Position paper of diagnosis and treatment of post-extubation laryngitis in children: a multidisciplinary expert-based opinion.

OBJECTIVES: To make recommendations on the diagnosis and treatment of post-extubation laryngitis (PEL) in children with or without other comorbidities. METHODS: A three-iterative modified Delphi method was applied. Specialists were recruited representing pediatric otolaryngologists, pediatric and neonatal intensivists. Questions and statements approached topics encompassing definition, diagnosis, endoscopic airway evaluation, risk factors, comorbidities, management, and follow-up. A consensus was defined as a supermajority >70%. RESULTS: Stridor was considered the most frequent symptom and airway endoscopy was recommended for definitive diagnosis. Gastroesophageal reflux and previous history of intubation were considered risk factors. Specific length of intubation did not achieve a consensus as a risk factor. Systemic corticosteroids should be part of the medical treatment and dexamethasone was the drug of choice. No consensus was achieved regarding dosage of corticosteroids, although endoscopic findings help defining dosage and length of treatment. Non-invasive ventilation, laryngeal rest, and use of comfort sedation scales were recommended. Indications for microlaryngoscopy and bronchoscopy under anesthesia were symptoms progression or failure to improve after the first 72-h of medical treatment post-extubation, after two failed extubations, and/or suspicion of severe lesions on flexible fiberoptic laryngoscopy. CONCLUSIONS: Management of post-extubation laryngitis is challenging and can be facilitated by a multidisciplinary approach. Airway endoscopy is mandatory and impacts decision-making, although there is no consensus regarding dosage and length of treatment.

Radiotherapy-activated NBTXR3 nanoparticles promote ferroptosis through induction of lysosomal membrane permeabilization.

PURPOSE: Radiotherapy-activated NBTXR3 (NBTXR3 + RT) has demonstrated superior efficacy in cancer cell destruction and tumor growth control, compared to radiotherapy (RT), in preclinical and clinical settings. Previous studies highlighted the immunomodulatory properties of NBTXR3 + RT, such as modification of tumor cell immunogenicity/adjuvanticity, producing an effective local tumor control and abscopal effect, related to an enhanced antitumor immune response. Furthermore, NBTXR3 + RT has shown potential in restoring anti-PD1 efficacy in a refractory tumor model. However, the early events leading to these results, such as NBTXR3 endocytosis, intracellular trafficking and primary biological responses induced by NBTXR3 + RT remain poorly understood. METHODS: We analyzed by transmission electron microscopy endocytosis and intracellular localization of NBTXR3 nanoparticles after endocytosis in various cell lines, in vitro and in vivo. A kinetic of NBTXR3 endocytosis and its impact on lysosomes was conducted using LysoTracker staining, and a RNAseq analysis was performed. We investigated the ability of NBTXR3 + RT to induce lysosomal membrane permeabilization (LMP) and ferroptosis by analyzing lipid peroxidation. Additionally, we evaluated the recapture by cancer cells of NBTXR3 released from dead cells. RESULTS: NBTXR3 nanoparticles were rapidly internalized by cells mainly through macropinocytosis and in a less extend by clathrin-dependent endocytosis. NBTXR3-containing endosomes were then fused with lysosomes. The day following NBTXR3 addition, we measured a significant increase in LysoTracker lysosome labeling intensity, in vitro as in vivo. Following RT, a significant lysosomal membrane permeabilization (LMP) was measured exclusively in cells treated with NBTXR3 + RT, while RT had no effect. The day post-irradiation, a significant increase in lipid peroxidation, a biomarker of ferroptosis, was measured with NBTXR3 + RT compared to RT. Moreover, we demonstrated that NBTXR3 nanoparticles released from dead cells can be recaptured by cancer cells. CONCLUSIONS: Our findings provide novel insights into the early and specific biological effects induced by NBTXR3 + RT, especially LMP, not induced by RT in our models. The subsequent significant increase in lipid peroxidation partially explains the enhanced cancer cell killing capacity of NBTXR3 + RT compared to RT, potentially by promoting ferroptosis. This study improves our understanding of the cellular mechanisms underlying NBTXR3 + RT and highlights its potential as an agnostic therapeutic strategy for solid cancers treatment.

Nanoparticle-enhanced proton beam immunoradiotherapy drives immune activation and durable tumor rejection.

The combination of radiation therapy (RT) and immunotherapy has emerged as a promising treatment option in oncology. Historically, x-ray radiation (XRT) has been the most commonly used form of RT. However, proton beam therapy (PBT) is gaining recognition as a viable alternative, as it has been shown to produce similar outcomes to XRT while minimizing off-target effects. The effects of PBT on the antitumor immune response have only just begun to be described, and to our knowledge no studies to date have examined the effect of PBT as part of a combinatorial immunoradiotherapeutic strategy. Here, using a 2-tumor model of lung cancer in mice, we show that PBT in tandem with an anti-PD1 antibody substantially reduced growth in both irradiated and unirradiated tumors. This was accompanied by robust activation of the immune response, as evidenced by whole-tumor and single-cell RNA sequencing showing upregulation of a multitude of immune-related transcripts. This response was further significantly enhanced by the injection of the tumor to be irradiated with NBTXR3 nanoparticles. Tumors of mice treated with the triple combination exhibited increased infiltration and activation of cytotoxic immune cells. This triple combination eradicated both tumors in 37.5% of the treated mice and showed robust long-term immunity to cancer.

Identification and targeting of protein tyrosine kinase 7 (PTK7) as an immunotherapy candidate for neuroblastoma.

GD2-targeting immunotherapies have improved survival in children with neuroblastoma, yet on-target, off-tumor toxicities can occur and a subset of patients cease to respond. The majority of neuroblastoma patients who receive immunotherapy have been previously treated with cytotoxic chemotherapy, making it paramount to identify neuroblastoma-specific antigens that remain stable throughout standard treatment. Cell surface glycoproteomics performed on human-derived neuroblastoma tumors in mice following chemotherapy treatment identified protein tyrosine kinase 7 (PTK7) to be abundantly expressed. Furthermore, PTK7 shows minimal expression on pediatric-specific normal tissues. We developed an anti-PTK7 chimeric antigen receptor (CAR) and find PTK7 CAR T cells specifically target and kill PTK7-expressing neuroblastoma in vitro. In vivo, human/murine binding PTK7 CAR T cells regress aggressive neuroblastoma metastatic mouse models and prolong survival with no toxicity. Together, these data demonstrate preclinical efficacy and tolerability for targeting PTK7 and support ongoing investigations to optimize PTK7-targeting CAR T cells for neuroblastoma.

Percepção dos internos de Medicina acerca do atendimento destinado às pessoas com deficiência: modelo curricular insuficiente? / Perception of Medical interns about assisting people with disabilities: insufficient curriculum model?

Resumo Introdução: O conceito de deficiência relaciona-se às limitações socialmente impostas aos deficientes por seus corpos não corresponderem ao modelo aceito como normal. Em geral, o acesso à saúde dessa população é precário, em parte, pela falta de preparação voltada ao cuidado dos pacientes deficientes durante a formação médica, demonstrando que os currículos das faculdades necessitam de revisão. Objetivo: Este estudo teve como objetivo analisar a percepção do aluno interno do curso de Medicina da Universidade Federal do Ceará (UFC) acerca do modelo curricular atual no contexto da formação médica, especificamente voltado ao atendimento destinado às pessoas com deficiência. Método: Foi disponibilizado um formulário elaborado no Google Formulários, para coletar informações de internos da Faculdade de Medicina (Famed). Depois, realizaram-se entrevistas semiestruturadas via Google Meet mediante questões que objetivaram entender a relação entre futuros médicos e pessoas com deficiências, além da confiança do preparo para o atendimento a tais pacientes. Para exame do material empírico adquirido, utilizou-se a análise de discurso de Rueda. Resultado: Foram entrevistados 13 internos que relataram limitação no aprendizado do estabelecimento da relação médico-paciente em relação a pessoas com deficiência, durante o ciclo básico (do primeiro ao quarto semestre), evoluindo, principalmente, para o internato. Consideraram-se a educação, o entendimento das condições socioeconômicas e culturais do paciente, e a construção de um plano terapêutico executável as qualidades a serem desenvolvidas pelo interno. Quanto aos principais problemas relatados, destacou-se a dificuldade na realização do exame físico e na comunicação. Por sua vez, a ajuda de acompanhantes e o auxílio da equipe profissional foram apontados como aspectos positivos. Percepções referentes ao preparo para atender deficientes foram contrastantes: alguns relataram segurança por capacitações e conhecimentos empíricos, enquanto outros se sentiram inseguros ou incapazes. Percebe-se, também, importante consideração para haver adaptação curricular acerca desse tema, com intervenções nas disciplinas obrigatórias e optativas. Conclusão: Evidenciou-se que os entrevistados sentem dificuldades no atendimento destinado a deficientes, o que sugere alterações no currículo da Famed-UFC.

Abstract Introduction: The concept of disability is related to the socially-imposed limitations on the disabled because their bodies do not correspond to the model accepted as normal. Access to health care for this population is generally precarious, partly due to the lack of preparation for the care of disabled patients during medical school. This suggests that the school curricula need to be revised. Objective: To analyse the perception of the senior medical students at the Federal University of Ceará (UFC) about the current curriculum, specifically, in relation to the care of people with disabilities. Method: A form previously prepared on the Google Forms platform was made available to collect information from senior medical students at the School of Medicine (FAMED). Afterwards, semi-structured interviews were carried out via Google Meet using questions that aimed to understand the relations between future doctors and people with disabilities, as well as the confidence of the students in their preparation for caring for such patients. For analysis of the empirical material acquired, we used Rueda's speech analysis. Result: Thirteen interns were interviewed, who reported limitations in learning how to establish a doctor-patient relationship with patients with disabilities during the first two years of medical school, having effects on the future clinical rotations. Politeness, knowledge of the patient's socioeconomic and cultural conditions, and building an executable therapeutic plan were considered qualities to be developed by the student. The main problems reported was difficulty in performing the physical examination and communication with disabled patients. However, help from the patients' carers and from the professional team was considered positive by the students. Perceptions regarding the preparation to care for the disabled were contrasting: some reported confidence, due to training and empirical knowledge, while others felt insecure or incapable. It is also important to consider adapting the curriculum on this subject, with interventions in compulsory and elective subjects. Conclusion: Considering the responses and analyses, it was evident that the interviewees experience difficulties in caring for the disabled, suggesting changes to the FAMED-UFC curriculum.

Caracterização da droga vegetal e solução extrativa obtidos a partir de Syzygium cumini (l. ) skeels para futuro desenvolvimento de formas farmacêuticas / Characterization of the plant drug and extractive solution obtained from Syzygium cumini (l. ) skeels for future development of pharmaceutical forms / Caracterización de la droga vegetal y de la solución extractiva obtenida de Syzygium cumini (l. ) skeels para el futuro desarrollo de formas farmacéuticas

Syzygium cumini (L.) Skeels é uma árvore popularmente conhecida como "jamelão" ou "azeitona-roxa", é um vegetal frutífero de grande porte, pertencente à família Myrtaceae e de origem asiática, teve sua expansão em diversas regiões do mundo, no Brasil é encontrada no Nordeste, Norte e Sudeste. Quando aplicada na medicina tradicional apresenta diversas propriedades farmacológicas contidas em suas folhas, frutos, sementes e caule, com isso torna-se uma espécie bastante promissora para a indústria farmacêutica e alimentícia, pois além de conter características benéficas possui um alto poder econômico. Desta forma, este trabalho visa caracterizar físicoquimicamente a droga vegetal e solução extrativa obtidos a partir das partes aéreas de Syzygium cumini (L.) Skeels para futuro desenvolvimento de forma farmacêutica. O material vegetal de Syzygium cumini (L.) Skeels foi coletado na região de Araçoiaba-PE. Suas partes aéreas foram secas e trituradas. A solução extrativa foi obtida através da maceração da matéria-prima utilizando o etanol como solvente e as caracterizações foram realizadas de acordo com a Farmacopeia Brasileira 6° edição.

Syzygium cumini (L.) Skeels is a tree popularly known as "jamelão" or "olive-purple", is a large fruiting plant, belonging to the Myrtaceae family and of Asian origin, had its expansion in several regions of the world, in Brazil is found in the Nor- theast, North and Southeast. When applied in traditional medicine it presents several phar- macological properties contained in its leaves, fruits, seeds and stem, thus becoming a very promising species for the pharmaceutical and food industry, because besides contai- ning beneficial characteristics it has a high economic power. Thus, this work aims to physicochemically characterize the plant drug and extractive solution obtained from the aerial parts of Syzygium cumini (L.) Skeels for future development of pharmaceutical form. The plant material of Syzygium cumini (L.) Skeels was collected in the region of Araçoiaba-PE. Its aerial parts were dried and triturated. The extractive solution was obtained by maceration of the raw material using ethanol as solvent and the characteriza- tions were performed according to the Brazilian Pharmacopeia 6th edition.

Syzygium cumini (L.) Skeels es un árbol conocido popularmente como "ja- melão" o "oliva-púrpura", es una planta fructífera de gran tamaño, perteneciente a la fa- milia Myrtaceae y de origen asiático, tuvo su expansión en varias regiones del mundo, en Brasil se encuentra en el Nordeste, Norte y Sudeste. Cuando se aplica en la medicina tradicional tiene varias propiedades farmacológicas contenidas en sus hojas, frutos, se- millas y tallo, convirtiéndose así en una especie muy prometedora para la industria far- macéutica y alimentaria, ya que además de contener características beneficiosas tiene un alto poder económico. Así, este trabajo tiene como objetivo caracterizar fisicoquímica- mente la droga vegetal y la solución extractiva obtenida de las partes aéreas de Syzygium cumini (L.) Skeels para el futuro desarrollo de forma farmacéutica. El material vegetal de Syzygium cumini (L.) Skeels fue recolectado en la región de Araçoiaba-PE. Sus partes aéreas fueron secadas y trituradas. La solución extractiva se obtuvo por maceración de la materia prima utilizando etanol como solvente y las caracterizaciones se realizaron de acuerdo con la Farmacopea Brasileña 6ª edición.

NBTXR3 improves the efficacy of immunoradiotherapy combining nonfucosylated anti-CTLA4 in an anti-PD1 resistant lung cancer model.

The efficacy of immunoradiotherapy consisting of radiation therapy and immune checkpoint blockade relies on effectively promoting the systemic antitumor immune response's activation while simultaneously reducing local factors favoring immune suppression. We previously demonstrated that NBTXR3, a nanoparticle radioenhancer, significantly improved immune responses in a murine anti-PD1-resistant metastatic lung cancer model. We hypothesize that radioactivated-NBTXR3 addition to anti-PD1 and a second-generation anti-CTLA4 could improve treatment effectiveness. To test this hypothesis, we inoculated mice with 344SQR cells in the right and left legs to establish primary and secondary tumors. The primary tumors were intratumorally injected with NBTXR3 nanoparticles on day 7, followed by three fractions of 12 Gy radiation on days 8, 9, and 10. The secondary tumors received two fractions of 1Gy radiation on days 13 and 14. Multiple rounds of anti-PD1, anti-CTLA4 or nonfucosylated anti-CTLA4 were given to the mice. Immune profiling of the tumors revealed that the combination of NBTXR3 with immunoradiotherapy significantly upregulated the activities of a wide range of antitumor immune pathways and reduced the abundance of regulatory suppressor T cells. This combination effectively eradicated the primary and secondary tumors and increased animal survival to 75%. Remarkably, previously treated with NBTXR3-containing treatment, the survivor mice exhibited a long-lasting antitumor memory immune response. This data provides compelling evidence of the efficacy of NBTXR3 to synergize with the immunoradiotherapy approach when combined with an anti-PD1 and multiple checkpoints such as a second generation anti-CTLA4 and show the potential for clinical uses of antitumor immunomodulatory effects of NBTXR3.

Combining a nanoparticle-mediated immunoradiotherapy with dual blockade of LAG3 and TIGIT improves the treatment efficacy in anti-PD1 resistant lung cancer.

BACKGROUND: While improvements in immunoradiotherapy have significantly improved outcomes for cancer patients, this treatment approach has nevertheless proven ineffective at controlling the majority of malignancies. One of the mechanisms of resistance to immunoradiotherapy is that immune cells may be suppressed via the myriad of different immune checkpoint receptors. Therefore, simultaneous blockade of multiple immune checkpoint receptors may enhance the treatment efficacy of immunoradiotherapy. METHODS: We combined NBTXR3-enhanced localized radiation with the simultaneous blockade of three different checkpoint receptors: PD1, LAG3, and TIGIT, and tested the treatment efficacy in an anti-PD1-resistant lung cancer model in mice. 129 Sv/Ev mice were inoculated with fifty thousand αPD1-resistant 344SQR cells in the right leg on day 0 to establish primary tumors and with the same number of cells in the left leg on day 4 to establish the secondary tumors. NBTXR3 was intratumorally injected into the primary tumors on day 7, which were irradiated with 12 Gy on days 8, 9, and 10. Anti-PD1 (200 µg), αLAG3 (200 µg), and αTIGIT (200 µg) were given to mice by intraperitoneal injections on days 5, 8, 11, 14, 21, 28, 35, and 42. RESULTS: This nanoparticle-mediated combination therapy is effective at controlling the growth of irradiated and distant unirradiated tumors, enhancing animal survival, and is the only one that led to the destruction of both tumors in approximately 30% of the treated mice. Corresponding with this improved response is robust activation of the immune response, as manifested by increased numbers of immune cells along with a transcriptional signature of both innate and adaptive immunity within the tumor. Furthermore, mice treated with this combinatorial therapy display immunological memory response when rechallenged by the same cancer cells, preventing tumor engraftment. CONCLUSION: Our results strongly attest to the efficacy and validity of combining nanoparticle-enhanced radiotherapy and simultaneous blockade of multiple immune checkpoint receptors and provide a pre-clinical rationale for investigating its translation into human patients.

Radiotherapy-activated NBTXR3 nanoparticles modulate cancer cell immunogenicity and TCR repertoire.

BACKGROUND: Radiotherapy is a powerful and widely used technique for the treatment of solid tumors. Beyond its ability to destroy tumor cells, it has been demonstrated that radiotherapy can stimulate the anti-tumor immune response. Unfortunately, this effect is mainly restricted to the irradiated lesion, as tumor control outside the treated field (called the 'abscopal effect') is rarely obtained. In addition, many pro-tumoral factors prevent this anti-tumor immune response from being sustained and efficient. We previously reported that radiotherapy-activated NBTXR3 produced a significant CD8-dependent abscopal effect in immunocompetent mice bearing CT26.WT tumors, while radiotherapy failed to generate such a response. METHODS: To identify the mechanisms that may explain this response, we evaluated the capacity of radiotherapy-activated NBTXR3 to modulate the immunogenicity of tumor cells by analysis of immunogenic cell death biomarkers and immunopeptidome sequencing. In vivo, we analyzed treated tumors for CD4+, CD8 + and CD68 + cell infiltrates by immunohistochemistry and digital pathology and sequenced the T cell receptor (TCR) repertoire in both treated and untreated distant tumors. RESULTS: We showed that NBTXR3 activated by radiotherapy both increased immunogenic cell death biomarkers and modulated the immunopeptidome profile of CT26.WT cells. Immunohistochemistry analysis of treated tumors revealed a significant increase in CD4+, CD8 + and CD68 + cell infiltrates for NBTXR3 activated by radiotherapy group, compared to radiotherapy. We also measured significant modifications in TCR repertoire diversity in the radiotherapy-activated NBTXR3 group, both in treated and distant untreated tumors, compared to radiotherapy alone. CONCLUSIONS: These results indicate that radiotherapy-activated NBTXR3 can act as an effective immunomodulator, modifying tumor cell immunogenicity and impacting the lymphocyte population.

A method for rapid flow-cytometric isolation of endothelial nuclei and RNA from archived frozen brain tissue.

Endothelial cells are important contributors to brain development, physiology, and disease. Although RNA sequencing has contributed to the understanding of brain endothelial cell diversity, bulk analysis and single-cell approaches have relied on fresh tissue digestion protocols for the isolation of single endothelial cells and flow cytometry-based sorting on surface markers or transgene expression. These approaches are limited in the analysis of the endothelium in human brain tissues, where fresh samples are difficult to obtain. Here, we developed an approach to examine endothelial RNA expression by using an endothelial-specific marker to isolate nuclei from abundant archived frozen brain tissues. We show that this approach rapidly and reliably extracts endothelial nuclei from frozen mouse brain samples, and importantly, from archived frozen human brain tissues. Furthermore, isolated RNA transcript levels are closely correlated with expression in whole cells from tissue digestion protocols and are enriched in endothelial markers and depleted of markers of other brain cell types. As high-quality RNA transcripts could be obtained from as few as 100 nuclei in archived frozen human brain tissues, we predict that this approach should be useful for both bulk analysis of endothelial RNA transcripts in human brain tissues as well as single-cell analysis of endothelial sub-populations.

Implicações da covid-19 no cotidiano das famílias nordestinas e no cuidado infantil / Implications of covid-19 in the daily lives of northeastern families and childcare

Resumo Este estudo explora o cotidiano das famílias com filhos de 0 a 6 anos, residentes em Fortaleza, no Ceará, durante o período de distanciamento físico, estipulado pelo Governo do Estado do Ceará. Esta pesquisa qualitativa se utilizou do referencial da teoria de formação de vínculos na adversidade. Foram entrevistadas 30 mães, entre os meses de julho e agosto de 2020, utilizando videochamadas ou telefonemas. Para análise do material empírico, recorreu-se à análise de conteúdo de Bardin, possibilitando a criação de duas categorias temáticas: (1) o exercício do cuidado parental em tempos de covid-19; (2) o cotidiano das crianças diante da pandemia. A interpretação das narrativas revelou que alguns cuidadores buscaram realizar brincadeiras e atividades manuais com os filhos, e explicavam o que estava acontecendo no cenário mundial, exercendo a parentalidade positiva. Ademais, o distanciamento físico favoreceu o aumento da tolerância dos pais no tempo em que os filhos ficaram expostos às telas. Percebeu-se que os pais influenciaram a prática de hábitos alimentares não saudáveis entre as crianças e a manifestação de mudanças no comportamento dos filhos. Como conclusão, destaca-se a necessidade de um acompanhamento contínuo dos aspectos referentes ao desenvolvimento dessas crianças e do retorno delas às atividades presenciais.

Abstract This study sought to explore the daily life of families with children aged 0 - 6 years old, living in Fortaleza, Ceará - Brazil, during the period of social distancing stipulated by the State Government. This qualitative research was conducted with data collected by means of video or phone interviews with thirty mothers, between July and August 2020. The empirical material was analyzed in the light of Bardin's content analysis, generating two thematic categories: (1) the exercise of parental care in Covid-19 times; and (2) the daily life of children in the face of the pandemic. The narratives revealed that some caregivers tried to play games and practice arts and crafts with their children, besides explaining what was happening in the world scenario - thus exercising positive parenting. Moreover, the social distancing measures promoted parental tolerance during the time children were exposed to screens, and parents showed to influence the practice of unhealthy eating habits and the manifestation of behavior changes in their children. These findings highlight the need for a continuous monitoring of aspects related to these children's development, as well as for their return to face-to-face activities.

A novel method of screening combinations of angiostatics identifies bevacizumab and temsirolimus as synergistic inhibitors of glioma-induced angiogenesis.

Tumor angiogenesis is critical for the growth and progression of cancer. As such, angiostasis is a treatment modality for cancer with potential utility for multiple types of cancer and fewer side effects. However, clinical success of angiostatic monotherapies has been moderate, at best, causing angiostatic treatments to lose their early luster. Previous studies demonstrated compensatory mechanisms that drive tumor vascularization despite the use of angiostatic monotherapies, as well as the potential for combination angiostatic therapies to overcome these compensatory mechanisms. We screened clinically approved angiostatics to identify specific combinations that confer potent inhibition of tumor-induced angiogenesis. We used a novel modification of the ex ovo chick chorioallantoic membrane (CAM) model that combined confocal and automated analyses to quantify tumor angiogenesis induced by glioblastoma tumor onplants. This model is advantageous due to its low cost and moderate throughput capabilities, while maintaining complex in vivo cellular interactions that are difficult to replicate in vitro. After screening multiple combinations, we determined that glioblastoma-induced angiogenesis was significantly reduced using a combination of bevacizumab (Avastin®) and temsirolimus (Torisel®) at doses below those where neither monotherapy demonstrated activity. These preliminary results were verified extensively, with this combination therapy effective even at concentrations further reduced 10-fold with a CI value of 2.42E-5, demonstrating high levels of synergy. Thus, combining bevacizumab and temsirolimus has great potential to increase the efficacy of angiostatic therapy and lower required dosing for improved clinical success and reduced side effects in glioblastoma patients.

COVID-19 and mental health of pregnant women in Ceará, Brazil

ABSTRACT OBJECTIVE To assess the perceptions of pregnant women about COVID-19 and the prevalence of common mental disorders during the implemented social distancing period. METHODS This was an observational, cross-sectional study using digital media, of pregnant women exposed to social distancing due to the COVID-19 pandemic, in Fortaleza, Ceará, Northeastern Brazil. Common mental disorders were estimated using the modified Self-Report Questionnaire-20 (SRQ-20) scale, and the feelings towards COVID-19 were assessed using the Fear of COVID-19 scale through telephone calls made in May 2020. COX multivariate regression models were used to verify the associations. RESULTS Of the 1,041 pregnant women, 45.7% (95%CI: 42.7-48.8) had common mental disorders (CMD). All items of the Fear of COVID-19 Scale showed a significant association with the prevalence of CMD (p < 0.001). A CMD risk gradient was observed, going from a prevalence ratio of 1.52 (95%CI: 1.13-2.04) in pregnant women with two positive items to 2.70 (95%CI: 2.08-3.51) for those with four positive items. Early gestational age and the lack of prenatal care were also associated with CMD. CONCLUSIONS The prevalence of common mental disorders in pregnant women was high during the period of social distancing and was aggravated by negative feelings towards COVID-19.

Nucleophilicity of cysteine and related biothiols and the development of fluorogenic probes and other applications.

Biothiols such as l-cysteine, l-homocysteine, and glutathione play essential roles in many biological processes, and are directly associated with several health conditions. Therefore, the development of fast, selective, sensitive, and inexpensive methods for quantitatively analyzing biothiols in aqueous solution, but especially in biological samples, is a very attractive research field. In this feature review, we have approached the relevance of biothiols' nucleophilicity to develop selective fluorogenic probes. Since biothiols have considerable structural similarity, relevant strategies are in full development, including several fluorescent molecular platforms, specific receptor sites, reaction conditions, and optical responses. All of these features are properly presented and discussed. Biothiol sensing protocols are based on traditional organic chemistry reactions such as (hetero)aromatic nucleophilic substitution, addition, and substitution at carbonyl carbon, conjugate addition, and nucleophilic substitution at saturated carbon, amongst others including combined processes; furthermore, mechanistic aspects are detailed herein, including some interesting historical contexts. The feasibility of related fluorogenic probes is illustrated by analysis in complex matrices such as serum, cells, tissues, and animal models. Applications of these reactions in more complex systems such as sulfhydryl-based peptides and proteins are also presented, aiming at functionalizing and detecting these nucleophiles. Most literature cited in this review is recent; however, some other prominent works are also detailed. It is believed that this review may be accessible for many academic levels and may efficiently contribute not only to popularizing science but also to the rational development of fluorogenic probes for biothiol sensing.

FHITlow /pHER2high signature in non-small cell lung cancer is predictive of anti-HER2 molecule efficacy.

Despite its efficacy in solid tumours, in particular HER2+ breast cancer, HER2-targeted therapy has given rise to disappointing results in non-small cell lung cancer (NSCLC). With the aim of refining the target population for anti-HER2 therapies in NSCLC, we investigated the relationships between HER2 and the tumour suppressor fragile histidine triad (FHIT) in lung tumour cells. First, we observed a negative correlation between FHIT expression and the activated form of HER2 (pHER2) in NSCLC samples and in lung tumour cell lines. Moreover, the silencing or overexpression of FHIT in lung cell lines led to an increase or decrease of HER2 activity, respectively. We also demonstrated that two anti-HER2 drugs, irbinitinib and trastuzumab, restore a more epithelial phenotype and counteract cell invasiveness and growth of FHIT-silenced tumour cell lines. Finally, we showed that the FHITlow /pHER2high phenotype predicts sensitivity to an anti-HER2 therapy in primary tumour cells from NSCLC patients. Our results show that FHIT regulates the activity of HER2 in lung tumour cells and that FHIT-inactivated tumour cells are sensitive to HER2 inhibitors. A new subclass of patients with NSCLC may be eligible for an anti-HER2 therapy. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Inflammatory response and macrophage polarization using different physicochemical biomaterials for oral and maxillofacial reconstruction.

Knowledge about the action of immune system in the recognition of biomaterials has been extremely helpful when it comes about understanding host response and biomaterials' fate in human body. This study aimed to investigate inflammatory response and macrophage polarization during bone healing process of rat's calvaria critical defects using different bone materials in order to evaluate their influence on bone repair and on the quality of the newly formed bone tissue. Eighty male albinus Wistar rats underwent surgical procedure for the confectioning of a 5-mm diameter bone defect in their right parietal bone, and divided in four groups (n = 20 each), according the biomaterial: AG - Control, particulate intramembranous autogenous bone graft, HA/TCP - particulate biphasic calcium phosphate with HA/TCP (60/40), DBB - particulate deproteinized bovine bone, VC - particulate bioactive vitroceramic. After 3, 7, 21, and 45 days, the specimens were removed and prepared for microcomputed tomography (microCT), light and polarized microscopy, immunohistochemical analysis, and histomorphometry. No significant differences were detected considering percentage of leukocytes among the groups and periods, as well as in relation to immunolabeling for inflammatory (M1) and reparative (M2) macrophages. However, immunolabeling for bone marker indicated a delayed osteoblast differentiation in VC group, resulting in a decrease in mineralized bone matrix parameters in this group, revealed by microCT. In addition, AG and HA/TCP presented a satisfactory bone collagenous content. Despite the distinct origins and physicochemical properties of the tested biomaterials, they presented similar immune-inflammatory responses in the present experimental model, influencing bone-related proteins and bone quality, which must be considered according to their use.

Noncovalent Interactions Drive the Efficiency of Molybdenum Imido Alkylidene Catalysts for Olefin Metathesis.

High-throughput experimentation and multivariate modeling allow identification of noncovalent interactions (NCIs) in monoaryloxy-pyrrolide Mo imido alkylidene metathesis catalysts prepared in situ as a key driver for high activity in a representative metathesis reaction (homodimerization of 1-nonene). Statistical univariate and multivariate modeling categorizes catalytic data from 35 phenolic ligands into two groups, depending on the substitution in the ortho position of the phenol ligand. The catalytic activity descriptor TON1h correlates predominantly with attractive NCIs when phenols bear ortho aryl substituents and, conversely, with repulsive NCIs when the phenol has no aryl ortho substituents. Energetic span analysis is deployed to relate the observed NCI and the cycloreversion metathesis step such that aryloxide ligands with no ortho aryls mainly impact the energy of metallacyclobutane intermediates (SP/TBP isomers), whereas aryloxides with pendant ortho aryls influence the transition state energy for the cycloreversion step. While the electronic effects from the aryloxide ligands also play a role, our work outlines how NCIs may be exploited for the design of improved d0 metathesis catalysts.

Tumour cell blebbing and extracellular vesicle shedding: key role of matrikines and ribosomal protein SA.

BACKGROUND: Carcinogenesis occurs in elastin-rich tissues and leads to local inflammation and elastolytic proteinase release. This contributes to bioactive matrix fragment (Matrikine) accumulation like elastin degradation products (EDP) stimulating tumour cell invasive and metastatic properties. We previously demonstrate that EDPs exert protumoural activities through Hsp90 secretion to stabilised extracellular proteinases. METHODS: EDP influence on cancer cell blebbing and extracellular vesicle shedding were examined with a videomicroscope coupled with confocal Yokogawa spinning disk, by transmission electron microscopy, scanning electron microscopy and confocal microscopy. The ribosomal protein SA (RPSA) elastin receptor was identified after affinity chromatography by western blotting and cell immunolocalisation. mRNA expression was studied using real-time PCR. SiRNA were used to confirm the essential role of RPSA. RESULTS: We demonstrate that extracellular matrix degradation products like EDPs induce tumour amoeboid phenotype with cell membrane blebbing and shedding of extracellular vesicle containing Hsp90 and proteinases in the extracellular space. EDPs influence intracellular calcium influx and cytoskeleton reorganisation. Among matrikines, VGVAPG and AGVPGLGVG peptides reproduced EDP effects through RPSA binding. CONCLUSIONS: Our data suggests that matrikines induce cancer cell blebbing and extracellular vesicle release through RPSA binding, favouring dissemination, cell-to-cell communication and growth of cancer cells in metastatic sites.