Abstract Background Persistent headache attributed to past stroke (PHAPS) is a controversial entity, recently included in the third edition of the International Classification of Headache Disorders (ICHD-3) despite being described only in retrospective studies. Objective To determine the frequency and characteristics of PHAPS in patients admitted with acute stroke. Methods We selected all patients with headache associated with acute stroke (HAAS) from a prospective, single-center registry of patients with acute stroke admitted to a Neurology ward between November 2018 and December 2019. We analyzed demographic, clinical, and neuroimaging data. We assessed the follow-up with a phone call questionnaire at 6 to 12 months. Results Among 121 patients with acute stroke, only 29 (24.0%) had HAAS. From these, 6 (5.0%) were lost to follow-up. In total, 23 (20.0%) patients answered the 6- to 12-month follow-up questionnaire and were included in this study. The median age of the sample was 53 years (interquartile range [IQR]: 38–78 years), and there was no sex predominance. Of the 10 patients (8,3%) that had persistent headache, 8 (6.6%) suffered from previous chronic headaches; however, they all mentioned a different kind of headache, and 1 (0,8%) probably had headache secondary to medication. Conclusions In the present study, only 10 out of 121 stroke patients (8.3%) referred persistent headache at the 6- to 12-month follow-up, but the majority already suffered from previous chronic headache, which raises the question that the actual prevalence of PHAPS may be lower than previously reported.

Resumo Antecedentes A cefaleia persistente atribuída a acidente vascular cerebral (AVC) prévio é uma entidade controversa, recentemente incluída na terceira edição da Classificação Internacional de Transtornos da Cefaleia (International Classification of Headache Disorders, ICHD-3), apesar de descrita apenas em estudos restrospectivos. Objetivo Determinar a frequência e as características do acidente vascular cerebral prèvio em doentes admitidos com AVC agudo. Métodos Selecionamos todos os doentes com cefaleia associada a AVC agudo de um registro unicêntrico e prospectivo de doentes admitidos no serviço de Neurologia entre novembro de 2018 e dezembro de 2019. Analisamos dados demográficos, clínicos e imagiológicos. Reavaliamos aos 6 a 12 meses por questionário telefónico. Resultados Entre os 121 doentes admitidos com AVC agudo, apenas 29 (24,0%) tinham cefaleia associada a AVC agudo. Destes, 6 (5,0%) perderam-se no seguimento. Ao todo, 23 (20,0%) responderam ao questionário e foram incluídos. A idade mediana foi de 53 anos (intervalo interquartílico [IIQ]: 38–78 anos), e não houve predominância de gênero. Dos 10 doentes (8,3%) que apresentam cefaleia persistente, 8 (6,6%) tinham histórico de cefaleia; no entanto todos mencionaram um tipo diferente de cefaleia, e 1 (0,8%) provavelmente tinha cefaleia secundária a medicação. Conclusões Neste estudo, apenas 10 em 121 doentes (8,3%) mencionaram cefaleia persistente aos 6 a 12 meses de seguimento, mas a maioria sofria de cefaleia prévia, pelo que a real prevalência de acidente vascular cerebral prèvio pode ser inferior à descrita nos estudos realizados até a data.

Resumen Actualmente las herramientas biotecnológicas como la ingeniería genética y la biología molecular se encuentran muy avanzadas, permitiendo el desarrollo de sistemas que se basan en el uso de células vegetales para la producción de proteínas recombinantes, lo cual permite el desarrollo de plataformas para la elaboración de biofármacos, enzimas y metabolitos secundarios. El uso de metodologías para transformación transitoria y estable, permiten la producción de proteínas recombinantes de interés. En este trabajo se realizó una transformación estable de callo de N. tabacum de la línea comercial BY-2 y una silvestre (T1). Además se realizaron ensayos de agroinfiltración por la técnica al vacío y con la técnica por jeringa. En ambos casos, la transformación fue mediada por Agrobacterium tumefaciens LBA4404 la cual poseía el vector pCAMBIA 1303, conteniendo los genes marcadores GFP y GUS, y un gen de resistencia a Higromicina. Se evaluó la expresión de la proteína verde fluorescente por medio de microscopía de fluorescencia. La expresión de los genes de resistencia a Higromicina y β-glucoronidasa se evaluaron por medio de PCR convencional. Se obtuvieron porcentajes de transformación aceptables, en conjunto con una alta expresión de la proteína verde fluorescente.

Abstract Biotechnological tools such as genetic engineering and molecular biology are very advanced, allowing the development of systems based on the use of plant cells for the production of recombinant proteins, which allows the development of platforms for biopharmaceuticals, enzymes and secondary metabolites production. Transient and stable transformation, allow production of recombinant proteins. In this work a stable transformation of N. tabacum callus from the BY-2 commercial line and a wild one (T1) was performed. In addition, agroinfiltration assays were carried out using two techniques: by syringe and vacuum. In both cases, the transformation was mediated by Agrobacterium tumefaciens LBA4404 which carried the vector pCAMBIA 1303, containing the GFP and GUS marker genes, and a hygromycin resistance gene. The expression of green fluorescent protein was evaluated by fluorescence microscopy. Expression of the hygromycin and β-glucuronidase resistance genes were evaluated by PCR. As a result, high GFP expression and good transformation rates were obtained.