bluuebunny/crossref_metadata_2025 huggingface dataset:
P206 Cardiovascular risk knowledge in patients of South Asian origin living with rheumatoid arthritis: data from India and the UK
Authors: Kumar Kanta , Arya Suvrat , Nightingale Peter , Sheeran Tom , Aggarwal Amita
Published: April 2020

Background

South Asians have a higher risk of cardiovascular disease (CVD). Rheumatoid arthritis (RA) increases the risk of premature atherosclerosis. Therefore, we compared the level of CVD risk knowledge among patients of South Asian origin with RA in India and in the UK.

Methods

In this cross-sectional survey, patients of South Asian origin with RA from India and the UK were recruited from secondary care settings. Data were collected via Heart Disease Fact Questionnaire-Rheumatoid Arthritis (HDFQ-RA), a validated self-completion questionnaire. The HDFQ-RA was translated into Hindi and piloted among patients from South Asian background before use. Additionally, clinical and demographic data was collected.

Results

Among 118 patients from each country, 84% were female and they had similar age, education level, employment status and co-morbidities. Patients from India had longer disease duration (5.5 years versus 4.1 years (p = 0.012) whereas those from the UK had higher disease activity score (4.0 + 0.8 versus 3.1 + 0.7, p < 0.01). Regarding modifiable risk factors for CVD only 44.6% from India and 45.4% in the UK were aware of them (p=ns). However, awareness of the link between RA and increased risk of CVD was even more limited (38.1% in India and 40.7% in UK (p=ns).

Conclusion

Patients of South Asians origin with RA from both countries had limited knowledge about CVD risk. There is a need to educate them about CVD risk during consultation, as this will result in better outcomes.

Disclosures

K. Kumar None. S. Arya None. P. Nightingale None. T. Sheeran None. A. Aggarwal None.
P208 A multidisciplinary approach to reproductive healthcare in women with rheumatic disease
Authors: Murray Kieran , Moore Louise , O'Brien Celine , Clohessy Anne , Brophy Caroline , McAuliffe Fionnuala , Veale Douglas J
Published: April 2020

Background

Rheumatic disease (RD) patients when family planning must consider fertility, disease activity, and management from preconception to lactation. A clear understanding is necessary, especially for those receiving disease-modifying antirheumatic medications. Previous studies have highlighted unmet needs in the care of women with RDs with reproductive healthcare needs. This study describes, to our knowledge, the first published standardised reproductive care pathway for women with RDs and the outcomes of this approach.

Methods

We developed the care pathway with multidisciplinary input from rheumatologists, rheumatology nurse specialists, obstetricians, midwives, maternal medicine specialists, and pharmacists. We identified patients’ emotional and healthcare needs, ensured access to expert advice, maintenance of good disease control, and positive reproductive outcomes. We prospectively followed the patients and report the results of the service.

Results

Ninety-eight women with median age (range) of 35 years (19-48) were assessed. The majority had an inflammatory arthritis. Seventy-six babies were born to 62 mothers. There were 12 miscarriages and one perinatal death. Breastfeeding rates at 6 weeks were low (28%).

Conclusion

We describe the first published evidence-based integrated multidisciplinary reproductive care pathway for women with RDs and the results of this approach. Seventy percent of women successful in trying to conceive delivered a healthy baby, and 90% of patients were ‘very satisfied’ with the service.

Disclosures

K. Murray: None. L. Moore: None. C. O'Brien: None. A. Clohessy: None. C. Brophy: None. F. McAuliffe: None. D. Veale: None.
P223 Influence of ambient air pollution on RA disease activity scores
Authors: Alsaber Ahmad R , Pan Jiazhu , Al-Herz Adeeba , Alkandary Dhary S , Al-Hurban Adeeba , Setiya Parul , Group KRRD RA Study
Published: April 2020

Background

This study aims to estimate the influence of air pollution ambients using the Air Quality Index (AQI) to Rheumatoid Arthritis (RA) disease activity. Disease Activity Score with 28 examined joints (DAS-28) and Clinical Disease Activity Index (CDAI) considered as disease activity indices for RA patients in the state of Kuwait.

Methods

Data for patients with RA disease were collected from Kuwait Registry for Rheumatic Diseases (KRRD) from 2013 to 2017. Moreover, data on air pollution obtained from The Kuwait Environmental Public Authority (K-EPA) during the same period. Statistical analysis was conducted using STATA to highlight the significant association between study variables. Descriptive statistics, correlation analysis and linear regression model techniques were employed to estimate the significant associations between RA disease activity represented by DAS-28 and CDAI; with air pollution components (PM10, NO2, SO2, O3, and CO). Multiple Imputation by Chained Equations (MICE) algorithm was also employed to tackle the value of the missing data for air pollution data.

Results

Total of 9,875 patients visits included in the analysis that matching with air pollution information from K-EPA database according to date and patient living address governorate. The study found SO2 and NO2 were significantly associated with RA disease activity using DAS-28 index, also, for CDAI index as well. For the score of RA disease activity using DAS-28 index, the correlation results show a positive significant correlation with exposure of SO2 using AQI (rp = 0.07), also the same results with the with the exposure of NO2 using AQI (rp = 0.07). The final model is demonstrating the effect from air-pollutants gaseous with RA factors (Swollen, RF, anti-CCP, ESR, CRP) on RA disease activity. The AQI of NO2 and SO2 still showed positive associations with disease activity performance of RA. The linear regression model shows a positive effects of NO2 (beta = 0.003, 95% CI: 0.002-0.005) and (beta = 0.048, 95\% CI: 0.030-0.066) for DAS-28 and CDAI respectively, where for SO2, the results shows positive significant effect with (beta = 0.003, 95% CI: 0.0004-0.005) and (beta = 0.044, 95% CI: 0.018-0.070) for DAS-28 and CDAI respectively.

Conclusion

In conclusion, our study showed that air ambients were significantly correlated to RA disease activity scores and should be considered as a possible risk factor for RA activity.

Disclosures

A.R. Alsaber None. J. Pan None. A. Al-Herz None. D.S. Alkandary None. A. Al-Hurban None. P. Setiya None.
P231 B-cell depletion therapy with rituximab in RA: UCLH/UCL 20 years of experience
Authors: Norris-Grey Caitlin M , Moore Samantha , Cambridge Geraldine , Reedy Venkat , Leandro Maria
Published: April 2020

Background

B-cell depletion therapy based on rituximab in patients with seropositive rheumatoid arthritis (RA) was pioneered at UCLH/UCL in 1998. The positive results of the initial open-label UCLH/UCL trials led to the Phase IIa proof of concept trial confirming efficacy and safety for rituximab in the treatment of patients with RA. A Phase III trial followed and licensing in 2006. We report a service evaluation of the UCLH/UCL cohort of RA patients treated with rituximab over the last 20 years.

Methods

A retrospective review of electronic records from all patients with RA treated with rituximab followed up in a dedicated clinic at UCLH/UCL (1998-2019) were included. Data on gender, age at first treatment, previous DMARDs, concomitant therapy at the time of the first rituximab cycle, total number of rituximab cycles and length of follow-up while on rituximab were collected. For patients currently on rituximab, further information was collected at last follow-up, including concomitant therapy, swollen and tender joint count, global VAS, CRP and ESR. DAS28ESR was calculated.

Results

296 patients had received rituximab therapy since November 1998, with 105 remaining on rituximab at final data collection (14/10/2019). Median follow-up was 77.5 months. Clinical data is shown in Table 1. Information for previous DMARDs was only available for 196 patients in the whole cohort, and for 70 patients currently on rituximab. The most common concomitant DMARD at first cycle was methotrexate with a similar percentage receiving methotrexate at last follow-up. The median number of previous DMARDs were 4 and 3 respectively. Within both cohorts approximately 31% of patients received rituximab as their first biologic.

Conclusion

This retrospective service evaluation of 296 patients with RA who had undergone rituximab therapy since 1998 showed that more than one third remained on rituximab at last follow-up. Of these, 70.2% of patients were in clinical remission or with low disease activity. Most patients (94%) remaining on rituximab were receiving only 1 or no concomitant therapies. Multiple cycles of treatment with rituximab have thus proved to be an effective therapeutic strategy for patients with RA resulting in long-term and highly favourable control of their disease.

Disclosures

C.M. Norris-Grey None. S. Moore None. G. Cambridge None. V. Reedy None. M. Leandro Consultancies; Genentech: consultancy fees for attending meeting on obinutuzumab trial in lupus nephritis.
P239 Third generation APCA improve prediction of clinical arthritis in at-risk individuals with positive second generation ACPA
Authors: Matteo Andrea Di , Mankia Kulveer , Duquenne Laurence , Mahler Michael , Corscadden Diane , Mbara Katie , Garcia-Montoya Leticia , Nam Jacqueline , Emery Paul
Published: April 2020

Background

Anti-citrullinated peptide antibodies (ACPA) are important biomarkers in rheumatoid arthritis (RA) and can predict arthritis development in at-risk individuals. ACPA are conventionally detected using the second-generation or third generation IgG anti-CCP antibody (Ab) assays. Although the value of anti-CCP2 Ab for predicting progression to RA in at-risk individuals is well understood, very little knowledge exists for anti-CCP3.The objectives of this study were: i) To evaluate the prevalence of serum anti-CCP3 Ab in serum anti-CCP2 Ab positive at-risk individuals (CCP2+ at-risk); ii)) To study the incremental value of anti-CCP3 Ab in CCP2+ at-risk subjects for predicting progression to clinical inflammatory arthritis (IA).

Methods

Anti-CCP3 Ab were tested on stored serum samples obtained from 347 CCP2 + (BioRad, USA) at-risk individuals from the Leeds CCP study. Anti-CCP2 and anti-CCP3 tests positivity threshold was >2.99 IU/ml and >20 units, respectively. Moreover, anti-CCP2 and anti-CCP3 titres were categorised according to manufacturer’s cut-off: low (LT) and high (HT) for anti-CCP2, negative/weak/moderate/strong for anti-CCP3. Progression to IA was defined as the development of clinical synovitis in at least one joint. Only subjects with at least one follow-up visit were included in the progression analysis (n = 317).

Results

Anti-CCP3 Ab were negative in 138/347 (39.7%) CCP2+ individuals, and weakly, moderately, and strongly positive in 15/347 (4.3%), 5/347 (1.4%) and 189/347 (54.4%) individuals, respectively. LT and HT anti-CCP2 Ab were found in 103/347 (29.7%) and in 244/347 (70.3%) individuals, respectively. Eighty-eight/317 subjects (28%) developed IA. The rate of progression of LT and HT anti-CCP2, when anti-CCP3 was negative, decreased from 6.5% to 2.7%, and from 36.6% to 9.4%, respectively. Progression in anti-CCP2 HT increased from 36.6% to 45.6% when anti-CCP3 was positive (Table 1).

Conclusion

Our results support an important role for anti-CCP3 Ab in improving prediction of IA in CCP2+ at-risk individuals.

Disclosures

A. Di Matteo None. K. Mankia None. L. Duquenne None. M. Mahler Other; Inova Diagnostic. D. Corscadden None. K. Mbara None. L. Garcia-Montoya None. J. Nam None. P. Emery Honoraria; AbbVie, BMS, Bristol-Myers Squibb, Gilead, Lilly, MSD, Novartis, Pfizer, Roche, Samsung, Samsung Bioepis Co., Ltd.
P265 Burden of psoriatic nail disease and response to biologic therapy in a PsA cohort
Authors: Nair Preeti , Anthony Anna , Lovell Christopher , Korendowych Eleanor , Cavill Charlotte , McHugh Neil , Tillett William
Published: April 2020

Background

Psoriatic nail disease is one of the six disease manifestations of psoriatic arthritis (PsA) in addition to peripheral arthritis, psoriasis, enthesitis, dactylitis and axial disease. Nail pitting is associated with an increased risk of developing PsA and onycholysis is associated with radiographic damage at the distal interphalangeal joints. Quantitative assessment of psoriatic nail disease is rarely performed in clinical practice therefore little is known about the burden of nail disease and effect of treatment in observational cohorts. We set out to quantify the burden of nail disease and response to b/tsDMARD treatment in a secondary care cohort.

Methods

Data were taken from the Bath PsA cohort. Patients fulfil CASPAR criteria and have clinical and patient reported outcomes at routine clinic follow-up visits. Patients commencing a b/tsDMARD with at least one follow up visit were included for analysis. Nail involvement was assessed using the Bath Nail score; pitting (0-10), onycholysis (0-10), hyperkeratosis (0-10) and severe deformity (0-10).

Results

157 of 485 (32%) of the cohort commencing b/tsDMARDS had nail disease at baseline and were included in the analysis. At baseline nail pitting (65%) and onycholysis (57%) were the most frequent nail manifestations followed by hyperkeratosis (14%) and severe nail deformity (7%) (Table 1). At the first follow-up visit (mean duration 20 weeks), 39% of patients achieved clear nails. At the final follow-up visit (mean duration 59 weeks) 56% had clear nails and 44% of patients had residual nail disease with a median (IQR) nail score of 0 (0.0-2.0). Earliest improvements were seen in nail bed manifestations (onycholysis/ hyperkeratosis) whilst improvement in nail matrix features (pitting) occurred later (Table 1) (p < 0.001).

Conclusion

Psoriatic nail disease affects one third of those commencing b/tsDMARDs in this cohort. Earliest treatment responses are seen in features of psoriatic nail dystrophy that affect the nail distally. At 20 weeks 39% of those with nail disease at baseline achieve clear nails. At one year, 44% of patients have ongoing nail manifestations, but few patients have persistently high nail scores.

Disclosures

P. Nair None. A. Anthony None. C. Lovell None. E. Korendowych None. C. Cavill None. N. McHugh None. W. Tillett None.
P266 IL-4/13 inhibitor dupilumab associated with new onset peripheral axial spondyloarthritis in patients with atopic dermatitis
Authors: Hughes Catherine D , Menon Bina , Willsmore Zena , Woolf Richard , Smith Catherine , Pink Andrew , Kirkham Bruce W
Published: April 2020

Background

Dupilumab is a recombinant human monoclonal antibody that inhibits signalling of interleukin-4 and interleukin-13. It has NICE approval for use in patients with moderate to severe atopic eczema who have failed at least one standard therapy. The main reported side effects in clinical trials were allergic conjunctivitis (1-5%), injection site reactions (8-19%) and herpes viral infections (3-7%).

Methods

Here we report symptoms and signs of a new peripheral axial spondyloarthritis/ psoriatic arthritis type presentation in 8 patients within 16 weeks of commencing dupilumab. This has not been described previously in the clinical trials patient population.

Results

These patients had a long history of atopy with severe eczema and raised baseline IgE levels of > 12000. A positive response to dupilumab was seen in all patients as evidenced by marked improvements in the Eczema Area and Severity Index (EASI). Musculoskeletal symptom onset began between 2 and 16 weeks after commencement of drug. Patients typically complained of inflammatory type pain in the small joints and entheseal sites. 2/8 patients had inflammatory sounding spinal pain. Early morning stiffness was common. All patients had normal acute phase reactants and one patient had a positive rheumatoid factor. 6/8 patients had radiologically evident enthesitis as seen on MRI or ultrasound (power doppler ultrasound signal-PDUS). 3/8 patients had such severe symptoms that they had to discontinue dupilumab. All patients were commenced on non-steroidal anti inflammatories. One patient was commenced on a low dose JAK inhibitor and exhibited improvement in musculoskeletal symptoms.

Conclusion

IL-4 has been shown to suppress delayed type hypersensitivity reactions (DTHRs) in both human and mice studies. It has been demonstrated previously that IL-4 can prevent bone erosion and disease progression in animal models of inflammatory arthritis. Guenova et al (2015) showed that IL-4 can selectively suppress IL-23 transcription and secretion with reduced Th17 function. They suggest that administration of IL-4 may be of therapeutic benefit in Th17 mediated inflammatory conditions. Conversely, dupilumab through the inhibition of IL-4 and IL-13 and possible resultant increased IL-17 and IL-23 levels, has perhaps unmasked a tendency towards development of a peripheral axial SpA phenotype in these patients.

Disclosures

C.D. Hughes None. B. Menon None. Z. Willsmore None. R. Woolf None. C. Smith Grants/research support; CS is a PI/CoPI on a number of commercially supported studie (Abbvie, Janssen, Leo, Sanofi). A. Pink None. B.W. Kirkham Consultancies; BK has worked as a consultant for Eli Lilly, Gilead, Janssen and Novartis. Grants/research support; BK has received research support from Eli Lilly and Novartis.
P269 Longitudinal patterns of pain distribution in axSpA patients: a retrospective cohort study
Authors: Nishtala Revathi , Barnett Rosie , Chyou Teyuan , Soni Anushka , Sengupta Raj
Published: April 2020

Background

Chronic pain is an important and debilitating symptom experienced by patients with axial Spondyloarthritis (axSpA). The patterns of pain and their associations in this patient group is poorly understood. The overall aim of the study is to investigate patterns of pain distribution in patients with axSpA using Margolis Pain Diagrams, and any associations with clinical and demographic measures.

Methods

We analysed data collected from individuals attending the axSpA outpatient clinic at the Royal National Hospital for Rheumatic Diseases in Bath. All participants were asked to record their pain on a pre-printed Margolis Pain diagram at each clinical visit. The number and distribution of painful areas were assessed and then categorised into regional or widespread pain, using pre-defined anatomical criteria. Descriptive analyses and any associations between pain distribution and demographic and clinical variables were assessed. Changes in pain distribution over time, using data from up to four clinic visits, were further investigated using a Sankey diagram.

Results

Of the 187 participants who had a baseline pain assessment, their mean age at diagnosis was 31.6 (11.9) years and 31.6% were female. 89.3% of patients reported pain at baseline, and of these 21.4%, 29.4 % and 38.5% reported pain in 1-2, 3-4, and 5-6 regions respectively. The most common sites of pain are in the Trunk (68.4%), followed by Head and Cervical (65.8%) and Left lower limb regions (58.8%). Univariate analyses did not reveal any significant associations between the presence of widespread pain and age at diagnosis, sex, smoking or HLA-B27 status. Disease activity, measured using Bath Ankylosing Spondylitis Disease Activity Index, was significantly higher in participants with widespread pain compared to those with regional pain OR 1.60 (1.30-1.96), and a trend towards higher sleep disturbance, was demonstrated OR 1.05 (1.00-1.10). Longitudinal analyses demonstrated considerable flux over time in the number and distribution of pain reported.

Conclusion

The Margolis Pain diagram is a practical instrument for assessing pain in axSpA patients. Several pain patterns were noted in patients with axSpA, and future research should focus on the potential impact on quality of life measures, response to treatment, and radiological findings.

Disclosures

R. Nishtala None. R. Barnett None. T. Chyou None. A. Soni None. R. Sengupta None.
P285 Using FITBITR for fitness monitoring in patients attending the AS residential course at RNHRD: it does make a difference!
Authors: Jamal Muhammad Safwan , Saeed Sadaf , Clarke Emily , Sengupta Raj
Published: April 2020

Background

Wearable technology has become a very popular tool in fitness training. It empowers people to be in charge of their own health and encourages them to exercise more. At the Royal National Hospital for Rheumatic Diseases we have been offering dedicated inpatient courses for management of spondyloarthropathy patients. Historically this course has helped patients with chronic arthritis to improve their physical well being while also focusing on different extra articular aspects of their condition like fatigue, sleep and pain issues. As part of this programme, we provided patients with Fitbit devices to evaluate whether regular recording of their activity and sleep tracking had an impact on their overall physical activity.

Methods

Patients attending the 2 weeks’ residential course from July 2018 to June 2019 were provided with Fitbit devices and were given a paper chart to log their data in. The patients gave verbal permission for their data to be included for audit purposes. The demographic data regarding the patients was collected through electronic patient record system. Bath indices were collected from electronic patient record system and results were compiled through Microsoft Excel.

Results

Data from 42 patients was collected. Data from 38 patients was analysed. Data for sleep indices was incomplete in 16 patients. 67% participants on the course were male and 33% were female. 90% patients had a diagnosis of ankylosing spondylitis, 5% had nonradiographic SpA and the rest had PsA or undifferentiated SpA. Majority of patients (97%) had improvement in their BASMI scores with a mean improvement of 37%. 51% patients improved their physical activity with an average increase in their step count by 13.3% compared to baseline. The participants on average spent 55.67 minutes being active on the course. While resting, majority of patients spent more time being awake or in light sleep (mean 14.3% and 54% respectively) although it might have been confounded by their stay in hospital. The verbal feedback from patients regarding the use of these devices has been mostly positive and it encouraged them to be more active.

Conclusion

Majority of patients attending the residential course noticed an improvement in their arthritis. The verbal feedback on using Fitbit devices to track daily activity and sleep was positive as it encouraged patients to increase their activity and improve sleep hygiene in future. Further refinement in data collection process and including other variables like pain, stiffness and fatigue scores can be valuable in a more focused discussion regarding management strategies of spondyloarthritis.

Disclosures

M. Jamal None. S. Saeed None. E. Clarke None. R. Sengupta None.
Asylum-Seekers Prosecuted for Human Smuggling: A Case Study of Scafisti in Italy
Authors: Patanè Flavia , Bolhuis Maarten P , van Wijk Joris , Kreiensiek Helena
Published: April 2020

States increasingly prosecute irregular migrants – asylum-seekers included – for their (alleged) involvement in human smuggling during their own migration journey. Based on a literature review and interviews with lawyers, prosecutors, judges, and migrants on Sicily, this article provides insight into the nature and scale of this phenomenon in Italy and discusses the effects of criminal prosecution on these migrants’ asylum procedures. From 2015–2018, as a standard operating procedure, roughly 1,300 “captains” and navigators – scafisti (literally: smugglers by boat) – of small dinghies with migrants arriving in Italy have been arrested for suspicion of “aiding clandestine (or irregular) immigration”. Most scafisti are migrants themselves and there are strong indications that they were forced to steer or navigate the boat. These prosecuted migrants face many difficulties in proving duress and are often inadequately advised about the consequences of a criminal conviction on their subsequent immigration procedures. After a conviction, as well as after an acquittal, they are often excluded from official reception centres and have difficulties accessing asylum procedures. When they manage to apply for asylum, they will be denied international protection if they have been convicted. When they cannot be expelled, they may end up in a legal limbo, having to rely on a temporary humanitarian status with strict limitations.
Transient brain networks underlying interpersonal strategies during synchronized action
Authors: Heggli Ole Adrian , Konvalinka Ivana , Cabral Joana , Brattico Elvira , Kringelbach Morten L , Vuust Peter
Published: April 2020

Interpersonal coordination is a core part of human interaction, and its underlying mechanisms have been extensively studied using social paradigms such as joint finger-tapping. Here, individual and dyadic differences have been found to yield a range of dyadic synchronization strategies, such as mutual adaptation, leading–leading, and leading–following behaviour, but the brain mechanisms that underlie these strategies remain poorly understood. To identify individual brain mechanisms underlying emergence of these minimal social interaction strategies, we contrasted EEG-recorded brain activity in two groups of musicians exhibiting the mutual adaptation and leading–leading strategies. We found that the individuals coordinating via mutual adaptation exhibited a more frequent occurrence of phase-locked activity within a transient action–perception-related brain network in the alpha range, as compared to the leading–leading group. Furthermore, we identified parietal and temporal brain regions that changed significantly in the directionality of their within-network information flow. Our results suggest that the stronger weight on extrinsic coupling observed in computational models of mutual adaptation as compared to leading–leading might be facilitated by a higher degree of action–perception network coupling in the brain.
Channelopathy and Carbohydrates: Bad Mix for Migraines
Authors: Stanton Angela A.
Published: April 2020

Migraine is among the top 20 most disabling diseases
1. Despite the significant progress in pain management, migraine‐cause is still not understood. Most research is focused on pain, making it difficult to see the cause of migraine. How nutrition affects ionic channels and electrolyte homeostasis have not yet been connected to migraines.

Migraine is caused by channelopathy
2 the following way: hyper‐sensory brain
3; increased sensory‐neuron connections
4; extra voltage used by extra connections
5,
6; extra voltage uses more sodium
7; glucose entering cells causes sodium efflux
8; kidneys waste potassium
9,
10 and water to spare sodium; reduced sodium in circulation triggers increased aldosterone, cortisol, and adrenaline
11; cortisol and adrenaline increase glucose in the blood
12; glucose entering cells causes sodium and water efflux (as noted earlier), initiating the previous steps with adrenaline and cortisol
8. This cycle leads to migraine.

In this presentation, I derive the migraine‐process, forming a coherent picture and show how the proper nutrition may prevent migraines. There is a considerable interplay between nutrition high in carbohydrates, low in salt, and migraines. Carbohydrates have a detrimental effect on electrolytes, which may be the most important factor in migraine cause.

Support or Funding Information

Stanton Migraine Protocol Inc.,
Unique Substrate Specificity of Human Phospholipase A2s
Authors: Hayashi Daiki , Mouchlis Varnavas D. , Dennis Edward A.
Published: April 2020

Phospholipase A2 (PLA2) constitutes a superfamily of enzymes that hydrolyze the sn‐2 acyl‐chain of phospholipids which are the major components of cell membranes. PLA2 produces free fatty acids and lysophospholipids as products which serve as lipid mediators and precursors of bioactive lipids. Among them, the release of ω‐6 and ω‐3 fatty acids including arachidonic acid (AA, ω‐6), eicosapentaenoic acid (EPA, ω‐3), and docosahexaenoic acid (DHA, ω‐3) is a pivotal role of PLA2s because the fatty acids are precursors of various eicosanoids and related bioactive oxylipins responsible for inflammation and resolution. Additionally, in contrast to conventional ester‐linked phospholipids, we also investigated PLA2 specificity toward sn‐1 vinyl ether‐linked phospholipids and phosphatidylinositol phosphates (PIPs) which are involved in various signaling processes.

Cytosolic PLA2 (cPLA2), calcium‐independent PLA2 (iPLA2), and secreted PLA2 (sPLA2) are the three major PLA2s that are well studied, and a significant number of PLA2‐related disorders have been reported. Therefore, investigating the substrate specificity of PLA2 toward different phospholipids, can be a key to understanding the etiology of PLA2‐related disorders and developing novel therapeutic targets. Recently, we developed a high‐throughput lipidomics‐based assay to measure PLA2 activity and successfully employed it to investigate the general specificity of PLA2s
(1). In the present study, we investigated the specificity of the three major PLA2s toward phospholipids containing ω‐6 and ω‐3 fatty acids in the sn‐2 position, sn‐1 vinyl ethers, and PIPs using this lipidomics‐based assay. Also, we determined the optimal binding of the substrate in the active site of each PLA2 by molecular dynamics (MD) simulations to explain the observed specificity.

cPLA2 showed high activity toward AA and EPA, and prefers sn‐1 vinyl ether phospholipids over ester phospholipids. iPLA2 showed the highest activity toward EPA, and no preference was observed between a vinyl ether and an ester. sPLA2 showed a preference for DHA and ester phospholipids. In addition, we determined that although additional phosphates on the inositol headgroup attenuated the activity of all three PLA2s, at least cPLA2 and iPLA2 could hydrolyze PI(4,5)P2. MD simulations revealed that the active site of cPLA2 could precisely bind AA and EPA in the sn‐2 position but not DHA. In summary, the present study revealed a unique and distinct substrate specificity for each of the PLA2s and explained the observed preference of cPLA2 for AA and EPA.

Support or Funding Information

Supported by NIH grant GM20501‐43
HUMAN ALLOGRAFTS IMPROVE HEALING OUTCOMES IN CHRONIC WOUNDS
Authors: Tabor Aaron J. , Kellar Robert
Published: April 2020

Background

Chronic wounds affect an estimated 8 million people worldwide and are often a result of underlying health conditions, such as obesity, diabetes, malnutrition, etc. The related morbidity and mortality places an increased burden on the healthcare system. Regenerative approaches, which engages and stimulates the body’s innate healing capabilities is addressing the limited successes in current standard of care treatments. Axolotl Biologix is a regenerative medicine company that produces allograft products from the human amnion. Axolotl Ambient™ and Shot™, conditioned media rich in cytokines and growth factors, are derived from human amniotic mesenchymal stromal cells. Axolotl DualGraft™ is a dehydrated, acellular, double layer, human amniotic membrane that encourages soft tissue repair by serving as a barrier, extracellular matrix scaffold, and provides a source of matrix‐bound growth factors.

Methods

Both allografts were used to facilitate repair and regeneration in a patient with difficult to close or intractable full‐thickness wounds. Axolotl Ambient™ was injected into the wound margin and Axolotl DualGraft™ was placed within the exposed wound bed. Wounds were bandaged per standard of care and imaged regularly to monitor wound healing progression. A range of bioassays were performed to ascertain the biological activity of the allografts.

Results

Multiple applications and injections of Ambient and DualGraft resulted in the formation of granulation tissue, restoration of blood flow, pain sensation, epithelialization, and wound closure. Bioassays confirmed the allografts contain measurable amounts of growth factors that promote angiogenesis, proliferation, tissue remodeling and reduction in inflammation.

Conclusions

Clinical application of the allografts in intractable wounds yielded dramatic improvement in wound repair by stimulating the normal physiological wound healing processes by the addition of exogenous growth factors, providing a scaffold in the wound bed, promoting normal blood flow, re‐epithelialization, and tissue remodeling. Longitudinal case studies are ongoing, concurrent with benchtop research to further elucidate the mechanisms of action responsible for reported clinical benefits in difficult to heal wounds.

Support or Funding Information

N/A
Hyperstable Cell‐Penetrating Peptides from Medicinal Plants
Authors: Tam James P. , Kam Antony , Loo Shining
Published: April 2020

A longstanding interest of our laboratory is to study cysteine‐rich peptides (CRPs) from medicinal plants as an inspiration to design orally‐active compounds. Plants produce CRPs as part of their host‐defense mechanism against microbes and insects. Most CRPs contain 15 to 25% of cysteine per molecule and are characterized by their cystine scaffold
[1–
4]. CRPs can display both hydrophilic and hydrophobic surface patches due to the cystine dense core which causes hydrophobic side chains to flip outwards, resulting in a unique “inside‐out” feature. As hydrophobicity is a major driving force for cell penetration
[5], we hypothesized that CRPs penetrate cells and bind to intracellular targets to exert their biological effects. Here, we report the discovery of hyperstable and cell‐penetrating CRPs, roseltide rT1 and rT7, from Hibiscus sabdariffa of the Malvaceae family. We show that both roseltide rT1 and rT7 are highly resistant against proteolytic degradation. Using live‐cell confocal microscopy, we show that cationic roseltide rT1 and anionic roseltide rT7 are cell‐penetrating. Functionally, we demonstrate that roseltide rT1 targets the mitochondria via TOM20 and increases cellular ATP production
[4]. Roseltide rT7, on the other hand, inhibits the 20S proteasome, which in turn, reduces tumor necrosis factor‐induced IκB□ degradation, and decreases intercellular adhesion molecule‐1 expressions. Taken together, our results demonstrate that hyperstable CRPs are cell‐penetrating and targets intracellularly to elucidate their biological effects.

Support or Funding Information

This research was supported in part by Nanyang Technological University Internal Funding ‐ Synzymes and Natural Products (SYNC) and the AcRF Tier 3 funding (MOE2016‐T3‐1‐003).
NMI and IFP35 are key DAMP molecules in inflammatory bowel diseases
Authors: Cheng Sijing , Jiao Na , Lin Xutao , Li Yichen , Huang Yibo , Vermeire Severine , Zou Yifeng , Zhu Ruixin , Liang Huanhuan , Liu Yingfang , Zhu Lixin , Lan Ping
Published: April 2020

Background & Aim

N‐myc and STAT interactor (NMI) and interferon‐induced protein 35 (IFP35) are damage‐associated molecular patterns (DAMPs), that mediate pro‐inflammatory reactions upon cell or tissue damage. Excessive signaling mediated by DAMP molecules can lead to several chronic inflammatory diseases. Here we investigated potential roles for NMI and IFP35 in the pathogenesis of inflammatory bowel diseases (IBD) including Crohn’s disease and ulcerative colitis.

Method

Differentially expressed genes (DEG) were identified in the colonic mucosa of CD and UC in comparison with controls by the gene expression microarray dataset that was validated by quantitative real‐time polymerase chain reaction and immunohistochemistry.

Result

NMI and IFP35 were highly elevated in the colonic mucosa of both CD and UC patients, compared to healthy controls. The fold changes of NMI in the colonic epithelium of CD and UC were 1.78 (P < 0.05) and 1.79 (P <0.05), respectively; while the fold changes of IFP35 were 1.73(P<0.05 ) and 1.74 (P<0.05), respectively in CD and UC. More importantly, the expression of NMI and IFP35 decreased to normal levels in the responders after infliximab treatment, but remained elevated in the non‐responders of the infliximab treatment. Compared to the controls, the fold changes of NMI in CD and UC non‐responder was 1.92 (P<0.05) and 1.65(P<0.05), respectively; the fold changes of IFP35 in CD and UC non‐responders were 1.64(P<0.05) and 1.4(P<0.05), respectively. Consistently, the expression levels of the downstream molecules including TLR4, TNF and IL‐6 paralleled and positively associated with those of the NMI/IFP35 (P < 0.05).

Conclusions

Our observations suggest that DAMP molecules NMI/IFP35 may mediate the early events of the IBD pathogenesis. We are confirming these observations at RNA and protein levels. Future plans include intervention studies using therapeutic antibodies to block NMI/IFP35 activities in IBD animal models. Considering that many DAMP molecules are effective targets in the treatment of chronic inflammatory diseases, we are optimistic about the future clinical application of NMI/IFP35 for the treatment of IBD.

Support or Funding Information

1.National Natural Science Foundation of China 81774152, 31200986, 41530105, 81770571,81970452. 2.Guangzhou laboratory clinical innovation project 2018GZR0201005. 3.National Postdoctoral Program for Innovative Talents of China BX20190393. 4.Natural Science Foundation of Shanghai 16ZR1449800. 5.the University at Buffalo Community of Excellence in Genome, Environment and Microbiome (GEM).
Hyaluronic Acid Receptor‐RHAMM Mediates Renal Carcinoma Metastasis
Authors: Moran Heriberto , Cancel Limary M , Huang Jing , Rodriguez Nora , Munn Lance L , Tarbell John M
Published: April 2020

The surface proteoglycan/glycoprotein layer (glycocalyx) on tumor cells has been associated with cellular functions that can potentially enable invasion and metastasis. In addition, aggressive renal carcinoma cells (SN12L1) with high metastatic potential have enhanced invasion rates compared to low metastatic (SN12C) cells in response to interstitial flow stimuli in vitro. Our previous studies suggest that heparan sulfate (HS) and hyaluronic acid (HA) in the glycocalyx play important roles in this flow mediated mechanotransduction and upregulation of invasive and metastatic potential. We examined HA receptor‐RHAMM protein levels in both SN12L1 and SN12C cell lines. We observed significantly higher RHAMM levels, but not CD44 levels, in flow dependent SN12L1 cells compared to SN12C cells suggesting that RHAMM may be transmitting the mechanical signal from hyaluronic acid to the cell. To examine this, we knocked down the expression of RHAMM by siRNA in SN12L1 cells. RHAMM knock‐down cells displayed suppression of flow induced migration suggesting that RHAMM receptor transmits the flow signal to the cell. SAHA (Suberoylanilide Hydroxamic Acid) is a small molecule drug (Vorinostat) that, in addition to functioning as an HDAC inhibitor, has been reported to inhibit the HS synthetic enzyme NDST1. We have previously demonstrated that NDST1 knockdown in highly metastatic SN12L1 cells blocked flow‐induced migration in vitro and reduced metastasis from the kidney by 95% in a mouse model
[1]. We conducted flow induced migration assays in highly metastatic SN12L1 cells in the presence of 1.0 uM SAHA. SN12L1 cells treated with 1.0 uM SAHA displayed suppression of flow induced migration consistent with previous results showing NDST1 knockdown in highly metastatic SN12L1 cells blocked flow induced migration in vitro. We also examined the effect of flow on the glycocalyx and found that SN12L1 cells exposed to 24 hours of flow increased their HS expression by 3‐fold compared to static control. We have previously shown that flow sensed by HS is transmitted to the cell by glypican‐1
[2]. All of our data are consistent with the hypothesis that RHAMM is transmitting the mechanical signal from hyaluronic acid to the cell and HS is a flow sensor linked to glypican‐1.

Support or Funding Information

Funding information

Grant: RO1CA204949 Targeting glycocalyx mediated mechanisms of tumor metastasis
SPARC Distribution of Intrinsic Cardiac Neurons Projecting to the Sinoatrial Node in Male and Female Rats: A Comparison
Authors: Chen Jin , Tappan Susan , Vadigepalli Rajanikanth , Schwaber James S. , Cheng Zixi J.
Published: April 2020

Multiple intrinsic cardiac ganglionated plexuses (ICGPs) in the intrinsic cardiac nervous system (ICN) are located on the posterior surface of the left and right atria. However, it is not clear which ICGPs project to the sinoatrial node (SAN) in male and female rats. Our goal is to compare the distribution of ICN neurons projecting to SAN in male and female rats. In this study, the SAN cells were first identified by immunostaining with Hyperpolarization Activated Cyclic Nucleotide Gated Potassium Channel 4 (HCN4), a pacemaker channel marker. The SAN were focally injected with fluorescent tracer Fast DiI to retrogradely label ICN neurons which projected to the SAN region (2 %; 4–5 μl/SAN injection; n=5/sex). After DiI injection, the animals were returned to their cages for 14 days for tracer transportation. Four to five days before sacrifice, Fluorogold (FG) was injected intraperitoneally to counterstain all ICN neurons, and at the end of the time point, animals were perfused and fixed through the heart. The left and right atria were cut open and prepared as whole mounts for analysis. The distribution of tracer‐labeled ICN neurons in the whole mounts of the left and right atria were imaged, overlaid, and analyzed using the Neurolucida Tracing and Digitization System (n=5/sex). We found that: 1) Consistent with our previous finding, there are multiple ICGPs. In each animal, there is a large ICGP near SAN. 2) DiI injection into the SAN region can retrogradely label ICN neurons in both male and female rats. These labeled‐ICN neurons are mainly located in this ICGP in both male and female, with some DiI‐labeled neurons in other ICGPs. Thus, the male and female may have a similar distribution pattern of ICN neurons projecting to the SAN. Our work may contribute to the understanding of the topographical organization of ICN innervation of the heart in the male and female rats.

Support or Funding Information

Supported by NIH SPARC OT2OD023848 (PD/PI: K. Shivkumar; Subaward PI: Z.J. Cheng, J. S. Schwaber, R. Vadigepalli).
Progression of Chemoreflex (Dys−)function in the Streptozotocin‐Induced Rat Model for Alzheimer’s Disease
Authors: Thapa Mahima , Trivedi Mansi , Ostrowski Tim Daniel
Published: April 2020

Alzheimer’s disease (AD) is associated with gradual memory decline and impairment of cognitive ability. The majority of AD patients also suffer from multiple apneas during sleep. The sleep‐disordered breathing entails a reduced ability to adjust respiration during low oxygen (hypoxia) and/or increased carbon dioxide (hypercapnia) conditions, indicating dysfunction of the respiratory system.

In this study, we measured respiratory responses of the Streptozotocin (STZ) ‐induced rat model of AD to hypoxia (10% O2 ‐ peripheral chemoreflex) and hypercapnia (7% CO2 ‐ central chemoreflex). Measurements were taken at three weeks (early stage) and three months (late stage) following STZ injection into the lateral ventricles of the brain (2 mg/kg STZ; 6‐week‐old rats, N = 6/group).

Three weeks following AD induction, hypoxic exposure increased respiratory rate (RR) of vehicle control (CTL) and STZ‐AD rats. However, this increase was significantly blunted by ~25% in STZ‐AD. The blunted response was partial compensation by increased tidal volume (TV) resulting in a non‐significant decrease of minute ventilation (MV; RR x TV). Three months following AD induction, STZ‐AD rats still had a blunted response of RR to hypoxia, which was similar in magnitude to that observed at 3 weeks. However, at this late time point, baseline RR and MV at normoxia (21% O2) appeared higher in STZ‐AD than those in CTL. Comparing data between time points (3 weeks AD vs. 3 months AD), both groups showed the expected age‐dependent increase in TV and decrease of RR. This decrease in RR was less in STZ‐AD than that in CTL, but the increase in TV was similar in both groups. Thus, STZ‐AD rats had a ~15% higher MV at the 3 months’ time point than at 3 weeks. CTL rats, on the other hand, had similar MV at both ages (~0% change).

With exposure to 7% CO2, responses for RR, TV, and MV at 3 weeks were similar between CTL and STZ‐AD rats. Also at 3 months responses did not differ between groups. However, within the STZ‐AD group, MV at 3 months was significantly greater in response to hypercapnia than that at the younger age. This increase was due to the absent age‐dependent drop of RR in STZ‐AD. CTL rats, in contrast, had significantly reduced RR at 3 months, leading to comparable MV at both ages.

In summary, major changes were observed in the peripheral chemoreflex of STZ‐AD rats throughout our 3‐months observation period. Both, responses to hypoxia (short‐term) and baseline values of respiration under normoxia (long‐term) were altered in these animals. Central chemoreflex function, on the other hand, remained largely unaltered.

Support or Funding Information

Conference Travel Scholarship by Truman State University to M Thapa and seed money from A.T. Still University to TDO
The Role of Variable Repetitive Protein Domainsin Azf1p Function and Prion Formation
Authors: Stewart Taylor , Parmenter Mackenzie , Fuchs Stephen M.
Published: April 2020

Prions are naturally occurring, self‐propagating isoforms of proteins that represent a heritable mechanism for tuning protein function. The yeast transcription factor Azf1p was recently demonstrated to form a prion, [AZF1+], that confers resistance to the drug radicicol but reduces Azf1p target gene expression. Using the Prion‐Like Amino Acid Composition tool, we identified regions of Azf1p that are predicted to contribute to prion formation. Two notable candidates are the polyglutamine (polyQ) and polyasparagine (polyN) domains. Through a bioinformatic analysis of the genomes of 93 lab and wild isolates of S. cerevisiae, we determined that these domains vary in repeat copy number across strains of yeast. The polyQ domain has two alleles, while the polyN domain has six alleles ranging from 17 to 25 N residues. In this work, we are investigating the extent to which the variable polyQ and polyN domains of Azf1p play a role in its transcription factor activity and contribute to [AZF1+] prion formation. This research determines the effects of repeat‐length variation on protein localization and stability, prion activity, and target gene expression.

Support or Funding Information

This work is supported by the Army Research Office (W911NF‐19‐1‐0299)