bluuebunny/crossref_metadata_2025_split huggingface dataset:
P191 To investigate the mechanisms of action of novel compounds ex vivo in cells obtained from the inflammatory synovium of patients with rheumatoid arthritis
Authors: Turner Madeleine P 
Published: April 2022

Abstract

Background/Aims
To investigate the ex vivo effect of JAK1/2 inhibitor baricitinib on the expression of pro-inflammatory mediators in normal and RA diseased fibroblasts stimulated with oncostatin M (OSM) as well as in cells derived from RA inflamed synovial membrane cells (RA SMC).

Methods
RA SMC and fibroblast like synoviocytes (RA FLS) were isolated from synovial membranes of patients undergoing surgery using enzymatic digestion and cell propagation. Normal fibroblast-like synoviocytes (FLS) and normal human skin fibroblasts (HSF) were purchased from commercial sources. All fibroblasts were cultured in media supplemented foetal bovine serum and antibiotics in incubators using established laboratory procedures, and serum-starved prior to stimulation with OSM with or without baricitinib. RA SMC were cultured in the presence of baricitinib without OSM as they spontaneously release pro-inflammatory cytokines. Levels of mRNA and proteins of the various inflammatory cytokines (IL-6, IL-8, MCP-1, CCL5 and IP-10) were determined by qPCR, ELISA and MSD. Western blot was used to measure levels of phospho-JAK specific components.

Results
qPCR analysis found that baricitinib inhibited OSM induced IP-10 and MCP-1 mRNA expression but not RANTES mRNA expression in normal human skin fibroblasts and RA FLS. In normal FLS however, baricitinib was found to inhibit RANTES and MCP-1 mRNA expression, but not IP-10 mRNA expression. Expression levels of IL-8 were too low for accurate conclusions to be drawn in all cell types apart from the RA SMCs, and in this case baricitinib did not inhibit IL-8 expression. ELISA results showed that baricitinib inhibited OSM induced MCP-1 expression in all cell types investigated. OSM induced IL-6 expression was inhibited in RA FLS but no other cell type.

Conclusion
Baricitinib inhibits OSM stimulated pro-inflammatory cytokine production through JAK1, 2. It has a varying inhibitory effect on the production of different cytokines: it is shown to particularly inhibit MCP-1. Baricitinib was found to have no effect on the production of spontaneously released pro-inflammatory cytokines in RA mixed cell populations.

Disclosure
M.P. Turner: None.
P208 A retrospective survey of arthritis among inpatients at a consultant hospital in Northern Tanzania from 2017 to 2019
Authors: Yongolo Nateiya M , Krauth Stefanie J , Biswaro Sanjura M , Moshi Baraka , Mwanswila Manasseh , Hemed Asia , Njau Rosalia , Ng’unda Neema Allen , Walker Richard , Kilonzo Kajiru , Siebert Stefan , Halliday Jo , McIntosh Emma , Kelly Clive , Mmbaga Blandina , Temu Gloria 
Published: April 2022

Abstract

Background/Aims
Musculoskeletal (MSK) disorders affect many people worldwide, but there is minimal access to medical care in low and middle-income countries (LMICs). Few hospital-based studies on MSK disorders have been conducted and there are limited prevalence data on MSK diseases in sub-Saharan Africa (SSA). Arthritis is the most common MSK condition, both reducing physical activity and increasing the risk of disability and other non-communicable diseases (NCDs). Given the high prevalence of MSK disorders and the associated significant clinical and economic burden in developed countries, an estimate of the frequency of these conditions in SSA is overdue. We aimed to determine the recorded prevalence of arthritis and related clinical features among patients admitted to a tertiary care hospital in Northern Tanzania from 2017-19.

Methods
Ethical approval was obtained to review the medical records retrospectively for all patients aged over 13 admitted to medical wards at Kilimanjaro Christian Medical Centre, a zonal tertiary care hospital, over this 3-year period. We collected demographic characteristics including age, gender, diagnoses and outcome. Diagnoses specifying arthritis were specifically sought. The prevalence of arthritis among inpatients over this period estimated. Analysis was conducted using STATA version 15 with categorical variables summarized using frequency and percentage.

Results
A total of 8023 patient files were examined with a slight majority of males 4181 (52.1%). An arthritis was recorded in 129 (1.6%) with 71 (55%) aged ≥60 years and 74 (57.4%) being female. The commonest presenting symptoms were joint pain in 69 (53.5%), swelling in 38 (29.5%) and stiffness in 18 (14%). A precise diagnosis for the arthritis was rarely recorded, although rheumatoid arthritis and gout were specified in a minority. Overall mortality was 12.4% among patients with arthritis. In addition, a diagnosis of systemic lupus erythematosus was recorded in a further 16 (0.2%) patients.

Conclusion
The recorded prevalence of arthritis among hospital inpatients in this LMIC study was low but the associated mortality appeared high. The retrospective nature of the data is highly likely to have underestimated the true prevalence of MSK disease and emphasizes the need for improved awareness of MSK diagnostic and management strategies. A teaching and training program has been developed to assist with this.

Disclosure
N.M. Yongolo: None. S.J. Krauth: None. S.M. Biswaro: None. B. Moshi: None. M. Mwanswila: None. A. Hemed: None. R. Njau: None. N. Allen Ng’unda: None. R. Walker: None. K. Kilonzo: None. S. Siebert: None. J. Halliday: None. E. McIntosh: None. C. Kelly: None. B. Mmbaga: None. G. Temu: None.
P218 Drug repurposing screen of Food and Drug Administration-approved medicines identifies novel anti-inflammatory and NF-kB inhibitory activity of the tyrosine kinase inhibitor sunitinib malate
Authors: Bowman Amelia C , Chaffey Laura , Greaves David 
Published: April 2022

Abstract

Background/Aims
Macrophage-mediated inflammation is a driver of both acute inflammatory responses and chronic inflammatory conditions such as rheumatoid arthritis. Drug repurposing is an attractive strategy for the development of novel anti-inflammatory therapeutics. Currently used anti-inflammatories for rheumatoid arthritis have their own risk factors, the development of small molecule anti-inflammatory drugs with decreased side effect profiles is thus an ongoing process. The aim of this study was to identify FDA-approved medicines with novel anti-inflammatory effects in macrophages.

Methods
A murine macrophage NF-κB reporter cell-line was used to screen a library of 159 FDA-approved medicines for NF-κB inhibition upon LPS stimulation. To investigate sunitinib malate in vitro, cytokine production in LPS-stimulated bone marrow derived macrophages treated with sunitinib malate was measured (n = 5-7) and IC50 values were calculated from resulting concentration response curves. To investigate NF-κB inhibitory effects of sunitinib malate in vivo, mice (n = 10-11 per group) dosed orally with vehicle or sunitinib malate (30mg kg-1) were exposed to a model of acute endotoxemia (1-hour intraperitoneal injection with LPS, 10mg kg-1). Statistical significance was assessed by one-way ANOVA followed by Tukey’s post-hoc test.

Results
Our initial screen identified 18 medicines with NF-κB inhibitory activity. Among these, the type III tyrosine kinase inhibitor sunitinib malate was further investigated and found to reduce pro-inflammatory mediator production in BMDMs (TNFα IC50=0.97μM, IL-1β IC50= 1.74μM, IL-6 IC50=1.14μM, CCL2 IC50=1.19μM, Nitrite IC50=0.59μM; n = 5-7) without compromising cell viability. In vivo, sunitinib malate significantly reduces serum TNFα (p &lt; 0.05; one-way ANOVA; n = 10) and liver tnf mRNA expression (p &lt; 0.01; one-way ANOVA; n = 10) in a murine model of acute endotoxemia, and reduces nuclear translocation of NF-κB compared to vehicle-treated mice (p &lt; 0.01; one-way ANOVA; n = 7).

Conclusion
Screening of existing medicines in macrophages allows previously unexplored anti-inflammatory properties to be identified. Here, we have found that the tyrosine kinase inhibitor sunitinib malate has potent anti-inflammatory activity in macrophages in vitro and in vivo, mediated via an NF-κB-dependent mechanism. These results justify further exploration of sunitinib malate in macrophages and its potential as a repurposed anti-inflammatory therapeutic in arthritis.

Disclosure
A.C. Bowman: None. L. Chaffey: None. D. Greaves: None.
P226 Bortezomib-induced muscle toxicity mimicking myositis
Authors: Ansari Ghazal , Alkutobi Ziad , Nandagudi Anupama 
Published: April 2022

Abstract

Background/Aims
Novel drugs such as bortezomib are proteasome inhibitors that are used to treat multiple myeloma. Bortezomib induces apoptosis of rapidly dividing, metabolically active cells with extensive protein synthesis. Literature review shows two case reports of bortezomib-induced reversible muscle toxicity and motor neuropathy, used to treat multiple myeloma (MM) with persistently normal creatine kinase. In a single-centre prospective study, 14 out of 24 patients with symptomatic MM were treated with bortezomib and, among these, seven developed proximal lower limb weakness with complete resolution of symptoms after discontinuation of bortezomib as depicted in our case.
We present a case of myositis mimic which was triggered by the use of immunotherapy Bortezomib to treat multiple myeloma.

Methods
An 81-year-old Caucasian gentleman was admitted with proximal lower limb weakness. He had a background history of multiple myeloma, treated with melphalan, prednisolone and bortezomib commenced in September 2019. Following his fourth cycle in January 2020, he noted proximal lower limb weakness and was referred to Rheumatology. He had MRI of the spine which did not reveal any cord compression. His prednisolone was stopped. On further questioning, there were no features to suggest connective tissue disease, inflammatory arthritis or vasculitis. On examination power was reduced to 4/5 in the proximal muscles of lower limb affecting his mobility. Rest of the examination was unremarkable.

Results
All bloods including creatinine kinase, autoimmune profile and inflammatory markers were normal. Magnetic resonance imaging (MRI) of the thighs showed diffuse fatty atrophy within all muscle groups of the thighs with low grade oedema within the vastus lateralis and intermedius particularly of the left thigh when compared to the right. The appearances were in keeping with inflammatory myositis, without any abscess formation. Electromyography showed abnormal signals with neurogenic and myopathic changes. Nerve conduction studies showed moderate length-dependent axonal peripheral neuropathy. Following his review, investigations and discussion between Rheumatology and Haematology, it was believed that his lower limb weakness was likely related to his bortezomib and the decision was made to stop bortezomib therapy. He was followed up in Rheumatology and Haematology clinic after stopping bortezomib with improvement of lower limb weakness and mobility. His creatinine kinase and inflammatory markers remained persistently normal. Muscle biopsy was not considered as there was enough evidence to suggest it was bortezomib induced muscle toxicity.

Conclusion
We present a case of myositis mimic whose symptoms completely resolved on stopping bortezomib. As this is one of the rare and early complications of bortezomib we suggest that patients treated with bortezomib should be monitored for muscle weakness.

Disclosure
G. Ansari: None. Z. Alkutobi: None. A. Nandagudi: None.
P227 Molecular and functional characterisation of distinct resident and migratory skin fibroblast populations in systemic sclerosis
Authors: Clark Kristina E. N , Cole Alice , Xu Shiwen , Ong Voon H , Buckley Christopher D , Denton Christopher P 
Published: April 2022

Abstract

Background/Aims
Modern single-cell analysis has confirmed functional heterogeneity amongst fibroblast populations within normal and diseased skin. To better understand differences between fibroblast subpopulations, we have developed a novel technique to selectively isolate “migratory” fibroblasts, and non-migratory “resident” fibroblasts from heathy control (HC) and systemic sclerosis (SSc) skin and used functional assays of fibrotic potential, and gene expression to compare these novel fibroblasts populations.

Methods
Forearm skin punch biopsies were collected from dcSSc (n = 3), and HC skin (n = 3). Migratory fibroblasts were isolated by standard explant culture. The remaining biopsy fragments then underwent collagenase digestion to yield a population of resident fibroblasts. These populations were expanded and used at passage 3-6. Functional characterisation included 3-D collagen gel contraction, and migratory scratch-wound assays. Expression of pro-collagen I, CTGF and alphaSMA was compared by western blot. Bulk RNAseq on each fibroblast population was performed. Statistical analysis was carried out using Rsoftware “tidyverse”. Significant differences in gene expression were those with a fold change of ≥ 1.5, and adjusted p-value (FDR) of &lt; 0.05.

Results
SSc fibroblasts showed a hallmark fibrotic phenotype with increased gel contraction, faster migration, and overexpression of Col1, CTGF and alphaSMA compared with HC. Conversely, resident fibroblasts showed reduced contraction, migration, and reduced secretion of alphaSMA compared to migratory fibroblasts in both SSc and HCs. RNAseq confirmed 1483 significantly differentially expressed genes between SSc resident and migratory fibroblasts, whereas no genes were significantly differentially expressed between HC resident and migratory fibroblasts. To further understand disease-related differences between the two fibroblast populations, we compared gene expression between SSc and HC migratory fibroblasts, and resident fibroblasts. 644 genes were upregulated in the migratory fibroblasts of SSc compared to HC (including ACTA2, C1QTNF3). 914 genes were upregulated in SSc resident fibroblasts compared to HC (including IL6, CCL2). Of these 375 genes were upregulated in both SSc fibroblast populations compared to HC (Table 1) including IL11 and COMP.

Conclusion
There are clear functional and transcriptional differences between migratory and resident fibroblasts, which is more marked in SSc compared to HC. Further work is required to understand their role in SSc pathogenesis and contribution to disease phenotype that may facilitate a more targeted approach to treatment

Disclosure
K.E.N. Clark: None. A. Cole: None. S. Xu: None. V.H. Ong: None. C.D. Buckley: Corporate appointments; founder of Mestag Therapeutics. C.P. Denton: None.
P280 Maternal use of hydroxychloroquine with anti-Ro and or anti-La during pregnancy: a systematic review
Authors: Nokhatha Shamma A. Al , Maguire Sinead , Harrington Robert 
Published: April 2022

Abstract

Background/Aims
Anti-Ro antibody and, to a lesser extent, anti-La antibody have been associated with fetal heart block during pregnancy. Use of hydroxychloroquine has previously been shown to reduce this risk in pregnancy. According to ACR 2020, all women with positive anti-Ro and or anti-La antibodies are conditionally advised to commence HCQ during pregnancy, but this is based on limited data. We performed a systematic review to evaluate the prevalence of hydroxychloroquine prescription in women with anti-Ro or La antibodies and examine the prevalence of congenital heart block.

Methods
This was a systematic review of case controlled and observational studies. A PICO model was used to design the research question. Literature searches were initiated based on five keywords: hydroxychloroquine, pregnancy(antenatal), maternal, anti-Ro(SSA), anti-La(SSB). Studies were retrieved from Pubmed, MEDLINE, Mendeley, Cochrane, University College Dublin Library using relevant search strategies from inception to May 2021. The search was restricted to English language articles only. Data were extracted and cross-checked by two reviewers.

Results
Sixteen studies involving 1,706 anti-Ro and 454 anti-La positive females, resulting in 2,279 pregnancies, were eligible for inclusion in this review. Mean age was 30.7 years at gestation. The prevalence of HCQ prescription in pregnancy in Ro+ women was 32.7%. The most common diagnoses were systemic lupus erythematosus (n = 990) and Sjögren’s syndrome (n = 452), while the remaining women had rheumatoid arthritis, antiphospholipid syndrome, undifferentiated connective tissue disease, scleroderma, autoimmune thyroiditis, dermatomyositis, polychondritis and overlap conditions. Eight studies included asymptomatic antibody carriers (n = 192). The prevalence of congenital heart block in neonates of women on HCQ in pregnancy was 1.8%. Due to issues with data reporting in the included studies, it was not possible to separate out Ro+ women on HCQ from those not on HCQ. For this reason, we could not compare and draw conclusions on the prevalence of CHB in pregnancies of Ro+ women.

Conclusion
This systematic review highlights the need for additional studies to examine use of HCQ in women positive for Ro or La antibodies. While these antibodies are commonly encountered in SLE and Sjögren’s syndrome, they can occur in other rheumatic diseases and should be screened for to limit risk of CHB.

Disclosure
S.A. Al Nokhatha: None. S. Maguire: None. R. Harrington: None.
P281 In vitro evaluation of the response of human tendon-derived stromal cells to a novel electrospun suture
Authors: Nezhentsev Andrey , Abhari Roxanna E , Baldwin Mathew J , Mimpen Jolet Y , Augustyniak Edyta , Isaacs Mark , Mouthuy Pierre-Alexis , Carr Andrew J , Snelling Sarah J. B 
Published: April 2022

Abstract

Background/Aims
Recurrent tears after surgical tendon repair remain common, with 40% of rotator cuff repairs failing within one year. Repair failures can be partly attributed to the use of sutures not designed for the tendon cellular niche nor designed for the promotion of repair processes. Clinically-used sutures have high tensile strength to mechanically support injured tendon. However, these sutures do not support cell integration into the material, but instead can stimulate a chronic inflammatory response. Synthetic electrospun materials can mechanically support the tendon while providing topographical cues that modulate the immune response to promote wound healing. Electrospun material has an increased surface area and porosity which can mimic that of tendon tissue, thus promoting tendon cell infiltration and proliferation. Here, a novel electrospun suture made from twisted polydioxanone (PDO) polymer filaments is compared to PDS II, a clinically used PDO suture currently utilised in tendon repair.

Methods
We evaluated the ability of electrospun suture and PDS II to support the attachment and proliferation of human tendon-derived stromal cells using PrestoBlue cell viability assays and scanning electron microscopy. Suture surface chemistry was analysed using X-ray photoelectron spectroscopy. Bulk RNA-Seq interrogated the transcriptional response of primary tendon-derived stromal cells to sutures after 14 days.

Results
The electrospun suture showed increased initial cell attachment compared to PDS II. X-ray photoelectron spectroscopy revealed that the local surface chemistry of electrospun suture and PDS II is similar, indicating that the tendon-like architecture of electrospun suture was responsible for the greater cell attachment. Furthermore, electrospun suture elicited a stronger transcriptional response compared with PDS II, with relative enrichment of pathways including mTorc1 signalling and depletion of epithelial-to-mesenchymal transition and extracellular matrix gene sets. When compared to baseline, neither suture induced transcriptional upregulation of inflammatory pathways such as IL6-JAK-STAT3 signalling and TNF-α signalling via NF-κB.

Conclusion
Twisted electrospun sutures demonstrated a tendon-derived stromal cell response indicative of cell proliferation and wound healing, without significant fibrosis, when compared to clinically-used and safe PDS II suture. Twisted electrospun sutures therefore show promise in improving outcomes in surgical tendon repair by allowing increased cell attachment while maintaining an appropriate tissue response.

Disclosure
A. Nezhentsev: None. R.E. Abhari: None. M.J. Baldwin: None. J.Y. Mimpen: None. E. Augustyniak: None. M. Isaacs: None. P. Mouthuy: None. A.J. Carr: None. S.J.B. Snelling: None.
P297 Takayasu arteritis presents at an older age in a northern European population
Authors: Ecclestone Tegwen , Coath Fiona , Ducker Georgina , Fordham Sarah , Mukhtyar Chetan 
Published: April 2022

Abstract

Background/Aims
The definition of Takayasu arteritis includes the age of onset of disease as being &lt;50 years. It has been reported most frequently in young females from Asian populations. We investigated whether the epidemiological features are the same in Norfolk, where the population is predominantly of northern European ancestry.

Methods
n our centre, cases are classified using validated international criteria. The electronic records of our vasculitis clinic were reviewed to identify all individuals with a new diagnosis of large vessel vasculitis from 2011-2020 who met a modified ACR 1990 criteria for Takayasu arteritis. The original criteria include decreased brachial artery pulse, bruit over subclavian arteries or aorta, and arteriogram abnormality. The respective modifications of these three criteria were to accept decreased radial pulse as well as the brachial pulse, bruit over the axillary artery as well as the subclavian artery, and abnormality of positron emission tomography and/or ultrasonography instead of arteriograms.

Results
14 individuals met the modified ACR 1990 criteria for Takayasu arteritis. 13 were female. The mean (standard deviation) age at diagnosis was 68.2 (7.7) years. None of the individuals were &lt;40 years old. On presentation, 8 had upper limb claudication, 8 had pulse discrepancy, 4 had a blood pressure difference &gt;10 mm Hg systolic, and 8 had an audible bruit over either subclavian or axillary artery. The diagnosis was confirmed on positron emission tomography in 7, ultrasonography of extracranial arteries in 5, and 2 by both modalities. The mean (standard deviation) ESR was 66.7 (48.1) and CRP was 49.2 (33.8). 2/14 also met the ACR 1990 criteria for GCA.

Conclusion
Takayasu arteritis can present at an older age in Norfolk where the population predominantly has northern European ancestry. Some of these individuals also fulfil the criteria for GCA. This has implications for treatment, identification of immunogenetic factors and disease classification.

Disclosure
T. Ecclestone: None. F. Coath: None. G. Ducker: None. S. Fordham: None. C. Mukhtyar: None.
Impact of NP follow-up calls on reducing 30-day readmissions in patients with stroke
Authors: Hwang Pauline J. 
Published: April 2022

Abstract:
Hospital readmissions are major contributors to increased healthcare costs and lower quality of life. Despite advanced stroke care, patients who have experienced a stroke require ongoing follow-up care to prevent complications and hospital readmissions. We evaluated the impact of NP follow-up calls, providing another level of expertise to promptly identify new symptoms and complications, on readmission rates in patients who have experienced stroke.