Binding of the five multistate species of the anthocyanin analog 7- β -D-glucopyranosyloxy-4′-hydroxyflavylium to the β -cyclodextrin derivative captisol

The host-guest chemistry of the anthocyanin analog 7-β-D-glucopyranosyloxy-4'-hydroxyflavylium (GHF) was studied in the presence of the β-cyclodextrin derivative captisol by stopped flow, UVevisible spectroscopy, flash photolysis, circular dichroism and isothermal titration calorimetry. The equilibrium and rate constants of the multistate of chemical species derived from the flavylium ion were calculated and compared with those in the absence of the host. A new procedure to obtain the host-guest association constants of the multistate (including the transient species) by superimposing the two energy level diagrams, in the presence and absence of the cyclodextrin, was developed. The results indicate that the magnitude of the association constants follows the order, trans-chalcone = cis-chalcone = hemiketal > quinoidal base > flavylium cation. The hydration equilibrium constant increases ca. 42 times in the presence of captisol as the hydration and dehydration rate constants respectively increases and decreases. The other equilibrium constants are modestly affected: the rate constants of ring closure and opening are significantly decreased in the complex and the isomerization rate constants increase in both directions. The quantum yield of the photochromic system in the presence of captisol is0.3, i.e. 3 times higher than in the absence of the host

Aplicação de Membranas Amnióticas na Reconstrução da Superfície Ocular Externa em Idade Pediátrica

Introdução: Vários estudos comprovam os benefícios da aplicação de membranas amnióticas na reconstrução da superfície ocular em idade adulta, pelos seus efeitos anti-inflamatorios, anti-adesivos e anti-apoptóticos. Em idade pediátrica, a reconstrução da superfície ocular carece de especial atenção e este procedimento encontra-se ainda pouco estudado. Não foi encontrado nenhum estudo especificamente dirigido à aplicação de membranas amnióticas na reconstrução da superfície ocular externa desta faixa etária, pelo que procurámos elucidá-lo. Material e métodos: Estudo retrospetivo englobando todos os doentes em idade pediátrica (7,1 anos +/- 4,17) submetidos a transplante de membrana amniótica no Centro Hospitalar de Lisboa Central entre 2008 e 2015, para reconstrução da superfície ocular externa. Entre os doentes (n=6 olhos de 6 crianças), quatro apresentavam patologia do foro neoformativo ou inflamatório e dois apresentavam queimaduras extensas da superfície ocular. Foi realizada divisão em dois grupos, com base na presença ou não de insuficiência de células limbares. Foram avaliadas características clínicas e demográficas, MAVC antes e após a cirurgia, tempo de reepitelização, amplitude de movimentos oculares antes e após a cirurgia, presença de recidiva ou complicações e resultado estético. O tempo de seguimento foi de 4 a 83 meses. Resultados: Verificaram-se sucessos completos em todos os doentes sem insuficiência limbar (50% do total de doentes), sucessos parciais em dois dos doentes com insuficiência limbar (33,3%) e um caso de falência terapêutica (16.7%). Nos doentes em que se observou recidiva, o tempo médio para esta ocorrência foi de 8,3 +/- 6,8 meses. Não se verificaram complicações em nenhum dos casos. Observou-se melhoria pós-operatória em um dos dois casos que tinham diminuição da acuidade visual pré-operatória (aumento da MAVC em 6 linhas). Verificou-se ainda uma melhoria da motilidade ocular e aspeto estético em todos os doentes com alterações prévias destes parâmetros. Conclusão: O transplante de membrana amniótica mostrou ser muito benéfico também em idade pediátrica. Pode ser realizado como tratamento isolado ou coadjuvante, sendo os resultados superiores nos casos de células limbares funcionantes. Não foi detetada maior incidência de complicações ou rejeições comparativamente ao descrito na literatura para a idade adulta.info:eu-repo/semantics/publishedVersio

Coloboma: Chave Ocular Para Patologia Sistémica

Introdução: Um coloboma é uma anomalia do desenvolvimento que se caracteriza, na maioria dos casos, por um deficiente encerramento da fissura embrionária na 6ª semana da gestação. Trata-se de um defeito que pode afectar diferentes estruturas do globo ocular, nomeadamente, a íris, corpo ciliar, coróide, retina ou nervo óptico. A eventual associação com patologia sistémica e alterações genéticas faz com que o diagnóstico oftalmológico seja fundamental para a orientação e seguimento dos doentes. Material e métodos:Estudo retrospectivo de 26 doentes da Consulta de Oftalmologia Pediátrica do Hospital de São José, do Centro Hospitalar de Lisboa Central, através da consulta do processo, exames complementares de diagnóstico e registo fotográfico. Foram caracterizados os doentes de acordo com o sexo, idade, antecedentes familiares de anomalias oculares, antecedentes gestacionais, tipo de coloboma, localização, lateralidade, melhor acuidade visual corrigida, presença de outras anomalias oculares concomitantes e associação com doenças sistémicas. Com este estudo pretendemos avaliar e comparar o perfil dos doentes portadores de colobomas, comparando-o com o descrito na literatura, bem como salientar a importância para o seu seguimento, despiste de doenças sistémicas associadas e tratamento das complicações. Resultados: Dos 26 doentes avaliados, 14 (54%) eram do sexo feminino, e 12 (46%) do sexo masculino. A idade variou entre 1 e 25 anos, com média de 11,23 anos. A média da idade do diagnóstico foi de 2,6 anos, variando entre 1 mês até 8 anos de idade. Nenhum doente apresentava história familiar de coloboma, existindo história de prematuridade em 2 casos (8%). Quanto à localização, 16 casos (62%) apresentavam coloboma do disco óptico, 14 (54%) da íris e 13 (50%) colobomas coriorretinianos. Em 13 casos (50%) o coloboma atingia mais do que uma estrutura anatómica e era bilateral em 11 casos (42%). As acuidades visuais variaram entre ausência de percepção luminosa e 20/20. Apenas 3 casos (12%) apresentavam um coloboma isolado, sendo os restantes associados a outras alterações oftalmológicas, nomeadamente estrabismo em 13 casos (50%), microftalmia em 8 casos (31%), catarata em 3 casos (12%), descolamento de retina em 3 casos (12%) e nistagmus em 4 casos (15%). Da série de 26 doentes, 19 (73%) não apresentavam doenças sistémicas associadas ao coloboma. 4 doentes (15%) foram classificados como tendo síndrome CHARGE, 1 aguardando confirmação molecular (4%), 1 síndrome de Joubert, 1 síndrome de Kabuki e 1 síndrome de DiGeorge. Foram detectadas anomalias do desenvolvimento psicomotor em 8 casos (31%). Conclusões:O exame oftalmológico completo é importante no diagnóstico, prognóstico e vigilância de doentes com coloboma. O coloboma é frequentemente o primeiro achado diagnóstico nestas crianças, sendo que a referenciação pela Oftalmologia a outras especialidades é fundamental, devendo incluir o despiste das várias anomalias sistémicas que podem estar associadas.info:eu-repo/semantics/publishedVersio

Ocular Disease in Children with Type 1 Diabetes

Specialized care provided to diabetic patients, in developed countries, has contributed to the reduced number of ocular complications in pediatric age. The effectiveness of annual eye examinations in this group is unclear. The authors intend to determine the prevalence and onset of ocular disease in this population and adapt the results to the dynamics of a tertiary center, based on a retrospective study of type 1 diabetes patients’ medical records registered and monitored in Pediatric Endocrinology Department of Centro Hospitalar de Lisboa Central, between January of 2008 and July of 2015. Demographic characteristics, underlying and ocular disease, referral and follow-up of these patients in the Pediatric Ophthalmology Department were evaluated. Medical records of 304 patients with type 1 diabetes were analyzed. 110 patients referred to Ophthalmology Department had been included in the study. The mean age was 13.2 years (4-20 years), 57 were male and 53 female. Type 1 diabetes was diagnosed at 7.5 years on average (0.75-13 years) and mean disease duration was 6.0 years (1-14 years). The average of the last hemoglobin A1c value was 8.6% (4.9-14.4%). The first eye exam was performed at 9.4 years on average (2-19 years), in 55 cases the patients had less than 10 years and, of those, 29 were referred in the first year after diagnosis (mean duration diabetes of 2.0 years; 0-7 years). Regarding the 55 patients older than 9 years, 27 were referred in the first year after diagnosis (mean duration of diabetes 2.3 years, 0-9 years). In 54 patients ophthalmological exam was repeated after 1 year, 15 after 2 years, 1 after 3 years and 1 after 5 years. No diabetic retinopathy cases were found. 28 astigmatism, 12 hyperopia, 16 myopia, 5 strabismus, 2 blepharoptosis and 1 polar cataract were diagnosed. International Society for Pediatric and Adolescent Diabetes recommend ophthalmological initial screening 2 years after the diagnosis of type 1 diabetes, in patients above 10 years, and 5 years after, if lower ages. American Academy of Ophthalmology 2014 guidelines recommend that the first eye examination should begin 5 years after the diagnosis and repeated annually. A previous study conducted at Hospital Dona Estefânia, between 1999 and 2000, by Rodrigues P et al has described a low incidence of ocular lesions (6.4%) in the studied population, being in agreement with our results. Recent published studies are also consistent and demonstrate that the rare incidence of ocular complications of diabetes until puberty can justify new guidelines in the coming years.info:eu-repo/semantics/publishedVersio

XEN® Implant and Trabeculectomy Medium-Term Quality of Life Assessment and Comparison of Results

AIM: To evaluate and compare the quality of life of patients submitted to XEN® implant or trabeculectomy and the relationship with potentially involved variables. METHODS: A cross-sectional study of patients with advanced open-angle glaucoma who underwent implantation of XEN® (group 1) and trabeculectomy (group 2) between October 2015 and February 2017. The studied variables were: age, gender, follow-up time, need of topical anti-hypertensive therapy, visual acuity and intraocular pressure (IOP). The quantification of the quality of life was attained through the Glaucoma Symptom Scale (GSS) questionnaire. RESULTS: Totally 34 eyes (34 patients) were included, 17 in each group. The mean GSS scores for group 1 were 42.6±6.8 (median, 47; p25, 36.5; p75, 48.5) and for group 2 it was 41.6±7.0 (median, 43; p25, 36.5; p75, 47.0; P=0.34). There was a strong negative correlation between the need for topical anti-hypertensive drugs and the GSS result in both groups (r=-0.88, P<0.01, r=-0.59, P=0.01, respectively) and a moderate negative correlation with IOP in group 1 (r=-0.50, P=0.03). CONCLUSION: The analysis demonstrates the non-inferiority of medium-term quality of life of one group in relation to the other (XEN® implant and trabeculectomy). The number of topical anti-hypertensive drugs and IOP negatively influenced the quality of life.info:eu-repo/semantics/publishedVersio

Late Onset Neuromyelitis Optica Spectrum Disorders (LONMOSD) from a Nationwide Portuguese Study: Anti-AQP4 Positive, Anti-MOG Positive and Seronegative Subgroups

Introduction: Several neuroimmunological disorders have distinct phenotypes according to the age of onset, as in multiple sclerosis or myasthenia gravis. It is also described that late onset NMOSD (LONMOSD) has a different phenotype. Objective: To describe the clinical/demographic characteristics of the LONMOSD and distinguish them from those with early onset (EONMOSD). Methods: From a nationwide Portuguese NMOSD study we analyzed the clinical/demographic characteristics of the LONMOSD. Results: From the 180 Portuguese patients 45 had disease onset after 50 years old, 80% were female. 23 had anti-AQP4 antibodies (51.1%), 13 anti-MOG antibodies (28.9%) and 9 were double seronegative (20.0%). The most common presenting phenotypes in LONMOSD were transverse myelitis (53.3%) and optic neuritis (26.7%), without difference from EONMOSD (p = 0.074). The mean EDSS for LONMOSD was 6.0 (SD=2.8), after a mean follow-up time of 4.58 (SD=4.47) years, which was significantly greater than the mean EDSS of EONMOSD (3.25, SD=1.80)(p = 0.022). Anti-AQP4 antibodies positive LONMOSD patients had increased disability compared to anti-MOG antibodies positive LONMOSD (p = 0.022). The survival analysis showed a reduced time to use a cane for LONMOSD, irrespective of serostatus (p<0.001). Conclusions: LONMOSD has increased disability and faster progression, despite no differences in the presenting clinical phenotype were seen in our cohort.info:eu-repo/semantics/publishedVersio

Neuromyelitis Optica Spectrum Disorders: a Nationwide Portuguese Clinical Epidemiological Study

Introduction: Neuromyelitis optica spectrum disorder (NMOSD) is a rare disorder in which astrocyte damage and/or demyelination often cause severe neurological deficits. Objective: To identify Portuguese patients with NMOSD and assess their epidemiological/clinical characteristics. Methods: This was a nationwide multicenter study. Twenty-four Portuguese adult and 3 neuropediatric centers following NMOSD patients were included. Results: A total of 180 patients met the 2015 Wingerchuk NMOSD criteria, 77 were AQP4-antibody positive (Abs+), 67 MOG-Abs+, and 36 seronegative. Point prevalence on December 31, 2018 was 1.71/100,000 for NMOSD, 0.71/100,000 for AQP4-Abs+, 0.65/100,000 for MOG-Abs+, and 0.35/100,000 for seronegative NMOSD. A total of 44 new NMOSD cases were identified during the two-year study period (11 AQP4-Abs+, 27 MOG-Abs+, and 6 seronegative). The annual incidence rate in that period was 0.21/100,000 person-years for NMOSD, 0.05/100,000 for AQP4-Abs+, 0.13/100,000 for MOG-Abs+, and 0.03/100,000 for seronegative NMOSD. AQP4-Abs+ predominated in females and was associated with autoimmune disorders. Frequently presented with myelitis. Area postrema syndrome was exclusive of this subtype, and associated with higher morbidity/mortality than other forms of NMOSD. MOG-Ab+ more often presented with optic neuritis, required less immunosuppression, and had better outcome. Conclusion: Epidemiological/clinical NMOSD profiles in the Portuguese population are similar to other European countries.info:eu-repo/semantics/publishedVersio

The NS1 Glycoprotein Can Generate Dramatic Antibody-Enhanced Dengue Viral Replication in Normal Out-Bred Mice Resulting in Lethal Multi-Organ Disease

Antibody-enhanced replication (AER) of dengue type-2 virus (DENV-2) strains and production of antibody-enhanced disease (AED) was tested in out-bred mice. Polyclonal antibodies (PAbs) generated against the nonstructural-1 (NS1) glycoprotein candidate vaccine of the New Guinea-C (NG-C) or NSx strains reacted strongly and weakly with these antigens, respectively. These PAbs contained the IgG2a subclass, which cross-reacted with the virion-associated envelope (E) glycoprotein of the DENV-2 NSx strain, suggesting that they could generate its AER via all mouse Fcγ-receptor classes. Indeed, when these mice were challenged with a low dose (<0.5 LD50) of the DENV-2 NSx strain, but not the NG-C strain, they all generated dramatic and lethal DENV-2 AER/AED. These AER/AED mice developed life-threatening acute respiratory distress syndrome (ARDS), displayed by diffuse alveolar damage (DAD) resulting from i) dramatic interstitial alveolar septa-thickening with mononuclear cells, ii) some hyperplasia of alveolar type-II pneumocytes, iii) copious intra-alveolar protein secretion, iv) some hyaline membrane-covered alveolar walls, and v) DENV-2 antigen-positive alveolar macrophages. These mice also developed meningo-encephalitis, with greater than 90,000-fold DENV-2 AER titers in microglial cells located throughout their brain parenchyma, some of which formed nodules around dead neurons. Their spleens contained infiltrated megakaryocytes with DENV-2 antigen-positive red-pulp macrophages, while their livers displayed extensive necrosis, apoptosis and macro- and micro-steatosis, with DENV-2 antigen-positive Kuppfer cells and hepatocytes. Their infections were confirmed by DENV-2 isolations from their lungs, spleens and livers. These findings accord with those reported in fatal human “severe dengue” cases. This DENV-2 AER/AED was blocked by high concentrations of only the NG-C NS1 glycoprotein. These results imply a potential hazard of DENV NS1 glycoprotein-based vaccines, particularly against DENV strains that contain multiple mutations or genetic recombination within or between their DENV E and NS1 glycoprotein-encoding genes. The model provides potential for assessing DENV strain pathogenicity and anti-DENV therapies in normal mice