Tuberculosis inequalities and socio-economic deprivation in Portugal

OBJECTIVE: To analyse the geographical distribution of tuberculosis (TB) in Portugal and estimate the association between TB and socio-economic deprivation. METHODS: An ecological study at the municipality level using TB notifications for 2010–2014 was conducted. Spatial Bayesian models were used to calculate smoothed standardised notification rates, identify high- and low-risk areas and estimate the association between TB notification and the European Deprivation Index (EDI) for Portugal and its component variables. RESULTS: Standardised notification rates ranged from 4.41 to 76.44 notifications per 100 000 population. Forty-one high-risk and 156 low-risk municipalities were identified. There was no statistically significant association between TB notification rate and the EDI, but some of its variables, such as the proportion of manual workers and the percentage unemployed, were significantly and directly associated with TB notification, whereas the variable ‘proportion of residents with low education level' showed an inverse relationship. CONCLUSION: Wide inequalities in TB notification rates were observed, and some areas continued to exhibit high TB notification rates. We found significant associations between TB and some socio-economic factors of the EDI.This work was supported by contributions from Iceland, Liechtenstein and Norway through the European Economic Area Grants, under the Public Health Initiatives Programme (PT 06, grant number 138DT1)

Sentinel lymph node biopsy in rats. Comparison between paraffin and frozen section analysis

PURPOSE:To simulate a lymph node metastasis in an animal model using activated carbon, assess their identification in frozen section analysis and compare with histopathological examination in paraffin.METHODS:Thirty two adult female rats were used. They received the carbon injection on its hind legs. Half of the rats was sacrificed on day one, and the other half after 21 days. Thus, 64 lymph nodes were dissected and split longitudinally. One half of the lymph node was sent immediately to frozen section analysis. The other half was fixed in 10% formaldehyde to be cut in paraffin. Slides were divided into quadrants and classified by the presence of carbon in these four quadrants_ They were also classified by the carbon staining intensity.RESULTS:Comparing the slides obtained in the first day and 21 days, there was a tendency of carbon to spread over time, but without statistical significance. The intensity did not alter over time.CONCLUSION:There was no concordance between the two methods of pathological analysis, however the actived carbon was seen in all lymph nodes.UNIFESPUNIFESP Pathology DepartmentUniversidade Federal de São Paulo (UNIFESP) Department of SurgeryUNIFESP, Pathology DepartmentUNIFESP, Department of SurgerySciEL

Selection of hydrotropes for enhancing the solubility of artemisinin in aqueous solutions

Artemisinin is an antimalarial substance very sparingly soluble in water. In the attempt to identify environmental-friendly and non-toxic aqueous-based solvents to extract it from Artemisia annua L., the solubility of artemisinin in aqueous solutions of different hydrotropes was measured at 303.2 K, for hydrotrope concentrations up to 5 M. The ability of the studied hydrotropes for enhancing the artemisinin solubility increases in the following order: Na[N(CN)2] < Na[SCN] < [Chol][Van] < [Chol][Gal] < [N4,4,4,4]Cl < [Chol][Sal] < [P4,4,4,4]Cl < Na[Sal], with Na[Sal] allowing an increase in the solubility of 750 fold compared to pure water. The COSMO-RS model and experimental Kamlet-Taft solvatochromic parameters were applied to connect the solubility enhancement with solvent properties. At low hydrotrope concentration, the solubility increases with the decreasing of the difference between the Apolar Factors of the hydrotrope and artemisinin, while for higher hydrotrope concentration, the hydrogen-bond acceptor character of the hydrotrope seems to have an impact on the solubility enhancement. Even if some mechanistic understanding is still to unfold, quantitatively the empirical correlations of solubility enhancement with the hydrotrope concentration and the solvatochromic parameters show very high accuracy. In particular, 93% of the change on the artemisinin solubility enhancement could be explained using the hydrotrope concentration and two combined solvatochromic parameters (αβ and π∗2) as explaining variables.This work was developed within the scope of the projects CICECOAveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, CIMO-Mountain Research Center, UIDB/00690/2020, and Green Health (Norte-01-0145-FEDER-000042) all financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 and NORTE 2020 Partnership Agreement. Isabela Sales and Silvana Mattedi thanks the finantial support from CAPES and CNPq/ Brazil (CAPES: Proc. 88881.189075/2018-01 and 88887.494428/2020- 00. CNPq: Grant 303089/2019-9 and Proc.438036/2018-2).info:eu-repo/semantics/publishedVersio

Tuberculosis among the homeless: should we change the strategy?

BACKGROUND: Tuberculosis (TB) is a major concern among high-risk populations such as the homeless. OBJECTIVES: To evaluate TB incidence and treatment outcomes among homeless patients in Portugal and to identify predictors of unsuccessful TB treatment outcomes among the homeless. DESIGN: This was a retrospective cohort study of all TB patients notified in Portugal from 2008 to 2014. Characteristics of homeless TB patients were assessed and predictors of unsuccessful TB treatment were determined using logistic regression. RESULTS: TB incidence among the homeless was 122/100 000 homeless persons and was positively correlated with TB incidence among non-homeless persons. Homeless TB patients had a higher prevalence of alcohol and/or drug use, human immunodeficiency virus (HIV) co-infection, cavitary TB and smear positivity. The rate of unsuccessful treatment outcomes among the homeless was 28.6%, and was significantly associated with increased age, injection drug use (IDU) and HIV co-infection. CONCLUSION: TB incidence among homeless persons was five times that among the non-homeless, and higher in regions with greater TB incidence among non homeless persons. The successful treatment outcome rate was lower. Predictors of unsuccessful treatment were age, IDU and HIV co-infection. Integrated TB programmes targeting homeless and non-homeless patients, with measures targeting specific characteristics, may contribute to TB elimination in Portugal.CONTEXTE : La tuberculose (TB) est un souci majeur dans les populations à haut risque comme les personnes sans domicile fixe. OBJECTIFS : Evaluer le taux d’incidence de la TB et les resultats du traitement parmi des patients sans domicile fixe au Portugal et identifier les facteurs de préediction d’ échec du traitement de la TB parmi ces patients. SCHÉMA : Etude rétrospective de cohorte incluant tous les patients TB notifies au Portugal entre 2008 et 2014. Les caractéristiques des patients sans domicile fixe ont été ́évaluées et les facteurs de prédiction d’ échec du traitement de la TB ont été déterminés par ŕegression logistique. RESULTATS : Le taux d’incidence de la TB parmi les personnes sans domicile fixe a été de 122/100 000, et il a été positivement corrélé avec l’incidence de la TB parmi le reste de la population. Les patients tuberculeux sans domicile fixe avaient une prévalence plus élevée de consommation d’alcool et/ou de drogues, de co- infection au virus de l’immunodéficience humaine (VIH), de forme caverneuse et de frottis positif. Le taux d’ ́echec du traitement a ́et ́e de 28,6% ; l’ ́echec a ́ et ́esignificativement associé à un âge plus avancé, à la consommation de drogues injectables et à la co-infection par le VIH. CONCLUSION : L’incidence dela TB parmi les personnes sans domicile fixe a été cinq fois plus élevée que celle du reste de la population et plus haute dans les régions ou l’incidence dans le reste de la population est egalement plus élevée. Leur taux d’ échec du traitement à été plus faible. Les facteurs de prédiction d’ échec du traitement ont été l´âge, la consommation de drogues injectables et la co-infection `a VIH. Des programmes de TB intégrés ciblant les patients sans domicile fixe et les autres, avec des mesures spécifiques adaptées à leurs caractéristiques particulières, pourrait contribuer à l’ élimination de la TB au Portugal.MARCO DE REFERENCIA: La tuberculosis (TB) constituye una gran preocupación en las poblaciones muy vulnerables como las personas sin hogar. OBJETIVOS: Evaluar la tasa de incidencia de TB y los desenlaces terapéuticos en las personas sin domicilio en Portugal y definir los factores pronósticos de fracaso terapéutico en este grupo de la población. MÉTODO: Fue este un estudio retrospectivo de cohortes de todos los pacientes con diagnóstico de TB notificados del 2008 al 2014 en Portugal. Mediante un análisis de regresión logística se analizaron las características de los pacientes tuberculosos sin hogar y los factores pronósticos de fracaso terapéutico. RESULTADOS: La tasa de incidencia de TB en la población sin hogar fue 122 por 100 000 personas y exhibió una correlación positiva con la incidencia de TB en las personas con domicilio. Los pacientes con diagnóstico de TB y sin hogar presentaron una prevalencia más alta de consumo de alcohol y/o de drogas, de coinfección por el virus de la inmunodeficiencia humana (VIH), de lesiones cavernosas y de resultados positivos de la baciloscopia. La tasa de fracaso terapéutico en esta población fue 28,6% y se asoción de manera significativa con una mayor edad, el consumo de drogas intravenosas y la coinfección por el VIH. CONCLUSIÓN: La incidencia de TB en las personas sin hogar fue cinco veces mayor que en las personas con domicilio y fue más alta en las regiones con una mayor incidencia de TB en las personas con domicilio. La tasa de éxito terapéutico en las personas sin hogar fue más baja. Los factores pronósticos de fracaso terapéutico fueron la edad, el consumo de drogas intravenosas y la coinfecció non por el VIH. La ejecución de programas integrados de atención de la TB dirigidos a las personas sin hogar y con domicilio, que comporten medidas específicas que aborden sus características particulares, podrıa contribuir a la eliminación de la TB en PortugalStudy Group for Infectious Diseases of Instituto de Saúde Púublica da Universidade do Porto who collaborated on this project: B Miranda, C Carvalho, C Matos, C Carvalho, G Rodrigues, J Goncalves, L Maio and T Rito. This work was supported by contributions from Iceland, Liechtenstein and Norway through the European Economic Area Grants under the Public Health Initiatives Programme (PT 06, grant number 138DT1). RG was also partially supported by Centro de Matemática da Universidade do Porto (UID/MAT/00144/2013), which is funded by Fundação do Ministério de Ciência e Tecnologia(Portugal) with national (MEC) and European structural funds (Fonds europeen de d ́eveloppement economique et regional) under the PT2020 Partnership Agreementinfo:eu-repo/semantics/publishedVersio

The impact of the counterion in the performance of ionic hydrotropes

The efficiency of an ionic hydrotrope is shown to increase with the hydrophobicity of its counterion, challenging the common view that ionic hydrotropes should possess a small, densely charged counterion such as sodium or chloride.publishe

Enhancing artemisinin solubility in aqueous solutions: searching for hydrotropes based on ionic liquids

Artemisinin is a sesquiterpenoid lactone peroxide, known for its potent antimalarial activity that can be extracted from Artemisia annua L. This compound is only sparingly soluble in water, making its extraction using environmental-friendly and non-toxic aqueous solvents difficult. In the attempt to overcome this limitation, hydrotropes, which are a class of compounds that can assist in increasing the solubility of hydrophobic solutes in water, were investigated in this work. In particular, the hydrotropic capability of ionic liquids (ILs) on the aqueous solubility of artemisinin was studied. The effects of IL concentration and anion nature of 1-butyl-3-methylimidazolium-based ILs on the solubility of artemisinin at 303.2 K in water were evaluated. It is here shown the excellent capacity of ILs containing thiocyanate or dicyanamide anions to enhance the solubility of artemisinin in aqueous media, with a magnitude comparable to that obtained with the best organic solvents. Furthermore, solvatochromic parameters of the ILs aqueous solutions were also measured and combined with COSMO-RS and the cooperative hydrotropy model to establish relations between the artemisinin solubility enhancement and the solvent characteristics. The solubility enhancement of artemisinin is favored by the apolarity of the medium and the lower hydrogen-bond acceptor character of the hydrotrope.This work was developed within the scope of the projects CICECO-Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, CIMO-Mountain Research Center, UIDB/00690/2020, all financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. Isabela Sales and Silvana Mattedi thanks the finantial support from CAPES and CNPq/Brazil (CAPES: Proc. 88881.189075/2018-01 and 88887.494428/2020-00. CNPq: Grant 303089/2019-9 and Proc.438036/2018-2).info:eu-repo/semantics/publishedVersio

Sentinel lymph node in children with melanoma: case report

Objective: to present a case of a child who was subjected to sentinel lymph node biopsy for cutaneous melanoma. Description: a 12 year-old child with Dysplastic Nevus Syndrome developed melanoma on the lumbar region. The excision biopsy revealed a melanoma with depth of 1.5 mm. The patient was submitted to amplification of the margins 2 cm in all directions and the sentinel node was also excised. The histopathological exam did not show residual disease. Sentinel on exam did not show metastases either under hematoxylin-eosin stain or immunohistochemistry (S-100 and HMB45). Therefore, RT-PCR for tyrosinase mRNA was positive. The patient has been followed for twelve months without evidence of recurrence. Comments: childhood melanoma is rare, corresponding to less than 1% of malignant tumors in children. Data point to a worldwide increase in its incidence. Melanoma occurs in melanocytic lesions in 70% and in the remaining 30% it occurs de novo. Melanoma is very aggressive, so the survival depends on an early diagnosis. Sentinel lymph node biopsy has selected patients to complete lymphadenectomy. Some authors have been using this technique in childhood melanoma.Objetivo: apresentar um caso de melanoma cutâneo na infância, submetido à biópsia de linfonodo sentinela. Descrição: criança de 12 anos de idade, portadora da síndrome do nevo displásico, desenvolveu melanoma em dorso. A biópsia excisional revelou melanoma (Breslow = 1,5 mm), sendo submetida a tratamento cirúrgico da lesão, com ampliação de margem de 2cm e biópsia de linfonodo sentinela. O exame anatomopatológico não mostrou doença residual. O exame do linfonodo sentinela não mostrou metástases ao exame histopatológico por hematoxilina eosina, nem à imunohistoquímica (S100 e HMB45). No entanto, a pesquisa de RNA mensageiro da tirosinase por RT-PCR se mostrou positiva. A paciente não mostrou sinais de metástase ou recidiva local nestes doze meses iniciais de seguimento. Comentários: o melanoma é raro em crianças, corresponde a menos de 1% dos tumores da infância. Inúmeros trabalhos mostram o aumento da incidência mundial do melanoma. Em cerca de 70%, se originam de nevos melanocíticos pré-existentes, e nos 30% restantes, eles surgem de novo. Dada a agressividade do melanoma, a sobrevida depende do diagnóstico precoce. A biópsia do linfonodo sentinela tem contribuído na decisão de se realizar ou não a linfadenectomia completa, e alguns autores já estão utilizando a mesma em crianças.Escola Paulista de Medicina Programa de Pós-graduação em Cirurgia PlásticaUniversidade Federal de São Paulo (UNIFESP) Disciplina de Cirurgia PlásticaUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de PatologiaUNIFESP, EPM, Programa de Pós-graduação em Cirurgia PlásticaEPM, Disciplina de Cirurgia PlásticaEPM, Escola Paulista de Medicina Depto. de PatologiaSciEL

Is intra-operative gamma probe detection really necessary for inguinal sentinel lymph node biopsy?

CONTEXT: Sentinel node (SN) biopsy has changed the surgical treatment of malignant melanoma. The literature has emphasized the importance of gamma probe detection (GPD) of the SN. OBJECTIVE: Our objective was to evaluate the efficacy of patent blue dye (PBD) and GPD for SN biopsy in different lymphatic basins. DESIGN: Patients with cutaneous malignant melanoma in stages I and II were submitted to biopsy of the SN, identified by PBD and GPD, as part of a research project. SETTING: Patients were seen at Hospital São Paulo by a multidisciplinary group (Plastic Surgery Tumor Branch, Nuclear Medicine and Pathology). PATIENTS: 64 patients with localized malignant melanoma were studied. The median age was 46.5 years. The primary tumor was located in the neck, trunk or extremities. INTERVENTIONS: Preoperative lymphoscintigraphy, lymphatic mapping with PBD and intraoperative GPD was performed on all patients. The SN was examined by conventional and immunohistochemical staining. If the SN was not found or contained micrometastases, only complete lymphadenectomy was performed. MAIN MEASUREMENTS: The SN was identified by PBD if it was blue-stained, and by GPD if demonstrated activity five times greater than the adipose tissue of the neighborhood. RESULTS: Seventy lymphatic basins were explored. Lymphoscintigraphy showed ambiguous drainage in 7 patients. GPD identified the SN in 68 basins (97%) and PBD in 53 (76%). PBD and GPD identified SN in 100% of the inguinal basins. For the remaining basins both techniques were complementary. A metastatic SN was found in 10 basins. Three patients with negative SN had recurrence (median follow-up = 11 months). CONCLUSION: Although both GPD and PBD are useful and complementary, PBD alone identified the SN in 100% of the inguinal lymphatic basins.CONTEXTO: A biópsia de linfonodo sentinela (LS) mudou a abordagem cirúrgica do melanoma maligno. A literatura tem enfatizado a importância da detecção gama intra-operatória (DG) do LS. OBJETIVO: Nosso objetivo é avaliar a eficácia do corante azul patente (AP) e da DG na biópsia de LS em diferentes bases linfáticas. TIPO DE ESTUDO: Pacientes portadores de melanoma maligno cutâneo foram submetidos à biópsia do LS, usando AP e DG como parte de um projeto de pesquisa. LOCAL: Hospital São Paulo, grupo multidisciplinar (cirurgião oncológico, médico nuclear e patologista). PACIENTES: Foram estudados 64 pacientes portadores de melanoma maligno localizado, com idade mediana de 46,5 anos. O sítio primário estava localizado no pescoço, tronco e nos membros. INTERVENÇÕES: Linfocintilografia pré-operatória, mapeamento linfático com AP e DG foram realizados em todos os pacientes. O LS foi examinado por histopatologia convencional e imunohistoquímica. Quando o LS não foi encontrado ou continha micrometástases, linfadenectomia completa da base linfática foi realizada. VARIÁVEIS ESTUDADAS: O LS foi considerado como identificado pelo AP se corado em azul e pela DG quando demonstrou pelo menos 5 vezes mais atividade do que o tecido gorduroso vizinho. RESULTADOS: Foram exploradas 70 bases linfáticas. A linfocintilografia mostrou drenagem ambígua em 7 pacientes. DG identificou o LS em 68 bases linfáticas (97%) e o AP o fez em 53 bases (76%). Os dois métodos identificaram separadamente 100% dos LS inguinais. Nas demais bases, as técnicas foram complementares. O LS estava invadido por células tumorais em 10 bases. Três pacientes com LS negativo apresentaram recorrência (seguimento mediano de 11 meses). CONCLUSÃO: Embora o emprego de AP e GP na pesquisa de LS sejam complementares, o AP demonstrou ser um método suficiente para a localização do LS inguinal.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Discipline of Plastic Surgery/Tumor branchUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Department of DermatologyUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Department of Nuclear MedicineUNIFESP, EPM, Discipline of Plastic Surgery/Tumor branchUNIFESP, EPM, Department of DermatologyUNIFESP, EPM, Department of Nuclear MedicineSciEL

Diagnóstico diferencial no melanoma primário e metastático por espectroscopia FT-Raman

PURPOSE: To qualify the FT-Raman spectral data of primary and metastatic cutaneous melanoma in order to obtain a differential diagnosis. METHODS: Ten normal human skin samples without any clinical or histopathological alterations, ten cutaneous melanoma fragments, and nine lymph node metastasis samples were used; 105, 140 and 126 spectra were obtained respectively. Each sample was divided into 2 or 3 fragments of approximately 2 mm³ and positioned in the Raman spectrometer sample holder in order to obtain the spectra; a monochrome laser light Nd:YAG at 1064 nm was used to excite the inelastic effect. RESULTS: To differentiate the three histopathological groups according to their characteristics extracted from the spectra, data discriminative analysis was undertaken. Phenylalanine, DNA, and Amide-I spectral variables stood out in the differentiation of the three groups. The percentages of correctly classified groups based on Phenylalanine, DNA, and Amide-I spectral features was 93.1%. CONCLUSION: FT-Raman spectroscopy is capable of differentiating melanoma from its metastasis, as well as from normal skin.OBJETIVO: Qualificar os dados espectrais FT-Raman do melanoma cutâneo primário e metastático e assim realizar o diagnóstico diferencial. MÉTODOS: Foram utilizadas amostras de 10 fragmentos de pele sem alterações clínicas ou histopatológicas, 10 de melanomas cutâneos e 9 de metástases linfonodais; 105, 140 and 126 espectros foram obtidos respectivamente. Cada amostra foi dividida em 2 ou 3 frações de 2 mm³ e posicionada no porta amostras do espectrômetro Raman para obtenção dos espectros, por meio da excitação do espalhamento inelástico pelo laser de Nd:YAG em 1064 nm incididos na amostra. RESULTADOS: Para diferenciar os três grupos formados de acordo com as características fornecidas pelos espectros, realizamos a análise discriminante dos dados. As variáveis espectrais Fenilalanina, DNA e Amida-I se destacaram na capacidade de diferenciação dos três grupos histológicos. A porcentagem de classificação correta utilizando estes critérios foi de 93,1%; o que mostra a eficiência da análise realizada. CONCLUSÃO: A espectroscopia FT-Raman é capaz de diferenciar o melanoma de sua metástase, assim como da pele normal.UNIFESPUniversidade Federal de São Paulo (UNIFESP) Department of SurgeryPathology DepartmentPathology Department Federal University of ABC Head of Center for Human and Natural Sciences (CCNH)UNIVAP Institute of Research and Development Head of Biomedical Vibrational Spectroscopy LaboratoryUniversidade Federal de São Paulo (UNIFESP) Department of Surgery Head of Division of Plastic SurgeryUNIFESP, Department of SurgeryUNIFESP, Department of Surgery Head of Division of Plastic SurgerySciEL

Distributed automated manufacturing of pluripotent stem cell products

Establishing how to effectively manufacture cell therapies is an industry-level problem. Decentralised manufacturing is of increasing importance, and its challenges are recognised by healthcare regulators with deviations and comparability issues receiving specific attention from them. This paper is the first to report the deviations and other risks encountered when implementing the expansion of human pluripotent stem cells (hPSCs) in an automated three international site–decentralised manufacturing setting. An experimental demonstrator project expanded a human embryonal carcinoma cell line (2102Ep) at three development sites in France, Germany and the UK using the CompacT SelecT (Sartorius Stedim, Royston, UK) automated cell culture platform. Anticipated variations between sites spanned material input, features of the process itself and production system details including different quality management systems and personnel. Where possible, these were pre-addressed by implementing strategies including standardisation, cell bank mycoplasma testing and specific engineering and process improvements. However, despite such measures, unexpected deviations occurred between sites including software incompatibility and machine/process errors together with uncharacteristic contaminations. Many only became apparent during process proving or during the process run. Further, parameters including growth rate and viability discrepancies could only be determined post-run, preventing ‘live’ corrective measures. The work confirms the critical nature of approaches usually taken in Good Manufacturing Practice (GMP) manufacturing settings and especially emphasises the requirement for monitoring steps to be included within the production system. Real-time process monitoring coupled with carefully structured quality systems is essential for multiple site working including clarity of decision-making roles. Additionally, an over-reliance upon post-process visual microscopic comparisons has major limitations; it is difficult for non-experts to detect deleterious culture changes and such detection is slow