Mpox outbreak in Rio de Janeiro, Brazil: A translational approach.

Mpox is a zoonotic disease historically reported in Africa. Since 2003, limited outbreaks have occurred outside Africa. In 2022, the global spread of cases with sustained interhuman transmission and unusual disease features raised public health concerns. We explore the mpox outbreak in Rio de Janeiro (RJ) state, Brazil, in an observational study of mpox-suspected cases from June to December 2022. Data collection relied on a public healthcare notification form. Diagnosis was determined by MPXV-PCR. In 46 confirmed cases, anti-OPXV IgG was determined by ELISA, and seven MPXV genomes were sequenced. A total of 3095 cases were included, 816 (26.3%) with positive MPXV-PCR results. Most positive cases were men in their 30 s and MSM. A total of 285 (34.9%) MPXV-PCR+ patients live with HIV. Eight were coinfected with varicella-zoster virus. Anogenital lesions and adenomegaly were associated with the diagnosis of mpox. Females and individuals under 18 represented 9.4% and 5.4% of all confirmed cases, respectively, showing higher PCR cycle threshold (Ct) values and fewer anogenital lesions compared to adult men. Anti-OPXV IgG was detected in 29/46 (63.0%) patients. All analyzed sequences belonged to clade IIb. In RJ state, mpox presented a diverse clinical picture, represented mainly by mild cases with low complication rates and prominent genital involvement. The incidence in females and children was higher than usually reported. The observation of a bimodal distribution of Ct values, with few positive results, may suggest the need to review the diagnostic criteria in these groups.

Discontinuing absorbent pants in children with bedwetting: a randomized controlled trial.

The objective of this study is to examine the effect of discontinuing wearing protective garments (absorbent pyjama pants - APP) in children with severe childhood nocturnal enuresis (NE). The study employs a multicenter, parallel, randomized controlled trial. Following a 4-week run-in period, participants were randomly allocated in a 2:1 group allocation to discontinue or continue using APP. The research was conducted across seven European pediatric incontinence centers. The study included treatment-naïve children aged 4-8 years with severe (7/7 wet nights per week) mono-symptomatic NE, who had used nighttime protection for at least 6 months prior to the study. The study consisted of a 4-week run-in period (± 7 days), where all children slept wearing APP (DryNites®). At week 4 (± 7 days), if meeting randomization criteria (7/7 wet nights during the last week of run-in), participants were randomized to continue to sleep in APP or to discontinue their use for a further 4 weeks, with the option of another 4 weeks in the extension period. The primary outcome was the difference between groups of wet nights during the last week of intervention. Quality of life (QoL) and sleep were secondary endpoints. In total, 105 children (43 girls and 62 boys, mean age 5.6 years [SD 1.13]) were randomized (no-pants group n = 70, pants group n = 35). Fifteen children (21%) in the no-pants group discontinued early due to stress related to the intervention. Children in the no-pants group experienced fewer wet nights compared to the pants group during the last week (difference 2.3 nights, 95% CI 1.54-3.08; p < 0.0001). In the no-pants group, 20% responded to the intervention, of whom 13% had a full response. Clinical improvement was detected within 2 weeks. Sleep and QoL were reported as negatively affected by APP discontinuation in the extension period but not in the core period.    Conclusion: A ~ 10% complete resolution rate was associated with discontinuing APP. While statistically significant, the clinical relevance is debatable, and the intervention should be tried only if the family is motivated. Response was detectable within 2 weeks. Discontinuing APP for 4-8 weeks was reported to negatively affect QoL and sleep quality. No severe side effects were seen.Trial registration: Clinicaltrials.gov Identifier: NCT04620356; date registered: September 23, 2020. Registered under the name: "Effect of Use of DryNites Absorbent Pyjama Pants on the Rate of Spontaneous Resolution of Paediatric Nocturnal Enuresis (NE)."

The larvicidal effect of the supernatant of Lactobacillus acidophilus ATCC 4356 on Toxocara canis.

Human toxocariasis is a parasitic anthropozoonosis that is difficult to treat and control. A previous study carried out with Lactobacillus acidophilus ATCC 4356 revealed that the cell free supernatant (CFS) of this probiotic killed 100% of Toxocara canis larvae in vitro. The present study aimed to investigate the characteristics of the CFS of L. acidophilus ATCC 4356, which may be involved in its larvicidal effects on T. canis. L. acidophilus ATCC 4356 was cultured, and lactic and acetic acids present in the CFS were quantified by high performance liquid chromatography (HPLC). The levels of pH and H2O2 were also analyzed. To assess the larvicidal effect of the CFS, this was tested pure and diluted (1:2 to 1:128) on T. canis larvae. High concentrations of lactic and acetic acids were detected in the CFS. The acidity of the pure CFS was observed at pH 3.8, remaining acidic at dilutions of 1:2 to 1:16. Regarding the in vitro larvicidal effect, 100% death of T. canis larvae was observed using the pure CFS and 1:2 dilution. Based on these results, it can be inferred that the presence of higher concentrations of organic acids and low pH of the medium contributed to the larvicidal activity of the CFS of L. acidophilus ATCC 4356. In addition, the maintenance of the larvicidal effect, even after dilution, suggests a greater chance of the larvicidal effect of this CFS against T. canis in vivo.

Screening with urine Histoplasma antigen test in asymptomatic patients starting TNF-alpha inhibitor therapy: a cohort study.

Background: Histoplasmosis is the second most frequent granulomatous disease in patients treated with tumor necrosis factor (TNF)-α inhibitors, second only to tuberculosis. However, there is limited information about pre-therapy screening procedures and the need for preventive treatments for patients who will start immunobiologicals. Methods: This is a cohort study that evaluated the prevalence of histoplasmosis in asymptomatic HIV-negative patients before initiation of TNF-α inhibitors by testing for Histoplasma antigen in urine samples. The patients included completed a 180-day follow-up after the initiation of the biologics to assess the onset of symptoms suggestive of histoplasmosis. Results: From January 2021 to December 2022, 54 patients who were prescribed a TNF-α inhibitor agent for treating autoimmune diseases in centers in southern Brazil were included. In the screening before therapy, the prevalence of a positive urinary Histoplasma antigen test was 14.8%. None of the 54 patients developed histoplasmosis after 6 months of immunobiological therapy, including the eight patients who tested positive. Conclusion: The prevalence of Histoplasma capsulatum infection in chronic patients may be higher than expected, but the impact of latent infection in asymptomatic patients is still uncertain, including those starting treatment with immunobiological drugs such as TNF-α inhibitors. Our study did not identify risk factors for the diagnosis of disseminated histoplasmosis in this group, including a positive result in an antigen test performed before immunobiological therapy. To date, there is no evidence to recommend routine antigen-based screening or preventive therapy for histoplasmosis before initiating a TNF-α inhibitor.

Using a urine test for fungal infection to screen people without symptoms who are about to start taking immunobiologic medications This study looked at the prevalence of histoplasmosis, a fungal infection, in asymptomatic patients who were about to start treatment with TNF-α inhibitors, which are medications used for autoimmune diseases. The researchers tested urine samples for Histoplasma antigen before the patients started the treatment and followed them for 180 days after starting the medication to see if they developed any symptoms of histoplasmosis. The study included 54 patients in southern Brazil, and they found that 14.8% of the patients tested positive for the Histoplasma antigen before starting the treatment. However, none of the patients, including those who tested positive, developed histoplasmosis during the 6-month follow-up. The researchers concluded that histoplasmosis infection may be more common in these patients than previously thought, but it's still not clear if asymptomatic patients with a positive antigen test will develop the infection when starting TNF-α inhibitor treatment. The study did not find any specific risk factors for developing histoplasmosis in this group of patients, and based on their findings, they did not recommend routine screening or preventive therapy for histoplasmosis before starting TNF-α inhibitor treatment.

Boron-doped graphene topological defects: unveiling high sensitivity to NO molecule for gas sensing applications.

Global air quality has deteriorated significantly in recent years due to large emissions from the transformation industry and combustion vehicles. This issue requires the development of portable, highly sensitive, and selective gas sensors. Nanostructured materials, including defective graphene, have emerged as promising candidates for such applications. In this work, we investigated the B-doped topological line defect in graphene as a sensing material for various gas molecules (CO, CO2, NO, and NH3) based on a combination of density functional theory and the non-equilibrium Green's function method. The electronic transport calculations reveal that the electric current can be confined to the line defect region by gate voltage control, revealing highly reactive sites. The B-doped topological line defect is metallic, favoring the adsorption of NO and NH3 over CO and CO2 molecules. We notice changes in the conductance after gas molecule adsorption, producing a sensitivity of 50% (16%) for NO (NH3). In addition, the recovery time for nitride gases was calculated for different temperatures and radiation frequencies. At 300 K the ultraviolet (UV) has a fast recovery time compared to the visible (VIS) one by about two orders of magnitude. This study gives an understanding of how engineering transport properties at the microscopic level (by topological line defect and chemical B-doping) leads to promising nanosensors for detecting nitride gas.

Reversible Tricuspid Regurgitation Due to Beriberi.

We report a case of cardiac beriberi in a 76-year-old man who was hospitalized with a congestive condition of subacute onset, diagnosed as high-output heart failure associated with severe tricuspid regurgitation and indication for caval valve implantation, which, after thiamine replacement, resulted in improvement of all conditions. (Level of Difficulty: Beginner.).

Incidence of histoplasmosis in patients receiving TNF-alpha inhibitors: A systematic review and meta-analysis.

BACKGROUND: Immunobiological drugs such as TNF-α inhibitors are valuable in rescue therapy for autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease (IBD), but they increase the risk of infectious complications. Histoplasmosis is a significant concern in patients living in endemic regions, however, few studies have assessed the incidence of Histoplasma infection during therapy, and classic estimates may underestimate the risk. This study aimed to produce an updated risk estimate of histoplasmosis in patients on TNF-α blocking therapy. METHODS: This is a systematic review and meta-analysis of studies that contain parameters for calculating the risk of histoplasmosis in people who use TNF-α inhibitors, to produce a risk estimate. RESULTS: We identified 11 studies with the necessary parameters for inclusion in the meta-analysis, most of which were from North America. The incidence rate of histoplasmosis found was 33.52 cases per 100,000 patients treated with TNF-ɑ inhibitors (95% CI 12.28-91.46). Considering only studies evaluating monoclonal antibodies, the calculated incidence was 54.88/100,000 patients treated (95%CI 23.45-128.34). In subgroup analysis, the incidence was much higher in patients with IBD compared to rheumatic diseases. There was significant heterogeneity among the studies. CONCLUSION: The risk of histoplasmosis during TNF-α inhibitory therapy may be considerably higher than that found in classical estimates, especially in patients with IBD. There is a lack of studies evaluating histoplasmosis in large endemic areas, such as Central and South America.

Brazilian Green Propolis' Artepillin C and Its Acetylated Derivative Activate the NGF-Signaling Pathways and Induce Neurite Outgrowth in NGF-Deprived PC12 Cells.

Artepillin C is the most studied compound in Brazilian Green Propolis and, along with its acetylated derivative, displays neurotrophic activity on PC12 cells. Specific inhibitors of the trkA receptor (K252a), PI3K/Akt (LY294002), and MAPK/ERK (U0126) signaling pathways were used to investigate the neurotrophic mechanism. The expression of proteins involved in axonal and synaptic plasticity (GAP-43 and Synapsin I) was assessed by western blotting. Additionally, physicochemical properties, pharmacokinetics, and drug-likeness were evaluated by the SwissADME web tool. Both compounds induced neurite outgrowth by activating the NGF-signaling pathways but through different neuronal proteins. Furthermore, in silico analyses showed interesting physicochemical and pharmacokinetic properties of these compounds. Therefore, these compounds could play an important role in axonal and synaptic plasticity and should be further investigated.

Acute exercise modulates the inflammatory response in adipose tissue in both lean and obese mice.

OBJECTIVES: Acute physical exercise acts as a metabolic stressor, promoting activation of the immune system, and this response could be relevant in the adipose tissue remodeling process. In addition, some cytokines have important functions in lipolysis. Because chronic exercise improves obesity-related metabolic and inflammatory dysfunction, herein we investigated the effect of acute exercise on the inflammatory responses in the adipose tissues of lean and obese mice. METHODS: Lean mice were fed a standard chow diet, whereas obese mice were fed a high-refined carbohydrate diet for 8 wk. Both groups were subjected to 60 min of moderate-intensity exercise. RESULTS: In the epididymal adipose tissue of lean mice, exercise enhanced interleukin (IL)-6 and tumor necrosis factor-α levels, which correlated positively with increased serum free fatty acid concentrations. In vivo confocal imaging of epididymal adipose tissue vessels revealed higher recruitment of neutrophils after exercise. Also, the number of leukocytes expressing CD11b+F480- was elevated 6 h after exercise. Similarly, the chemokine (C-X-C motif) ligand 1 level increased at 6 h and remained high until 24 h after exercise. Myeloperoxidase activity was increased at 6, 12, and 24 h after exercise. Surprisingly, however, no changes were observed in epididymal adipose tissue from obese mice, considering proinflammatory cytokines (IL-6 and tumor necrosis factor-α). On the other hand, IL-13, IL-4, and IL-10 levels were higher in obese mice after exercise. CONCLUSIONS: These data suggest that acute exercise promotes an inflammatory response in the adipose tissue of lean mice that is observed as part of its role in adipose tissue remodeling. In contrast, acute exercise promotes an antiinflammatory response in adipose tissue from obese mice, likely as an important tool for restoring homeostasis.

Diversity of fungal microbiome obtained from plant rhizoplanes.

Microbial communities, and their ecological importance, have been investigated in several habitats. However, so far, most studies could not describe the closest microbial interactions and their functionalities. This study investigates the co-occurring interactions between fungi and bacteria in plant rhizoplanes and their potential functions. The partnerships were obtained using fungal-highway columns with four plant-based media. The fungi and associated microbiomes isolated from the columns were identified by sequencing the ITS (fungi) and 16S rRNA genes (bacteria). Statistical analyses including Exploratory Graph and Network Analysis were used to visualize the presence of underlying clusters in the microbial communities and evaluate the metabolic functions associated with the fungal microbiome (PICRUSt2). Our findings characterize the presence of both unique and complex bacterial communities associated with different fungi. The results showed that Bacillus was associated as exo-bacteria in 80 % of the fungi but occurred as putative endo-bacteria in 15 %. A shared core of putative endo-bacterial genera, potentially involved in the nitrogen cycle was found in 80 % of the isolated fungi. The comparison of potential metabolic functions of the putative endo- and exo-communities highlighted the potential essential factors to establish an endosymbiotic relationship, such as the loss of pathways associated with metabolites obtained from the host while maintaining pathways responsible for bacterial survival within the hypha.

Brazilian Brown Propolis: an Overview About Its Chemical Composition, Botanical Sources, Quality Control, and Pharmacological Properties.

Brazil is one of the largest propolis producers in the world. Propolis is produced by bees from plant exudates and tissues, leading to many variations in the types of propolis. Generally, Brazilian propolis types are green, brown, and red. Despite not being the main research focus as the green and red propolis, brown propolis is the second most produced propolis type in Brazil and has tremendous economic and medicinal importance. Propolis has drawn attention with the rise in the search for healthier lifestyles, functional foods, biocosmetics, and natural products as therapeutic sources. This review covers the main chemical constituents identified in different types of Brazilian brown propolis, and their botanical sources, chemistry, and biological activities. The economic aspect of brown propolis is also presented. There are many gaps to be filled for brown propolis regarding the development of analytical methods, and quality control to allow its standardization, limiting its applicability in the food and pharmaceutical industries. Future perspectives regarding brown propolis research were discussed, especially biological activities, to support the medicinal uses of different types of brown propolis. Supplementary Information: The online version contains supplementary material available at 10.1007/s43450-023-00374-x.

Novel synthetic derivatives of cinnamic and p-coumaric acids with antiproliferative effect on breast MCF-7 tumor cells.

p-Coumaric acid is derived from cinnamic acid and is one of the major compounds in the Brazilian green propolis extract. Studies have shown that both p-coumaric acid and cinnamic acid have promising antiproliferative effects. In this context, aiming to increase the complexity of these active natural products and their activities, we performed coupling reactions with propargylamine and benzylamine, as well as with threonine, phenylalanine and lysine amino acids, aiming to enhance their antiproliferative effects towards the hormone-dependent breast cancer MCF-7 cells. Overall, the p-coumaric acid coupling with L-threonine amino acid (compound 15) had the best selectivity index (SI = 5.1), with half-maximal inhibitory concentration of 39.6 ± 1 µM, showing a high selectivity against hormone-dependent breast cancer cell lines MCF-7 and low cytotoxicity against the normal breast cell lines MCF-10A. Thus, this new natural product derivative may represent a prototype for the future development of antiproliferative agents, especially against hormone-dependent breast cancer.

Brazilian green propolis extracts modulate cholesterol homeostasis in a preclinical guinea pig model: an in vitro and in vivo study.

Green propolis produced by Apis melífera bees, having Baccharis dracunculifolia D.C. (Asteraceae) as the primary botanical source, has been used in traditional medicine to treat numerous disorders. However, studies evaluating propolis' potential in treating cardiovascular diseases via its effects on cholesterol metabolism are lacking. Therefore, this study investigated the effects of green propolis extracts on lipid metabolism in hypercholesterolemic guinea pigs. Chemical characterization of ethanolic extracts of green propolis samples was undertaken using HPLC. The in vitro characterization included an evaluation of the antioxidant capacity of the hydroalcoholic extract of green propolis (DPPH and FRAP assays) and its ability to act as an inhibitor of the HMG-CoA reductase enzyme. In vivo, we investigated the effect of the hydroalcoholic extract of green propolis on lipid metabolism in hypercholesterolemic guinea pigs. Results obtained validated previous reports of significant antioxidant activity. HPLC analysis confirmed that coumaric acid, artepillin C, and baccharin were the most common and abundant compounds in green propolis samples among the studied compounds. Furthermore, the compounds in these extracts acted as effective HMG-CoA reductase inhibitors in vitro. In vivo assays demonstrated that a hypercholesterolemic diet significantly reduced serum levels of the HDL cholesterol fraction. Simvastatin and propolis hydroalcoholic extracts promoted a significant increase in HDL cholesterol, suggesting that these extracts can improve the serum lipid profile of hypercholesterolemic guinea pigs. Results obtained in this study provide a perspective on the possible hypocholesterolemic effect of green propolis, suggesting that it can improve the serum lipid profile in hypercholesterolemic guinea pigs.

Eucalyptus botryoides' resin and its new 2-O-galloyl-1,6-O-di-trans-p-coumaroyl-ß-D-glycopyranoside compound display good antimicrobial activity.

Fungal resistance to different therapeutic drugs has become a growing challenge. This crucial health problem requires new effective drug alternatives. Herein, we report the study of Eucalyptus botryoides' resin used in folk medicine as antimicrobial. Thus, E. botryoides' resin was extracted with aqueous-ethanol and fractionated using Sephadex chromatography, furnishing its major compounds. The crude extracts and the isolated compounds were evaluated for their in vitro antimicrobial activity against bacteria and yeasts. The crude extract displayed MIC of 25 µg/mL against S. salivarius, and for C. albicans, C. glabrata, and C. tropicalis the MIC were between 2.9 and 5.9 µg/mL. The 7-O-Methyl-aromadendrin was the most effective against C. glabrata and C. krusei (MIC = 1.6 µg/mL). 2-O-Galloyl-1,6-O-di-trans-p-coumaroyl-ß-D-glycopyranoside, first time reported, showed MIC of 3.1 µg/mL against C. glabrata and C. krusei. Overall, this work gave promising results, indicating that Eucalyptus botryoides' resin and its compounds have the potential for developing anti-yeast products.

Avaliação dos efeitos do curativo compressivo após procedimentos intervencionistas por via radial / Evaluation of the effects of compressive dressing after interventional procedures via radial approach

Introdução: Dispositivos externos compressivos após a punção radial para cateterismo cardíaco e intervenção coronária percutânea visam à hemostasia, ao conforto para o paciente e à segurança. O estudo objetivou avaliar pacientes submetidos a procedimentos coronários invasivos por via radial, nos quais se utilizou um curativo compressivo e de baixo custo, desenvolvido no serviço, avaliando a segurança dele e a ocorrência de complicações. Métodos: Indivíduos submetidos à coronariografia e à intervenção coronária percutânea, avaliados no momento de retirada do curativo compressivo e após 7 dias, por meio de questionário abrangendo comorbidades, quadro clínico, tempo do procedimento e de compressão, avaliação física de hematoma e dor, e Doppler vascular sonoro para avaliação de oclusão da artéria radial. Resultados: Foram avaliados 144 pacientes, sendo 138 com seguimento em 7 dias. Os eventos não diferiram entre procedimentos diagnósticos e terapêuticos. Na avaliação imediata à retirada do curativo, revelou-se incidência de 4,2% de oclusão da artéria radial, com dor referida em 23,6% dos pacientes, graduada em 2,9±1,7 pela Escala Visual Analógica (intensidade de zero a dez) e sem sangramentos graves. Hematoma ocorreu em um paciente (0,9%), com classificação III pelo critério EASY. Na avaliação de 7 dias, a incidência de oclusão da artéria radial foi de 2,2%, a dor foi referida em 11,1% da amostra (intensidade 1,8±0,8), e o hematoma foi evidenciado em 3,5%. Conclusão: O curativo compressivo mostrou-se um procedimento seguro, com baixa taxa de complicações e baixa taxa de dor local nos pacientes submetidos a procedimentos coronários invasivos pela via radial.

Background: External compressive devices after radial puncture for cardiac catheterization and percutaneous coronary intervention aim at hemostasis, patient comfort, and safety. The objective of the study was to evaluate patients undergoing invasive coronary procedures by radial approach, in which a low-cost compressive dressing developed at the service was used, assessing its safety and the occurrence of complications. Methods: Patients undergoing coronary angiography and percutaneous coronary intervention, evaluated at the time of removal of compressive dressing and after 7 days, by means of a questionnaire addressing comorbidities, clinical picture, procedure and compression time, physical assessment of hematoma and pain, and a vascular Doppler ultrasound to evaluate radial artery occlusion. Results: A total of 144 patients were evaluated, 138 of whom were followed up within 7 days. Events did not differ among diagnostic and therapeutic procedures. In the immediate evaluation after removal of dressing, an incidence of 4.2% of radial artery occlusion was observed, with pain reported by 23.6% of patients, graded at 2.9±1.7 points in the Visual Analogue Scale (intensity of zero to ten), and no major bleeding. Hematoma occurred in one patient (0.9%), classified as type III according to the EASY criteria. In the 7-day evaluation, the incidence of radial artery occlusion was 2.2%, pain was reported in 11.1% of sample (intensity 1.8±0.8), and hematoma was evident in 3.5%. Conclusion: The compressive dressing proved to be a safe procedure, with a low rate of complications and a low rate of local pain in patients undergoing invasive coronary procedures via radial approach.

Elevated Adsorption of Lead and Arsenic over Silver Nanoparticles Deposited on Poly(amidoamine) Grafted Carbon Nanotubes.

An efficient adsorbent, CNTs-PAMAM-Ag, was prepared by grafting fourth-generation aromatic poly(amidoamine) (PAMAM) to carbon nanotubes (CNTs) and successive deposition of Ag nanoparticles. The FT-IR, XRD, TEM and XPS results confirmed the successful grafting of PAMAM onto CNTs and deposition of Ag nanoparticles. The absorption efficiency of CNTs-PAMAM-Ag was evaluated by estimating the adsorption of two toxic contaminants in water, viz., Pb(II) and As(III). Using CNTs-PAMAM-Ag, about 99 and 76% of Pb(II) and As(III) adsorption, respectively, were attained within 15 min. The controlling mechanisms for Pb(II) and As(III) adsorption dynamics were revealed by applying pseudo-first and second-order kinetic models. The pseudo-second-order kinetic model followed the adsorption of Pb(II) and As(III). Therefore, the incidence of chemisorption through sharing or exchanging electrons between Pb(II) or As(III) ions and CNTs-PAMAM-Ag could be the rate-controlling step in the adsorption process. Further, the Weber-Morris intraparticle pore diffusion model was employed to find the reaction pathways and the rate-controlling step in the adsorption. It revealed that intraparticle diffusion was not a rate-controlling step in the adsorption of Pb(II) and As(III); instead, it was controlled by both intraparticle diffusion and the boundary layer effect. The adsorption equilibrium was evaluated using the Langmuir, Freundlich, and Temkin isotherm models. The kinetic data of Pb(II) and As(III) adsorption was adequately fitted to the Langmuir isotherm model compared to the Freundlich and Temkin models.

Distinguishing nanoparticle drug release mechanisms by asymmetric flow field-flow fractionation.

This research demonstrates the development, application, and mechanistic value of a multi-detector asymmetric flow field-flow fractionation (AF4) approach to acquire size-resolved drug loading and release profiles from polymeric nanoparticles (NPs). AF4 was hyphenated with multiple online detectors, including dynamic and multi-angle light scattering for NP size and shape factor analysis, fluorescence for drug detection, and total organic carbon (TOC) to quantify the NPs and dissolved polymer in nanoformulations. The method was demonstrated on poly(lactic-co-glycolic acid) (PLGA) NPs loaded with coumarin 6 (C6) as a lipophilic drug surrogate. The bulk C6 release profile using AF4 was validated against conventional analysis of drug extracted from the NPs and complemented with high performance liquid chromatography - quadrupole time-of-flight (HPLC-QTOF) mass spectrometry analysis of oligomeric PLGA species. Interpretation of the bulk drug release profile was ambiguous, with several release models yielding reasonable fits. In contrast, the size-resolved release profiles from AF4 provided critical information to confidently establish the release mechanism. Specifically, the C6-loaded NPs exhibited size-independent release rate constants and no significant NP size or shape transformations, suggesting surface desorption rather than diffusion through the PLGA matrix or erosion. This conclusion was supported through comparative experimental evaluation of PLGA NPs carrying a fully entrapped drug, enrofloxacin, which showed size-dependent diffusive release, along with density functional theory (DFT) calculations indicating a higher adsorption affinity of C6 onto PLGA. In summary, the development of the size-resolved AF4 method and data analysis framework fulfills salient analytical gaps to determine drug localization and release mechanisms from nanomedicines.

Complexity of high-risk pregnancy care in the health care network.

OBJECTIVE: To understand the complexity of high-risk care for pregnant women in the health care network. METHOD: Qualitative study with theoretical framework of Edgar Morin's Complex Thought and Grounded Theory methodology, Strauss's version. Data collection by theoretical sampling, including twelve health professionals and seven women users of the care network in a municipality in the south of Brazil from July to October 2018. Analysis by open and axial coding and selective integration. RESULTS: The phenomenon "Caring for high-risk pregnant women in the health care network", comprises four categories: Noticing autonomous decision making; Promoting care; Developing multiprofessional work; and Accessing the health care network. CONCLUSION: Every high-risk pregnant woman should be seen as a singular and multidimensional being with comprehensive and continuous care, considering the complexity of local, regional, and global reality.

Antimitotic and toxicogenetic action of Stevia urticifolia aerial parts on proliferating vegetal and mammalian cells: in vitro and in vivo traditional and replacement methods.

Stevia urticifolia Thunb. is an underexploited herb possessing bioactive flavonoids, saponins, and terpenoids. The aim of this study was to examine the antiproliferative and toxicogenetic properties of the ethyl acetate extract from Stevia urticifolia aerial parts (EtAcSur) upon Artemia salina, erythrocytes, Allium cepa and sarcoma 180 cells and fibroblasts, as well as in vivo studies on mice to determine systemic, macroscopic, and behavioral alterations and bone marrow chromosomal damage. The assessment using A. salina larvae and mouse blood cells revealed LC50 and EC50 values of 68.9 and 113.6 µg/ml, respectively. Root growth and mitosis were inhibited by EtAcSur, and chromosomal aberrations were detected only at 100 µg/ml. EtAcSur exhibited potent concentration-dependent viability reduction of S180 and L-929 cells and antioxidant capacity employing ABTS• and DPPH•. No previous in vivo studies were performed before with the EtAcSur. Signals of acute toxicity were not observed at 300 mg/kg. Physiological and toxicological investigations at 25 and 50 mg/mg/day i.p. for 8 days did not markedly change body or organ relative weights, nor patterns of spontaneous locomotor and exploratory activities. In contrast, clastogenic effects on bone marrow were found at 50 mg/mg/day. EtAcSur was found to (1) produce toxicity in microcrustaceans, (2) capacity as free radical scavenger, (3) antimitotic, cytotoxic and clastogenic activties upon vegetal and mammalian cells, and (4) lethality on both tumor and normal murine cells indistinctly. In vivo damage systemic effects were not remarkable and clinical signals of toxicity were not observed, suggesting the significant pharmacological potential of S. urticifolia for the development of antineoplastic agents.Abbreviations: ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); DMSO: dimethylsulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EC50: effective concentration 50%; EtAcSur: ethyl acetate extract from Stevia urticifolia aerial parts; Hb, hemoglobin; IC50: inhibitory concentration 50%; LC50,: lethal concentration 50%; MI: mitotic index; RBC, red blood cells; Trolox: 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid.