Single ion mobility monitoring (SIM2) stitching method for high-throughput and high ion mobility resolution chiral analysis.

Chiral analysis is of high interest in many fields such as chemistry, pharmaceuticals and metabolomics. Mass spectrometry and ion mobility spectrometry are useful analytical tools, although they cannot be used as stand-alone methods. Here, we propose an efficient strategy for the enantiomer characterization of amino acids (AAs) using non-covalent copper complexes. A single ion mobility monitoring (SIM2) method was applied on a TIMS-ToF mass spectrometer to maximize the detection and mobility separation of isomers. Almost all of the 19 pairs of proteinogenic AA enantiomers could be separated with at least one combination with the chiral references L-Phe and L-Pro. Furthermore, we extended the targeted SIM2 method by stitching several mobility ranges, in order to be able to analyze complex mixtures in a single acquisition while maintaining high mobility resolution. Most of the enantiomeric pairs of AAs separated with the SIM2 method were also detected with this approach. The SIM2 stitching method thus opens the way to a more comprehensive chiral analysis with TIMS-ToF instruments.

Aprimoramento da Estrutura Normativa: a consolidação de normas do SUS e as atualizações do Decreto nº 12.002, de 2024 / Enhancement of the Regulatory Framework: The Consolidation of SUS Standards and updates to Decree No. 12,002, of 2024 / Mejora de la Estructura Normativa: La Consolidación de Normas del SUS y actualizaciones al Decreto N° 12.002, de 2024

O processo de consolidação depende de uma boa redação e elaboração dos atos normativos e é consequência inerente do aprimoramento de normas. Nesse sentido, apresenta-se as inovações do Decreto n.º 12.002, de 2024, relativas à consolidação, que dizem respeito, sobretudo, à obrigatoriedade de manutenção da consolidação normativa por meio da realização de alteração da norma consolidada e de medidas periódicas de revisão dessas normas. Essa manutenção constante dos atos consolidados preserva a necessidade de melhoria normativa, essencial para garantir segurança jurídica, transparência no ambiente democrático e cumprir as determinações da Organização para a Cooperação e o Desenvolvimento Econômico voltadas para política regulatória. A saúde, como pioneira no processo de consolidação dos seus atos normativos infralegais, tem a vanguarda no aprimoramento de normas e deve ser exemplo para outros órgãos.

The process of consolidation depends on good drafting and preparation of normative acts and is an inherent consequence of the improvement of regulations. In this regard, the innovations of Decree No. 12,002 of 2024 concerning Consolidation are presented, which mainly pertain to the mandatory maintenance of normative consolidation through amendments to the consolidated norms and periodic reviews of these norms. This constant maintenance of consolidated acts preserves the need for regulatory improvement, which is essential to ensure legal certainty, transparency in the democratic environment, and compliance with OECD regulatory policy directives. The health sector, as a pioneer in the consolidation of its infralegal normative acts, leads in the improvement of regulations and should serve as an example for other bodies.

El proceso de consolidación depende de una buena redacción y elaboración de los actos normativos y es una consecuencia inherente de la mejora de las normas. En este sentido, se presentan las innovaciones del Decreto nº 12.002 de 2024 relativas a la Consolidación, que se refieren principalmente a la obligatoriedad de mantener la consolidación normativa mediante la realización de modificaciones a la norma consolidada y de medidas periódicas de revisión de estas normas. Este mantenimiento constante de los actos consolidados preserva la necesidad de mejora normativa, esencial para garantizar seguridad jurídica, transparencia en el ambiente democrático y cumplir con las determinaciones de la OCDE dirigidas a la política regulatoria. La salud, como pionera en el proceso de consolidación de sus actos normativos infralegales, está a la vanguardia en la mejora de las normas y debe ser un ejemplo para otros órganos.

Caso Moradores de La Oroya contra o Peru: Corte Interamericana de Direitos Humanos atenta ao meio ambiente / Caso Habitantes de La Oroya contra Perú: Corte Interamericana de Derechos Humanos atenta al medio ambiente / Residents of La Oroya case against Peru: Inter-American Court of Human Rights attentive to the environment

Abordam-se aspectos da decisão da Corte Interamericana de Direitos Humanos, no caso Moradores de La Oroya contra o Peru, publicada em março de 2024, afirmando a justiciabilidade do direito ao meio ambiente como um direito difuso e reconhecendo a responsabilidade internacional do Peru (Estado parte) pela omissão em relação às medidas de prevenção e na prestação de informações à população exposta. Trata-se de precedente de relevância emitido pela Corte Internacional para a defesa do meio ambiente e dos direitos humanos, abrindo alvissareiras possibilidades de um novo e efetivo espaço para a afirmação de princípios caros ao Direito Sanitário e a preservação da vida.

Aspects of the decision of the Inter-American Court of Human Rights are discussed, in the case of Residents of La Oroya against Peru, affirming the justiciability of the right to the environment as a diffuse right and recognizing the international responsibility of Peru (State party) for omission in relation to prevention measures and the provision of information to the exposed population.This is a highly relevant precedent for the defense of the environment and human rights, opening up promising possibilities for a new and effective space for the affirmation of principles dear to Health Law and the preservation of life.

Se discuten aspectos de la decisión de la Corte Interamericana de Derechos Humanos, en el caso de Residentes de La Oroya contra Perú, afirmando la justiciabilidad del derecho al medio ambiente como un derecho difuso y reconociendo la responsabilidad internacional del Perú (Estado parte) por omisión en relación con las medidas de prevención y suministro de información a la población expuesta. Se trata de un precedente de gran relevancia para la defensa del medio ambiente y de los derechos humanos, abriendo posibilidades prometedoras para un nuevo y eficaz espacio de afirmación de principios queridos por el Derecho de la Salud y la preservación de la vida.

Parental Perceptions of the Impact of a Child's Complex Chronic Condition: A Validation Study of the Impact on Family Scale.

The diagnosis of a child's complex chronic illness may impact family relationships and cohesion. The Impact on Family Scale (IFS) is an instrument used to assess the parental perception of the effects of children's chronic illness on family life. With a sample of 110 mothers and fathers between the ages of 29 and 50 who have a child with a complex chronic illness, we examine evidence of the validity of the IFS for use in Portugal within this specific family configuration, (1) comparing its factor structure with the original one; (2) assessing its reliability; and (3) evaluating its relationship with life satisfaction and family cohesion/acceptance. As expected, CFA analysis showed that IFS is a one-factor reliable instrument with 12 items (Cronbach's alpha = 0.910), which are negatively correlated with satisfaction with life (r = -0.229, p = 0.016) and positively correlated with family acceptance and cohesion (r = 0.363; p < 0.001). The results support the validity of the IFS in families with children and adolescents with a complex chronic illness. The implications of the use of this instrument for research and professional practice is analyzed.

Interplatform comparison between three ion mobility techniques for human plasma lipid collision cross sections.

The implementation of ion mobility spectrometry (IMS) in liquid chromatography-high-resolution mass spectrometry (LC-HRMS) workflows has become a valuable tool for improving compound annotation in metabolomics analyses by increasing peak capacity and by adding a new molecular descriptor, the collision cross section (CCS). Although some studies reported high repeatability and reproducibility of CCS determination and only few studies reported good interplatform agreement for small molecules, standardized protocols are still missing due to the lack of reference CCS values and reference materials. We present a comparison of CCS values of approximatively one hundred lipid species either commercially available or extracted from human plasma. We used three different commercial ion mobility technologies from different laboratories, drift tube IMS (DTIMS), travelling wave IMS (TWIMS) and trapped IMS (TIMS), to evaluate both instrument repeatability and interlaboratory reproducibility. We showed that CCS discrepancies of 0.3% (average) could occur depending on the data processing software tools. Moreover, eleven CCS calibrants were evaluated yielding mean RSD below 2% for eight calibrants, ESI Low concentration tuning mix (Tune Mix) showing the lowest RSD (< 0.5%) in both ion modes. Tune Mix calibrated CCS from the three different IMS instruments proved to be well correlated and highly reproducible (R2 > 0.995 and mean RSD ≤ 1%). More than 90% of the lipid CCS had deviations of less than 1%, demonstrating high comparability between techniques, and the possibility to use the CCS as molecular descriptor. We highlighted the need of standardized procedures for calibration, data acquisition, and data processing. This work demonstrates that using harmonized analytical conditions are required for interplatform reproducibility for CCS determination of human plasma lipids.

Energy Resolved Mass Spectrometry for Interoperable Non-resonant Collisional Spectra in Metabolomics.

In untargeted metabolomics, the unambiguous identification of metabolites remains a major challenge. This requires high-quality spectral libraries for reliable metabolite identification, which is essential for translating metabolomics data into meaningful biological information. Several attempts have been made to generate reproducible product ion spectra (PIS) under a low collision energy (ELab) regime and nonresonant collisional conditions but have not fully succeeded. We examined the ERMS (energy-resolved mass spectrometry) breakdown curves of two lipo-amino acids and showed the possibility to highlight "singular points", called descriptors hereafter (linked to respective ELab depending on the instrument), for each of the monomodal product ion profiles. Using several instruments based on different technologies, the PIS recorded at these specific ELab sites shows remarkable similarities. The descriptors appeared as being independent of the fragmentation mechanisms and can be used to overcome the main instrumental effects that limit the interoperability of spectral libraries. This proof-of-concept study, performed on two particular lipo-amino acids, demonstrates the high potential of ERMS-derived information to determine the instrument-specific ELab at which PIS recorded in nonresonant conditions become highly similar and instrument-independent, thus comparable across platforms. This innovative but straightforward approach could help remove some of the obstacles to metabolite identification in nontargeted metabolomics, putting an end to a challenging chimera.

Innovative direct introduction-ion mobility-mass spectrometry (DI-IM-MS) approach for fast and robust isomer-specific quantification in a complex matrix: Application to 2'-fucosyllactose (2'-FL) in breast milk.

Identification and specific quantification of isomers in a complex biological matrix by mass spectrometry alone is not an easy task due to their identical chemical formula and therefore their same mass-to-charge ratio (m/z). Here, the potential of direct introduction combined with ion mobility-mass spectrometry (DI-IM-MS) for rapid quantification of isomers as human milk oligosaccharides (HMOs) was investigated. Differences in HMO profiles between various analyzed breast milk samples were highlighted using the single ion mobility monitoring (SIM2) acquisition for high ion mobility resolution detection. Furthermore, the Se+ (secretor) or Se- (non-secretor) phenotype could be assigned to breast milk samples studied based on their HMO contents, especially on the response of 2'-fucosyllactose (2'-FL) and lacto-N-fucopentaose I (LNFP I). The possibility of quantifying a specific isomer in breast milk by DI-IM-MS was also investigated. The standard addition method allowed the determination of the 2'-FL despite the presence of other oligosaccharides, including 3-fucosyllactose (3-FL) isomer in breast milk. This proof-of-concept study demonstrated the high potential of such an approach for the rapid and convenient quantification of isomers in complex mixtures.

Impactos do lockdown sobre os direitos dos cidadãos na cidade de São Luís do Maranhão / Impacts of the lockdown on citizens' rights in the municipality of São Luís do Maranhão / Impactos del confinamiento sobre los derechos de los ciudadanos en la ciudad de São Luís do Maranhão

O objetivo do presente estudo foi analisar os impactos do período do lockdown ocorrido em 2020 na cidade de São Luís, no Maranhão, de acordo com as percepções de cinco categorias profissionais. Trata-se de um estudo observacional descritivo de caráter qualitativo, realizado por meio de entrevista semiestruturada aplicada aos profissionais selecionados sobre os impactos do lockdown no direito constitucional de ir e vir, na vida profissional e no sistema de saúde brasileiro. Foram entrevistados 130 profissionais, entre médicos, professores universitários, gestores de saúde, empresários e autônomos. Para a análise da pergunta aberta, foi utilizado o método do discurso do sujeito coletivo (DSC). Foram também aplicados Testes de qui-quadrado ou exato de Fisher, adotando-se p < 0,05 como hipótese de nulidade, para verificar a associação entre variáveis categóricas. A maioria dos entrevistados considerou como negativos os impactos do lockdown sobre o direito de ir e vir dos cidadãos (51,6%) e nas atividades de vida diária de suas respectivas profissões (67,7%). O Sistema Único de Saúde não conseguiu conter os avanços da doença, segundo opiniões da maioria (59,2%) dos entrevistados. Os resultados qualitativos mostraram que o lockdown foi desfavorável para 66 profissionais, seguido de 33 que o classificaram como favorável. Os participantes destacaram que o lockdown causou impactos negativos no direito de ir e vir e na vida profissional dos cidadãos, tendo sido considerada uma medida política equivocada e ineficiente na contenção da disseminação da covid-19.

Objective: to analyze the impacts of the lockdown period in the municipality of São Luís, Maranhão (2020), according to the perceptions of five professional categories. Methodology: this descriptive, qualitative, and observational study was carried out by a semi-structured interview applied to selected professionals about the impacts of the lockdown on their constitutional right to free movement, their professional life, and on the Brazilian health system. Results: Overall, 130 professionals were interviewed, including physicians, university professors, health managers, businesspeople, and self-employed people. To analyze the open question, the Collective Subject Discourse-DSC method was used. Chi-squared or Fisher's exact tests were also applied, adopting p < 0.05 as the null hypothesis, to attest to the association between categorical variables. Most interviewees considered the impacts of the lockdown on citizens' right to free movement (51.6%) and on the daily life activities of their respective professions (67.7%) as negative. The Brazilian Unified Health System was unable to contain the spread of the disease, according to the opinions of most (59.2%) interviewees. Qualitative results showed that the lockdown was unfavorable for 66 professionals, followed by 33 professionals who classified it as favorable. Conclusion: the lockdown negatively impacted the right to free movement and professional lives and was considered a mistaken and inefficient political measure to contain the spread of COVID-19.

El objetivo de este estudio fue analizar los impactos del período de confinamiento en 2020 en la ciudad de São Luís, Maranhão, según las percepciones de cinco categorías profesionales. Se trata de un estudio observacional descriptivo, de carácter cualitativo, realizado mediante una entrevista semiestructurada aplicada a profesionales seleccionados sobre los impactos del confinamiento en el derecho constitucional de ir y venir, en la vida profesional y en el sistema de salud brasileño. Se entrevistaron a 130 profesionales, entre médicos, profesores universitarios, gestores de salud, empresarios y autónomos. Para analizar la pregunta abierta se utilizó el método discurso del sujeto colectivo (DSC). También se aplicaron las pruebas de Chi-cuadrado o exacta de Fisher, adoptando p < 0,05 como hipótesis nula, para verificar la asociación entre variables categóricas. La mayoría de los entrevistados consideraron negativos los impactos del confinamiento en el derecho de ir y venir de los ciudadanos (51,6%) y en las actividades de la vida diaria de sus respectivas profesiones (67,7%). El Sistema Único de Salud (SUS) no pudo contener la propagación de la enfermedad, según la opinión de la mayoría (59,2%) de los entrevistados. Los resultados cualitativos mostraron que el confinamiento fue desfavorable para 66 profesionales, seguido de 33 profesionales que lo calificaron como favorable. Los participantes destacaron que el confinamiento provocó impactos negativos en el derecho a ir y venir y en la vida profesional, además, fue considerado una medida política equivocada e ineficaz para contener la propagación del covid-19.

Experiences and strategies of people with sickle cell disease in the Federal District: the biographical rupture. / Experiências e estratégias de pessoas com doença falciforme no Distrito Federal: a ruptura biográfica.

This is a study on sickle cell disease, a chronic illness that affects many Brazilians, that aims to understand and analyze how people address the adversities arising from the diagnosis and the biographical rupture. The description of people's experiences and strategies conjures a picture that expresses the respondents' habitus, built in a dialectical relationship with the vulnerability determined by the disease. We adopted a qualitative approach and focused interviews as proposed by Merton, combined with the snowball technique, applied to groups related to sickle cell disease on social networks. Seven participants were selected because they were privileged informants with the disease, were over eighteen, lived in the Federal District, and were non-exclusive users of the Unified Health System. The interview material was categorized from the focal groups employed. The results indicated the following categories: biographical rupture, experience and coping strategies, and healthcare. It is necessary to sensitize professionals and the population about the challenging living conditions of people with sickle cell disease and the consolidation of public policies and care networks to accommodate this population.

Trata-se de um estudo sobre a doença falciforme, enfermidade crônica que afeta muitas pessoas no Brasil. O objetivo foi compreender como as pessoas lidam com as adversidades oriundas do diagnóstico e a ruptura biográfica. A descrição das experiências e estratégias empregadas pelas pessoas compuseram um quadro que expressa os habitus dos entrevistados, construídos em relação dialética com a vulnerabilidade determinada pela doença. A abordagem foi qualitativa e empregou entrevistas focadas como propostas por Merton, a técnica bola de neve aplicada em grupos relacionados à doença falciforme em redes sociais. Sete participantes foram selecionados por serem informantes privilegiados, portadores da doença, maiores de 18 anos, moradores no Distrito Federal e usuários não exclusivos do Sistema Único de Saúde. O material das entrevistas foi categorizado a partir dos núcleos focais empregados. Os resultados apontaram as categorias: ruptura biográfica, experiência e estratégias de enfrentamento, atenção à saúde. Concluímos que é necessária uma sensibilização dos profissionais e da população sobre as dificuldades da condição de vida das pessoas com doença falciforme e a consolidação das políticas públicas e das redes de atendimento para acolher essa população.

Knowledge about type 2 diabetes: its impact for future management.

Diabetes can cause several long-term complications. Knowledge about this disease can play an important role in reducing diabetes-related complications. In addition, the lack of awareness leads to misconceptions, which joined with inadequate knowledge, are relevant barriers to proper diabetes management. In this study, we aimed to assess the diabetes knowledge of a type 2 diabetes (T2D) population and identify major knowledge gaps, in order to prevent complications and to increase quality of life. In a cross-sectional, observational study in a convenience sample, we identified individuals diagnosed with T2D attending ambulatory visits from five health settings, older than 18 years, with a time diagnosis of at least 1 year, and attending multidisciplinary visits for at least 3 months. To assess the knowledge of T2D individuals, we applied the Portuguese version of the Diabetes Knowledge Test. The sample included a total of 1,200 persons, of whom almost half were female. The age range of the participants varied from 24 to 94 years old, and the mean age was 65.6 ± 11.4 years. Most of the sample had a level of education under secondary and lived with someone. In our sample, 479 (39.9%) were insulin-treated. The percentage of correct answers was 51.8% for non-insulin vs. 58.7% for insulin treated (p < 0.05). There were three items with a percentage of correct answers lower than 15%; the item with the lower value of correct answers was the one related to the identification of signs of ketoacidosis with only 4.4% of correct answers, the errors presented a random pattern; the item related to the identification of which food should not be used to treat low blood glucose with 11.9%, where 56.9% of the sample's participants considered that one cup of skim milk would be the correct answer (53.1% in non-insulin patients and 62.6% in insulin treated patients; p < 0.001). The item regarding the knowledge of free food presented a 13.3% of correct answers (10.8% non-insulin group vs. 17.1% insulin group; p < 0.01). Two of the three items with lower value of correct answers were related to glycemic control and health status monitoring, the other was related to diet and food.

Modeling Lysosomal Storage Disorders in an Innovative Way: Establishment and Characterization of Stem Cell Lines from Human Exfoliated Deciduous Teeth of Mucopolysaccharidosis Type II Patients.

Among the many lysosomal storage disorders (LSDs) that would benefit from the establishment of novel cell models, either patient-derived or genetically engineered, is mucopolysaccharidosis type II (MPS II). Here, we present our results on the establishment and characterization of two MPS II patient-derived stem cell line(s) from deciduous baby teeth. To the best of our knowledge, this is the first time a stem cell population has been isolated from LSD patient samples obtained from the dental pulp. Taking into account our results on the molecular and biochemical characterization of those cells and the fact that they exhibit visible and measurable disease phenotypes, we consider these cells may qualify as a valuable disease model, which may be useful for both pathophysiological assessments and in vitro screenings. Ultimately, we believe that patient-derived dental pulp stem cells (DPSCs), particularly those isolated from human exfoliated deciduous teeth (SHEDs), may represent a feasible alternative to induced pluripotent stem cells (iPSCs) in many labs with standard cell culture conditions and limited (human and economic) resources.

Bedside preoperative predictors of difficult laryngeal exposure in microlaryngeal surgery.

PURPOSE: Successful microlaryngeal surgery relies on an adequate laryngeal exposure. Recognizing the likelihood of challenging exposure prior to microlaryngeal surgery may assist in selecting the appropriate surgical approach and even prompt consideration of alternative treatment options. We aim to apply the mini-Laryngoscore, a preoperative assessment tool, to our study population and incorporate novel variables to optimize the prediction model. METHODS: This single-center prospective cohort study included 80 consecutive patients undergoing elective microlaryngeal surgery, from January 1, to June 30, 2023. Each patient underwent a presurgical evaluation of 15 parameters and an intraoperative scoring of the anterior commissure visualization. These parameters were assessed for their association with difficult laryngeal exposure, using multiple logistic regression analysis. We created a novel prediction model for DLE and compared it with the existing model, the mini-Laryngoscore. RESULTS: Out of 80 patients, 24 (30%) patients had difficult laryngeal exposure, including 3 cases (3.8%) in which visualization of the anterior commissure was not possible. A large neck diameter (OR, 1.4; CI 1.1-1.9) and the presence of upper teeth (OR, 8.9; CI 1.3-62.8) were independent risk factors for a difficult laryngeal exposure, while a larger interincisors gap was the only independent protector factor (OR, 0.3; CI 0.1-0.8). The logistic regression model combining these three independent risk factors displayed a high discriminative value AUC = 0.89 (CI 0.81-0.97). The predictive performance of the mini-Laryngoscore was 0.73 (CI 0.62-0.85). CONCLUSION: Combining two parameters from the mini-Laryngoscore (upper jaw dental state and interincisors gap distance) with neck circumference measurement can accurately predict the risk of difficult laryngeal exposure.

Publicação em inglês: um novo marco editorial dos Cadernos Ibero-Americanos de Direito Sanitário / Publication in English: a new editorial milestone for Cadernos Ibero-Americanos de Direito Sanitário / Publicación en Inglés: un nuevo hito editorial de los Cadernos Ibero-Americanos de Direito Sanitário

Os avanços tecnológicos impactaram diretamente a forma como a ciência é comunicada, reduzindo fronteiras e conectando indivíduos, mas a barreira linguística ainda persiste como um obstáculo significativo para a colaboração entre pesquisadores e a disseminação do conhecimento. Publicações em inglês têm alcance ampliado, viabilizando a difusão dos resultados dos estudos e pesquisas dos periódicos nacionais. Os Cadernos Ibero-Americanos de Direito Sanitário (CIADS), na esteira dessa vanguarda, passam a publicar seus artigos originais, resenhas e comunicações breves em inglês, sem custo para o autor, além da manutenção do idioma original. Este importante passo marca uma inovadora trajetória editorial do periódico, rumo à sua internacionalização.

Los avances tecnológicos han impactado directamente la forma en que se comunica la ciencia, reduciendo fronteras y conectando individuos, pero la barrera lingüística aún persiste como un obstáculo significativo para la colaboración entre investigadores y la difusión del conocimiento. Las publicaciones en inglés han ampliado su alcance, facilitando la difusión de los resultados de estudios e investigaciones en revistas nacionales. Siguiendo esta vanguardia, los Cadernos Ibero-Americanos de Direito Sanitário (CIADS) ahora publicarán sus artículos originales, reseñas y comunicaciones breves en inglés, sin costo para el autor, además de mantener el idioma original. Este importante paso marca una trayectoria editorial innovadora del periódico hacia su internacionalización.

Technological advances have directly impacted the way science is communicated, reducing borders and connecting individuals, but the language barrier still persists as a significant obstacle to collaboration between researchers and the dissemination of knowledge. Publications in English have a wider reach, making it possible to disseminate the results of studies and research in national journals. Cadernos Ibero-Americanos de Direito Sanitário (CIADS), in the wake of this vanguard, is now publishing its original articles, reviews and short communications in English, at no cost to the author, in addition to maintaining the original language. This important step marks the journal's innovative editorial path towards internationalization.

Enantioselective Reduction of Noncovalent Complexes of Amino Acids with CuII via Resonant Collision-Induced Dissociation: Collision Energy, Activation Duration Effects, and RRKM Modeling.

Formation of noncovalent complexes is one of the approaches to perform chiral analysis with mass spectrometry. Enantiomeric distinction of amino acids (AAs) based on the relative rate constants of competitive fragmentations of quaternary copper complexes is an efficient method for chiral differentiation. Here, we studied the complex [CuII,(Phe,PhG,Pro-H)]+ (m/z 493) under resonant collision-induced dissociation conditions while varying the activation time. The precursor ion can yield two main fragments through the loss of the non-natural AA phenylglycine (PhG): the expected product ion [CuII,(Phe,Pro-H)]+ (m/z 342) and the reduced product ion [CuI,(Phe,Pro)]+ (m/z 343). Enantioselective reduction describes the difference in relative abundance of these ions, which depends on the chirality of the precursor ion: the formation of the reduced ion m/z 343 is favored in homochiral complexes (DDD) compared to heterochiral complexes (such as LDD). Energy-resolved mass spectrometry data show that reduction, which arises from rearrangement, is favored at a low collision energy (CE) and long activation time (ActT), whereas direct cleavage preferentially occurs at a high CE and short ActT. These results were confirmed with kinetic modeling based on RRKM theory. For this modeling, it was necessary to set a pre-exponential factor as a reference, so that the E0 values obtained are relative values. Interestingly, these simulations showed that the critical energy E0 required to form the reduced ion is comparable in both homochiral and heterochiral complexes. However, the formation of product ion m/z 342 through direct cleavage is associated with a lower E0 in heterochiral complexes. Consequently, enantioselectivity would not be caused by enhanced reduction in homochiral complexes but rather by direct cleavage being favored in heterochiral complexes.

Very-early detection of atrial fibrillation after ablation evaluated by a wearable ECG-patch predicts late blanking period recurrence: Preliminary data from a prospective registry.

Introduction: Atrial fibrillation (AF) ablation represents a safe and effective procedure to restore sinus rhythm. The idea that post-procedural AF episodes - during the blanking period - are not considered treatment failure has been increasingly challenged. The E-Patch, a single-use adhesive electrode, facilitates extended continuous ECG monitoring for 120 h. This pilot study aims to assess the effectiveness of this ambulatory monitoring device and investigate whether very-early AF recurrence correlates with delayed blanking period ablation outcomes. Methods: We conducted a single-center, prospective, longitudinal study, including consecutive post-ablation patients monitored with the E-patch. The ability of the device to continuously record was analyzed, as well as the occurrence of AF episodes during external 7-day loop-recorder in the 2nd-month post-ablation. Results: We included 40 patients, median age 62 years (IQR 56-70). E-Patch monitoring was obtained for a median of 118 h (IQR 112-120), with no discomfort nor interpretation artefacts. Very-early AF recurrence was detected in 11 (27.5 %) patients, with a median AF burden of 7 % (IQR 6 %-33 %). Late-blanking period AF was detected in 13 (33 %) of the external 7-day loop recordings. Of the 11 patients that had very-early AF recurrence, 10 (91 %) had late-blanking AF. Very-early AF detection showed 77 % (95 % CI 64 %-90 %) sensitivity and 96 % (95 % CI 90-100 %) specificity in predicting late-blanking AF, with a non-parametric ROC curve AUC of 0.903 (95 % 0.797--1.0). Conclusion: The E-Patch was able to detect very-early AF during an extended period. Very-early AF detection emerges as a predictor of AF recurrence during the late blanking period post-ablation.

Exploring the role of ChatGPT in clinical decision-making in otorhinolaryngology: a ChatGPT designed study.

PURPOSE: Since the beginning of 2023, ChatGPT emerged as a hot topic in healthcare research. The potential to be a valuable tool in clinical practice is compelling, particularly in improving clinical decision support by helping physicians to make clinical decisions based on the best medical knowledge available. We aim to investigate ChatGPT's ability to identify, diagnose and manage patients with otorhinolaryngology-related symptoms. METHODS: A prospective, cross-sectional study was designed based on an idea suggested by ChatGPT to assess the level of agreement between ChatGPT and five otorhinolaryngologists (ENTs) in 20 reality-inspired clinical cases. The clinical cases were presented to the chatbot on two different occasions (ChatGPT-1 and ChatGPT-2) to assess its temporal stability. RESULTS: The mean score of ChatGPT-1 was 4.4 (SD 1.2; min 1, max 5) and of ChatGPT-2 was 4.15 (SD 1.3; min 1, max 5), while the ENTs mean score was 4.91 (SD 0.3; min 3, max 5). The Mann-Whitney U test revealed a statistically significant difference (p < 0.001) between both ChatGPT's and the ENTs's score. ChatGPT-1 and ChatGPT-2 gave different answers in five occasions. CONCLUSIONS: Artificial intelligence will be an important instrument in clinical decision-making in the near future and ChatGPT is the most promising chatbot so far. Despite needing further development to be used with safety, there is room for improvement and potential to aid otorhinolaryngology residents and specialists in making the most correct decision for the patient.

Neutrophil extracellular traps in rheumatoid arthritis and periodontitis: Contribution of PADI4 gene polymorphisms.

AIM: We sought to investigate the release of neutrophil extracellular traps (NETs) in neutrophils from individuals with rheumatoid arthritis (RA) and controls and compare the presence of NETs in gingival tissues according to periodontal status. Also, the association between single nucleotide polymorphisms (SNPs) of the peptidyl arginine deaminase type 4 (PADI4) gene and the GTG haplotype with RA, periodontitis and NETs was evaluated in vitro. MATERIALS AND METHODS: Peripheral neutrophils were isolated by density gradient, and NET concentration was determined by the PicoGreen method. Immunofluorescence was studied to identify NETs by co-localization of myeloperoxidase (MPO)-citrullinated histone H3 (H3Cit). Genotyping for SNPs (PADI4_89; PADI4_90; PADI4_92; and PADI4_104) was performed in 87 individuals with RA and 111 controls. RESULTS: The release of NETs in vitro was significantly higher in individuals with RA and periodontitis and when stimulated with Porphyromonas gingivalis. Gingival tissues from subjects with RA and periodontitis revealed increased numbers of MPO-H3Cit-positive cells. Individuals with the GTG haplotype showed a higher release of NETs in vitro and worse periodontal parameters. CONCLUSIONS: The release of NETs by circulating neutrophils is associated with RA and periodontitis and is influenced by the presence of the GTG haplotype.

Experiências e estratégias de pessoas com doença falciforme no Distrito Federal: a ruptura biográfica / Experiences and strategies of people with sickle cell disease in the Federal District: the biographical rupture

Resumo Trata-se de um estudo sobre a doença falciforme, enfermidade crônica que afeta muitas pessoas no Brasil. O objetivo foi compreender como as pessoas lidam com as adversidades oriundas do diagnóstico e a ruptura biográfica. A descrição das experiências e estratégias empregadas pelas pessoas compuseram um quadro que expressa os habitus dos entrevistados, construídos em relação dialética com a vulnerabilidade determinada pela doença. A abordagem foi qualitativa e empregou entrevistas focadas como propostas por Merton, a técnica bola de neve aplicada em grupos relacionados à doença falciforme em redes sociais. Sete participantes foram selecionados por serem informantes privilegiados, portadores da doença, maiores de 18 anos, moradores no Distrito Federal e usuários não exclusivos do Sistema Único de Saúde. O material das entrevistas foi categorizado a partir dos núcleos focais empregados. Os resultados apontaram as categorias: ruptura biográfica, experiência e estratégias de enfrentamento, atenção à saúde. Concluímos que é necessária uma sensibilização dos profissionais e da população sobre as dificuldades da condição de vida das pessoas com doença falciforme e a consolidação das políticas públicas e das redes de atendimento para acolher essa população.

Abstract This is a study on sickle cell disease, a chronic illness that affects many Brazilians, that aims to understand and analyze how people address the adversities arising from the diagnosis and the biographical rupture. The description of people's experiences and strategies conjures a picture that expresses the respondents' habitus, built in a dialectical relationship with the vulnerability determined by the disease. We adopted a qualitative approach and focused interviews as proposed by Merton, combined with the snowball technique, applied to groups related to sickle cell disease on social networks. Seven participants were selected because they were privileged informants with the disease, were over eighteen, lived in the Federal District, and were non-exclusive users of the Unified Health System. The interview material was categorized from the focal groups employed. The results indicated the following categories: biographical rupture, experience and coping strategies, and healthcare. It is necessary to sensitize professionals and the population about the challenging living conditions of people with sickle cell disease and the consolidation of public policies and care networks to accommodate this population.

Help Comes from Unexpected Places: How a Tiny Fairy and a Tropical Fish may help us Model Mucopolysaccharidoses.

INTRODUCTION: When it comes to disease modeling, countless models are available for Lysosomal Storage Diseases (LSD). Historically, two major approaches are well-established: in vitro assessments are performed in patient fibroblasts, while in vivo pre-clinical studies are performed in mouse models. Still, both platforms have a series of drawbacks. Thus, we implemented two alternative and innovative protocols to mimic a particular sub-group of LSDs, the Mucopolysaccharidoses both in vitro and in vivo. METHODS: The first one relies on a non-invasive approach using dental pulp stem cells from deciduous teeth (SHEDs). SHEDs are multipotent neuronal precursors that can easily be collected. The second uses a state-of-the-art gene editing technology (CRISPR/Cas9) to generate zebrafish disease models. RESULTS: Even though this is an ongoing project, we have already established and characterized two MPS II and one MPS VI SHED cell models. These cells self-maintain through several passages and can give rise to a variety of cells including neurons. Furthermore, all MPS-associated sub-cellular phenotypes we have assessed so far are easily observable in these cells. Regarding our zebrafish models, we have successfully knocked down both naglu and hgsnat and the first results we got from the behavioral analysis are promising ones, as we can observe altered activity and sleep patterns in the genetically modified fish. For this particular approach we chose MPS III forms as our target disorders, since their neurological features (hyperactivity, seizures and motor impairment) and lifespan decrease would be easily recognizable in zebrafish. CONCLUSION: Now that these methods are well-established in our lab, their potential is immense. On one hand, the newly developed models will be of ultimate value to understand the mechanisms underlying MPS sub-cellular pathology, which have to be further elucidated. On the other hand, they will constitute an optimal platform for drug testing in house. Also noteworthy, our models will be published as lab resources and made available for the whole LSD community.

Challenges in the Definitive Diagnosis of Niemann-Pick Type C-Leaky Variants and Alternative Transcripts.

Niemann-Pick type C (NPC, ORPHA: 646) is a neuro-visceral, psychiatric disease caused predominantly by pathogenic variants in the NPC1 gene or seldom in NPC2. The rarity of the disease, and its wide range of clinical phenotypes and ages of onset, turn the diagnosis into a significant challenge. Other than the detailed clinical history, the typical diagnostic work-up for NPC includes the quantification of pathognomonic metabolites. However, the molecular basis diagnosis is still of utmost importance to fully characterize the disorder. Here, the authors provide an overview of splicing variants in the NPC1 and NPC2 genes and propose a new workflow for NPC diagnosis. Splicing variants cover a significant part of the disease-causing variants in NPC. The authors used cDNA analysis to study the impact of such variants, including the collection of data to classify them as leaky or non-leaky pathogenic variants. However, the presence of naturally occurring spliced transcripts can misdiagnose or mask a pathogenic variant and make the analysis even more difficult. Analysis of the NPC1 cDNA in NPC patients in parallel with controls is vital to assess and detect alternatively spliced forms. Moreover, nonsense-mediated mRNA decay (NMD) analysis plays an essential role in evaluating the naturally occurring transcripts during cDNA analysis and distinguishing them from other pathogenic variants' associated transcripts.