey Genomics, serves on the boards of directors at Spotlight Therapeutics and Survey Genomics, is a board observer (and former member of the board of directors) at Arsenal Biosciences, is a member of the scientific advisory boards of Arsenal Biosciences, Spotlight Therapeutics, Survey Genomics, NewLimit, Amgen, Tenaya, and Lightcast, owns stock in Arsenal Biosciences, Spotlight Therapeutics, NewLimit, Survey Genomics, PACT Pharma, Tenaya, and Lightcast, and has received fees from Arsenal Biosciences, Spotlight Therapeutics, NewLimit, 23andMe, PACT Pharma, Juno Therapeutics, Tenaya, Lightcast, Trizell, Vertex, Merck, Amgen, Genentech, AlphaSights, Rupert Case Management, Bernstein, and ALDA. A.M. is an investor in and informal advisor to Offline Ventures and a client of EPIQ. The Marson laboratory has received research support from Juno Therapeutics, Epinomics, Sanofi, GlaxoSmithKline, Gilead, and Anthem. All other authors declare no competing interests. The ChIP-seq and ATAC-seq raw and processed data has been submitted to GEO and can be accessed with the following super-series accession number: GSE243064., (Copyright © 2023. Published by Elsevier Inc.)

Somatic mutations in T cells can cause cancer but also have implications for immunological diseases and cell therapies. The mutation spectrum in nonmalignant T cells is unclear. Here, we examined somatic mutations in CD4 + and CD8 + T cells from 90 patients with hematological and immunological disorders and used T cell receptor (TCR) and single-cell sequencing to link mutations with T cell expansions and phenotypes. CD8 + cells had a higher mutation burden than CD4 + cells. Notably, the biggest variant allele frequency (VAF) of non-synonymous variants was higher than synonymous variants in CD8 + T cells, indicating non-random occurrence. The non-synonymous VAF in CD8 + T cells strongly correlated with the TCR frequency, but not age. We identified mutations in pathways essential for T cell function and often affected lymphoid neoplasia. Single-cell sequencing revealed cytotoxic T EMRA phenotypes of mutated T cells. Our findings suggest that somatic mutations contribute to CD8 + T cell expansions without malignant transformation.

Single-cell transcriptomics allows unbiased characterization of cell heterogeneity in a sample by profiling gene expression at single-cell level. These profiles capture snapshots of transient or steady states in dynamic processes, such as cell cycle, activation, or differentiation, which can be computationally ordered into a "flip-book" of cell development using trajectory inference methods. However, prediction of more complex topology structures, such as multifurcations or trees, remains challenging. In this chapter, we present two user-friendly protocols for inferring tree-shaped single-cell trajectories and pseudotime from single-cell transcriptomics data with Totem. Totem is a trajectory inference method that offers flexibility in inferring both nonlinear and linear trajectories and usability by avoiding the cumbersome fine-tuning of parameters. The QuickStart protocol provides a simple and practical example, whereas the GuidedStart protocol details the analysis step-by-step. Both protocols are demonstrated using a case study of human bone marrow CD34+ cells, allowing the study of the branching of three lineages: erythroid, lymphoid, and myeloid. All the analyses can be fully reproduced in Linux, macOS, and Windows operating systems (amd64 architecture) with >8 Gb of RAM using the provided docker image distributed with notebooks, scripts, and data in Docker Hub (elolab/repro-totem-ti). These materials are shared online under open-source license at https://elolab.github.io/Totem-protocol ., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The serine/threonine-specific Moloney murine leukemia virus (PIM) kinase family (i.e., PIM1, PIM2, and PIM3) has been extensively studied in tumorigenesis. PIM kinases are downstream of several cytokine signaling pathways that drive immune-mediated diseases. Uncontrolled T helper 17 (Th17) cell activation has been associated with the pathogenesis of autoimmunity. However, the detailed molecular function of PIMs in human Th17 cell regulation has yet to be studied. In the present study, we comprehensively investigated how the three PIMs simultaneously alter transcriptional gene regulation during early human Th17 cell differentiation. By combining PIM triple knockdown with bulk and scRNA-seq approaches, we found that PIM deficiency promotes the early expression of key Th17-related genes while suppressing Th1-lineage genes. Further, PIMs modulate Th cell signaling, potentially via STAT1 and STAT3. Overall, our study highlights the inhibitory role of PIMs in human Th17 cell differentiation, thereby suggesting their association with autoimmune phenotypes., Competing Interests: Declaration of interests A.M. is a co-founder of Arsenal Biosciences, Spotlight Therapeutics, and Survey Genomics; serves on the boards of directors at Spotlight Therapeutics and Survey Genomics; is a board observer (and former member of the board of directors) at Arsenal Biosciences; is a member of the scientific advisory boards of Arsenal Biosciences, Spotlight Therapeutics, Survey Genomics, NewLimit, Amgen, and Tenaya; owns stock in Arsenal Biosciences, Spotlight Therapeutics, NewLimit, Survey Genomics, PACT Pharma, and Tenaya; and has received fees from Arsenal Biosciences, Spotlight Therapeutics, NewLimit, 23andMe, PACT Pharma, Juno Therapeutics, Trizell, Vertex, Merck, Amgen, Genentech, AlphaSights, Rupert Case Management, Bernstein, and ALDA. A.M. is an investor in and informal advisor to Offline Ventures and a client of EPIQ. The Marson laboratory has received research support from Juno Therapeutics, Epinomics, Sanofi, GlaxoSmithKline, Gilead, and Anthem., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Motivation: Single-cell RNA-sequencing enables cell-level investigation of cell differentiation, which can be modelled using trajectory inference methods. While tremendous effort has been put into designing these methods, inferring accurate trajectories automatically remains difficult. Therefore, the standard approach involves testing different trajectory inference methods and picking the trajectory giving the most biologically sensible model. As the default parameters are often suboptimal, their tuning requires methodological expertise., Results: We introduce Totem, an open-source, easy-to-use R package designed to facilitate inference of tree-shaped trajectories from single-cell data. Totem generates a large number of clustering results, estimates their topologies as minimum spanning trees, and uses them to measure the connectivity of the cells. Besides automatic selection of an appropriate trajectory, cell connectivity enables to visually pinpoint branching points and milestones relevant to the trajectory. Furthermore, testing different trajectories with Totem is fast, easy, and does not require in-depth methodological knowledge., Availability and Implementation: Totem is available as an R package at https://github.com/elolab/Totem., (© The Author(s) 2023. Published by Oxford University Press.)

Single-cell RNA-sequencing (scRNA-seq) enables researchers to quantify transcriptomes of thousands of cells simultaneously and study transcriptomic changes between cells. scRNA-seq datasets increasingly include multisubject, multicondition experiments to investigate cell-type-specific differential states (DS) between conditions. This can be performed by first identifying the cell types in all the subjects and then by performing a DS analysis between the conditions within each cell type. Naïve single-cell DS analysis methods that treat cells statistically independent are subject to false positives in the presence of variation between biological replicates, an issue known as the pseudoreplicate bias. While several methods have already been introduced to carry out the statistical testing in multisubject scRNA-seq analysis, comparisons that include all these methods are currently lacking. Here, we performed a comprehensive comparison of 18 methods for the identification of DS changes between conditions from multisubject scRNA-seq data. Our results suggest that the pseudobulk methods performed generally best. Both pseudobulks and mixed models that model the subjects as a random effect were superior compared with the naïve single-cell methods that do not model the subjects in any way. While the naïve models achieved higher sensitivity than the pseudobulk methods and the mixed models, they were subject to a high number of false positives. In addition, accounting for subjects through latent variable modeling did not improve the performance of the naïve methods., (© The Author(s) 2022. Published by Oxford University Press.)

Motivation: Computational models are needed to infer a representation of the cells, i.e. a trajectory, from single-cell RNA-sequencing data that model cell differentiation during a dynamic process. Although many trajectory inference methods exist, their performance varies greatly depending on the dataset and hence there is a need to establish more accurate, better generalizable methods., Results: We introduce scShaper, a new trajectory inference method that enables accurate linear trajectory inference. The ensemble approach of scShaper generates a continuous smooth pseudotime based on a set of discrete pseudotimes. We demonstrate that scShaper is able to infer accurate trajectories for a variety of trigonometric trajectories, including many for which the commonly used principal curves method fails. A comprehensive benchmarking with state-of-the-art methods revealed that scShaper achieved superior accuracy of the cell ordering and, in particular, the differentially expressed genes. Moreover, scShaper is a fast method with few hyperparameters, making it a promising alternative to the principal curves method for linear pseudotemporal ordering., Availability and Implementation: scShaper is available as an R package at https://github.com/elolab/scshaper. The test data are available at https://doi.org/10.5281/zenodo.5734488., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2021. Published by Oxford University Press.)

Data from the Swedish Renal Registry (SRR) show that during the period March 16, 2020 to March 15, 2021 0.4% of all renal transplant recipients and 3% of all dialysis patients died due to COVID-19 in Sweden. Of all registered deaths, 20% were attributed to COVID-19.  In the age group 50-59 years the risk ratio for COVID-19 related mortality was 16 (95% CI 6.5-38) among transplant recipients and 22 (95% CI 7.1-69) among dialysis patients, compared to the background population in the same age group. Excess mortality, compared to the five years preceding the pandemic, was 30% for transplant recipients and 8.7% for dialysis patients, compared to 7.7% for the entire Swedish population. Detailed reports were sent to SRR for 864 patients with confirmed COVID-19 infection representing 5.0% of all transplant recipients and 13% of all dialysis patients. The case fatality rate was 7.0% and 21% respectively.

Motivation: Single-cell RNA-seq allows researchers to identify cell populations based on unsupervised clustering of the transcriptome. However, subpopulations can have only subtle transcriptomic differences and the high dimensionality of the data makes their identification challenging., Results: We introduce ILoReg, an R package implementing a new cell population identification method that improves identification of cell populations with subtle differences through a probabilistic feature extraction step that is applied before clustering and visualization. The feature extraction is performed using a novel machine learning algorithm, called iterative clustering projection (ICP), that uses logistic regression and clustering similarity comparison to iteratively cluster data. Remarkably, ICP also manages to integrate feature selection with the clustering through L1-regularization, enabling the identification of genes that are differentially expressed between cell populations. By combining solutions of multiple ICP runs into a single consensus solution, ILoReg creates a representation that enables investigating cell populations with a high resolution. In particular, we show that the visualization of ILoReg allows segregation of immune and pancreatic cell populations in a more pronounced manner compared with current state-of-the-art methods., Availability and Implementation: ILoReg is available as an R package at https://bioconductor.org/packages/ILoReg., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2020. Published by Oxford University Press.)

Background: Detection of copy number variations (CNVs) from high-throughput next-generation whole-genome sequencing (WGS) data has become a widely used research method during the recent years. However, only a little is known about the applicability of the developed algorithms to ultra-low-coverage (0.0005-0.8×) data that is used in various research and clinical applications, such as digital karyotyping and single-cell CNV detection., Result: Here, the performance of six popular read-depth based CNV detection algorithms (BIC-seq2, Canvas, CNVnator, FREEC, HMMcopy, and QDNAseq) was studied using ultra-low-coverage WGS data. Real-world array- and karyotyping kit-based validation were used as a benchmark in the evaluation. Additionally, ultra-low-coverage WGS data was simulated to investigate the ability of the algorithms to identify CNVs in the sex chromosomes and the theoretical minimum coverage at which these tools can accurately function. Our results suggest that while all the methods were able to detect large CNVs, many methods were susceptible to producing false positives when smaller CNVs (< 2 Mbp) were detected. There was also significant variability in their ability to identify CNVs in the sex chromosomes. Overall, BIC-seq2 was found to be the best method in terms of statistical performance. However, its significant drawback was by far the slowest runtime among the methods (> 3 h) compared with FREEC (~ 3 min), which we considered the second-best method., Conclusions: Our comparative analysis demonstrates that CNV detection from ultra-low-coverage WGS data can be a highly accurate method for the detection of large copy number variations when their length is in millions of base pairs. These findings facilitate applications that utilize ultra-low-coverage CNV detection.

Rheumatoid arthritis (RA) is a complex autoimmune disease targeting synovial joints. Traditionally, RA is divided into seropositive (SP) and seronegative (SN) disease forms, the latter consisting of an array of unrelated diseases with joint involvement. Recently, we described a severe form of SN-RA that associates with characteristic joint destruction. Here, we sought biological characteristics to differentiate this rare but aggressive anti-citrullinated peptide antibody-negative destructive RA (CND-RA) from early seropositive (SP-RA) and seronegative rheumatoid arthritis (SN-RA). We also aimed to study cytotoxic CD8+ lymphocytes in autoimmune arthritis. CND-RA, SP-RA and SN-RA were compared to healthy controls to reveal differences in T-cell receptor beta (TCRβ) repertoire, cytokine levels and autoantibody repertoires. Whole-exome sequencing (WES) followed by single-cell RNA-sequencing (sc-RNA-seq) was performed to study somatic mutations in a clonally expanded CD8+ lymphocyte population in an index patient. A unique TCRβ signature was detected in CND-RA patients. In addition, CND-RA patients expressed higher levels of the bone destruction-associated TNFSF14 cytokine. Blood IgG repertoire from CND-RA patients recognized fewer endogenous proteins than SP-RA patients' repertoires. Using WES, we detected a stable mutation profile in the clonally expanded CD8+ T-cell population characterized by cytotoxic gene expression signature discovered by sc-RNA-sequencing. Our results identify CND-RA as an independent RA subset and reveal a CND-RA specific TCR signature in the CD8+ lymphocytes. Improved classification of seronegative RA patients underlines the heterogeneity of RA and also, facilitates development of improved therapeutic options for the treatment resistant patients., (Copyright © 2020 Kelkka, Savola, Bhattacharya, Huuhtanen, Lönnberg, Kankainen, Paalanen, Tyster, Lepistö, Ellonen, Smolander, Eldfors, Yadav, Khan, Koivuniemi, Sjöwall, Elo, Lähdesmäki, Maeda, Nishikawa, Leirisalo-Repo, Sokka-Isler and Mustjoki.)

Acute kidney injury (AKI), albuminuria and hematuria are common in Covid-19 and have been shown to increase mortality. Assessment with a urinary dipstick and creatinine at admission should be completed with a urinary sediment and quantification of albuminuria if positive. SARS-Cov-2 seems to enter and infect the endothelium and kidney cells, and contributes to damage in addition to hypercoagulability, multi organ failure and hyperinflammation. Underhydration and rhabdomyolysis can contribute to acute tubular necrosis. Anti-inflammatory treatment may be considered and discussed with a nephrologist. Treatment with ACEi/ARBs should be continued if possible.

There are many important sport events that are organized in environments with a very hot ambient temperature (Summer Olympics, FIFA World Cup, Tour de France, etc.) and in hot locations (e.g., Qatar). Additionally, in the context of global warming and heat wave periods, athletes are often subjected to hot ambient temperatures. It is known that exercising in the heat induces disturbances that may provoke premature fatigue and negatively affects overall performance in both endurance and high intensity exercises. Deterioration in several cognitive functions may also occur, and individuals may be at risk for heat illnesses. To train, perform, work and recover and in a safe and effective way, cooling strategies have been proposed and have been routinely applied before, during and after exercise. However, there is a limited understanding of the influences of per-cooling on performance, and it is the subject of the present review. This work examines the influences of per-cooling of different areas of the body on performance in terms of intense short-term exercises ("anaerobic" exercises), endurance exercises ("aerobic" exercises), and cognitive functioning and provides detailed strategies that can be applied when individuals train and/or perform in high ambient temperatures.

Background: Deciphering the meaning of the human DNA is an outstanding goal which would revolutionize medicine and our way for treating diseases. In recent years, non-coding RNAs have attracted much attention and shown to be functional in part. Yet the importance of these RNAs especially for higher biological functions remains under investigation., Methods: In this paper, we analyze RNA-seq data, including non-coding and protein coding RNAs, from lung adenocarcinoma patients, a histologic subtype of non-small-cell lung cancer, with deep learning neural networks and other state-of-the-art classification methods. The purpose of our paper is three-fold. First, we compare the classification performance of different versions of deep belief networks with SVMs, decision trees and random forests. Second, we compare the classification capabilities of protein coding and non-coding RNAs. Third, we study the influence of feature selection on the classification performance., Results: As a result, we find that deep belief networks perform at least competitively to other state-of-the-art classifiers. Second, data from non-coding RNAs perform better than coding RNAs across a number of different classification methods. This demonstrates the equivalence of predictive information as captured by non-coding RNAs compared to protein coding RNAs, conventionally used in computational diagnostics tasks. Third, we find that feature selection has in general a negative effect on the classification performance which means that unfiltered data with all features give the best classification results., Conclusions: Our study is the first to use ncRNAs beyond miRNAs for the computational classification of cancer and for performing a direct comparison of the classification capabilities of protein coding RNAs and non-coding RNAs.

Background: Alzheimer's disease results from a neurodegenerative process that starts well before the diagnosis can be made. New prognostic or diagnostic markers enabling early intervention into the disease process would be highly valuable. Environmental and lifestyle factors largely modulate the disease risk and may influence the pathogenesis through epigenetic mechanisms, such as DNA methylation. As environmental and lifestyle factors may affect multiple tissues of the body, we hypothesized that the disease-associated DNA methylation signatures are detectable in the peripheral blood of discordant twin pairs., Results: Comparison of 23 disease discordant Finnish twin pairs with reduced representation bisulfite sequencing revealed peripheral blood DNA methylation differences in 11 genomic regions with at least 15.0% median methylation difference and FDR adjusted p value ≤ 0.05. Several of the affected genes are primarily associated with neuronal functions and pathologies and do not display disease-associated differences in gene expression in blood. The DNA methylation mark in ADARB2 gene was found to be differentially methylated also in the anterior hippocampus, including entorhinal cortex, of non-twin cases and controls. Targeted bisulfite pyrosequencing of the DNA methylation mark in ADARB2 gene in 62 Finnish and Swedish twin pairs revealed that, in addition to the disease status, DNA methylation of this region is influenced by gender, age, zygosity, APOE genotype, and smoking. Further analysis of 120 Swedish twin pairs indicated that this specific DNA methylation mark is not predictive for Alzheimer's disease and becomes differentially methylated after disease onset., Conclusions: DNA methylation differences can be detected in the peripheral blood of twin pairs discordant for Alzheimer's disease. These DNA methylation signatures may have value as disease markers and provide insights into the molecular mechanisms of pathogenesis. We found no evidence that the DNA methylation marks would be associated with gene expression in blood. Further studies are needed to elucidate the potential importance of the associated genes in neuronal functions and to validate the prognostic or diagnostic value of the individual marks or marker panels.

Genomics data provide great opportunities for translational research and the clinical practice, for example, for predicting disease stages. However, the classification of such data is a challenging task due to their high dimensionality, noise, and heterogeneity. In recent years, deep learning classifiers generated much interest, but due to their complexity, so far, little is known about the utility of this method for genomics. In this paper, we address this problem by studying a computational diagnostics task by classification of breast cancer and inflammatory bowel disease patients based on high-dimensional gene expression data. We provide a comprehensive analysis of the classification performance of deep belief networks (DBNs) in dependence on its multiple model parameters and in comparison with support vector machines (SVMs). Furthermore, we investigate combined classifiers that integrate DBNs with SVMs. Such a classifier utilizes a DBN as representation learner forming the input for a SVM. Overall, our results provide guidelines for the complex usage of DBN for classifying gene expression data from complex diseases., (© 2019 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)

Background: Approximately 30% of adult patients with immunoglobulin A (IgA) nephropathy (IgAN) or IgA vasculitis with nephritis (IgAVN) develop end-stage renal disease during long-term follow-up. In particular, patients with nephritic-nephrotic syndrome have an increased risk of rapid progression. Conventional immunosuppressive therapy with corticosteroids (CSs) may be insufficient for disease control and is associated with a number of side effects. Rituximab (RTX) has been shown to be well tolerated and effective in a range of glomerular diseases, but there is little information on its therapeutic potential in IgAN. The humanized anti-CD20 monoclonal antibody ofatumumab (OFAB) may be an alternative drug for patients intolerant or unresponsive to RTX, but so far there is no report on its use in IgAVN or IgAN., Methods: We describe clinical outcomes after 17-22 months in four adult patients with biopsy-confirmed IgAVN or IgAN treated with RTX or OFAB as well as CS soon after diagnosis. All presented with nephritic-nephrotic syndrome and one had crescentic IgAN. Rebiopsy was performed in two cases., Results: RTX and OFAB were well tolerated. Albuminuria was <250 mg/day in three patients at last evaluation and two regained normal renal function. In all cases, renal function improved after therapy. In one patient with severe IgA vasculitis, rebiopsy showed disappearance of subendothelial but not mesangial immune complexes. In the case with crescentic IgAN, rebiopsy after 9 months showed no active necrotic lesions., Conclusions: B cell-depleting therapy may be an alternative treatment for patients with IgAN or IgAVN and nephritic-nephrotic syndrome. A possible CS-sparing effect should be further evaluated in randomized controlled clinical trials.

The arteriovenous anastomoses (AVAs) in the distal parts of the extremities play a significant role in the heat exchange with the environment. The aim of the study was to examine the thermal responses to whole-body cooling in air, and especially the behaviour of finger skin temperature (T(f) , rich in AVAs). Eight young men sat in minimal clothing at 32°C air temperature (T(a) ), which was then lowered gradually to 13°C in 100 min. In the beginning of cooling, T(f) was high and fluctuating, and then suddenly exhibited a rapid fall, while temperatures in other skin sites fell fairly linearly along decreasing T(a) to the end of cooling. During the period from start to the rapid fall in T(f) , rectal temperature decreased from 37·4°C (SD 0.2) to 37·2°C (0·2), mean skin temperature (T(sk) ) from 34·6°C (0·5) to 31·2°C (2·0) and whole-body thermal sensation from 'slightly warm/warm' to 'slightly cool/cold'. The start of the steep fall in T(f) varied considerably between individuals in terms of time (2-75 min), T(a) (16·7 - 32·0°C) and T(sk) (28·8 - 34·7°C). On the other hand, the range of T(f) at that point was narrower (32·1 - 35·8°C). The findings stress the importance of taking into account the distal skin temperatures in thermoregulatory studies in addition to the ordinarily used more proximal and central skin sites. Also, it might be advisable to start such experiments with relatively high and fluctuating T(f) to guarantee that the thermal state of the subject is well defined., (© 2012 The Authors Clinical Physiology and Functional Imaging © 2012 Scandinavian Society of Clinical Physiology and Nuclear Medicine.)

Heart rate (HR) as an estimator of oxygen consumption (VO(2) ) usually requires HR to be individually calibrated in a separate test. This study examined the validity of a new HR - and HR variability-based method (Firstbeat PRO heartbeat analysis software) in the estimation of VO(2) in real-life tasks. The method takes into account the respiration rate determined from HR variability and the differences in the on/off dynamics of HR and VO(2) , and no calibration tests are needed. Ten men and nine women performed 25 tasks representing different types of daily activities. Portable devices were used to measure R-to-R intervals (ECG), VO(2) and respiration rate. In pooled regression analysis, the estimated VO(2) accounted for 87% of the variability in the actual VO(2) , SEE 3·5 ml min(-1) kg(-1) (1 MET). At group level, the method underestimated slightly the measured VO(2) (mean difference - 1·5 ml min(-1) kg(-1) or - 0·4 METs). Some of the values at low exercise intensities were markedly underestimated, but the agreement was better during light and heavy activities. The limits of agreement for the data were from -8·4 to 5·4 ml min(-1) kg(-1) or from -2·4 to 1·5 METs. At individual level, the average deviations of the predicted VO(2) ranged from -1·0 to 0·6 METs and R(2) from 0·77 to 0·94, respectively. The present data indicate that the prediction method may be considered sufficiently accurate to determine the average VO(2) in field use, but it does not allow precise estimation of VO(2) ., (© 2011 The Authors. Clinical Physiology and Functional Imaging © 2011 Scandinavian Society of Clinical Physiology and Nuclear Medicine.)

Background: Limited data exist on the associations between muscle performance and work ability measures in working age adults., Aims: To evaluate how the results of simple muscle performance tests correlate with self-reported work ability and physical functioning in middle-aged men., Methods: Muscle performance was measured with handgrip, sit-up, arch-up, squatting and lifting tests. The Physical Functioning scale and the Role Limitations due to Physical Health Problems scale of the RAND-36 health questionnaire were used to assess functioning and the Work Ability Index (WAI) questionnaire to assess perceived work ability in a subgroup of 51 subjects., Results: A total of 104 men aged 45-55 years employed in physically active work participated. Muscle performance was weakly associated with WAI and functioning measures, accounting for 10% of the variance at most. Dynamic lifting test results for both arms correlated with WAI scores (right: r = 0.31, left: r = 0.34). Scores of the dynamic lifting test for both arms correlated with the Physical Functioning scores (right: r = 0.23, left: r = 0.28) and with the Role Limitations due to Physical Health Problems scores (right: r = 0.25, left: r = 0.28). Results of the squatting test were associated with the Physical Functioning scores (r = 0.24)., Conclusions: The study failed to provide support for the use of muscle performance tests in work-related fitness evaluations in middle-aged men employed in physical jobs.

C.E.R.A., a continuous erythropoietin receptor activator, is a new third-generation erythropoiesis-stimulating agent (ESA) that has recently been linked with abuse in endurance sports. In order to combat this new form of doping, we examined an enzyme-linked immunosorbent assay (ELISA) designed to detect the presence of C.E.R.A. in serum samples. The performance of the assay was evaluated using a pilot excretion study that involved six subjects receiving C.E.R.A. Validation data demonstrated an excellent reproducibility and ensured the applicability of the assay for anti-doping purposes. To maximize the chances of detecting the drug in serum samples, we propose the use of this specific ELISA test as a high-throughput screening method, combined with a classic isoelectric focusing test as a confirmatory assay. This strategy should make C.E.R.A. abuse relatively easy to detect, thereby preventing the future use of this drug as a doping agent.

The endothelin receptor antagonist avosentan may cause fluid overload at doses of 25 and 50 mg, but the actual mechanisms of this effect are unclear. We conducted a placebo-controlled study in 23 healthy subjects to assess the renal effects of avosentan and the dose dependency of these effects. Oral avosentan was administered once daily for 8 days at doses of 0.5, 1.5, 5, and 50 mg. The drug induced a dose-dependent median increase in body weight, most pronounced at 50 mg (0.8 kg on day 8). Avosentan did not affect renal hemodynamics or plasma electrolytes. A dose-dependent median reduction in the fractional renal excretion of sodium was found (up to 8.7% at avosentan 50 mg); this reduction was paralleled by a dose-related increase in proximal sodium reabsorption. It is suggested that avosentan dose-dependently induces sodium retention by the kidney, mainly through proximal tubular effects. The potential clinical benefits of avosentan should therefore be investigated at doses of

Cold therapy is used to relieve pain and inflammatory symptoms. Humoral changes may account for the pain alleviation related to the cold exposures. The aim of the present study was to examine the effects of two types of cold therapy, winter swimming in ice-cold water (WS) and whole body cryotherapy (WBC), on the serum levels of the growth hormone, prolactin, thyrotropin and free fractions of thyroid hormones (fT3, fT4). One group of healthy females (n = 6) was exposed to WS (water 0-2 degrees C) for 20 s and another group (n = 6) to WBC (air 110 degrees C) for 2 min, three times a week for 12 weeks. Blood samples used for the hormone measurements were taken on weeks 1, 4 and 12 before and 35 min after the cold exposures and on the days of the respective weeks, when the cold exposures were not performed. During the WS treatments, serum thyrotropin increased significantly at 35 min on weeks 1 (p < 0.01) and 4 (p < 0.05), but the responses were within the health-related reference interval. During the WS, the serum prolactin measured at 35 min on week 12 was lower than during the control treatment, and no changes in fT3 or fT4 were observed. During the WBC, no changes in the serum levels of the studied hormones were observed during the 12 weeks. In conclusion, repeated WS and WBC treatments for healthy females do not lead to disorders related to altered secretions of the growth hormone, prolactin, thyrotropin, or thyroid hormones.

The Work ability of ageing work force is a matter of major concern in many countries. The aim of this study was to examine the relationship between perceived work ability and health-related quality of life (HRQoL), and to investigate their associations with age, physical activity and physical fitness in middle-aged men working in blue-collar occupations. The study population consisted of 196 middle-aged (aged 40-60 years) men (construction and industrial work) attending occupationally orientated early medical rehabilitation. They were mostly healthy having only symptoms of musculoskeletal or psychological strain. Perceived work ability was assessed with the work ability index (WAI) and HRQoL with the Rand, 36-item health survey (Rand-36). Information on physical activity was obtained with a structured questionnaire. Cardiorespiratory fitness was estimated with a submaximal exercise test on a cycle-ergometer. The WAI was significantly (p<0.001) associated with the total score of Rand-36, and with all its domains. Age, physical activity and cardiorespiratory fitness were neither associated with the WAI, nor did physical activity predict any of the dimensions of Rand-36. Cardiorespiratory fitness was associated with the physical functioning dimension of the Rand-36 whilst age was positively associated with the dimensions of the energy, emotional well being and social functioning of the Rand-36. The present study on middle-aged men showed a close relationship between perceived work ability and the HRQoL. It is suggested that the promotion of work ability may have beneficial effects on quality of life.

Traditionally, the estimation of oxygen consumption (VO2) at work using heart rate (HR) has required the determination of individual HR/VO2 calibration curves in a separate exercise test in a laboratory (VO2-TRAD). Recently, a new neural network-, and heart rate variability-based method has been developed (Firstbeat PRO heartbeat analysis software) for the estimation of VO2 without individual calibration (VO2-HRV). In the present study, the VO2-values by the VO2-HRV were compared with the values by VO2-TRAD in 22 postal workers. Within individuals the correlation between the two methods was high (range 0.80-0.99). The VO2-TRAD gave higher values of VO2 compared to VO2-HRV (19%) especially during low physical activity work when non-metabolic factors may increase HR. When assessed in different HR categories, the smallest difference (11%), and highest correlations (range 0.83-0.99) in VO2 between the methods were observed at higher HR levels. The results indicate that the VO2-HRV is a potentially useful method to estimate VO2 in the field without laboratory calibration.

Objective: Cold therapy is used to relieve pain and inflammatory symptoms. The present study was designed to determine the influence of long-term regular exposure to acute cold temperature. Two types of exposure were studied: winter swimming in ice-cold water and whole-body cryotherapy. The outcome was investigated on humoral factors that may account for pain alleviation related to the exposures., Material and Methods: During the course of 12 weeks, 3 times a week, a group of healthy females (n = 10) was exposed to winter swimming (water 0-2 degrees C) for 20 s and another group (n = 10) to whole-body cryotherapy (air -110 degrees C) for 2 min in a special chamber. Blood specimens were drawn in weeks 1, 2, 4, 8 and 12, on a day when no cold exposure occurred (control specimens) and on a day of cold exposures (cold specimens) before the exposures (0 min), and thereafter at 5 and 35 min., Results: Plasma ACTH and cortisol in weeks 4-12 on time-points 35 min were significantly lower than in week 1, probably due to habituation, suggesting that neither winter swimming nor whole-body cryotherapy stimulated the pituitary-adrenal cortex axis. Plasma epinephrine was unchanged during both experiments, but norepinephrine showed significant 2-fold to 3-fold increases each time for 12 weeks after both cold exposures. Plasma IL-1-beta, IL-6 or TNF alpha did not show any changes after cold exposure., Conclusions: The main finding was the sustained cold-induced stimulation of norepinephrine, which was remarkably similar between exposures. The frequent increase in norepinephrine might have a role in pain alleviation in whole-body cryotherapy and winter swimming.

Objectives: In Finland, testing cardiorespiratory fitness (VO2max) is popular in health promotion programs and work ability evaluations. The most common instruments used for this purpose are the submaximal cycle ergometer test, and the 2-km walking test. However, limited data exist on the associations between VO2max and wellbeing in working age adults. The aim of the study was to evaluate how the measured (cycle ergometer) and the estimated (walking test) VO2max and walking performance are associated with health-related quality of life and work ability., Materials and Methods: The subjects were 104 middle-aged men workers (45-55 years old), mostly from the construction and manufacturing industries. VO2max was directly measured by a maximal exercise test on a cycle ergometer. The 2-km walking test parameters were the walking time, predicted VO2max, and fitness index. Health-related quality of life was assessed with the RAND-36 questionnaire which was further divided into physical dimensions (P-RAND-36) and mental dimensions (M-RAND-36). Perceived work ability was assessed with the work ability index (WAI) in a subgroup of 51 subjects., Results: The 2-km walking test parameters significantly predicted the score on P-RAND-36 (r2=0.18, p=0.001), and correlated significantly with WAI. The directly measured VO2max was not associated with P-RAND-36, M-RAND-36 or WAI., Conclusions: The inexpensive 2-km walking test may be more useful when evaluating the quality of life and work ability, compared to the more expensive direct measurement of one's cardiorespiratory fitness in a laboratory.

Whole-body cryotherapy (WBC) is one mode of cold therapy, during which rheumatic patients are exposed to very cold air (-110 degrees C) in minimal clothing. It is also proposed to have a bronchodilatory effect. The aim was to examine the effects of WBC on lung function in healthy humans after acute and repeated exposures. Twenty-five healthy, non-smoking subjects participated in the study. They were exposed to WBC for 2 min three times per week for 12 weeks. The peak expiratory flow rate (PEF) and forced expiratory volume in 1 s (FEV1) were measured before and after (at 2 and 30 min) the first WBC, and then similarly at 4, 8 and 12 weeks. At all time points, after 30 min of the WBC the PEF values were slightly lower compared with values before the WBC, and the reductions reached statistical significance at 1 month (5.1 +/- 1.2%), and at 3 months (3.2 +/- 1.7%). After 30 min of the first WBC, the FEV1 was significantly reduced by 2.3 +/- 0.8%, but no other changes were observed during the study. In conclusion, the WBC induced minor bronchoconstriction in healthy humans instead of proposed bronchodilatation. The WBC seems not to be harmful for lung function, but should be used with caution in susceptible individuals.

The influence of an upper-limb interval-training program after total hip arthroplasty (THA) in elderly patients was studied during a 1 yr follow-up on health status and walking ability. After surgery, 14 patients were randomly assigned to the control group that started a 6 wk general rehabilitation program or the training group that combined it with an interval exercise program on an arm ergometer. A Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index was completed 1 mo before and 2 mo and 1 yr after surgery. A 6 min walking test was performed at 2 mo and 1 yr after surgery. The training group covered a significantly longer distance than the control group in the 6 min walk test and obtained significantly lower WOMAC scores both at 2 mo and 1 yr after surgery. However, both groups significantly improved in WOMAC and in their performance during the survey. Endurance-type upper-body aerobic training in a rehabilitation program might be important after THA.

The effects of severe cold stress on total peroxyl radical trapping antioxidant capacity of plasma (TRAP) were studied in two groups of healthy women: a whole-body cryotherapy group (WBC, n = 10) and a winter swimming group (WS, n = 10). The biovariability of TRAP values was also analysed. The WBC group was exposed to -110 degrees C for 2 min, whereas the exposure for the WS group lasted 20 s in ice-cold water. Sessions were organized three times per week for 12 weeks. Blood specimens were collected at 2, 4, 8 and 12 weeks at rest, 2 and 35 min after the cold exposures and at the corresponding times without cold exposure on a separate day. Conventional methods were used to determine TRAP values. The between-subject variation was 13.6% and the within-subject variation 6.4%. The index of individuality was 0.46, and the index of heterogeneity was 0.079. These results indicate a marked heterogeneity among subjects. During the first 4 weeks, the mean TRAP value significantly increased at 2 min after cold exposure in the WBC group, returning to baseline 35 min after the exposure. Similar changes were observed in the WS group. However, all changes due to cold were relatively mild (<5%). After 4 weeks no changes in TRAP values after the cold exposures were noticed and no long-term changes in basal TRAP values were observed. In the main, regular WBC and WS do not seem to be harmful as far as plasma antioxidative capacity is concerned.