Search

Summary: In this study, we optimized the dissociation of synovial tissue biopsies for single-cell omics studies and created a single-cell atlas of human synovium in inflammatory arthritis. The optimized protocol allowed consistent isolation of highly viable cells from tiny fresh synovial biopsies, minimizing the synovial biopsy drop-out rate. The synovium scRNA-seq atlas contained over 100,000 unsorted synovial cells from 25 synovial tissues affected by inflammatory arthritis, including 16 structural, 11 lymphoid, and 15 myeloid cell clusters. This synovial cell map expanded the diversity of synovial cell types/states, detected synovial neutrophils, and broadened synovial endothelial cell classification. We revealed tissue-resident macrophage subsets with proposed matrix-sensing (FOLR2+COLEC12high) and iron-recycling (LYVE1+SLC40A1+) activities and identified fibroblast subsets with proposed functions in cartilage breakdown (SOD2highSAA1+SAA2+SDC4+) and extracellular matrix remodeling (SERPINE1+COL5A3+LOXL2+). Our study offers an efficient synovium dissociation method and a reference scRNA-seq resource, that advances the current understanding of synovial cell heterogeneity in inflammatory arthritis.

AimTo determine characteristics associated with more severe outcomes in a global registry of people with systemic lupus erythematosus (SLE) and COVID-19.MethodsPeople with SLE and COVID-19 reported in the COVID-19 Global Rheumatology Alliance registry from March 2020 to June 2021 were included. The ordinal outcome was defined as: (1) not hospitalised, (2) hospitalised with no oxygenation, (3) hospitalised with any ventilation or oxygenation and (4) death. A multivariable ordinal logistic regression model was constructed to assess the relationship between COVID-19 severity and demographic characteristics, comorbidities, medications and disease activity.ResultsA total of 1606 people with SLE were included. In the multivariable model, older age (OR 1.03, 95% CI 1.02 to 1.04), male sex (1.50, 1.01 to 2.23), prednisone dose (1-5 mg/day 1.86, 1.20 to 2.66, 6-9 mg/day 2.47, 1.24 to 4.86 and ≥10 mg/day 1.95, 1.27 to 2.99), no current treatment (1.80, 1.17 to 2.75), comorbidities (eg, kidney disease 3.51, 2.42 to 5.09, cardiovascular disease/hypertension 1.69, 1.25 to 2.29) and moderate or high SLE disease activity (vs remission; 1.61, 1.02 to 2.54 and 3.94, 2.11 to 7.34, respectively) were associated with more severe outcomes. In age-adjusted and sex-adjusted models, mycophenolate, rituximab and cyclophosphamide were associated with worse outcomes compared with hydroxychloroquine; outcomes were more favourable with methotrexate and belimumab.ConclusionsMore severe COVID-19 outcomes in individuals with SLE are largely driven by demographic factors, comorbidities and untreated or active SLE. Patients using glucocorticoids also experienced more severe outcomes.

Idiopathic Inflammatory Myopathies are rare conditions with several heterogeneous disease subtypes. They can range from limited muscle or skin involvement to severe, systemic, life-threatening disease. Although the etiology is unknown, some evidence suggests a role for external agents, particularly drugs. Herein, we present a case of a 71-year-old woman with chronic myeloid leukemia who developed imatinib-induced dermatomyositis sine dermatitis. The presentation was predominantly muscular, characterized by proximal muscle weakness and myalgia of the lower limbs, with positive anti-Mi2a antibodies. Spontaneous recovery was observed after drug discontinuation, without the need for immunosuppressive therapy. This is the first confirmed description of an imatinib-induced dermatomyositis sine dermatitis. It reflects the importance of a high awareness from rheumatologists and hematologists to accurately anticipate and identify similar situations.

IntroductionMaturation in seawater-adapted (smoltified) male Atlantic salmon is undesirable yet frequently reported in fish weighing up to 1 kg reared under intensive conditions in land-based facilities. Although the photoperiod regulates both smoltification and maturation, the optimal post-smoltification regimen for the prevention of maturation is unclear.MethodsIn this study, mixed-sex Atlantic salmon weighing 95 g were exposed to three different photoperiod regimes: continuous daylength, gradually decreasing daylength, and short-to-long daylength after the induction of smoltification by the “winter signal” (12 h:12 h light:dark). Fish were maintained under these conditions in triplicate for 18 weeks in a brackish water flow-through system at 12°C and with a salinity of 20 ppt. This initial phase was followed by a grow-out period (1 kg–2.5 kg) in seawater (32-ppt salinity) at 13°C for 11 weeks, with exposure to decreasing daylength. Throughout the 29-week study, body condition, gonad development, and morphological traits were assessed. Additionally, during exposure to brackish water, serum 11-ketotestosterone (11-KT) levels were quantified, and the testes were examined histologically. Upon termination of the study, the maturation rate was determined.ResultsThis study exclusively detected maturation among male salmon. Under continuous light exposure, the number of fish initiating maturity steadily increased from week 8 and continued to increase in the subsequent samplings. In the short-to-long daylength group, a subset of male fish showed elevated 11-KT levels at week 14, although their gonadal development and morphological changes were not evident until week 18. Male fish exposed to decreasing daylength showed no substantial increase in 11-KT levels, although a few fish exhibited advanced gonad development by week 18. At the end of the trial, markedly lower percentages of maturing and fully mature male fish were observed among those fish previously exposed to decreasing daylength. The maturation rates at study termination were 44.4% for the short-to-long daylength group, 29.4% for the continuous daylength group, and 4.3% for the decreasing daylength group.DiscussionThese results show that photoperiod can be utilized to regulate maturation in male Atlantic salmon. In addition, decreasing the photoperiod after smoltification seems to limit early sexual maturation without compromising body size development. Thus, decreasing the photoperiod could be utilized to limit maturation in the production of post-smolt salmon or salmon in land-based facilities.

IntroductionThe use of recirculating aquaculture systems (RAS) to produce Atlantic salmon smolts has resulted in exceptionally high fish growth rates. However, there are potential negative trade-offs between fast growth and key physiological processes, such as inadequate smoltification and early sexual maturation, which can both be linked to body energy reserves.MethodsThis study determined the effect of i) dietary fat levels on Atlantic salmon whole-body fat and fatty acids composition, growth performance and smoltification and ii) a previous dietary regime on seawater growth performance and male early sexual maturation. In freshwater RAS, salmon parr (~19g) were fed 3 fat levels (20, 24, and 28%) over a 14-week period at 12°C. Subsequently, in seawater flow-through systems, smolt (~96g) were fed a control diet (26%) for 12 weeks at 2 temperatures (12 and 16°C).ResultsDietary fat levels resulted in differences in k-factor, fish whole-body fat and fatty acids composition; the low fat diet resulted in fish with a 23% lower whole-body fat when compared to the other two dietary groups at the end of the freshwater phase. These differences in whole-body fat faded at the end of the seawater phase. During the freshwater phase, all three feeds resulted in a comparable growth performance and smoltification indicator values, including k-factor, gill Na+, K+-ATPase, blood serum chloride and smolt index score. In contrast, water temperature resulted in fish body weight differences, where groups reared at 16°C were larger than those reared at 12°C. Nevertheless, both temperature regimes supported a similar thermal growth coefficient. Smolts grown at 16°C showed a higher level of the sex steroids androstenedione compared to those at 12°C. However, 11-ketotestosterone and testosterone levels did not differ despite a trend for higher levels at 16°C. Furthermore, testis histology at the final sampling indicated that some individuals showed initial signs of maturation (stage 3).DiscussionIn conclusion, varying dietary fat levels (20 - 28%) during the freshwater phase did not influence smoltification or male early sexual maturation during the subsequent grow-out phase. However, a temperature increase from 12 to 16°C resulted in larger fish and appeared to stimulate early male maturation in some fish individuals.

Background Hepatitis B virus (HBV) vaccination is recommended for non-immunised patients with rheumatic diseases starting biological disease-modifying antirheumatic drugs (bDMARDs). There is some evidence that HBV vaccination is effective in patients under conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs), but it is currently unclear whether this also applies to bDMARDs.Objectives To assess the efficacy and safety of HBV vaccination in patients with inflammatory arthritides treated with bDMARDs.Methods A prospective cohort with inflammatory arthritides treated with bDMARDs, negative for anti-HBs and anti-HBc and never vaccinated for HBV was recruited. Engerix B was administered at 0, 1 and 6 months and anti-HBs was reassessed ≥1 month after last dose. Response was defined as anti-HBs≥10 IU/L and compared against vaccinated healthy controls. Disease flare, serious adverse events and immune-related disorders not previously present were recorded.Results 62 patients, most treated with TNF inhibitors (TNFi), and 38 controls were recruited. Most patients were taking csDMARDs (67.7%) and were in remission/low disease activity (59.4%). Only 20/62 patients (32.3%) had a positive response to vaccination, in comparison to 36/38 age-matched controls (94.7%, p

Abstract Autoantibodies are produced within germinal centers (GC), in a process regulated by interactions between B, T follicular helper (Tfh), and T follicular regulatory (Tfr) cells. The GC dysregulation in human autoimmunity has been inferred from circulating cells, albeit with conflicting results due to diverse experimental approaches. We applied a consistent approach to compare circulating Tfr and Tfh subsets in patients with different autoimmune diseases. We recruited 97 participants, including 72 patients with Hashimoto’s thyroiditis (HT, n = 18), rheumatoid arthritis (RA, n = 16), or systemic lupus erythematosus (SLE, n = 32), and 31 matched healthy donors (HD). We found that the frequency of circulating T follicular subsets differed across diseases. Patients with HT had an increased frequency of blood Tfh cells (p = 0.0215) and a reduced Tfr/Tfh ratio (p = 0.0338) when compared with HD. This was not observed in patients with systemic autoimmune rheumatic diseases (RA, SLE), who had a reduction in both Tfh (p = 0.0494 and p = 0.0392, respectively) and Tfr (p = 0.0003 and p = 0.0001, respectively) cells, resulting in an unchanged Tfr/Tfh ratio. Activated PD-1+ICOS+Tfh and CD4+PD-1+CXCR5–Tph cells were raised only in patients with SLE (p = 0.0022 and p = 0.0054), without association with disease activity. Our data suggest that GC dysregulation, assessed by T follicular subsets, is not uniform in human autoimmunity. Specific patterns of dysregulation may become potential biomarkers for disease and patient stratification.

Ultrafiltration (UF) membranes are used to successfully remove waterborne virus and bacteria from wastewater and drinking water. However, UF membrane application in aquaculture water treatment is limited. In this study we evaluate the performance of a capillary polyethersulfone UF membrane to remove two benchmark waterborne fish pathogens: i) the infectious pancreatic necrosis virus - IPNV, which is an unenveloped icosahedral virus, and ii) the bacterium Aeromonas salmonicida, which is a Gram-negative, facultative anaerobic bacilli. Moreover, the UF membrane bench-scale unit was tested at two temperatures according to salmonid aquaculture: low (4 - 7°C) and high (16 - 19°C). Sterilised natural seawater was spiked with laboratory cultured pathogens, the water was filtrated, and the membrane permeate collected. Both pathogen solution and permeate were evaluated using a cell culture method to estimate the colony-forming units (CFU/ml) for bacteria presence, a median tissue culture infectious dose (TCID50/ml) assay for virus presence, and real-time quantitative polymerase chain reaction (RT-qPCR) for both bacteria and virus presence. The membrane permeate was negative for both virus and bacteria for all analysis and for both low and high temperatures. The results from this bench-scale study are encouraging for the application of UF membrane technology in aquaculture water treatment to prevent virus and bacteria outbreaks. Further studies should validate this UF membrane technology results in commercial aquaculture conditions.

In the last decades, the concept of preclinical rheumatoid arthritis (RA) has become established. In fact, the discovery that disease mechanisms start years before the onset of clinical RA has been one of the major recent insights in the understanding of RA pathogenesis. In accordance with the complex nature of the disease, preclinical events extend over several sequential phases. In a genetically predisposed host, environmental factors will further increase susceptibility for incident RA. In the initial steps of preclinical disease, immune disturbance mechanisms take place outside the joint compartment, namely in mucosal surfaces, such as the lung, gums or gut. Herein, the persistent immunologic response to altered antigens will lead to breach of tolerance and trigger autoimmunity. In a second phase, the immune response matures and is amplified at a systemic level, with epitope spreading and widening of the autoantibody repertoire. Finally, the synovial and bone compartment are targeted by specific autoantibodies against modified antigens, initiating a local inflammatory response that will eventually culminate in clinically evident synovitis. In this review, we discuss the elaborate disease mechanisms in place during preclinical RA, providing a broad perspective in the light of current evidence.

ObjectiveTo identify risk factors for SARS-CoV-2 infection and for severe/critical COVID-19, and to assess the humoral response after COVID-19 in these patients.MethodsNationwide study of adult patients with inflammatory RMDs prospectively followed in the Rheumatic Diseases Portuguese Register—Reuma.pt—during the first 6 months of the pandemic. We compared patients with COVID-19 with those who did not develop the disease and patients with mild/moderate disease with those exhibiting severe/critical COVID-19. IgG antibodies against SARS-CoV-2 were measured ≥3 months after infection and results were compared with matched controls.Results162 cases of COVID-19 were registered in a total of 6,363 appointments. Patients treated with TNF inhibitors (TNFi; OR = 0.160, 95% CI 0.099–0.260, P < 0.001) and tocilizumab (OR 0.147, 95% CI 0.053–0.408, P < 0.001) had reduced odds of infection. Further, TNFi tended to be protective of severe and critical disease. Older age, major comorbidities, and rituximab were associated with an increased risk of infection and worse prognosis. Most patients with inflammatory RMDs (86.2%) developed a robust antibody response. Seroconversion was associated with symptomatic disease (OR 13.46, 95% CI 2.21–81.85, P = 0.005) and tended to be blunted by TNFi (OR 0.17, 95% CI 0.03–1.05; P = 0.057).ConclusionsTNFi and tocilizumab reduced the risk of infection by SARS-CoV-2. Treatment with TNFi also tended to reduce rates of severe disease and seroconversion. Older age, general comorbidities and rituximab were associated with increased risk for infection and worse prognosis, in line with previous reports. Most patients with RMDs developed a proper antibody response after COVID-19, particularly if they had symptomatic disease.

Peracetic acid (PAA) is an oxidative disinfectant with a broad spectrum of antimicrobial activity and low environmental impact. In this study, we investigated the physiological impacts of PAA application in Atlantic salmon (Salmo salar) parr reared in freshwater recirculating aquaculture systems over a 4-week period. PAA at a target concentration of 1 mg/L was administered either in pulse (every 3 days) or continuous. The group that did not receive PAA served as a control. Fish tissue samples were collected for histology, gene expression, and biochemical analyses at day 0 and after 2 and 4 weeks of exposure. The expression of genes encoding for antioxidant defence in the olfactory organs, skin, and gills changed during the trial, but the temporal effects were more pronounced than inter-treatment impacts. The glutathione group of antioxidant genes was more responsive to PAA. In most cases, an upregulation was observed. Significantly lower levels of reactive oxygen species were identified in the plasma and skin mucus of the two PAA-exposed groups at week 4; nonetheless, significantly increased levels of total antioxidant capacity were only observed in the skin mucus of fish from the continuous treatment group. Additional markers of oxidative stress (i.e., 8-oxo-2′-deoxyguanosine and o,o'-dityrosine) were analysed in the skin, gills, liver, and dorsal fins. These markers were unaffected by the two PAA treatments. Sporadic reversible structural alterations were observed in the three mucosal organs; the changes were time-dependent, and the effects of PAA treatment were minimal. The number of mucous cells varied over time but not within treatments except in the skin of the pulse group at week 4 where a reduction was observed. The ratio of acidic and neutral mucous cells in the skin and gills were affected by PAA treatments especially in the pulse group. Overall, this study revealed that Atlantic salmon parr mobilised mucosal and systemic antioxidant defences against the oxidative disinfectant PAA, but it was evident that the mode of application did not impose a strong influence. The minimal effects of PAA application on the indicators of health and welfare underscore the potential use of PAA as a routine disinfectant in recirculating aquaculture systems.

A better understanding of recirculating aquaculture system (RAS) biosecurity is crucial for the sustainable and ethical production of Atlantic salmon smolt and post-smolt in these systems. This study described and evaluated the performance of a RAS facility for fish infection research with Atlantic salmon as the main animal model. Fish body weight, length, water quality, and system metrics from five independent experimental trials conducted between September 2020 and July 2021 were used to analyze the variation within and between treatments. Statistical power analysis was performed to determine the minimum number of fish required. The fish parameters variability showed that the inter-class correlation coefficient was on average low (0.1) and that the variation within tanks was larger than the variation between the tanks. The power analysis showed that 15 fish were required to be sampled per tank under these study conditions. Variation of water quality and system management metrics among the five experimental trials was higher compared to the variation within the five experimental trials. Moreover, the variation of the water quality parameters controlled by sensors was relatively low, whereas the parameters depending on biofilter maturation level and performance presented a very high variation. Water exchange rate-dependent quality parameters showed a similar variation value, i.e., nitrate and water turbidity. The established baseline for variability and performance presents an important reference for the design and realization of future experiments in RAS facilities. It is foreseen that the current research facility will develop new knowledge to improve the RAS biosecurity in the Atlantic salmon aquaculture industry.

Introduction: The use of recirculating aquaculture systems (RAS) to produce Atlantic salmon smolts has resulted in exceptionally high fish growth rates. However, there are potential negative trade-offs between fast growth and key physiological processes, such as inadequate smoltification and early sexual maturation, which can both be linked to body energy reserves. Methods: This study determined the effect of i) dietary fat levels on Atlantic salmon whole-body fat and fatty acids composition, growth performance and smoltification and ii) a previous dietary regime on seawater growth performance and male early sexual maturation. In freshwater RAS, salmon parr (~19g) were fed 3 fat levels (20, 24, and 28%) over a 14-week period at 12℃. Subsequently, in seawater flow-through systems, smolt (~96g) were fed a control diet (26%) for 12 weeks at 2 temperatures (12 and 16℃). Results: Dietary fat levels resulted in differences in k-factor, fish whole-body fat and fatty acids composition; the low fat diet resulted in fish with a 23% lower whole-body fat when compared to the other two dietary groups at the end of the freshwater phase. These differences in whole-body fat faded at the end of the seawater phase. During the freshwater phase, all three feeds resulted in a comparable growth performance and smoltification indicator values, including k-factor, gill Na+, K+-ATPase, blood serum chloride and smolt index score. In contrast, water temperature resulted in fish body weight differences, where groups reared at 16℃ were larger than those reared at 12℃. Nevertheless, both temperature regimes supported a similar thermal growth coefficient. Smolts grown at 16℃ showed a higher level of the sex steroids androstenedione compared to those at 12℃. However, 11-ketotestosterone and testosterone levels did not differ despite a trend for higher levels at 16℃. Furthermore, testis histology at the final sampling indicated that some individuals showed initial signs of maturation (stage 3). Discussion: In conclusion, varying dietary fat levels (20 - 28%) during the freshwater phase did not influence smoltification or male early sexual maturation during the subsequent grow-out phase. However, a temperature increase from 12 to 16℃ resulted in larger fish and appeared to stimulate early male maturation in some fish individuals. [ABSTRACT FROM AUTHOR]

Introduction: Maturation in seawater-adapted (smoltified) male Atlantic salmon is undesirable yet frequently reported in fish weighing up to 1 kg reared under intensive conditions in land-based facilities. Although the photoperiod regulates both smoltification and maturation, the optimal post-smoltification regimen for the prevention of maturation is unclear. Methods: In this study, mixed-sex Atlantic salmon weighing 95 g were exposed to three different photoperiod regimes: continuous daylength, gradually decreasing daylength, and short-to-long daylength after the induction of smoltification by the "winter signal" (12 h:12 h light:dark). Fish were maintained under these conditions in triplicate for 18 weeks in a brackish water flow-through system at 12℃ and with a salinity of 20 ppt. This initial phase was followed by a grow-out period (1 kg-2.5 kg) in seawater (32-ppt salinity) at 13℃ for 11 weeks, with exposure to decreasing daylength. Throughout the 29-week study, body condition, gonad development, and morphological traits were assessed. Additionally, during exposure to brackish water, serum 11-ketotestosterone (11-KT) levels were quantified, and the testes were examined histologically. Upon termination of the study, the maturation rate was determined. Results: This study exclusively detected maturation among male salmon. Under continuous light exposure, the number of fish initiating maturity steadily increased from week 8 and continued to increase in the subsequent samplings. In the short-to-long daylength group, a subset of male fish showed elevated 11-KT levels at week 14, although their gonadal development and morphological changes were not evident until week 18. Male fish exposed to decreasing daylength showed no substantial increase in 11-KT levels, although a few fish exhibited advanced gonad development by week 18. At the end of the trial, markedly lower percentages of maturing and fully mature male fish were observed among those fish previously exposed to decreasing daylength. The maturation rates at study termination were 44.4% for the short-to-long daylength group, 29.4% for the continuous daylength group, and 4.3% for the decreasing daylength group. Discussion: These results show that photoperiod can be utilized to regulate maturation in male Atlantic salmon. In addition, decreasing the photoperiod after smoltification seems to limit early sexual maturation without compromising body size development. Thus, decreasing the photoperiod could be utilized to limit maturation in the production of post-smolt salmon or salmon in landbased facilities. [ABSTRACT FROM AUTHOR]

Breast cancer (BC) is one of the most frequent cancers in females across the globe. Treatment recommendations for BC patients are primarily driven by patient age, staging and tumor molecular subtype. Thus, we updated the general overview of BC staging, molecular surrogates, and treatment choices for women >70 years based on a systematic study encompassing the years 2013-2023. A PRISMA guidelines and PICO framework were followed, and relevant research articles were searched using different data bases (Web of Sciences, PubMed, MEDLINE, and Scopus). Mixed Methods Appraisal Tool was used for studies quality assessment. The research articles that made it into the systematic review were compiled using qualitative criteria. In the meanwhile, heterogeneity was determined using meta-analysis with RevMan 5.4. We applied a random effects model with a 0.05 significance level. Overall, there were 4151 research articles, after screening only 17 articles with 39,906 patients were included. Conclusion: Elderly patients with breast cancer should be treated differently in an adapted way. The treatment should not be the same worldwide due to different health systems. Molecular surrogates are different in geriatric patients. Surgery is the best option for treatment in this subset of patients. We need to have therapeutic decision appointments for elderly patients with breast cancer. The guidelines and medical authority should be used in the best decision. [ABSTRACT FROM AUTHOR]