mRNAs are emerging modalities for vaccination and protein replacement therapy. Increasing the amount of protein produced by stabilizing the transcript or enhancing translation without eliciting a ...strong immune response are major steps towards overcoming the present limitations and improving their therapeutic potential. The 5′ cap is a hallmark of mRNAs and non-natural modifications can alter the properties of the entire transcript selectively. Here, we developed a versatile enzymatic cascade for regioselective benzylation of various biomolecules and applied it for post-synthetic modification of mRNA at the 5′ cap to demonstrate its potential. Starting from six synthetic methionine analogues bearing (hetero-)benzyl groups, S -adenosyl- l -methionine analogues are formed and utilized for N7G-cap modification of mRNAs. This post-synthetic enzymatic modification exclusively modifies mRNAs at the terminal N7G, producing mRNAs with functional 5′ caps. It avoids the wrong orientation of the 5′ cap—a problem in common co-transcriptional capping. In the case of the 4-chlorobenzyl group, protein production was increased to 139% during in vitro translation and to 128–150% in four different cell lines. This 5′ cap modification did not activate cytosolic pathogen recognition receptors TLR3, TLR7 or TLR8 significantly more than control mRNAs, underlining its potential to contribute to the development of future mRNA therapeutics.
To analyse the demographics, main clinical and laboratory features and subtype distribution of juvenile idiopathic arthritis (JIA) in an eastern Mediterranean country, based on a multicentre ...registry.
Between March 2008 and February 2009 with this cross-sectional study, consecutive patients seen with JIA in selected centres were registered through a web-based registry. All patients were classified according to the International League of Associations for Rheumatology (ILAR) criteria.
There were 634 patients with a mean age of 11.84 ± 4.66 years and the female/male ratio was 1.2. The distributions of JIA patients according to onset of disease were as follows: systemic 92 (14.5%), oligoarticular extended 26 (4.1%), oligoarticular persistent 234 (36.9%), rheumatoid factor (RF) positive polyarthritis 20 (3.2%), RF negative polyarthritis 129 (20.3%), enthesitis-related 120 (18.9%), psoriatic 13(2.1%). The frequency of uveitis was 15.7% among all of the oligoarthritis patients. Anti-nuclear antibody (ANA) was positive mainly among the oligoarticular onset patients. Twenty-one patients also had Familial Mediterranean fever (FMF). Among systemic JIA patients, the frequency of macrophage activation syndrome (MAS) was 15.2% (n=14). At the end of the mean follow-up of 7.6 ± 4.4 years, 305 (48.1%) patients were defined to have inactive disease on medication, and 106 (16.7%) were completely free of any disease symptoms without medication.
Enthesitis related arthritis had a high frequency whereas psoriatic arthritis was very rare compared to other series. We suggest that there are certain differences in the characteristics of JIA in our eastern Mediterranean population. Thus, genetic studies need to be assessed in these populations separately and findings of genome wide association studies need to be confirmed in different populations.
Evidence from the literature has shown that Saccharomyces boulardii provides a clinically significant benefit in the treatment of acute infectious diarrhoea in children. In this multicentre, ...randomised, prospective, controlled, single blind clinical trial performed in children with acute watery diarrhoea, we aimed to evaluate the impact of S. boulardii CNCM I-745 in hospitalised children, in children requiring emergency care unit (ECU) stay and in outpatient settings. The primary endpoint was the duration of diarrhoea (in hours). Secondary outcome measures were duration of hospitalisation and diarrhoea at the 3rd day of intervention. In the whole study group (363 children), the duration of diarrhoea was approximately 24 h shorter in the S. boulardii group (75.4±33.1 vs 99.8±32.5 h, P<0.001). The effect of S. boulardii (diarrhoea-free children) was observed starting at 48 h. After 72 h, only 27.3% of the children receiving probiotic still had watery diarrhoea, in contrast to 48.5% in the control group (P<0.001). The duration of diarrhoea was significantly reduced in the probiotic group in hospital, ECU and outpatient settings (P<0.001, P<0.01 and P<0.001, respectively). The percentage of diarrhoea-free children was significantly larger after 48 and 72 h in all settings. The mean length of hospital stay was shorter with more than 36 h difference in the S. boulardii group (4.60±1.72 vs 6.12±1.71 days, P<0.001). The mean length of ECU stay was shorter with more than 19 h difference in the probiotic group (1.20±0.4 vs 2.0±0.3 days, P<0.001). No adverse effects related to the probiotic were noted. Because treatment can shorten the duration of diarrhoea and reduce the length of ECU and hospital stay, there is likely a social and economic benefit of S. boulardii CNCM I-745 in adjunction to oral rehydration solution in acute infectious gastroenteritis in children.
Background Canakinumab (CAN), a selective, fully-human anti-IL-1β monoclonal antibody is approved for SJIA in over 30 countries. Efficacy and safety of CAN over 12 weeks have been demonstrated in 2 ...phase III trials.1 In these trials, >60% of the pts received a previous biologic and were switched to CAN due to lack of efficacy or for safety reasons, and may be more refractory to another biologic therapy. Objectives To present a post-hoc evaluation of CAN efficacy in biologic-naïve (BN) pts and those previously exposed to biologics (BE) during the first 12-weeks. Methods Pooled data from CAN naïve pts, enrolled in two phase III trials1 and an extension phase (up to interim data lock 10 August 2012) were considered. Pts (2–19 yrs) with active SJIA were enrolled and received CAN 4 mg/kg or placebo sc every 4 weeks for 12 weeks. CAN naïve pts who entered the trials and received at least one dose of CAN were included in this analysis (N=178 CAN naïve pts). Descriptive efficacy analyses of adapted ACR-JIA responses at Week 12 are provided for the BN and BE pts groups. Results At baseline, there were 66 (37%) BN pts whereas anakinra (ANA), tocilizumab (TCZ), etanercept (ETN) and adalimumab (ADA), were the biologics received by 78 (44%), 10 (6%), 58 (33%) and 9 (5%) pts, respectively. The main reasons for discontinuation of biologics in BE group (n=112) was lack of efficacy (ANA, n=32; TCZ, n=7; ETN, n=56; ADA, n=9) or safety/tolerability (ANA, n=20; TCZ, n=4, ETN, n=0). At Week 12, the BN and BE groups were similar in aACR-JIA 30 and 50 response rates. Numerically higher aACR-JIA 70 and 90 response rates were achieved in BN vs. BE pts (Table). aACR-JIA 70 and 90 response rates were similar in pts previously exposed to ANA vs those not exposed to ANA at 12 weeks (aACR-JIA70: 58% vs.63% and aACR-JIA 90:47% vs 50%). Compared to pts who discontinued ANA due to lack of efficacy, there was a trend towards higher aACR-JIA 70 and 90 response rates at Week 12 in pts who stopped ANA for other reasons (aACR-JIA70: 34% vs.74%; aACR-JIA90: 25% vs. 63%). A higher aACR-JIA 30, 50, 70 and 90 response rates were observed in TCZ naïve pts vs. those pts exposed to TCZ (n=10) aACR-JIA30: 71% vs.50%; aACR-JIA50: 70% vs. 50%; aACR-JIA70: 61% vs.50%; aACR-JIA90: 49% vs. 40%. Higher aACR-JIA 70 and 90 responses were observed for ETN naïve pts vs. those exposed to ETN aACR-JIA70: 67% vs. 48%; aACR-JIA90: 58% vs. 31%; while ADA- naïve pts had similar responses to CAN as ADA-exposed pt (aACR-JIA 70: 61% vs 56%) and they had higher aACR-JIA 90 response (aACR-JIA90: 50% vs. 22%). Conclusions In general, BE pts achieved aACR-JIA 50,70 and 90 responses to CAN quickly in the first 2 weeks, and maintained their response up to Week 12; albeit at a numerically lower level than BN pts. These data support the consistent efficacy of CAN across different subgroups of pts. References Ruperto N, et al. N Engl J Med 2012;367(25). Disclosure of Interest P. Quartier Grant/research support: Abbvie, BMS, Chugai-Roche, Novartis, Pfizer and SOBI, Consultant for: Abbvie, Chugai-Roche, Novartis, Pfizer, Servier and SOBI, Speakers bureau: Chugai-Roche, MEDIMMUNE, Novartis, Pfizer, A. Grom Consultant for: Novartis, Roche, NovImmune, N. Ruperto Grant/research support: To Gaslini Hospital: Abbott, Astrazeneca, BMS, Centocor Research & Development, Eli Lilly and Company, “Francesco Angelini”, Glaxo Smith & Kline, Italfarmaco, Novartis, Pfizer Inc., Roche, Sanofi Aventis, Schwarz Biosciences GmbH, Xoma, Wyeth Pharmaceuticals Inc., Speakers bureau: Astrazeneca, Bristol Myers and Squibb, Janssen Biologics B.V., Roche, Wyeth/Pfizer, H. Brunner Consultant for: Novarits, Roche, Astrazeneca, Celgene, Pfizer, Janssen, UCB, GSK, BMS, Speakers bureau: Novartis, K. Schikler Grant/research support: Pfizer, Novartis, Abbvie, Roche, Genentech, Speakers bureau: Abbvie, Novartis, M. Erguven: None declared, L. Goffin Consultant for: Novartis, Pfizer, M. Hofer Grant/research support: Novartis, Pfizer, Abbvie, T. Kallinich Grant/research support: Novartis, Speakers bureau: Roche, Novartis, ALK, K. Marzan Grant/research support: Novartis, C. Gaillez Employee of: Novartis, K. Lheritier Shareholder of: Novartis, Employee of: Novartis, K. Abrams Shareholder of: Novartis, Employee of: Novartis, A. Martini Grant/research support: The Gaslini Hospital, which is the public Hospital where I work as full time employee, has received contributions to support the PRINTO research activities from the following companies: Bristol Myers and Squibb, Centocor Research & Development, Glaxo Smith & Kline,Novartis,Pfizer Inc, Roche, Sanofi Aventis, Schwarz Biosciences GmbH., Speakers bureau: Abbott, Bristol MyersSquibb, Astellas, Behringer, Italfarmaco, MedImmune, Novartis, NovoNordisk, Pfizer,Sanofi, Roche, Servier, D. Lovell Grant/research support: National Institutes of Health- NIAMS, Consultant for: Astra-Zeneca, Centocor, Amgen, Bristol Meyers Squibb, Abbott, Pfizer, Regeneron, Roche, Novartis, UBC, Forest Research Institute, Horizon, Johnson & Johnson, Speakers bureau: Novartis, Roche DOI 10.1136/annrheumdis-2014-eular.1099
Background IL-1β, a key inflammatory cytokine, appears to play a prominent role in systemic juvenile idiopathic arthritis (SJIA). Canakinumab (CAN), a fully human, anti-IL-1β monoclonal antibody ...selectively binds to IL-1β and inactivates its signaling, thus suppressing inflammation and associated pain. Objectives To assess the efficacy and safety of CAN in SJIA patients (pts) with active systemic features at enrollment. Methods In this 4-week randomized, controlled, double-blind study (β-SPECIFIC 1), pts with active signs and symptoms of SJIA were randomized (1:1) to receive a single dose of CAN (4mg/kg) or placebo sc at Day 1. The primary endpoint, proportion of pts achieving an adapted ACRpedi30 criteria (plus absence of fever) at Day 15, has been reported earlier1. Secondary endpoints included higher adapted ACRpedi response both at Day 15 and 29, Physician global assessment (PGA) of disease activity (0-100 mm VAS), number of joints with active arthritis, C-reactive protein (CRP), pain intensity (0-100 mm VAS) and overall safety. Results A total of 84 pts (CAN, n=43; placebo, n=41) were randomized. High discontinuation rate in the placebo group (90.2%) was due to unsatisfactory therapeutic response vs CAN group (14%). CAN provided statistically significant superior response rates vs placebo at all visits for all ACR response levels: at Day 15, ACRpedi 30, 83.7% vs 9.8%; ACRpedi 50, 67.4% vs 4.9%; ACRpedi 70, 60.5% vs 2.4%; ACRpedi 100, 32.6% vs 0, respectively (all p≤0.0001). Furthermore, CAN provided clinically meaningful improvement vs placebo for, PGA of disease activity; number of active joints and CRP as early as Day 3 (Table). Least square means in overall pain intensity were significantly lower in the CAN vs placebo group at Days 15 and 29 (both, p<0.0001). 55.8% pts in the CAN vs 39.0% placebo group had AEs with the most frequently reported AEs being upper respiratory tract infections. No discontinuations occurred due to AEs. 4 pts had serious AEs, 2 in each treatment group: CAN: varicella, bronchopneumonia (this patient also had MAS); placebo: gastroenteritis, MAS. Both pts with MAS recovered. No deaths were reported in either treatment group. Conclusions A single dose of CAN has superior efficacy to placebo in SJIA pts, providing rapid onset of action and robust initial response in majority of pts, while demonstrating an acceptable safety and tolerability profile. References Ruperto N et al, abstract Arthritis Rheum 2011;63 S1030 Disclosure of Interest P. Quartier Grant/Research support from: Novartis, Consultant for: Novartis, M. Erguven Grant/Research support from: Novartis, G. Horneff Grant/Research support from: Abbott, Pfizer, Consultant for: Abbott, Pfizer, Novartis, Roche, Y. Berkun: None Declared, K. Lheritier Shareholder of: Novartis, Employee of: Novartis, D. Kim Shareholder of: Novartis, Employee of: Novartis, K. Abrams Shareholder of: Novartis, Employee of: Novartis, T. Constantin: None Declared
mRNAs are emerging modalities for vaccination and protein replacement therapy. Increasing the amount of protein produced by stabilizing the transcript or enhancing translation without eliciting a ...strong immune response are major steps towards overcoming the present limitations and improving their therapeutic potential. The 5′ cap is a hallmark of mRNAs and non-natural modifications can alter the properties of the entire transcript selectively. Here, we developed a versatile enzymatic cascade for regioselective benzylation of various biomolecules and applied it for post-synthetic modification of mRNA at the 5′ cap to demonstrate its potential. Starting from six synthetic methionine analogues bearing (hetero-)benzyl groups,
S
-adenosyl-
l
-methionine analogues are formed and utilized for N7G-cap modification of mRNAs. This post-synthetic enzymatic modification exclusively modifies mRNAs at the terminal N7G, producing mRNAs with functional 5′ caps. It avoids the wrong orientation of the 5′ cap-a problem in common co-transcriptional capping. In the case of the 4-chlorobenzyl group, protein production was increased to 139% during
in vitro
translation and to 128-150% in four different cell lines. This 5′ cap modification did not activate cytosolic pathogen recognition receptors TLR3, TLR7 or TLR8 significantly more than control mRNAs, underlining its potential to contribute to the development of future mRNA therapeutics.
Novel
S
-adenosyl-
l
-methionine analogues were generated enzymatically and used for regioselective benzylation of biomolecules. Applied to the mRNA 5′ cap, protein production in cells can be increased and immunogenicity altered.
The October 23, 2011 M7.2 Tabanli - Van and November 9, 2011 M5.2 Edremit - Van earthquakes caused damage in a widespread area across the Van province in Turkey. The ground motions, the damage caused ...by these earthquakes and the recent progress related to recovery efforts are presented herein. First, the key properties of the recorded strong ground motions like spectral amplitudes and directionality are evaluated. The observed damage in the affected reinforced concrete and masonry structures are discussed. The set of common structural damage mechanisms （i.e., soft story failure, torsional response due to plan irregularity, short column failure, pull out failure, pounding） observed in the damaged buildings were identified. The relationship between the key structural properties and the extent of damage is investigated. The primary loss drivers across the region were identified to be the poor quality of workmanship and improper use of building materials. The results from the investigation suggest that a large portion of the loss could have been prevented if sufficient attention and care were given to the implementation of the design regulations and in particular to the construction practice. Lastly, the recent progress in the ongoing rebuildin~ activities is presented.
Background Systemic juvenile idiopathic arthritis (SJIA) is an IL1β-mediated autoinflammatory disease, characterized by spiking intermittent fever, rash, and arthritis. Canakinumab (CAN), a ...selective, fully human, anti-IL1β monoclonal antibody has shown promising efficacy and safety data in SJIA patients (pts). Objectives To demonstrate CAN allows tapering of corticosteroids (CS) and to evaluate time to flare in CAN vs placebo (PCB) pts with SJIA and active systemic features. Methods This Phase 3 trial had two parts. In Part 1 (P1), all patients (2-20 yrs) received open label CAN to identify responders and to taper CS. In Part 2 (P2), pts who successfully tapered their CS were randomized to receive CAN or PCB. In P1, pts received CAN (4mg/kg; max 300 mg) sc injection every 4 wks. Primary objectives were to determine if ≥25% of pts on CS at study entry could successfully taper the CS dose (P1); if time to flare was longer for CAN vs PCB (P2), and safety. Results 177 pts (128 on CS; 49 steroid-free) entered P1, of which 100 pts (CAN, n=50; PCB, n=50) entered P2; most of the remaining 77 pts were eligible (>ACR30 at Day 15) to directly enter an open-label extension study following P1. In P1, 92 pts attempted CS tapering and 57 (62%) were successful, representing 44.5% (57/128; p<0.0001) of pts on CS at study entry. 46% (42/92) of pts who attempted CS reduction and 33% (42/128) of pts on CS at study entry discontinued the CS. In P2, the median time to flare was 236 days (95%CI: 141-449 days) for PCB vs not determinable for CAN as <50% had flared, corresponding to a 63% relative risk reduction in flare for CAN vs PCB (HR 0.37; 95%CI: 0.17-0.78; p=0.0043). In P1, 78% (138/177) of pts had AEs, of which infections of the upper respiratory tract were the most common. Five pts discontinued due to an AE and 14 pts had ≥1 SAE; mostly infections, MAS, or flare-associated events. 1 pt on CAN had died with MAS. In P2, a direct comparison of safety between the 2 groups was difficult as all pts were exposed to CAN in P1. 80% of pts on CAN and 70% on PCB had ≥1 AE, with infections of the upper respiratory tract being the most common in both groups. 4 pts on PCB discontinued due to an AE, and SAEs were reported in 6 pts in each group; mostly due to an infection, MAS, or flare-associated event. 1 fatal MAS case occurred in 1 pt after 6 months of PCB. Conclusions For SJIA patients with active features, canakinumab sc injection every 4 weeks allowed successful steroid tapering, with one third being able to discontinue their CS. Canakinumab significantly decreased the risk of SJIA flare, while demonstrating an acceptable safety and tolerability profile. Disclosure of Interest N. Ruperto Grant/Research support from: Abbott, Astrazeneca, Bristol Myers and Squibb, Centocor Research & Development, Eli Lilly and Company, “Francesco Angelini” s.p.a, Glaxo Smith & Kline, Italfarmaco, Merck Serono, Novartis, Pfizer Inc., Regeneron, Roche, Sanofi Aventis, Schwarz Biosciences GmbH, Xoma, Wyeth Pharmaceuticals Inc., Speakers Bureau: Astrazeneca, Novartis, Bristol Myers and Squibb, Roche, Janssen Bilogics B.V., H. Brunner Consultant for: Novartis, UCB, Genentech, Jansen, GSK and Medimmune, Paid Instructor for: Participant in Scientific Advisory Committee for Canakinumab program in SJIA, P. Quartier Speakers Bureau: Novartis, T. Constantin: None Declared, N. Wulffraat: None Declared, G. Horneff Grant/Research support from: Abbott, Pfizer, Consultant for: Abbott, Pfizer, Novartis, Roche, R. Brik: None Declared, L. McCann: None Declared, O. Kasapcopur: None Declared, L. Rutkowska-Sak: None Declared, R. Schneider Consultant for: Hoffman-La-Roche, Speakers Bureau: Hoffman-La-Roche, Y. Berkun: None Declared, I. Calvo: None Declared, M. Erguven Grant/Research support from: Novartis, L. Goffin: None Declared, M. Hofer Consultant for: Novartis, T. Kallinich Speakers Bureau: Novartis, S. Knupp: None Declared, Y. Uziel Consultant for: Novartis, S. Viola: None Declared, K. Nistala Grant/Research support from: Arthritis Research UK Career Progression Fellow, C. Wouters: None Declared, R. Cimaz: None Declared, M. Ferrandiz Grant/Research support from: Novartis, B. Flato: None Declared, M. Luz Gamir: None Declared, I. Kone-Paut: None Declared, A. Grom Consultant for: Novartis, J&J, B. Magnusson Grant/Research support from: Swedish Association of local Authorities and Regions, Consultant for: Roche AB, S. Ozen Consultant for: Novartis, F. Sztajnbok: None Declared, K. Lheritier Shareholder of: Novartis, Employee of: Novartis, D. Kim Shareholder of: Novartis, Employee of: Novartis, K. Abrams Shareholder of: Novartis, Employee of: Novartis, A. Martini Grant/Research support from: Abbott, Astrazeneca, Bristol Myers and Squibb, Centocor Research & Development, Eli Lilly and Company, “Francesco Angelini” s.p.a, Glaxo Smith & Kline, Italfarmaco, Merck Serono, Novartis, Pfizer Inc., Regeneron, Roche, Sanofi Aventis, Schwarz Biosciences GmbH, Xoma, Wyeth Pharmaceuticals Inc., Speakers Bureau: Astrazeneca, Novartis, Bristol Myers and Squibb, Glaxo Smith & Kline, D. Lovell Consultant for: Abbott Laboratories, Centocor Inc, Amgen, Astra Zeneca Pharmaceutical, Bristol-Myers Squibb, Hoffmann-La Roche, Inc., Novartis Pharmaceutical Corporation, Pfizer Inc, Regeneron, UBC, Wyeth Pharmaceuticals, Xoma Corporation, Speakers Bureau: Wyeth Pharmaceuticals
Tofacitinib is an oral Janus kinase inhibitor. This trial assessed the efficacy and safety of tofacitinib versus placebo in patients with polyarticular course juvenile idiopathic arthritis (JIA).
...This double-blind, withdrawal phase 3 trial enrolled patients with polyarticular course JIA (extended oligoarthritis, rheumatoid factor-positive or rheumatoid factor-negative polyarthritis, or systemic JIA without active systemic features) aged 2 years to younger than 18 years, and was done at 64 centres of the Paediatric Rheumatology International Trials Organisation and Pediatric Rheumatology Collaborative Study Group networks in 14 countries. Patients with psoriatic arthritis or enthesitis-related arthritis were enrolled for exploratory endpoints. During part 1 of the study, patients received oral open-label tofacitinib (weight-based doses; 5 mg twice daily or lower) for 18 weeks. Patients achieving at least JIA/American College of Rheumatology 30 response were randomly assigned (1:1) using an Interactive Response Technology system to continue tofacitinib or switch to placebo in part 2 of the study for 26 weeks. The primary endpoint was JIA flare rate by week 44 in part 2 in patients with polyarticular course JIA; the intention-to-treat principle was applied. Safety was evaluated throughout part 1 and part 2 of the study in all patients who received one dose or more of study medication. This trial is registered with ClinicalTrials.gov, NCT02592434.
Between June 10, 2016, and May 16, 2019, of 225 patients enrolled, 184 (82%) patients had polyarticular course JIA, 20 (9%) had psoriatic arthritis, and 21 (9%) had enthesitis-related arthritis. 147 (65%) of 225 patients received concomitant methotrexate. In part 2, 142 patients with polyarticular course JIA were assigned to tofacitinib (n=72) or placebo (n=70). Flare rate by week 44 was significantly lower with tofacitinib (21 29% of 72 patients) than with placebo (37 53% of 70 patients; hazard ratio 0·46, 95% CI 0·27–0·79; p=0·0031). In part 2 of the study, adverse events occurred in 68 (77%) of 88 patients receiving tofacitinib and 63 (74%) of 85 in the placebo group. Serious adverse events occurred in one (1%) and two (2%), respectively. In the entire tofacitinib exposure period, 107 (48%) of 225 patients had infections or infestations. There were no deaths during this study.
The results of this pivotal trial show that tofacitinib is an effective treatment in patients with polyarticular course JIA. New oral therapies are particularly relevant for children and adolescents, who might prefer to avoid injections.
Pfizer.
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