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Risdiplam is an oral, survival of motor neuron 2 (SMN2) pre-mRNA splicing modifier approved for the treatment of spinal muscular atrophy (SMA). SUNFISH (NCT02908685) Part 2, a Phase 3, randomized, double-blind, placebo-controlled study, investigated the efficacy and safety of risdiplam in type 2 and non‑ambulant type 3 SMA. The primary endpoint was met: a significantly greater change from baseline in 32-item Motor Function Measure (MFM32) total score was observed with risdiplam compared with placebo at month 12. After 12 months, all participants received risdiplam while preserving initial treatment blinding. We report 24-month efficacy and safety results in this population. Month 24 exploratory endpoints included change from baseline in MFM32 and safety. MFM‑derived results were compared with an external comparator. At month 24 of risdiplam treatment, 32% of patients demonstrated improvement (a change of ≥ 3) from baseline in MFM32 total score; 58% showed stabilization (a change of ≥ 0). Compared with an external comparator, a treatment difference of 3.12 (95% confidence interval [CI] 1.67-4.57) in favor of risdiplam was observed in MFM-derived scores. Overall, gains in motor function at month 12 were maintained or improved upon at month 24. In patients initially receiving placebo, MFM32 remained stable compared with baseline (0.31 [95% CI - 0.65 to 1.28]) after 12 months of risdiplam; 16% of patients improved their score and 59% exhibited stabilization. The safety profile after 24 months was consistent with that observed after 12 months. Risdiplam over 24 months resulted in further improvement or stabilization in motor function, confirming the benefit of longer-term treatment., (© 2023. The Author(s).)

Background: Azobenzene disperse dyes (azo DDs) are well-known as textile allergens, but the knowledge of their occurrence in garments is low. The numerous azo DDs and dye components found in textiles constitute a potential health risk, but only seven azo DDs are included in the European baseline patch test series (EBS)., Objectives: To investigate non-regulated azo DDs and dye components in synthetic garments on the Swedish market., Methods: High-performance liquid chromatography/mass spectrometry, gas chromatography/mass spectrometry and computerized data mining., Results: Sixty-two azo DDs were detected, with Disperse Red 167:1 occurring in 67%, and 14 other DDs each found in >20% of the garments. Notably, the EBS dyes were less common, three even not detected, while arylamines were frequently detected and exceeded 1 mg/g in several garments. Also, halogenated dinitrobenzenes were identified in 25% of the textiles., Conclusion: Azo DDs and dye components, in complex compositions and with large variations, occurred frequently in the synthetic garments. The arylamines were shown to occur at much higher levels compared to the azo DDs, suggesting the former constitute a potentially higher health risk. The role of arylamines and halogenated dinitrobenzenes in textile allergy has to be further investigated., (© 2022 The Authors. Contact Dermatitis published by John Wiley & Sons Ltd.)

Introduction: Spinal muscular atrophy (SMA) is a rare autosomal recessive neuromuscular disease which is characterised by muscle atrophy and early death in most patients. Risdiplam is the third overall and first oral drug approved for SMA with disease-modifying potential. Risdiplam acts as a survival motor neuron 2 ( SMN2 ) pre-mRNA splicing modifier with satisfactory safety and efficacy profile. This review aims to critically appraise the place of risdiplam in the map of SMA therapeutics., Areas Covered: This review gives an overview of the current market for SMA and presents the mechanism of action and the pharmacological properties of risdiplam. It also outlines the development of risdiplam from early preclinical stages through to the most recently published results from phase 2/3 clinical trials. Risdiplam has proved its efficacy in pivotal trials for SMA Types 1, 2, and 3 with a satisfactory safety profile., Expert Opinion: In the absence of comparative data with the other two approved drugs, the role of risdiplam in the treatment algorithm of affected individuals is examined in three different patient populations based on the age and diagnosis method (newborn screening or clinical, symptom-driven diagnosis). Long-term data and real-world data will play a fundamental role in its future.

Risdiplam (Evrysdi ® ) is the first oral drug developed to treat spinal muscular atrophy (SMA) and is approved in multiple countries worldwide. It is approved for the treatment of SMA in patients aged ≥ 2 months in the USA and the EU, with this approval further specified in the EU for the treatment of 5q-autosomal recessive SMA with a clinical diagnosis of SMA types 1, 2, or 3 or with one to four survival motor neuron 2 (SMN2) copies. As an SMN2 pre-mRNA splicing modifier, risdiplam increases the production of full-length SMN protein, the lack of which drives the pathophysiology of SMA. In phase 2/3 clinical trials, risdiplam significantly improved motor function in infants with SMA type 1 and in patients aged 2-25 years with SMA types 2 or 3. These motor improvements were maintained with up to 2 years of treatment with risdiplam. Risdiplam was generally well tolerated, with a favourable benefit to risk balance. As an oral drug, risdiplam provides a convenient and useful treatment option across a broad range of patient ages and subtypes of SMA., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Background: Risdiplam is an oral small molecule approved for the treatment of patients with spinal muscular atrophy, with approval for use in patients with type 2 and type 3 spinal muscular atrophy granted on the basis of unpublished data. The drug modifies pre-mRNA splicing of the SMN2 gene to increase production of functional SMN. We aimed to investigate the safety and efficacy of risdiplam in patients with type 2 or non-ambulant type 3 spinal muscular atrophy., Methods: In this phase 3, randomised, double-blind, placebo-controlled study, patients aged 2-25 years with confirmed 5q autosomal recessive type 2 or type 3 spinal muscular atrophy were recruited from 42 hospitals in 14 countries across Europe, North America, South America, and Asia. Participants were eligible if they were non-ambulant, could sit independently, and had a score of at least 2 in entry item A of the Revised Upper Limb Module. Patients were stratified by age and randomly assigned (2:1) to receive either daily oral risdiplam, at a dose of 5·00 mg (for individuals weighing ≥20 kg) or 0·25 mg/kg (for individuals weighing <20 kg), or daily oral placebo (matched to risdiplam in colour and taste). Randomisation was conducted by permutated block randomisation with a computerised system run by an external party. Patients, investigators, and all individuals in direct contact with patients were masked to treatment assignment. The primary endpoint was the change from baseline in the 32-item Motor Function Measure total score at month 12. All individuals who were randomly assigned to risdiplam or placebo, and who did not meet the prespecified missing item criteria for exclusion, were included in the primary efficacy analysis. Individuals who received at least one dose of risdiplam or placebo were included in the safety analysis. SUNFISH is registered with ClinicalTrials.gov, NCT02908685. Recruitment is closed; the study is ongoing., Findings: Between Oct 9, 2017, and Sept 4, 2018, 180 patients were randomly assigned to receive risdiplam (n=120) or placebo (n=60). For analysis of the primary endpoint, 115 patients from the risdiplam group and 59 patients from the placebo group were included. At month 12, the least squares mean change from baseline in 32-item Motor Function Measure was 1·36 (95% CI 0·61 to 2·11) in the risdiplam group and -0·19 (-1·22 to 0·84) in the placebo group, with a treatment difference of 1·55 (0·30 to 2·81, p=0·016) in favour of risdiplam. 120 patients who received risdiplam and 60 who received placebo were included in safety analyses. Adverse events that were reported in at least 5% more patients who received risdiplam than those who received placebo were pyrexia (25 [21%] of 120 patients who received risdiplam vs ten [17%] of 60 patients who received placebo), diarrhoea (20 [17%] vs five [8%]), rash (20 [17%] vs one [2%]), mouth and aphthous ulcers (eight [7%] vs 0), urinary tract infection (eight [7%] vs 0), and arthralgias (six [5%] vs 0). The incidence of serious adverse events was similar between treatment groups (24 [20%] of 120 patients in the risdiplam group; 11 [18%] of 60 patients in the placebo group), with the exception of pneumonia (nine [8%] in the risdiplam group; one [2%] in the placebo group)., Interpretation: Risdiplam resulted in a significant improvement in motor function compared with placebo in patients aged 2-25 years with type 2 or non-ambulant type 3 spinal muscular atrophy. Our exploratory subgroup analyses showed that motor function was generally improved in younger individuals and stabilised in older individuals, which requires confirmation in further studies. SUNFISH part 2 is ongoing and will provide additional evidence regarding the long-term safety and efficacy of risdiplam., Funding: F Hoffmann-La Roche., Competing Interests: Declaration of interests EM has received fees for serving on scientific advisory boards; speaker fees from F Hoffmann-La Roche, Biogen, AveXis/Novartis, Scholar Rock, and Cytokinetics; and grants from Biogen during the conduct of the study. ND served on scientific advisory boards for F Hoffmann-La Roche and Novartis pharmaceuticals; he has received personal fees from Biogen for congress and travel support. ESM reports that she has served on advisory boards for Biogen and Scholar Rock. She has received consulting fees, travel support, and speaker honoraria as an independent contractor from F Hoffmann-La Roche, Biogen, AveXis, and Scholar Rock. AN has received fees for serving on scientific advisory boards and speaker fees from F Hoffmann-La Roche. MO reports grants from F Hoffmann-La Roche during the conduct of the study and has received grants as a clinical trial investigator from Biogen. KS reports grants from F Hoffman-La Roche/Chugai Pharmaceutical during the conduct of the study and grants from Biogen Japan and lecture fees from Novartis Pharmaceuticals, outside the submitted work. CV reports personal fees and financial support to her institution from F Hoffmann-La Roche for activities outside the submitted work. GB received consultancy fees and speaker honoraria from F Hoffmann-La Roche and AveXis and speaker fees from PTC Therapeutics. NG reports fees for serving on advisory boards and presentations at symposia from F Hoffmann-La Roche, Biogen, and AveXis. JK received grants or contracts from Novartis Gene Therapies and Biogen. He has received consulting and speaker fees from F Hoffmann La Roche, Biogen, and Novartis Gene Therapies and consulting fees from Scholar Rock. AK-P reports that she received an institutional support grant from Biogen; serving on scientific advisory boards for and receiving speaker honoraria from Biogen, F Hoffmann-La Roche, Novartis, and PTC Therapeutics; and receiving personal fees from Biogen and F Hoffmann-La Roche for travel support. LS received grants and personal fees from F Hoffman-La Roche, Biogen, and AveXis/Novartis Gene Therapy and personal fees from Cytokinetics, outside the submitted work. OK reports that he was previously an employee and stockholder in F Hoffmann-La Roche during the submitted work and that he now holds the position of honorary chief medical officer at The Spinal Muscular Atrophy Foundation. JWD has received fees for serving on scientific advisory boards from Biogen, Novartis, Sarepta, and Avidity; has received consulting fees from Affinia Therapeutics and Shift Therapeutics for a therapeutic platform; and has received grant support for clinical trials from F Hoffman-La Roche, Biogen, Novartis Gene Therapies, Cytokinetics, Scholar Rock, and Sarepta. CM, PF, MG, KG, HK, HS, RSS, and WYY report that they are current employees and stockholders in F Hoffmann-La Roche. OB-T declared no competing interests., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Although it is known that textile wastewater contains highly toxic contaminants whose effects in humans represent public health problems in several countries, studies involving mammal species are scarce. This study was aimed to evaluate the toxicity profile of 90-days oral administration of textile dyeing effluent (TDE) on oxidative stress status and histological changes of male mice. The TDE was collected from the textile plant of Monastir, Tunisia and evaluated for the metals, aromatic amines, and textile dyes using analytical approaches. Metal analysis by ICP-MS showed that the tested TDE exhibited very high levels of Cr, As, and Sr, which exceeded the wastewater emission limits prescribed by WHO and Tunisian authority. The screening of TDE through UPLC-MS/MS confirmed the presence of two textile dyes: a triphenylmethane dye (Crystal violet) and a disperse azo dye (Disperse yellow 3). Exposure to TDE significantly altered the malondialdehyde (MDA), Conjugated dienes (CDs), Sulfhydryl proteins (SHP) and catalase levels in the hepatic and renal tissues. Furthermore, histopathology observation showed that hepatocellular and renal lesions were induced by TDE exposure. The present study concluded that TDE may involve induction of oxidative stress which ensues in pathological lesions in several vital organs suggesting its high toxicity. Metals and textile dyes may be associated with the observed toxicological effects of the TDE. These pollutants, which may have seeped into surrounding rivers in Monastir city, can cause severe health malaise in wildlife and humans.

Competing Interests: The authors have no funding or conflicts of interest to declare.

Background: Type 1 spinal muscular atrophy (SMA) is a progressive neuromuscular disease characterized by an onset at 6 months of age or younger, an inability to sit without support, and deficient levels of survival of motor neuron (SMN) protein. Risdiplam is an orally administered small molecule that modifies SMN2 pre-messenger RNA splicing and increases levels of functional SMN protein in blood., Methods: We conducted an open-label study of risdiplam in infants with type 1 SMA who were 1 to 7 months of age at enrollment. Part 1 of the study (published previously) determined the dose to be used in part 2 (reported here), which assessed the efficacy and safety of daily risdiplam as compared with no treatment in historical controls. The primary end point was the ability to sit without support for at least 5 seconds after 12 months of treatment. Key secondary end points were a score of 40 or higher on the Children's Hospital of Philadelphia Infant Test of Neuromuscular Disorders (CHOP-INTEND; range, 0 to 64, with higher scores indicating better motor function), an increase of at least 4 points from baseline in the CHOP-INTEND score, a motor-milestone response as measured by Section 2 of the Hammersmith Infant Neurological Examination (HINE-2), and survival without permanent ventilation. For the secondary end points, comparisons were made with the upper boundary of 90% confidence intervals for natural-history data from 40 infants with type 1 SMA., Results: A total of 41 infants were enrolled. After 12 months of treatment, 12 infants (29%) were able to sit without support for at least 5 seconds, a milestone not attained in this disorder. The percentages of infants in whom the key secondary end points were met as compared with the upper boundary of confidence intervals from historical controls were 56% as compared with 17% for a CHOP-INTEND score of 40 or higher, 90% as compared with 17% for an increase of at least 4 points from baseline in the CHOP-INTEND score, 78% as compared with 12% for a HINE-2 motor-milestone response, and 85% as compared with 42% for survival without permanent ventilation (P<0.001 for all comparisons). The most common serious adverse events were pneumonia, bronchiolitis, hypotonia, and respiratory failure., Conclusions: In this study involving infants with type 1 SMA, risdiplam resulted in higher percentages of infants who met motor milestones and who showed improvements in motor function than the percentages observed in historical cohorts. Longer and larger trials are required to determine the long-term safety and efficacy of risdiplam in infants with type 1 SMA. (Funded by F. Hoffmann-La Roche; FIREFISH ClinicalTrials.gov number, NCT02913482.)., (Copyright © 2021 Massachusetts Medical Society.)

In this study, alginate reinforced reduced graphene oxide@hydroxyapatite (rGO@HAP-Alg) hybrids have been fabricated via co-precipitation technique. The developed adsorbent was effectively utilized for the removal of Reactive Blue 4 (RB4), Indigo Carmine (IC) and Acid Blue 158 (AB158) azo dyes from aqueous solution, and found to have the adsorption efficiency of 45.56, 47.16 and 48.26 mg/g, respectively. The thermodynamic parameters demonstrated the endothermic and spontaneous nature of the adsorption process. The adsorption system was pH-dependent and showed maximum dye removal at pH 6-7, which was indicative of the electrostatic interactions, surface complexation and the hydrogen bonding mechanisms involved between the adsorbate and adsorbent during the adsorption process. Furthermore, the renewability studies demonstrated the reusability and stability of rGO@HAP-Alg hybrids up to five successive cycles. This study delivers a promising strategy for removing dye molecules and extends the potential application of rGO@HAP-Alg hybrids to treat practical dye contaminated water/wastewater., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Type 1 spinal muscular atrophy is a rare, progressive neuromuscular disease that is caused by low levels of functional survival of motor neuron (SMN) protein. Risdiplam is an orally administered, small molecule that modifies SMN2 pre-messenger RNA splicing and increases levels of functional SMN protein., Methods: We report the results of part 1 of a two-part, phase 2-3, open-label study of risdiplam in infants 1 to 7 months of age who had type 1 spinal muscular atrophy, which is characterized by the infant not attaining the ability to sit without support. Primary outcomes were safety, pharmacokinetics, pharmacodynamics (including the blood SMN protein concentration), and the selection of the risdiplam dose for part 2 of the study. Exploratory outcomes included the ability to sit without support for at least 5 seconds., Results: A total of 21 infants were enrolled. Four infants were in a low-dose cohort and were treated with a final dose at month 12 of 0.08 mg of risdiplam per kilogram of body weight per day, and 17 were in a high-dose cohort and were treated with a final dose at month 12 of 0.2 mg per kilogram per day. The baseline median SMN protein concentrations in blood were 1.31 ng per milliliter in the low-dose cohort and 2.54 ng per milliliter in the high-dose cohort; at 12 months, the median values increased to 3.05 ng per milliliter and 5.66 ng per milliliter, respectively, which represented a median of 3.0 times and 1.9 times the baseline values in the low-dose and high-dose cohorts, respectively. Serious adverse events included pneumonia, respiratory tract infection, and acute respiratory failure. At the time of this publication, 4 infants had died of respiratory complications. Seven infants in the high-dose cohort and no infants in the low-dose cohort were able to sit without support for at least 5 seconds. The higher dose of risdiplam (0.2 mg per kilogram per day) was selected for part 2 of the study., Conclusions: In infants with type 1 spinal muscular atrophy, treatment with oral risdiplam led to an increased expression of functional SMN protein in the blood. (Funded by F. Hoffmann-La Roche; ClinicalTrials.gov number, NCT02913482.)., (Copyright © 2021 Massachusetts Medical Society.)

Competing Interests: The authors have no funding or conflicts of interest to declare.

Background: The textile dye mix (TDM) 6.6% pet. contains Disperse Blue (DB) 35, Disperse Yellow 3, Disperse Orange (DO) 1 and 3, Disperse Red 1 and 17, and DB 106 and 124. The most frequent allergen in TDM-positive patients is DO 3. Around 85% of p-phenylenediamine (PPD)-allergic dermatitis patients have shown positive patch test reactions to DO 3. There has been a discussion to exclude DO 3 from TDM 6.6% because of frequent, strong reactions to TDM 6.6% and PPD., Objectives: To study if DO 3 can be omitted from a TDM., Methods: Patch tests were performed on 2250 dermatitis patients with TDM 6.6%, TDM 5.6% pet., TDM 7.0% pet., and PPD 1.0% pet.; 122 patients were also patch tested with DO 3 1.0% pet., Results: Among the 2250 patients patch tested, contact allergy prevalence to TDM 6.6% was 2.4%, to TDM 5.6% 1.8%, and to TDM 7.0% 2.0%. Of the 54 TDM 6.6%-positive patients, 55.6% reacted to PPD; as much as 42.2% of PPD-allergic women and 50% of PPD-allergic men reacted to TDM 6.6%. Of the 17 DO 3-positive patients, 94.1% showed a positive reaction to PPD., Conclusion: Results indicate that DO 3 can probably be omitted from TDM, but patch testing with TDM 6.6%, TDM 7.0%, DO 3 1.0%, and PPD 1.0% simultaneously is needed to finally decide whether it is possible or not., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This study examined the effect of Red-S3B textile dye on soil microbial activities, uptake of the dye by wheat plants and growth on the dye-contaminated soil. Moreover, pressmud (PM) application was investigated for its alleviative effect on wheat yield and dye uptake by plants. Preliminarily, soil was spiked with a wide concentration range (0, 100, 250, 500, 750 and 1000 mg kg -1 soil) of Red-S3B dye and wheat was grown for 42-days. The dye did not suppress the activities of soil enzymes and growth of wheat seedlings at 100 mg kg -1 ; however, beyond this level the dye had a linear negative effect on these attributes. With 1000 mg dye kg -1 soil, wheat seedling biomass, viable microbial count, soil respiration, dehydrogenase, phosphatase, and urease activities decreased by 84%, 33%, 45%, 69%, 24%, and 11%, respectively as compared to uncontaminated soil. Moreover, phosphorus and potassium content in wheat shoot decreased, while the nitrogen content increased in Red-S3B contaminated soil. In the subsequent pot experiment, PM application (12.5 g kg - 1 soil) was assessed to alleviate the adverse effect of moderately toxic level of Red-S3B dye (500 mg kg -1 soil) on wheat growth and yield. Root and straw biomass, and grain yield of wheat decreased by 13, 19 and 12%, respectively in Red-S3B contaminated soil as compared to uncontaminated soil. However, PM application to dye-contaminated soil retrieved the dye-induced reduction in root and straw biomass and grain yield to become statistically (p ≤ 0.05) at par with control plants. The color of Red-S3B was clearly visible in spikes depicting that plants absorbed Red-S3B but probably could not metabolize it. Amending the dye-contaminated soil with PM decreased Red-S3B content in awns from 78 to 37 mg kg -1 . Hence, it is concluded that Red-S3B textile dye is highly toxic to soil microbes and wheat plants at levels exceeding 100 mg kg -1 soil. Soil application of PM alleviates the adverse effect of Red-S3B dye on wheat growth through reducing its uptake by plants., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Food coloring is often used as a coloring agent in foods, medicines and cosmetics, and it was reported to have certain carcinogenic and mutagenic effects in living organisms. Investigation of physiological parameters using zebrafish is a promising methodology to understand disease biology and drug toxicity for various drug discovery on humans. Zebrafish (Danio rerio) is a well-acknowledged model organism with combining assets such as body transparency, small size, low cost of cultivation, and high genetic homology with humans and is used as a specimen tool for the in-vivo throughput screening approach. In addition, recent advances in microfluidics show a promising alternative for zebrafish manipulation in terms of drug administration and extensive imaging capability. This pilot work highlighted the design and development of a microfluidic detection platform for zebrafish larvae through investigating the effects of food coloring on cardiovascular functionality and pectoral fin swing ability. The zebrafish embryos were exposed to the Cochineal Red and Brilliant Blue FCF pigment solution in a concentration of (0.02‰, 0.2‰) cultured in the laboratory from the embryo stage to hatching and development until 9 days post fertilization (d.p.f.). In addition, zebrafish swimming behaviors in terms of pectoral fin beating towards the toxicity screening were further studied by visualizing the induced flow field. It was evidenced that Cochineal Red pigment at a concentration of 0.2‰ not only significantly affected the zebrafish pectoral fin swing behavior, but also significantly increased the heart rate of juvenile fish. The higher concentration of Brilliant Blue FCF pigment (0.2%) increased heart rate during early embryonic stages of zebrafish. However, zebrafish exposed to food coloring did not show any significant changes in cardiac output. The applications of this proposed platform can be further extended towards observing the neurobiological/hydrodynamic behaviors of zebrafish larvae for practical applications in drug tests.

Background: Allergic contact dermatitis (ACD) has become more frequent in children. The prevalence of contact sensitization varies with respect to age, sex, and geographic localization., Objective: The aim of the study was to investigate the experience of a tertiary health center regarding the patch test results of contact sensitization in children without atopic dermatitis., Methods: This is a retrospective review of 89 children (30 boys and 59 girls) who were aged between 3 and 18 years and who were diagnosed with ACD between July 2013 and July 2017. Children with a known history of atopic dermatitis were excluded. All patients were tested with the TRUE (Thin-layer Rapid Use Epicutaneous) test series., Results: The most frequently determined allergens by TRUE test were methylchloroisothiazolinone (n = 7 [16.3%]), disperse blue 106 (n = 5 [11.6%]), and bacitracin (n = 5 [11.6%]). Formaldehyde-related allergens produced 15 positives., Conclusions: Preservatives, such as methylchloroisothiazolinone, formaldehyde, and formaldehyde releasers, emerge as the most frequent allergens in children who undergo patch testing because of ACD. This finding might be attributed to the increase in the utilization of these chemical compounds in personal hygiene products for children.

Several environmental factors (e.g. food source, pesticides, toxins, parasites and pathogens) influence development and maturation of honey bees (Apis mellifera). Therefore, controlled experimental conditions are mandatory when studying the impact of environmental factors: particularly food quality and nutrient consumption. In vitro larval rearing is a standard approach for monitoring food intake of larvae and the labelling of food is necessary to quantify intake in controlled feeding experiments. Here, we tested the suitability of two food dyes, Allura Red and Brilliant Blue, in an experimental set up using in vitro reared honey bee larvae and freshly hatched adult workers. Absorbance of both dyes was measured, in food and dye-fed larvae, to determine the optimal dye concentrations for accurate detection and quantification. By quantifying relative dye concentrations in dye mixtures, relative concentrations of mixed dyes can be estimated independent of the total food consumed by the larvae. Survival assays were conducted to test the impact of both dyes on larval and worker bee survival. Worker bees showed no increase in adult mortality, when fed with dyed honey. Larval survival was not significantly different until the late pupal stage. The physiological impact of dye feeding was tested by measuring larval immune response. No changes in innate immune gene expression were detectable for larvae fed with dyed and non-dyed food. In conclusion, we established a non-invasive food labelling protocol for food intake quantification in in vitro reared honey bee larvae, using non-toxic, inexpensive, and easy to apply food dyes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aims: Risdiplam (RG7916, RO7034067) is an orally administered, centrally and peripherally distributed, survival of motor neuron 2 (SMN2) mRNA splicing modifier for the treatment of spinal muscular atrophy (SMA). The objectives of this entry-into-human study were to assess the safety, tolerability, pharmacokinetics (PK) and pharmacodynamics of risdiplam, and the effect of the strong CYP3A inhibitor itraconazole on the PK of risdiplam in healthy male volunteers., Methods: Part 1 had a randomized, double-blind, adaptive design with 25 subjects receiving single ascending oral doses of risdiplam (ranging from 0.6-18.0 mg, n = 18) or placebo (n = 7). A Bayesian framework was applied to estimate risdiplam's effect on SMN2 mRNA. The effect of multiple doses of itraconazole on the PK of risdiplam was also assessed using a two-period cross-over design (n = 8)., Results: Risdiplam in the fasted or fed state was well tolerated. Risdiplam exhibited linear PK over the dose range with a multi-phasic decline with a mean terminal half-life of 40-69 h. Food had no relevant effect, and itraconazole had only a minor effect on plasma PK indicating a low fraction of risdiplam metabolized by CYP3A. The highest tested dose of 18.0 mg risdiplam led to approximately 41% (95% confidence interval 27-55%) of the estimated maximum increase in SMN2 mRNA., Conclusions: Risdiplam was well tolerated and proof of mechanism was demonstrated by the intended shift in SMN2 splicing towards full-length SMN2 mRNA. Based on these data, Phase 2/3 studies of risdiplam in patients with SMA are now ongoing., (© 2018 The British Pharmacological Society.)

SMA is an inherited disease that leads to loss of motor function and ambulation and a reduced life expectancy. We have been working to develop orally administrated, systemically distributed small molecules to increase levels of functional SMN protein. Compound 2 was the first SMN2 splicing modifier tested in clinical trials in healthy volunteers and SMA patients. It was safe and well tolerated and increased SMN protein levels up to 2-fold in patients. Nevertheless, its development was stopped as a precautionary measure because retinal toxicity was observed in cynomolgus monkeys after chronic daily oral dosing (39 weeks) at exposures in excess of those investigated in patients. Herein, we describe the discovery of 1 (risdiplam, RG7916, RO7034067) that focused on thorough pharmacology, DMPK and safety characterization and optimization. This compound is undergoing pivotal clinical trials and is a promising medicine for the treatment of patients in all ages and stages with SMA.

Objectives: The aim of this study was to examine the occurrence of five azo food dyes-tartrazine, sunset yellow, carmoisine, allura red, and ponceau 4 R-in the food supply chain of Singapore and their effects on the in vitro synthesis of leukotriene B 4 (LTB 4 ) and F 2 -isoprostanes., Methods: Trained personnel recorded the names of foods and beverages sold in a local supermarket that contained at least one of the five azo dyes. The occurrence of the azo dyes in the local food supply was computed. The synthesis of LTB 4 and F 2 -isoprostanes from freshly isolated blood neutrophils were measured using gas chromatography-mass spectrometry., Results: Of the 1681 processed food items, 194 (11.54%) contained at least one of the five azo dyes. Tartrazine was most prevalent in food and beverage products sold in Singapore, followed by allura red, sunset yellow, ponceau 4 R, and carmoisine. The five azo dyes augmented the in vitro synthesis of LTB 4 and F 2 -isoprostanes from blood neutrophils. Tartrazine was significantly more potent in increasing LTB 4 synthesis than the other dyes, which exhibited similar potencies. The five food dyes increased the formation of F 2 -isoprostanes from blood neutrophils at all tested concentrations., Conclusion: The high prevalence of azo dyes in the food supply of Singapore and their ability to elicit proinflammatory responses in vitro suggest a potential health risk to the local population., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In vitro gene expression signatures to predict toxicological responses can provide mechanistic context for regulatory testing. We previously developed the TGx-28.65 genomic biomarker from a database of gene expression profiles derived from human TK6 cells exposed to 28 well-known compounds. The biomarker comprises 65 genes that can classify chemicals as DNA damaging or non-DNA damaging. In this study, we applied the TGx-28.65 genomic biomarker in parallel with the in vitro micronucleus (MN) assay to determine if two chemicals of regulatory interest at Health Canada, disperse orange (DO: the orange azo dye 3-[[4-[(4-Nitrophenyl)azo]phenyl] benzylamino]propanenitrile) and 1,2,4-benzenetriol (BT: a metabolite of benzene) are genotoxic or non-genotoxic. Both chemicals caused dose-dependent declines in relative survival and increases in apoptosis. A strong significant increase in MN induction was observed for all concentrations of BT; the top two concentrations of DO also caused a statistically significant increase in MN, but these increases were <2-fold above controls. TGx-28.65 analysis classified BT as genotoxic at all three concentrations and DO as genotoxic at the mid and high concentrations. Thus, although DO only caused a small increase in MN, this response was sufficient to induce a cellular DNA damage response. Benchmark dose modeling confirmed that BT is much more potent than DO. The results strongly suggest that follow-up work is required to assess whether DO and BT are also genotoxic in vivo. This is particularly important for DO, which may require metabolic activation by bacterial gut flora to fully induce its genotoxic potential. Our previously published data and this proof of concept study suggest that the TGx-28.65 genomic biomarker has the potential to add significant value to existing approaches used to assess genotoxicity., (Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.)

In this contribution, the azo dye (E)-1-(4-((4-(phenylamino)phenyl)diazenyl) phenyl)ethanone (DPA) was combined with reduced graphene oxide (RGO) for the electrochemical modification of a pencil graphite electrode (RGO/DPA/PGE) surface. A series of electrochemical measurements were used for the characterization of the modified electrode surfaces. At the modified electrode, nicotine was irreversibly reduced. An obvious increase was observed in the reductive peak current of nicotine at the modified electrode, indicating the capability of the RGO/DPA composite to increase the electron transfer rate. The current was found proportional to the nicotine concentration in a range of 31 to 1900 μM, and the limit of detection (LOD) was calculated as 7.6 μM.

Background: Recurrent aphthous stomatitis (RAS) is a common disease of the oral mucosa with an unknown etiology. This study aimed to determine if food additives play a role in the etiology of RAS as well as to determine if patch testing can be used to detect which allergens cause RAS., Methods: This prospective study included 24 patients with RAS and 22 healthy controls. All the participants underwent patch testing for 23 food additives., Results: In total, 21 (87.5%) RAS patients and 3 (13.6%) controls had positive patch test reactions to ≥1 allergens; the difference in the patch test positivity rate between groups was significant (P < 0.05). The most common allergen that elicited positive patch test results in the patient group was cochineal red (n = 15 [62.5%]), followed by azorubine (n = 11 [45.8%]) and amaranth (n = 6 [25%])., Conclusions: The present findings show that food additives might play a role in the etiology of RAS and that patch testing could be a method for determining the etiology of RAS., (© 2016 The International Society of Dermatology.)

Local venous pain caused by dacarbazine (DTIC) injection is due to its photodegradation product 5-diazoimidazole-4-carboxamide (Diazo-IC). The production of Diazo-IC can be decreased by protecting the drug from light. Furthermore, the production of Diazo-IC reportedly increases with time; however, there are no studies reporting the association between the injection preparation time and local venous pain caused by the DTIC injection. We evaluated the efficacy of the following: (1) method used to shorten the injection preparation time and (2) method used to change the diluting solution for DTIC. We found that shortening the injection preparation time tended to decrease the local venous pain expression due to DTIC, and Veen F decreased the production of Diazo-IC compared with the normal saline and 5% glucose solution. These results indicate that shortening the injection preparation time may be effective in preventing the local venous pain caused by the DTIC injection; moreover, using Veen F for DTIC may also reduce the pain.

The objective of this study was to evaluate the single and combined genotoxic effects of six food pollutants (Chrysoidine G, Sudan I, acid orange II, malachite green, acrylamide, and potassium bromate) on THP-1 cells through comet assay. The results of the single tests indicated that the pollutants increased the percentage of tail DNA (% tail DNA) in a dose-dependent manner. Moreover, the % tail DNA values induced by synthetic colorants (Chrysoidine G, Sudan I, acid orange II, and malachite green) were significantly higher than those by acrylamide or potassium bromate at most concentrations. In the combined tests, Chrysoidine G (422 μmol/L) or acrylamide (400 μmol/L) was mixed with different concentrations of the other five pollutants respectively. In the first combined tests, most mixtures significantly increased the % tail DNA values with the exception of Chrysoidine G and acid orange II. In the second tests, there were no significant differences in the % tail DNA values between the single and combined tests at most cases., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Dietary exposures to the seven food, drug, and cosmetic (FD&C) colour additives that are approved for general use in food in the United States were estimated for the US population (aged 2 years and older), children (aged 2-5 years) and teenage boys (aged 13-18 years) based on analytical levels of the FD&C colour additives in foods. Approximately 600 foods were chosen for analysis, based on a survey of product labels, for the levels of FD&C colour additives. Dietary exposure was estimated using both 2-day food consumption data from the combined 2007-10 National Health and Nutrition Examination Survey (NHANES) and 10-14-day food consumption data from the 2007-10 NPD Group, Inc. National Eating Trends - Nutrient Intake Database (NPD NET-NID). Dietary exposure was estimated at the mean and 90th percentile using three different exposure scenarios: low exposure, average exposure and high exposure, to account for the range in the amount of each FD&C colour additive for a given food. For all populations and all exposure scenarios, the highest cumulative eaters-only exposures in food were determined for FD&C Red No. 40, FD&C Yellow No. 5 and FD&C Yellow No. 6. In addition, the eaters-only exposure was estimated for individual food categories in order to determine which food categories contributed the most to the exposure for each FD&C colour additive. Breakfast Cereal, Juice Drinks, Soft Drinks, and Frozen Dairy Desserts/Sherbet (also referred to as Ice Cream, Frozen Yogurt, Sherbet (including Bars, Sticks, Sandwiches)) were the major contributing food categories to exposure for multiple FD&C colour additives for all three populations.

Decorative tattoo is a popular practice that is generally safe when performed in the professional setting but can be associated with a variety of inflammatory, infectious, and neoplastic complications, risks that may be increased with current trends in home tattooing. Modern tattoo inks contain azo dyes and are often of unknown composition and not currently regulated for content or purity. Biopsy of most (if not all) tattoo reactions presenting to the dermatologist is recommended, given recent clusters of nontuberculous mycobacterial infections occurring within tattoo, as well as associations between tattoo reactions and systemic diseases such as sarcoidosis., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Allura Red AC (Food Red No. 40) is a red azo dye that is used for food coloring in beverage and confectionary products. However, its genotoxic properties remain controversial. To clarify the in vivo genotoxicity, we treated mice with Allura Red AC and investigated the induction of DNA damage (liver, glandular stomach), clastogenicity/anuegenicity (bone marrow), and mutagenicity (liver, glandular stomach) using Comet assays, micronucleus tests, and transgenic gene mutation assays, respectively. All studies were conducted in accordance with the Organization for Economic Co-operation and Development (OECD) guideline. Although Allura Red AC was administered up to the maximum doses recommended by the OECD guideline, no genotoxic effect was observed in any of the genotoxic endpoints. These data clearly show no evidence of in vivo genotoxic potential of Allura Red AC administered up to the maximum doses in mice., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Disperse dyes are well-known contact sensitizers not included in the majority of commercially available baseline series., Objective: To investigate the outcome of patch testing to a textile dye mix (TDM) consisting of 8 disperse dyes., Methods: Two thousand four hundred ninety-three consecutive dermatitis patients in 9 dermatology clinics were patch tested with a TDM 6.6%, consisting of Disperse (D) Blue 35, D Yellow 3, D Orange 1 and 3, D Red 1 and 17, all 1.0% each, and D Blue 106 and D Blue 124, each 0.3%. 90 reacted positively to the TDM. About 92.2% of the patients allergic to the TDM were also tested with the 8 separate dyes., Results: Contact allergy to TDM was found in 3.6% (1.3-18.2) Simultaneous reactivity to p-phenylenediamine was found in 61.1% of the TDM-positive patients. Contact allergy to TDM and not to other p-amino-substituted sensitizers was diagnosed in 1.2%. The most frequent dye allergen in the TDM-positive patients was D Orange 3., Conclusions: Over 30% of the TDM allergic patients had been missed if only the international baseline series was tested. Contact allergy to TDM could explain or contribute to dermatitis in over 20% of the patients. Textile dye mix should be considered for inclusion into the international baseline series.

Background: Induction of delayed hypersensitivity reactions by red tattoos has been occasionally reported. Little is known about the inks used. Azo pigments have been implicated in some instances, but there is only one reported case involving quinacridones., Objectives: To describe the clinical and pathological features and outcome of skin reactions induced by red tattoo pigments., Patients, Materials, and Methods: Six patients with a cutaneous reaction induced by a red tattoo pigment underwent biopsy and prick and patch testing with the inks supplied., Results: We observed seven reactions in the 6 patients. Histology showed various patterns: three lichenoid, two eczematous, and two pseudolymphomatous. Five reactions occurred with azo pigments, and two with quinacridones, in both cases with Violet 19 and Red 122. Four inks were tested. Only one patch test gave a positive result at a late reading (day 7). Prick tests gave negative results. The reactions required various treatments, including laser treatment for 2 patients. Activation of the reaction in 1 case was transient., Conclusion: Azo pigments and quinacridones both triggered reactions with similar clinical aspects but with varying histological findings. Patch and prick test results were disappointing with both. Reactions occurred following laser use in 1 case., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Disperse dyes are well-known contact sensitizers. However, they are not included in the majority of commercially available baseline patch test series., Objectives: To investigate the outcome of patch testing with a textile dye mix (TDM) consisting of eight disperse dyes at dermatology clinics in various countries., Patients/materials/methods: Two thousand nine hundred and seven consecutive dermatitis patients at 12 dermatology clinics representing nine countries were tested with a TDM at 6.6%, consisting of Disperse Blue 35, Disperse Yellow 3, Disperse Orange 1 and 3, and Disperse Red 1 and 17, all at 1.0%, and Disperse Blue 106 and Disperse Blue 124, each at 0.3%, provisionally included in the baseline series. Eighty-seven per cent of the patients allergic to the TDM were also tested with the eight separate dyes., Results: Contact allergy to TDM was found in 108 patients (3.7%). The frequency of contact allergy varied from 2.1% to 6.9% in different centres. Simultaneous reactivity to p-phenylenediamine was found in 57 of the TDM-positive patients (53%). The most frequent dye allergen among the TDM-positive patients was Disperse Orange 3. The contact allergy could have explained or contributed to the dermatitis in approximately one-third of the patients for whom clinical relevance of the TDM contact allergy was recorded., Conclusions: The TDM should be considered for inclusion in the European baseline series., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Azo dyes in textiles may release aromatic amines after enzymatic cleavage by skin bacteria or after dermal absorption and metabolism in the human body. From the 896 azo dyes with known chemical structure in the available textile dyes database, 426 azo dyes (48%) can generate one or more of the 22 regulated aromatic amines in the European Union in Annex XVII of REACH. Another 470 azo dyes (52%) can be cleaved into exclusively non-regulated aromatic amines. In this study, a search for publicly available toxicity data on non-regulated aromatic amines was performed. For a considerable percentage of non-regulated aromatic amines, the toxicity database was found to be insufficient or non-existent. 62 non-regulated aromatic amines with available toxicity data were prioritized by expert judgment with objective criteria according to their potential for carcinogenicity, genotoxicity, and/or skin sensitization. To investigate the occurrence of azo dye cleavage products, 153 random samples of clothing textiles were taken from Swiss retail outlets and analyzed for 22 high priority non-regulated aromatic amines of toxicological concern. Eight of these 22 non-regulated aromatic amines of concern could be detected in 17% of the textile samples. In 9% of the samples, one or more of the aromatic amines of concern could be detected in concentrations >30 mg/kg, in 8% of the samples between 5 and 30 mg/kg. The highest measured concentration was 622 mg/kg textile. There is an obvious need to assess consumer health risks for these non-regulated aromatic amines and to fill this gap in the regulation of clothing textiles., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: The prevalence of occupational and non-occupational textile dermatitis seems to be increasing, probably because of changed textile manufacturing techniques., Objective: Evaluation of the clinical features and epidemiology of textile contact dermatitis (TCD) in Italy., Methods: Demographic data, clinical history aspects, atopy and positive patch test reactions to occupational and non-occupational allergens were investigated in 277 textile dermatitis patients., Results: Contact dermatitis was the most frequent clinical presentation (95.3%). TCD was more common in females, in the fourth to fifth decades of life, and in atopic dermatitis patients. The lesions were prevalently eczematous (74.2%), and mostly located on the trunk and lower limbs in non-occupational cases, and on the hands in textile workers. Allergic TCD (58.3%) was more frequent than irritant TCD. The dyes (Disperse Blue 124, Disperse Blue 106, and Disperse Yellow 3) were most frequently responsible (79.8%), especially in non-occupational TCD. Formaldehyde and resins were more important in occupational TCD. Concomitant reactions among textile dyes and/or finishing resins were observed in 50.0% of patients., Conclusions: Some strategies (sensitization potential of new textile chemicals, more stable dyes, reduced levels of formaldehyde in clothing, and collaboration with textile industry and trade associations) should be adopted to decrease the TCD incidence and update the textile patch testing series., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The safety of several azo colouring agents, used as food additives, has during the years been questioned. Allura Red AC (E129) has in some publications been classified as genotoxic. In fact, in the European Union, Allura Red is permitted as a food additive in human food, but, surprisingly, it was not acceptable as an additive for use in animal feed. In this study we have evaluated whether Allura Red is genotoxic using a flow cytometer-based micronucleus assay in peripheral blood of mice. Male FVB mice were given a single intra-peritoneal injection of various doses of Allura Red and sacrificed at 46 h after treatment. The tested doses were 0, 100, 200, 400, 600, 800, 1000, 1500, and 2000 mg/kg body weight (b.w.). Each dose group constituted three mice, except for in the dose group of 1000 mg/kg b. w., which constituted four mice. Blood samples were collected and the frequency of micronucleated polychromatic erythrocytes (fMNPCE) and the cell proliferation (%PCE) was determined. The analyses did not show any significant difference in the %PCE or in the fMNPCE. Consequently, under the testing circumstances one can conclude that Allura Red is not genotoxic., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: Simultaneous contact allergies to Disperse Orange 1, 4-nitroaniline and p-aminodiphenylamine (PADPA), as well as to other disperse azo dyes and to p-phenylenediamine (PPD), have been reported. Cross-reactivity is one of the possible explanations for simultaneous reactions between PPD and disperse azo dyes. Some metabolites from the azo reduction of these disperse azo dyes could be sensitizers, as human skin bacteria produce azo reductases., Objectives: To investigate the sensitizing capacity of Disperse Orange 1, PADPA and 4-nitroaniline, and the cross-reactivity between these substances and Disperse Yellow 3, its potential metabolites from azo reduction (4-aminoacetanilide and 2-amino-p-cresol), and PPD., Method: The guinea-pig maximization test was used., Results: It was found that both Disperse Orange 1 and PADPA are strong sensitizers and cross-react with each other. We were unable to sensitize guinea-pigs with 4-nitroaniline tested in equimolar concentrations to Disperse Orange 1., Conclusions: The results indicate that patients sensitized primarily to Disperse Orange 1 will also react to PADPA, which can be found mainly in hair dyes. PPD, 4-nitroaniline, 4-aminoacetanilide, 2-amino-p-cresol and Disperse Yellow 3 did not show any cross-reactivity with Disperse Orange 1 or PADPA., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Several disperse dyes (DDs) are still considered to be the most important allergens in textile dermatitis, but there are sparse data about their current use in textiles. The aim of this review was to evaluate published studies and reports on contact allergy to DDs published in PubMed during the last 22 years (1990-2012). Prevalence data are provided by study and by dye, as well as by the described clinical peculiarities of DD dermatitis. We reviewed 54 studies. In total, 26 DDs were tested. The average prevalence in screening studies was >1% for Disperse Blue 106, Disperse Blue 124, and Disperse Orange 3. There is a lack of data on patch testing with Disperse Blue 26, Disperse Blue 102, Disperse Orange 37, Disperse Orange 149, Disperse Yellow 23 and Disperse Yellow 49, which are listed as allergens by the EU Commission. It is necessary to check the purity and identity of dyes used for patch testing, confirm the clinical relevance of positive reactions by patch testing with suspected textiles, and, if the results are positive, determine the culprit dye., (© 2012 John Wiley & Sons A/S.)

Background: Recognition of allergic contact dermatitis attributed to textile dyes and resins is steadily increasing., Objective: This study aims to review the results of patch testing with a textile series at our institution and to compare with previously published reports., Methods: We performed a retrospective review of results in patients who underwent patch testing using a series of textile dyes and resins from January 1, 2000, through September 30, 2011., Results: A total of 671 patients (mean age, 56.5 years; female, 65.9%) were patch tested with the textile series (42 dyes and resins). These patients were also generally tested with the standard patch test series (n = 620). Of the patients, 219 (32.6%) demonstrated allergic reaction to 1 or more textile dyes and resins, and 71 (10.6%) manifested irritant reactions. The most frequent allergens were disperse blue 106 1% (8.3%), disperse blue 124 1% (8.0%), and melamine formaldehyde 7% (8.0%). Of patients tested with the standard series, 36 (5.8%) showed a positive reaction to the traditional textile screening allergen p-phenylenediamine 1%., Conclusions: Supplementing the standard series with the textile series increased detection of textile allergies. In patients suspected of textile allergy, addition of the textile series is necessary for appropriate diagnosis.

Background: It is known that, in vitro, human skin bacteria are able to split disperse azo dyes into the corresponding aromatic amines, some of which are sensitizers in the local lymph node assay. We hypothesize that the molecules of disperse dyes migrate onto the skin while garments are worn, and are metabolized and degraded by commensal skin bacteria. These molecules penetrate the skin and induce sensitization., Objectives: To evaluate the elicitation capacities of the possible azo-degradation products of the selected azo disperse dyes in patients allergic to them and to compare it with the elicitation capacities of other para-compounds., Methods: Ten patients allergic to Disperse Yellow 3 (DY3) and/or Disperse Orange 1 (DO1) were patch tested with a dilution series of the purified dyes 4-nitroaniline and p-aminodiphenylamine in concentrations equimolar to those of purified DO1 in the dilution series, as well as 4-aminoacetanilide and 2-amino-p-cresol in concentrations equimolar to those of purified DY3 in the dilution series. Results. Three patterns of patch test reactions could be seen. The 6 patients who were positive to DO1 and DY3 also reacted to p-aminodiphenylamine and 2-amino-p-cresol. Two patients were positive to DO1 only, and both reacted to p-aminodiphenylamine, but to neither 4-aminoacetanilide or 2-amino-p-cresol. Two patients did not react to DO1 or DY3 on this occasion., Conclusion: We show that it is possible that the major sensitizers in contact allergy to DO1 and DY3 are their metabolites, p-aminodiphenylamine and 2-amino-p-cresol, respectively, which might be formed by the azoreductase pathway of skin bacteria., (© 2012 John Wiley & Sons A/S.)