A transcriptional switch governs fibroblast activation in heart disease.

In diseased organs, stress-activated signalling cascades alter chromatin, thereby triggering maladaptive cell state transitions. Fibroblast activation is a common stress response in tissues that worsens lung, liver, kidney and heart disease, yet its mechanistic basis remains unclear1,2. Pharmacological inhibition of bromodomain and extra-terminal domain (BET) proteins alleviates cardiac dysfunction3-7, providing a tool to interrogate and modulate cardiac cell states as a potential therapeutic approach. Here we use single-cell epigenomic analyses of hearts dynamically exposed to BET inhibitors to reveal a reversible transcriptional switch that underlies the activation of fibroblasts. Resident cardiac fibroblasts demonstrated robust toggling between the quiescent and activated state in a manner directly correlating with BET inhibitor exposure and cardiac function. Single-cell chromatin accessibility revealed previously undescribed DNA elements, the accessibility of which dynamically correlated with cardiac performance. Among the most dynamic elements was an enhancer that regulated the transcription factor MEOX1, which was specifically expressed in activated fibroblasts, occupied putative regulatory elements of a broad fibrotic gene program and was required for TGFß-induced fibroblast activation. Selective CRISPR inhibition of the single most dynamic cis-element within the enhancer blocked TGFß-induced Meox1 activation. We identify MEOX1 as a central regulator of fibroblast activation associated with cardiac dysfunction and demonstrate its upregulation after activation of human lung, liver and kidney fibroblasts. The plasticity and specificity of BET-dependent regulation of MEOX1 in tissue fibroblasts provide previously unknown trans- and cis-targets for treating fibrotic disease.

Trypanosoma cruzi killing and immune response boosting by novel phenoxyhydrazine-thiazole against Chagas disease.

Trypanosoma cruzi (T. cruzi) causes Chagas, which is a neglected tropical disease (NTD). WHO estimates that 6 to 7 million people are infected worldwide. Current treatment is done with benznidazole (BZN), which is very toxic and effective only in the acute phase of the disease. In this work, we designed, synthesized, and characterized thirteen new phenoxyhydrazine-thiazole compounds and applied molecular docking and in vitro methods to investigate cell cytotoxicity, trypanocide activity, nitric oxide (NO) production, cell death, and immunomodulation. We observed a higher predicted affinity of the compounds for the squalene synthase and 14-alpha demethylase enzymes of T. cruzi. Moreover, the compounds displayed a higher predicted affinity for human TLR2 and TLR4, were mildly toxic in vitro for most mammalian cell types tested, and LIZ531 (IC50 2.8 µM) was highly toxic for epimastigotes, LIZ311 (IC50 8.6 µM) for trypomastigotes, and LIZ331 (IC50 1.9 µM) for amastigotes. We observed that LIZ311 (IC50 2.5 µM), LIZ431 (IC50 4.1 µM) and LIZ531 (IC50 5 µM) induced 200 µg/mL of NO and JM14 induced NO production in three different concentrations tested. The compound LIZ331 induced the production of TNF and IL-6. LIZ311 induced the secretion of TNF, IFNγ, IL-2, IL-4, IL-10, and IL-17, cell death by apoptosis, decreased acidic compartment formation, and induced changes in the mitochondrial membrane potential. Taken together, LIZ311 is a promising anti-T. cruzi compound is not toxic to mammalian cells and has increased antiparasitic activity and immunomodulatory properties.

Multiscale Analysis of Extracellular Matrix Remodeling in the Failing Heart.

RATIONALE: Cardiac ECM (extracellular matrix) comprises a dynamic molecular network providing structural support to heart tissue function. Understanding the impact of ECM remodeling on cardiac cells during heart failure (HF) is essential to prevent adverse ventricular remodeling and restore organ functionality in affected patients. OBJECTIVES: We aimed to (1) identify consistent modifications to cardiac ECM structure and mechanics that contribute to HF and (2) determine the underlying molecular mechanisms. METHODS AND RESULTS: We first performed decellularization of human and murine ECM (decellularized ECM) and then analyzed the pathological changes occurring in decellularized ECM during HF by atomic force microscopy, 2-photon microscopy, high-resolution 3-dimensional image analysis, and computational fluid dynamics simulation. We then performed molecular and functional assays in patient-derived cardiac fibroblasts based on YAP (yes-associated protein)-transcriptional enhanced associate domain (TEAD) mechanosensing activity and collagen contraction assays. The analysis of HF decellularized ECM resulting from ischemic or dilated cardiomyopathy, as well as from mouse infarcted tissue, identified a common pattern of modifications in their 3-dimensional topography. As compared with healthy heart, HF ECM exhibited aligned, flat, and compact fiber bundles, with reduced elasticity and organizational complexity. At the molecular level, RNA sequencing of HF cardiac fibroblasts highlighted the overrepresentation of dysregulated genes involved in ECM organization, or being connected to TGFß1 (transforming growth factor ß1), interleukin-1, TNF-α, and BDNF signaling pathways. Functional tests performed on HF cardiac fibroblasts pointed at mechanosensor YAP as a key player in ECM remodeling in the diseased heart via transcriptional activation of focal adhesion assembly. Finally, in vitro experiments clarified pathological cardiac ECM prevents cell homing, thus providing further hints to identify a possible window of action for cell therapy in cardiac diseases. CONCLUSIONS: Our multiparametric approach has highlighted repercussions of ECM remodeling on cell homing, cardiac fibroblast activation, and focal adhesion protein expression via hyperactivated YAP signaling during HF.

Prevalence and associated factors of aspiration and severe dysphagia in asymptomatic patients in the late period after open partial laryngectomy: a videofluoroscopic evaluation.

PURPOSE: This study aimed to evaluate late and asymptomatic patients after open partial horizontal laryngectomy (OPHL), investigating the clinical-surgical and socio-demographic factors associated with aspiration and severe dysphagia. METHODS: One-thousand videofluoroscopic swallowing studies were performed in 100 asymptomatic patients in the late period after OPHL(median 6.5 years). Aspiration and severe dysphagia were, respectively, assessed by the Penetration-Aspiration scale (PAS) and by the Dynamic Imaging Grade of Swallowing Toxicity (DIGEST) classification. Associated factors were investigated by multivariate logistic regressions. RESULTS: 34% (95% CI 24.3-47.6%) of patients presented aspiration and 23% (95% CI 15.3-34.6%) had severe or life-threatening dysphagia (DIGEST grades 3-4). On logistic regression, the presence of aspiration was associated with lower preoperative serum albumin (odds ratio [OR]: 0.22; 95% CI 0.07-0.64; p = 0.005, for each 1 g/dL increment); a greater weight loss in early postoperative period (OR: 1.19, 95% CI 1.05-1.35; p = 0.008, for each 1 kg loss); older age at surgery (OR: 1.08; 95% CI 1.01-1.17, for each 1-year older); and with the presence of diabetes (OR: 5.16; 95% CI 1.09-27.47; p = 0.039). CONCLUSION: Deglutition abnormalities are frequent in asymptomatic patients later after OPHL. Older patients, with lower preoperative serum albumin levels, with greater postoperative weight loss, and with diabetes compose the clinical profile at risk for having worse swallowing function in the late period after OPHL.

Using the quality by design (QbD) approach to optimize formulations of lipid nanoparticles and nanoemulsions: A review.

Quality-by-design (QbD) approach has been applied to optimize lipid-based nanosystems formulations, including solid lipid nanoparticles (SLN), nanostructured lipid carriers (NLC) and nanoemulsions, besides being increasingly requested by regulatory authorities. Different mathematical models and statistical tests have been used, with similar conclusions regarding the parameters that influence the physical features of the resulting nanosystems. These include, variations in composition (e.g. lipid(s) and/or emulsifier(s)) and manufacturing parameters (e.g. emulsification rate and/or time, sonication amplitude and/or time, and homogenization pressure and/or cycles). These are critical parameters that influence nanoparticle/globule mean size, polydispersity index, zeta potential, drug encapsulation efficiency and in vitro drug release. This review addresses the concepts and applications of QbD for the development of lipid-based nanosystems, reporting successful examples published in the last 2 years. Although, some limitations have been identified, it is expected that in the upcoming years the application of QbD in pharmaceutical development will be an established approach.

Pelvic floor disorders: what's the best test?

Pelvic floor dysfunctions represent a common health problem affecting particularly post-menopausal women impacting significantly the quality of life. A large number of these patients suffer for many years without proper treatment often due to the lack of objective findings necessary to plan proper treatment. Because abnormalities of the different pelvic compartments are frequently associated, thorough diagnostic characterization of how many compartments are affected is paramount in order to plan the management approach that can include a multidisciplinary surgical approach. This pictorial essay will review the different imaging methods used for the characterization of these disorders, how to do them and its rationale providing a clinically understandable interpretation with clinical correlates and a correlation between fluoroscopic and MR defecography in order to illustrate the strengths and shortcomings of each. The need to use a standardized, reliable, and clinically understandable method of quantification has become more obvious in the last decades with the increasing rate of scientific and professional interchanges. A review of the grading systems used to convey the imaging findings also highlights the importance of using a standardized tool for comparing and communicating clinical findings understandable to referring physicians with proven inter-observer and intra-observer agreement of the examinations.

Nanoparticles in Ocular Drug Delivery Systems.

Conventional ophthalmic formulations lack a prolonged drug release effect and mucoadhesive properties, decreasing their residence time in the precorneal area and, therefore, in drug penetration across ocular tissues, presenting low bioavailability with a consequent reduction in therapeutic efficacy [...].

Ocular Delivery of Therapeutic Proteins: A Review.

Therapeutic proteins, including monoclonal antibodies, single chain variable fragment (ScFv), crystallizable fragment (Fc), and fragment antigen binding (Fab), have accounted for one-third of all drugs on the world market. In particular, these medicines have been widely used in ocular therapies in the treatment of various diseases, such as age-related macular degeneration, corneal neovascularization, diabetic retinopathy, and retinal vein occlusion. However, the formulation of these biomacromolecules is challenging due to their high molecular weight, complex structure, instability, short half-life, enzymatic degradation, and immunogenicity, which leads to the failure of therapies. Various efforts have been made to overcome the ocular barriers, providing effective delivery of therapeutic proteins, such as altering the protein structure or including it in new delivery systems. These strategies are not only cost-effective and beneficial to patients but have also been shown to allow for fewer drug side effects. In this review, we discuss several factors that affect the design of formulations and the delivery of therapeutic proteins to ocular tissues, such as the use of injectable micro/nanocarriers, hydrogels, implants, iontophoresis, cell-based therapy, and combination techniques. In addition, other approaches are briefly discussed, related to the structural modification of these proteins, improving their bioavailability in the posterior segments of the eye without affecting their stability. Future research should be conducted toward the development of more effective, stable, noninvasive, and cost-effective formulations for the ocular delivery of therapeutic proteins. In addition, more insights into preclinical to clinical translation are needed.

Current Insights on Lipid-Based Nanosystems 2023.

Among the different types of nanosystems that have been investigated for therapeutic use, lipid-based ones are the most explored, as they have advantages over non-lipid nanosystems, especially for improving the transport and efficacy of drugs through different routes of administration, such as ocular, cutaneous, intranasal, and intravenous [...].

Intranasal Lipid Nanoparticles Containing Bioactive Compounds Obtained from Marine Sources to Manage Neurodegenerative Diseases.

Marine sources contain several bioactive compounds with high therapeutic potential, such as remarkable antioxidant activity that can reduce oxidative stress related to the pathogenesis of neurodegenerative diseases. Indeed, there has been a growing interest in these natural sources, especially those resulting from the processing of marine organisms (i.e., marine bio-waste), to obtain natural antioxidants as an alternative to synthetic antioxidants in a sustainable approach to promote circularity by recovering and creating value from these bio-wastes. However, despite their expected potential to prevent, delay, or treat neurodegenerative diseases, antioxidant compounds may have difficulty reaching the brain due to the need to cross the blood-brain barrier (BBB). In this regard, alternative delivery systems administered by different routes have been proposed, including intranasal administration of lipid nanoparticles, such as solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), which have shown promising results. Intranasal administration shows several advantages, including the fact that molecules do not need to cross the BBB to reach the central nervous system (CNS), as they can be transported directly from the nasal cavity to the brain (i.e., nose-to-brain transport). The benefits of using SLN and NLC for intranasal delivery of natural bioactive compounds for the treatment of neurodegenerative diseases have shown relevant outcomes through in vitro and in vivo studies. Noteworthy, for bioactive compounds obtained from marine bio-waste, few studies have been reported, showing the open potential of this research area. This review updates the state of the art of using SLN and NLC to transport bioactive compounds from different sources, in particular, those obtained from marine bio-waste, and their potential application in the treatment of neurodegenerative diseases.

An In Vitro Evaluation of the Potential Neuroprotective Effects of Intranasal Lipid Nanoparticles Containing Astaxanthin Obtained from Different Sources: Comparative Studies.

The intranasal route has been suggested as a promising alternative to improve the direct transport of molecules to the brain, avoiding the need to cross the blood-brain barrier (BBB). In this area, the use of lipid nanoparticles, namely solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), has been highlighted as a promising strategy to improve the treatment of neurodegenerative diseases. In this work, formulations containing SLN and NLC that were loaded with astaxanthin that was obtained from different sources (astaxanthin extract (AE) from the algae Haematococcus pluvialis and pure astaxanthin (PA) from the fungi Blakeslea trispora) were prepared for nose-to-brain administration, and comparative in vitro experiments were performed to evaluate the biocompatibility of the formulations with nasal (RPMI 2650) and neuronal (SH-SY5Y) cells. Afterwards, the antioxidant activity of the formulations was evaluated for its potential neuroprotective effects, using different chemical aggressors. Finally, the cellular uptake of the astaxanthin was evaluated for the formulations that showed the greatest neuroprotection of the neuronal cells against chemical-induced damage. On the production day, all the formulations showed a particle size, a high encapsulation efficiency (EE), the presence of nanoparticles with a typical spherical shape, and a polydispersity index (PDI) and zeta potential (ZP) that are suitable for nose-to-brain administration. After three months of storage at room temperature, no significant changes were observed in the characterization parameters, predicting a good long-term stability. Furthermore, these formulations were shown to be safe with concentrations of up to 100 µg/mL in differentiated SH-SY5Y and RPMI 2650 cells. Regarding neuroprotection studies, the PA-loaded SLN and NLC formulations showed an ability to counteract some mechanisms of neurodegeneration, including oxidative stress. Moreover, when compared with the PA-loaded SLN, the PA-loaded NLC showed greater neuroprotective effects against the cytotoxicity induced by aggressors. In contrast, the AE-loaded SLN and NLC formulations showed no significant neuroprotective effects. Although further studies are needed to confirm these neuroprotective effects, the results of this study suggest that the intranasal administration of PA-loaded NLC may be a promising alternative to improve the treatment of neurodegenerative diseases.

1,3-Thiazole derivatives as privileged structures for anti-Trypanosoma cruzi activity: Rational design, synthesis, in silico and in vitro studies.

Chagas disease is a deadly and centenary neglected disease that is recently surging as a potential global threat. Approximately 30% of infected individuals develop chronic Chagas cardiomyopathy and current treatment with the reference benznidazole (BZN) is ineffective for this stage. We presently report the structural planning, synthesis, characterization, molecular docking prediction, cytotoxicity, in vitro bioactivity and mechanistic studies on the anti-T. cruzi activity of a series of 16 novel 1,3-thiazoles (2-17) derived from thiosemicarbazones (1a, 1b) in a two-step and reproducible Hantzsch-based synthesis approach. The anti-T. cruzi activity was evaluated in vitro against the epimastigote, amastigote and trypomastigote forms of the parasite. In the bioactivity assays, all thiazoles were more potent than BZN against epimastigotes. We found that the compounds presented an overall increased anti-tripomastigote selectivity (Cpd 8 was 24-fold more selective) than BZN, and they mostly presented anti-amastigote activity at very low doses (from 3.65 µM, cpd 15). Mechanistic studies on cell death suggested that the series of 1,3-thiazole compounds herein reported cause parasite cell death through apoptosis, but without compromising the mitochondrial membrane potential. In silico prediction of physicochemical properties and pharmacokinetic parameters showed promising drug-like results, being all the reported compounds in compliance with Lipinski and Veber rules. In summary, our work contributes towards a more rational design of potent and selective antitripanosomal drugs, using affordable methodology to yield industrially viable drug candidates.

Editorial-Current Insights on Lipid-Based Nanosystems.

Lipid-based nanosystems, including solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), cationic lipid nanoparticles, nanoemulsions and liposomes, have been extensively studied to improve drug delivery through different administration routes [...].

Impact of COVID-19 Pandemic on Air Quality: A Systematic Review.

With the emergence of the COVID-19 pandemic, several governments imposed severe restrictions on socio-economic activities, putting most of the world population into a general lockdown in March 2020. Although scattered, studies on this topic worldwide have rapidly emerged in the literature. Hence, this systematic review aimed to identify and discuss the scientifically validated literature that evaluated the impact of the COVID-19 pandemic and associated restrictions on air quality. Thus, a total of 114 studies that quantified the impact of the COVID-19 pandemic on air quality through monitoring were selected from three databases. The most evaluated countries were India and China; all the studies intended to evaluate the impact of the pandemic on air quality, mainly concerning PM10, PM2.5, NO2, O3, CO, and SO2. Most of them focused on the 1st lockdown, comparing with the pre- and post-lockdown periods and usually in urban areas. Many studies conducted a descriptive analysis, while others complemented it with more advanced statistical analysis. Although using different methodologies, some studies reported a temporary air quality improvement during the lockdown. More studies are still needed, comparing different lockdown and lifting periods and, in other areas, for a definition of better-targeted policies to reduce air pollution.

Vine Cane Compounds to Prevent Skin Cells Aging through Solid Lipid Nanoparticles.

The long lifespan of the world's population has been raising interest in the research for new solutions to delay the aging process. With the aim of skin aging prevention, solid lipid nanoparticles (SLNs) were developed in this work for the encapsulation of three lipophilic natural compounds extracted from vine cane-epigallocatechin gallate (EGCG), resveratrol and myricetin. The developed loaded-SLNs proved to be stable, maintaining their adequate physicochemical characteristics for 30 days. In addition, the loaded-SLNs formulations exhibited high encapsulation efficiencies and loading capacities and high intracellular antioxidant activity. The mixture of EGCG-loaded SLNs with resveratrol-loaded SLNs proved to have the highest protection against induced oxidative stress. The in vitro cytotoxicity of the loaded SLNs was also evaluated, showing that the developed formulations are biocompatible for concentrations up to 50 µg/mL and could be safe for use in cosmetics. The encapsulation of EGCG, resveratrol and myricetin in SLNs seems to be a suitable strategy for the delivery of these antioxidants to the skin, improving their bioavailability.

Residue localization and risk for aspiration in partial laryngectomy: the relevance of assertive therapeutic strategies and resources.

OBJECTIVE: To describe the correlation between the residues, their anatomical location and the presence of laryngotracheal penetration and aspiration in patients after supracricoid laryngectomy undergoing cricohyoidoepiglotopexy reconstruction. METHODS: This study included 70 patients treated by supracricoid laryngectomy with cricohyoidoepiglotopexy reconstruction in a referral national cancer hospital. The patients were submitted to swallowing videofluoroscopy, and the findings were classified by the penetration and aspiration scale. The images were described observing the presence or absence of residues and their anatomical location. RESULTS: The prevalence of penetration in this study was 68.6% and aspiration was 34.3%. An association was found between the presence of residue on the tongue (p=0.005), posterior pharyngeal wall (p=0.013), pyriform recesses (p=0.002), valecula (p=0.061), and laryngeal penetration. The residue in the upper esophageal sphincter (p=0.039) was associated with the occurrence of laryngotracheal aspiration. CONCLUSION: Patients undergoing supracricoid laryngectomy with cricohioidoepiglotopexy reconstruction have food residues in different anatomical regions after swallowing. Penetration was associated with the presence of residues on the base of the tongue and posterior pharyngeal wall. Aspiration was associated with the presence of residues in the upper esophageal sphincter.

Can mental practice adjunct in the recovery of motor function in the upper limbs after stroke? A systematic review and meta-analysis.

BACKGROUND: Studies indicate that mental practice can be an adjuvant rehabilitation, improving motor functions. AIM: To synthesize the evidence on the intervention with the mental practice for the rehabilitation of the upper limb after stroke in the context of a dependent task. METHODS: The review was registered on the PROSPERO with protocol number: CRD42020166624. We searched the PubMed, Medline, Embase, Central, PEDro, and Web of Science from randomized clinical trials from 1975 to 2022. A literature review was conducted with 13 studies that synthesized findings on mental practice such as adjuvant rehabilitation in the recovery of the upper limb after stroke based on Fugl-Meyer Assessment (FMA) Motor and action research arm test (ARAT) scores. RESULTS: The sample size was 232 were part of the intervention group and 180 of the control group. The findings no showed results in favor of mental practice after stroke accordingly to ARAT and FMA Motor scores (P > 0.05). CONCLUSION: Current evidence does not support the use of the mental practice to increase the recovery of the upper limb after stroke, although the evidence is conflicting for some aspects of the technique.

DIGEST Scale Predictis More Quality of Life Than PAS: The Residue Influence on Supracricoid Laryngectomy.

Introduction Supracricoid laryngectomy (SCL CHEP) removes ∼ 70% of the larynx, resulting in structural rearrangement and modification of the swallowing mechanism, promoting chronic dysphagia. One of the consequences of this new physiology is the formation of pharyngeal residues that can increase the possibility of aspiration. The formation of residues after SCL CHEP, its functional consequences, and its influence on quality of life (QOL) is still poorly described in the literature. Objective To investigate and compare the association between self-reported QoL and objective assessments of swallowing function in patients undergoing SCL CHEP. Methods A cross-sectional study was performed from 2018 to 2020 in a reference service for head and neck surgery in Brazil. A total of 860 swallowing videofluoroscopy images were evaluated using the Penetration and Aspiration Scale (PAS) and Dynamic Imaging Grade of Swallowing Toxicity (DIGEST). Results In a group of 86 patients, there was a significant relationship between oncological staging and the global ( p < 0.001) and total ( p = 0.002) QoL domains. There was a negative correlation between the DIGEST scale and the emotional domain of the QoL protocol ( p = 0.045). The swallowing function proved to be relevant for QoL. Conclusion The PAS scale did not show any correlation with QoL. The functional performance of swallowing according to the DIGEST scale was coherent with the QOL scores. It is suggested that the residue may be a more relevant aspect for QoL than the aspiration, making DIGEST a promising tool in the assessment of dysphagic patients.

Preclinical Evaluation of Lipid-Based Nanosystems.

The use of lipid-based nanosystems, including lipid nanoparticles (solid lipid nanoparticles-SLN, and nanostructured lipid carriers-NLC), nanoemulsions, and liposomes, among others, is widespread [...].

Improving Drug Delivery for Alzheimer's Disease Through Nose-to-Brain Delivery Using Nanoemulsions, Nanostructured Lipid Carriers (NLC) and in situ Hydrogels.

Current treatments for Alzheimer's disease (AD) attenuate the progression of symptoms and aim to improve the patient's quality of life. Licensed medicines are mostly for oral administration and are limited by the difficulty in crossing the blood-brain barrier (BBB). Here in, the nasal route has been explored as an alternative pathway that allows drugs to be directly delivered to the brain via the nasal cavity. However, clearance mechanisms in the nasal cavity impair the delivery of drugs to the brain and limit their bioavailability. To optimize nose-to-brain delivery, formulations of lipid-based nanosystems, namely nanoemulsions and nanostructured lipid carriers (NLC), formulated in situ gelling hydrogels have been proposed as approaches for nose-to-brain delivery. These formulations possess characteristics that facilitate drug transport directly to the brain, minimizing side effects and maximizing therapeutic benefits. It has been recommended that the manufacture of these drug delivery systems follows the quality by design (QbD) approach based on nasal administration requirements. This review provides an insight into the current knowledge of the AD, highlighting the need for an effective drug delivery to the brain. Considering the mounting interest in the use of nanoemulsions and NLC for nose-to-brain delivery, a description of drug transport pathways in the nasal cavity and the application of these nanosystems and their in situ hydrogels through the intranasal route are presented. Relevant preclinical studies are summarised, and the future prospects for the use of lipid-based nanosystems in the treatment of AD are emphasized.