Cell type-specific regulation of CFTR trafficking-on the verge of progress.

Trafficking of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein is a complex process that starts with its biosynthesis and folding in the endoplasmic reticulum. Exit from the endoplasmic reticulum (ER) is coupled with the acquisition of a compact structure that can be processed and traffic through the secretory pathway. Once reaching its final destination-the plasma membrane, CFTR stability is regulated through interaction with multiple protein partners that are involved in its post-translation modification, connecting the channel to several signaling pathways. The complexity of the process is further boosted when analyzed in the context of the airway epithelium. Recent advances have characterized in detail the different cell types that compose the surface epithelium and shifted the paradigm on which cells express CFTR and on their individual and combined contribution to the total expression (and function) of this chloride/bicarbonate channel. Here we review CFTR trafficking and its relationship with the knowledge on the different cell types of the airway epithelia. We explore the crosstalk between these two areas and discuss what is still to be clarified and how this can be used to develop more targeted therapies for CF.

Ir(III) Half-Sandwich Photosensitizers with a π-Expansive Ligand for Efficient Anticancer Photodynamic Therapy.

One approach to reduce the side effects of chemotherapy in cancer treatment is photodynamic therapy (PDT), which allows spatiotemporal control of the cytotoxicity. We have used the strategy of coordinating π-expansive ligands to increase the excited state lifetimes of Ir(III) half-sandwich complexes in order to facilitate the generation of 1O2. We have obtained derivatives of formulas [Cp*Ir(C∧N)Cl] and [Cp*Ir(C∧N)L]BF4 with different degrees of π-expansion in the C∧N ligands. Complexes with the more π-expansive ligand are very effective photosensitizers with phototoxic indexes PI > 2000. Furthermore, PI values of 63 were achieved with red light. Time-dependent density functional theory (TD-DFT) calculations nicely explain the effect of the π-expansion. The complexes produce reactive oxygen species (ROS) at the cellular level, causing mitochondrial membrane depolarization, cleavage of DNA, nicotinamide adenine dinucleotide (NADH) oxidation, as well as lysosomal damage. Consequently, cell death by apoptosis and secondary necrosis is activated. Thus, we describe the first class of half-sandwich iridium cyclometalated complexes active in PDT.

Use of adenine base editing and homology-independent targeted integration strategies to correct the cystic fibrosis causing variant, W1282X.

Small molecule drugs known as modulators can treat ~90% of people with cystic fibrosis (CF), but do not work for premature termination codon variants such as W1282X (c.3846G>A). Here we evaluated two gene editing strategies, Adenine Base Editing (ABE) to correct W1282X, and Homology-Independent Targeted Integration (HITI) of a CFTR superexon comprising exons 23-27 (SE23-27) to enable expression of a CFTR mRNA without W1282X. In Flp-In-293 cells stably expressing a CFTR expression minigene bearing W1282X, ABE corrected 24% of W1282X alleles, rescued CFTR mRNA from nonsense mediated decay and restored protein expression. However, bystander editing at the adjacent adenine (c.3847A>G), caused an amino acid change (R1283G) that affects CFTR maturation and ablates ion channel activity. In primary human nasal epithelial cells homozygous for W1282X, ABE corrected 27% of alleles, but with a notably lower level of bystander editing, and CFTR channel function was restored to 16% of wild-type levels. Using the HITI approach, correct integration of a SE23-27 in intron 22 of the CFTR locus in 16HBEge W1282X cells was detected in 5.8% of alleles, resulting in 7.8% of CFTR transcripts containing the SE23-27 sequence. Analysis of a clonal line homozygous for the HITI-SE23-27 produced full-length mature protein and restored CFTR anion channel activity to 10% of wild-type levels, which could be increased three-fold upon treatment with the triple combination of CF modulators. Overall, these data demonstrate two different editing strategies can successfully correct W1282X, the second most common class I variant, with a concomitant restoration of CFTR function.

Ferrozoles: Ferrocenyl derivatives of letrozole with dual effects as potent aromatase inhibitors and cytostatic agents.

In the treatment of hormone-dependent cancers, aromatase inhibitors (AI) are receiving increased attention due to some undesirable effects such as the risk of endometrial cancer and thromboembolism of SERMs (selective estrogen receptor modulators). Letrozole is the most active AI with 99% aromatase inhibition. Unfortunately, this compound also exhibits some adverse effects such as hot flashes and fibromyalgias. Therefore, there is an urgent need to explore new types of AIs that retain the same-or even increased-antitumor ability. Inspired by the letrozole structure, a set of new derivatives has been synthesized that include a ferrocenyl moiety and different heterocycles. The derivative that contains a benzimidazole ring, namely compound 6, exhibits a higher aromatase inhibitory activity than letrozole and it also shows potent cytostatic behavior when compared to other well-established aromatase inhibitors, as demonstrated by dose-response, cell cycle, apoptosis and time course experiments. Furthermore, 6 promotes the inhibition of cell growth in both an aromatase-dependent and -independent fashion, as indicated by the study of A549 and MCF7 cell lines. Molecular docking and molecular dynamics calculations on the interaction of 6 or letrozole with the aromatase binding site revealed that the ferrocene moiety increases the van der Waals and hydrophobic interactions, thus resulting in an increase in binding affinity. Furthermore, the iron atom of the ferrocene fragment can form a metal-acceptor interaction with a propionate fragment, and this results in a stronger coupling with the heme group-a possibility that is consistent with the strong aromatase inhibition of 6.

Female reproductive health impacts of Long COVID and associated illnesses including ME/CFS, POTS, and connective tissue disorders: a literature review.

Long COVID disproportionately affects premenopausal women, but relatively few studies have examined Long COVID's impact on female reproductive health. We conduct a review of the literature documenting the female reproductive health impacts of Long COVID which may include disruptions to the menstrual cycle, gonadal function, ovarian sufficiency, menopause, and fertility, as well as symptom exacerbation around menstruation. Given limited research, we also review the reproductive health impacts of overlapping and associated illnesses including myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS), postural orthostatic tachycardia syndrome (POTS), connective tissue disorders like Ehlers-Danlos syndrome (EDS), and endometriosis, as these illnesses may help to elucidate reproductive health conditions in Long COVID. These associated illnesses, whose patients are 70%-80% women, have increased rates of dysmenorrhea, amenorrhea, oligomenorrhea, dyspareunia, endometriosis, infertility, vulvodynia, intermenstrual bleeding, ovarian cysts, uterine fibroids and bleeding, pelvic congestion syndrome, gynecological surgeries, and adverse pregnancy complications such as preeclampsia, maternal mortality, and premature birth. Additionally, in Long COVID and associated illnesses, symptoms can be impacted by the menstrual cycle, pregnancy, and menopause. We propose priorities for future research and reproductive healthcare in Long COVID based on a review of the literature. These include screening Long COVID patients for comorbid and associated conditions; studying the impacts of the menstrual cycle, pregnancy, and menopause on symptoms and illness progression; uncovering the role of sex differences and sex hormones in Long COVID and associated illnesses; and addressing historical research and healthcare inequities that have contributed to detrimental knowledge gaps for this patient population.

Mutation-class dependent signatures outweigh disease-associated processes in cystic fibrosis cells.

BACKGROUND: The phenotypic heterogeneity observed in Cystic Fibrosis (CF) patients suggests the involvement of other genes, besides CFTR. Here, we combined transcriptome and proteome analysis to understand the global gene expression patterns associated with five prototypical CFTR mutations. RESULTS: Evaluation of differentially expressed genes and proteins unveiled common and mutation-specific changes revealing functional signatures that are much more associated with the specific molecular defects associated with each mutation than to the CFTR loss-of-function phenotype. The combination of both datasets revealed that mutation-specific detected translated-transcripts (Dtt) have a high level of consistency. CONCLUSIONS: This is the first combined transcriptomic and proteomic study focusing on prototypical CFTR mutations. Analysis of Dtt provides novel insight into the pathophysiology of CF, and the mechanisms through which each mutation class causes disease and will likely contribute to the identification of new therapeutic targets and/or biomarkers for CF.

Comparison of Cas9 and Cas12a CRISPR editing methods to correct the W1282X-CFTR mutation.

BACKGROUND: W1282X-CFTR variant (c.3846G>A) is the second most common nonsense cystic fibrosis (CF)-causing mutation in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. Even though remarkable breakthroughs have been done towards CF treatment with the approval of four CFTR protein modulators, none of these are approved for patients with nonsense mutations. CRISPR gene editing tools can be of great value to permanently correct the genetic defects caused by these mutations. METHODS: We compared the capacity of homology-directed repair (HDR) mediated by Cas9 or Cas12a to correct W1282X CFTR mutation in the CFF-16HBEge W1282X CFTR cell line (obtained from CFF), using Cas9/gRNA and Cas12a/gRNA ribonucleoproteins (RNPs) and single strand DNA (ssODN) oligonucleotide donors. RESULTS: Cas9 shows higher levels of correction than Cas12a as, by electroporating cells with Cas9 RNPs and ssODN donor, nearly 18% of precise editing was achieved compared to just 8% for Cas12a. Such levels of correction increase the abundance of CFTR mRNA and protein, and partially restore CFTR function in the pool of edited cells to 18% of WT CFTR function. Moreover, homozygous corrected clones produced levels of mRNA, protein, and function comparable to those of cells expressing WT CFTR. CONCLUSION: Altogether, this work demonstrates the potential of gene editing as a therapeutic strategy for CF directly correcting the root cause of the disease.

Allopurinol for fibromyalgia pain in adults: A randomized controlled trial.

BACKGROUND: Allopurinol is a potent inhibitor of the enzyme xanthine oxidase used in the treatment of hyperuricemia and gout. Because it is well known that purines exert multiple affects on pain transmission, we hypothesized that the inhibition of xanthine oxidase by allopurinol could be a valid strategy to treat pain in humans. This study aimed to compare the analgesic efficacy of oral allopurinol versus placebo as an adjuvant therapy in patients displaying fibromyalgia. METHODS: This randomized, double-blinded, placebo-controlled study included 60 women with the diagnosis of fibromyalgia. Patients were randomly assigned to receive either oral allopurinol 300 mg (n = 31) or placebo (n = 29) twice daily during 30 days. The patients were submitted to evaluation for pain sensitivity, anxiety, depression, and functional status before treatment, and 15 and 30 days thereafter. RESULTS: Oral administration of allopurinol 300 mg twice daily was ineffective in improving pain scores measured by several tools up to 30 days of treatment (P > 0.05). Additionally, no significant effects of allopurinol over anxiety, depressive symptoms, and functional status of fibromyalgia patients were observed in the present study. CONCLUSIONS: Although previous findings indicated that allopurinol could present intrinsic analgesic effects in both animals and humans, this study showed no benefit of the use of oral allopurinol as an adjuvant strategy during 30 days in women displaying fibromyalgia. However, considering previous promising results, new prospective studies are still valid to further investigate allopurinol and more selective purine derivatives in the management of pain syndromes.

Protocol for visits of patients' pets in palliative care in a hospital.

OBJECTIVE: To report the experience of the multidisciplinary health team in the construction and implementation of the assistance protocol for pets visiting patients admitted to a palliative care unit. METHOD: This is an experience report about the construction and implementation of a protocol in a university hospital in the South of Brazil by a working group from the multidisciplinary team of the Palliative Care Program and composed of three nurses, a physician, a psychologist, a social worker, a manager, and a nutritionist. The period of construction and implementation of the protocol was from September 2017 to April 2019. RESULTS: The construction of the protocol by the multidisciplinary team followed three stages: planning, execution, and implementation, which enabled the release of visits of patients' pets in palliative care. CONCLUSION: The construction of the protocol allowed for the institutionalization of the visit of patients' pets in palliative care in a university hospital, bringing benefits to patients and their families.

Photodynamic Therapy as an adjunct in the Treatment of Medication-Related Osteonecrosis of the Jaw: A Case Report.

Introduction: Medication-related osteonecrosis of the jaw (MRONJ) corresponds to an adverse effect of the use of drugs such as bisphosphonates and denosumab. This condition is often associated with pain, infection, purulent secretion, paraesthesia, tooth mobility and halitosis, decreasing the patient's quality of life. The management of MRONJ tends to be conservative, through the guidance of oral hygiene, antibiotic therapy and mouthwashes. However, the use of antimicrobial photodynamic therapy (aPDT) has shown promise in the treatment of these injuries. The purpose of this article is to report a case of MRONJ treatment associated with aPDT. Case Report: A 75-year-old patient, with a history of breast cancer and use of intravenous Zoledronic Acid, presented with bilateral MRONJ lesions in tuberosity on the right and left sides. Treatment was conservatively instituted with the use of aPDT as an adjuvant. After 12 aPDT sessions, complete regression of the lesion was observed. However, after two weeks, the presence of a new lesion was noted, this time in the anterior region of the maxilla. The same protocol previously established was followed and after two aPDT sessions, the patient returned with complete lesion regression. Conclusion: The use of aPDT may represent an important adjuvant within a set of clinical protocols in the treatment of MRONJ.

Effect of the aniline fragment in Pt(II) and Pt(IV) complexes as anti-proliferative agents. Standard reduction potential as a more reliable parameter for Pt(IV) compounds than peak reduction potential.

The problems of resistance and side effects associated with cisplatin and other chemotherapeutic drugs have boosted research aimed at finding new compounds with improved properties. The use of platinum(IV) prodrugs is one alternative, although there is some controversy regarding the predictive ability of the peak reduction potentials. In the work described here a series of fourteen chloride Pt(II) and Pt(IV) compounds was synthesised and fully characterised. The compounds contain different bidentate arylazole heterocyclic ligands. Their cytotoxic properties against human lung carcinoma (A549), human breast carcinoma (MCF7) and human colon carcinoma (HCT116 and HT29) cell lines were studied. A clear relationship between the type of ligand and the anti-proliferative properties was found, with the best results obtained for the Pt(II) compound that contains an aniline fragment, (13), thus evidencing a positive effect of the NH2 group. Stability and aquation studies in DMSO, DMF and DMSO/water mixtures were carried out on the active complexes and an in-depth analysis of the two aquation processes, including DFT analysis, of 13 was undertaken. It was verified that DNA was the target and that cell death occurred by apoptosis in the case of 13. Furthermore, the cytotoxic derivatives did not exhibit haemolytic activity. The reduction of the Pt(IV) compounds whose Pt(II) congeners were active was studied by several techniques. It was concluded that the peak reduction potential was not useful to predict the ability for reduction. However, a correlation between the cytotoxic activity and the standard reduction potential was found.

Deriving valorization of phenolic compounds from olive oil by-products for food applications through microencapsulation approaches: a comprehensive review.

Nowadays, olive oil consumption is correlated to many health benefits, essentially due to the presence of antioxidants, especially phenolic compounds, which fostered its intensive production worldwide. During olive oil extraction, through continuous or discontinuous processes, many olive oil by-products are generated. These by-products constitute an environmental problem regarding its management and disposal. They are phytotoxic and biotoxic due to their high content of phenolic compounds, presenting contrastingly relevant health benefits due to their potent radical scavenging activities. In the framework of the disposal and management of olive oil by-products, treatment, and valorization approaches are found. As currently, the majority of the valorization techniques applied have a null market value, alternative strategies for the obtainment of innovative products as fortified foods are being investigated. The recovery and valorization strategies of olive oil by-products may comprise extraction and further encapsulation of bioactive compounds, as an innovative valorization blueprint of phenolic compounds present in these by-products. The majority of phenolic compounds present in olive oil by-products possess limited application on the food industry since they are promptly amended by environmental factors like temperature, pH, and light. Consequently, they must be protected previously ending in the final formulation. Prior to foods fortification with phenolic-rich extracts obtained from olive oil by-products, they should be protected through microencapsulation approaches, allowing a sustained release of phenolic compounds in the fortified foods, without losing their physicochemical properties. The combined strategies of extraction and microencapsulation will contribute to promoting the sustainability of the olive oil sector and aid the food industry to obtain reinvented added-value products.

Protocol for visits of patients' pets in palliative care in a hospital / Protocolo para la visita de mascotas de pacientes en cuidados paliativos en un hospital / Protocolo para visita do animal de estimação do paciente em cuidados paliativos em um hospital

ABSTRACT Objective To report the experience of the multidisciplinary health team in the construction and implementation of the assistance protocol for pets visiting patients admitted to a palliative care unit. Method This is an experience report about the construction and implementation of a protocol in a university hospital in the South of Brazil by a working group from the multidisciplinary team of the Palliative Care Program and composed of three nurses, a physician, a psychologist, a social worker, a manager, and a nutritionist. The period of construction and implementation of the protocol was from September 2017 to April 2019. Results The construction of the protocol by the multidisciplinary team followed three stages: planning, execution, and implementation, which enabled the release of visits of patients' pets in palliative care. Conclusion The construction of the protocol allowed for the institutionalization of the visit of patients' pets in palliative care in a university hospital, bringing benefits to patients and their families.

RESUMEN Objetivo Informar la experiencia del equipo de salud multiprofesional en la construcción e implementación de un protocolo de atención para visita de mascotas, para los pacientes internados en una unidad de cuidados paliativos. Método Se trata de un informe de experiencia sobre la construcción e implementación de un protocolo en un hospital universitario en el sur de Brasil, realizado por un grupo de trabajo originado a partir del equipo multidisciplinario del Programa de Cuidados Paliativos y compuesto por tres enfermeras, un médico, un psicólogo, un trabajador social, un administrador y un nutricionista. El período de construcción e implementación del protocolo abarca desde septiembre de 2017 hasta abril de 2019. Resultados La construcción del protocolo por la experiencia del equipo multiprofesional siguió tres etapas: planificación, ejecución e implementación, que permitieron la habilitación de las visitas de mascotas de pacientes en cuidados paliativos. Conclusión La construcción del protocolo permitió institucionalizar la visita de mascotas de pacientes en cuidados paliativos en un hospital universitario, brindando beneficios tanto a los pacientes como a sus familias.

RESUMO Objetivo Relatar a experiência da equipe multiprofissional em saúde na construção e implantação do protocolo assistencial para visita dos animais de estimação dos pacientes internados em uma Unidade de Cuidados Paliativos. Método Trata-se de um relato de experiência de uma ação realizada em um hospital universitário do sul do Brasil por um grupo de trabalho originado da equipe multiprofissional do Programa de Cuidados Paliativos, composto por três enfermeiros, um médico, um psicólogo, um assistente social, um administrador e uma nutricionista. O período de construção e implementação do protocolo foi de setembro de 2017 a abril de 2019. Resultados: A construção do protocolo pela equipe multiprofissional seguiu três etapas, planejamento, execução e implementação, e resultou na liberação para visita dos animais de estimação dos pacientes em cuidados paliativos. Conclusão A construção do protocolo permitiu a institucionalização da visita dos animais de estimação dos pacientes em cuidados paliativos em um hospital universitário, trazendo benefícios para os pacientes e seus familiares.

Programa Saúde na Escola: uma análise das ações de saúde* / Health in School Program: an analysis of health actions

Objetivo: analisar a implementação do Programa Saúde na Escola, a partir das ações de saúde na escola, valendo-se da perspectiva dos enfermeiros da Equipes de Saúde da Família e gestores de educação da cidade de Teresina, Piauí, Brasil. Método: Realizamos um estudo de análise de implementação com abordagem qualitativa, envolvendo os gestores das escolas e os enfermeiros das equipes de Saúde da Família. A seleção dos participantes deste estudo foi intencional. Para a coleta de dados, utilizamos a entrevista semiestruturada, realizada por uma única pesquisadora. Resultados: As narrativas indicam limites na evolução da atenção integral à saúde do escolar e no alcance dos objetivos vislumbrados pelo PSE. A presença de escolas ao lado das unidades de saúde é referida como um fator que facilita o diálogo, a criação de vínculos e a realização de ações entre os dois setores. Conclusão: As poucas ações existentes são desarticuladas, predominantemente centradas na dimensão biológica, preventivas e protagonizadas pelo setor saúde.

Objective: To analyze the implementation of the Health at School Program, based on health actions at school, using the perspective of nurses from the Family Health Teams and education managers of the city of Teresina, Piauí, Brazil. Method: We performed an analysis of implementation with a qualitative approach, involving the managers of the schools and the nurses of the Family Health teams. The selection of participants in this study was intentional. For data collection, we used the semi-structured interview, conducted by a single researcher. Results: The narratives indicate limits in the evolution of integral attention to the school health and in the achievement of the goals envisioned by the PSE. The presence of schools next to the health units is a factor that facilitates dialogue, the creation of bonds and the performance of actions between the two sectors. Conclusion: The few existing actions are disjointed, predominantly focused on the biological dimension, preventive and carried out by the health sector.

Analytical methodology to screen UV-filters and synthetic musk compounds in market tomatoes.

A Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) methodology followed by gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis was developed to extract thirteen synthetic musk compounds (SMCs: cashmeran, celestolide, phantolide, traseolide, galaxolide, tonalide, musk ambrette, musk xylene, musk ketone, musk tibetene, musk moskene, ethylene brassylate and exaltolide) and six ultraviolet-filters (UVFs: 2-ethylhexyl 4-dimethylaminobenzoate, 3-(4'-methylbenzylidene) camphor, 2-ethylhexyl 4-methoxycinnamate, 2-ethylhexyl 2-cyano-3,3-diphenylacrylate, benzophenone and drometrizole trisiloxane) from tomatoes. The proposed methodology was optimized: 2 g of freeze-dried tomato was extracted with 4 mL of water and 10 mL of ethyl acetate, adding 6 g of MgSO4 and 1.5 g of NaCl, then a dispersive solid-phase extraction was performed using 3 g of MgSO4, 300 mg of primary-secondary amino adsorbent (PSA) and 300 mg of octadecyl-silica (C18). Validation delivered recoveries between 81 (celestolide) and 119% (musk tibetene), with relative standard deviations <10%. The instrumental limit of detection varied from 0.02 (2-ethylhexyl 4-methoxycinnamate) to 3.00 pg (exaltolide and musk xylene). Regarding the method quantification limits, it ranged between 0.4 (celestolide) and 47.9 ng g-1 dw (exaltolide). The method was applied to different varieties of tomatoes (Solanum lycopersicum), revealing UVFs and SMCs between 1 and 210 ng g-1 dw. Higher concentrations were found for benzophenone (29-210 ng g-1 dw) and galaxolide (9-53 ng g-1 dw). The risk associated to the ingestion of contaminated tomatoes has also been estimated, showing that a potential health risk is unlikely.

Multi-layer pre-vascularized magnetic cell sheets for bone regeneration.

The lack of effective strategies to produce vascularized 3D bone transplants in vitro, hampers the development of thick-constructed bone, limiting the translational of lab-based engineered system to clinical practices. Cell sheet (CS) engineering techniques provide an excellent microenvironment for vascularization since the technique can maintain the intact cell matrix, crucial for angiogenesis. In an attempt to develop hierarchical vascularized 3D cellular constructs, we herein propose the construction of stratified magnetic responsive heterotypic CSs by making use of iron oxide nanoparticles previously internalized within cells. Magnetic force-based CS engineering allows for the construction of thick cellular multilayers. Results show that osteogenesis is achieved due to a synergic effect of human umbilical vein endothelial cells (HUVECs) and adipose-derived stromal cells (ASCs), even in the absence of osteogenic differentiating factors. Increased ALP activity, matrix mineralization, osteopontin and osteocalcin detection were achieved over a period of 21 days for the heterotypic CS conformation (ASCs/HUVECs/ASCs), over the homotypic one (ASCs/ASCs), corroborating our findings. Moreover, the validated crosstalk between BMP-2 and VEGF releases triggers not only the recruitment of blood vessels, as demonstrated in an in vivo CAM assay, as well as the osteogenesis of the 3D cell construct. The in vivo angiogenic profile also demonstrated preserved human vascular structures and human cells showed the ability to migrate and integrate within the chick vasculature.

Fluxos assistenciais intermunicipais de nascimentos e óbitos perinatais na Região Metropolitana do Rio de Janeiro, Brasil, em 2011 e 2014 / Intermunicipal healthcare flows and perinatal births and deaths in Greater Metropolitan Rio de Janeiro, Brazil, in 2011 and 2014 / Flujos asistenciales intermunicipales de nacimientos y óbitos perinatales en la región metropolitana de Río de Janeiro, Brasil, en 2011 y 2014

A regionalização do cuidado perinatal deve considerar as estruturas assistenciais existentes e facilitar o acesso. Este estudo identificou fluxos assistenciais intermunicipais de nascimentos e óbitos perinatais ocorridos na Região Metropolitana do Rio de Janeiro, Brasil, em 2011 e 2014, definiu parâmetros e sistematizou propostas para a organização da regionalização da assistência perinatal. Estudo ecológico espaço-temporal. As fontes de dados foram os Sistemas de Informações sobre Nascidos Vivos e sobre Mortalidade, o Cadastro Nacional de Estabelecimentos de Saúde e o Censo Demográfico de 2010. Foram identificadas relações existentes entre municípios de residência e ocorrência dos nascimentos e óbitos perinatais. Cada município foi analisado em separado e em pares - residência/ocorrência - segundo o evento vital, recursos assistenciais e critérios pragmáticos de ameaça à vida. Foram realizadas análises descritivas de fluxos dominantes, fatorial exploratória de componentes principais e cluster. Identificaram-se as redes assistenciais existentes, e as 47 variáveis analisadas foram resumidas em três fatores (dimensões analíticas) - disponibilidade de leitos, situação de ameaça à vida e condições socioeconômicas - responsáveis, respectivamente, por 60/80%, 20/30% e 13/22% da variância, relativas a cada ano analisado. Os fatores foram utilizados para a formação dos clusters, classificados de 3 a 5 estratos. Três propostas de regiões de saúde perinatal foram elaboradas. A principal contribuição deste estudo foi apresentar parâmetros para o acompanhamento da regionalização e a reavaliação desse processo sistematicamente com base nos registros administrativos.

The regionalization of perinatal care should consider existing healthcare structures for facilitating access. This spatial-temporal ecological study identified intermunicipal flows of perinatal births and deaths in Greater Metropolitan Rio de Janeiro, Brazil, in 2011 and 2014, defined parameters, and systematized proposals for organizing the regionalization of perinatal care. The data sources were the Brazilian Information System on Live Births, Mortality Information System, National Registry of Healthcare Establishments, and 2010 Population Census. The study identified existing relations between the mothers' municipalities of residence and the occurrence of perinatal births and deaths. Each municipality was analyzed singly and pairwise (residence/occurrence) according to the vital event, healthcare resources, and pragmatic criteria of life-threatening conditions at birth. We conducted descriptive analyses of dominant flows, exploratory principal components analysis, and cluster analysis. The existing healthcare networks were identified, and 47 variables were summarized in three factors (analytical dimensions) - availability of beds, risk of life-threatening conditions, and socioeconomic status - accounting for 60%/80%, 20%/30%, 13%/22%, respectively, of the variance pertaining to each year analyzed. The factors were used to form clusters, classified in 3 to 5 strata. Three proposals were drafted for perinatal health regions. The study's principal contribution was having presented parameters for monitoring the regionalization and systematic reevaluation of this process based on administrative records.

La regionalización del cuidado perinatal debe considerar las estructuras asistenciales existentes y facilitar el acceso a las mismas. Este estudio identificó flujos asistenciales intermunicipales de nacimientos y óbitos perinatales, ocurridos en la región metropolitana de Río de Janeiro, Brasil, en 2011 y 2014, definió parámetros y sistematizó propuestas para la organización de la regionalización de la asistencia perinatal. Se trata de un estudio ecológico espacio-temporal. Las fuentes de datos fueron: Sistema de Información sobre Nacidos Vivos y sobre Mortalidad, registro nacional de establecimientos de salud y Censo Demográfico de 2010. Se identificaron las relaciones existentes entre municipios de residencia y ocurrencia de nacimientos y óbitos perinatales. Cada municipio se analizó por separado y por pares -residencia/ocurrencia-, según el evento vital, recursos asistenciales y criterios pragmáticos de amenaza para la vida. Se realizaron análisis descriptivos de flujos dominantes, así como análisis factoriales exploratorios de componentes principales y clúster. Se identificaron las redes asistenciales existentes y las 47 variables analizadas se resumieron en tres factores (dimensiones analíticas): disponibilidad de camas, situación de amenaza para la vida y condiciones socioeconómicas; responsables respectivamente de un 60%/80%, 20%/30% y 13%/22%, de la variancia, relativas a cada año analizado. Los factores fueron utilizados para la formación de los clústeres, clasificados de entre 3 a 5 estratos. Se elaboraron tres propuestas de regiones de salud perinatal. La principal contribución de este estudio fue presentar parámetros para realizar un seguimiento de la regionalización, así como la reevaluación de este proceso sistemáticamente, a partir de registros administrativos.

Nivolumab-induced hypothyroidism followed by isolated ACTH deficiency.

Cancer immunotherapy has been used in several malignancies with clinical benefit. Despite the clinical success, immune-related adverse events are frequent and endocrinopathies can be particularly severe. We report a 55-year-old male patient with stage IV pulmonary carcinoma treated with nivolumab who presented with thyroid dysfunction after the sixth administration of the drug. One year after thyroid dysfunction, the patient complained of severe fatigue, asthenia and weight loss. Laboratory testing showed low morning cortisol with undetected adrenocorticotropic hormone; other pituitary hormones were normal and MRI showed homogeneous enhancement of the pituitary gland and no space-occupying lesions. The diagnosis of nivolumab-induced hypophysitis was made and replacement treatment with hydrocortisone was started with clinical improvement. This case demonstrates that patients under immunotherapy are at risk for a large spectrum of endocrine dysfunctions that may worsen their prognosis. Close monitoring of these patients is warranted.

Gene and Base Editing as a Therapeutic Option for Cystic Fibrosis-Learning from Other Diseases.

Cystic fibrosis (CF) is a monogenic autosomal recessive disorder caused by mutations in the CFTR gene. There are at least 346 disease-causing variants in the CFTR gene, but effective small-molecule therapies exist for only ~10% of them. One option to treat all mutations is CFTR cDNA-based therapy, but clinical trials to date have only been able to stabilise rather than improve lung function disease in patients. While cDNA-based therapy is already a clinical reality for a number of diseases, some animal studies have clearly established that precision genome editing can be significantly more effective than cDNA addition. These observations have led to a number of gene-editing clinical trials for a small number of such genetic disorders. To date, gene-editing strategies to correct CFTR mutations have been conducted exclusively in cell models, with no in vivo gene-editing studies yet described. Here, we highlight some of the key breakthroughs in in vivo and ex vivo gene and base editing in animal models for other diseases and discuss what might be learned from these studies in the development of editing strategies that may be applied to cystic fibrosis as a potential therapeutic approach. There are many hurdles that need to be overcome, including the in vivo delivery of editing machinery or successful engraftment of ex vivo-edited cells, as well as minimising potential off-target effects. However, a successful proof-of-concept study for gene or base editing in one or more of the available CF animal models could pave the way towards a long-term therapeutic strategy for this disease.

Simultaneous determination of synthetic musks and UV-filters in water matrices by dispersive liquid-liquid microextraction followed by gas chromatography tandem mass-spectrometry.

An analytical method was developed for the simultaneous analysis of 19 emergent compounds in water matrices, six UV-filters (UVFs), five nitro, six polycyclic and two macrocyclic musks. The target compounds were extracted by a dispersive liquid-liquid microextraction (DLLME) approach, using 2-propanol as the dispersive solvent and 1,1,2-trichloroethane as the extractant solvent. The extracts were then analysed by gas chromatography tandem mass-spectrometry (GC-MS/MS). This methodology was successfully validated for the analyses of the target compounds in five types of aqueous samples (tap, river and sea water and influent and effluent wastewater). Recoveries of the analytes based on the surrogate correction ranged from 80 to 120%, with a good repeatability (relative standard deviations less than 10%). The method limit of detection ranges from 0.1 ng L-1 (octocrylene (OC), celestolide (ADBI)) to 20.0 ng L-1 (benzophenone (BZ)). Both UVFs and synthetic musk compounds (SMCs) were detected in all matrices. Higher concentrations were found in wastewaters (total mean concentration in influents of 6248 ng L-1 and 3856 ng L-1 in effluents), followed by river water (159 ng L-1). Only BZ was detected in one of the analysed seawater samples and none of the compounds were detected in tap water. The most detected UVFs among all matrices were BZ and drometrizole (DTS) and tonalide (AHTN) and galaxolide (HHCB) within the SMCs class. Among all matrices, wastewater was the one with higher number of compounds found per sample (both UVFs and SMCs).