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BACKGROUND: Anxiety and depression are prevalent comorbidities in patients with COPD. Breathing techniques can improve anxiety and depression in patients hospitalized for COPD exacerbation. METHODS: We conducted a randomized clinical study with 46 male subjects, 67- 86 years old, hospitalized with acute COPD exacerbation. Subjects were randomly and equally divided into a control group and a controlled breathing intervention group. We measured baseline and post-intervention dyspnea, anxiety and depression, quality of life (with the St George's Respiratory Questionnaire and the European Quality of Life questionnaire), maximum inspiratory and expiratory pressure, hand-grip strength, and sleep quality. The cohort had high dyspnea and low overall quality of life. RESULTS: Controlled breathing techniques significantly improved dyspnea, anxiety, and mobility. All the measured variables improved in the intervention group. The control group had poorer values in all the variables after the hospitalization period. CONCLUSIONS: Controlled breathing exercises improve anxiety and depression in patients hospitalized for COPD exacerbation. [ABSTRACT FROM AUTHOR]

Life cycle assessment (LCA) is a promising tool in the pursuit of sustainable mining. However, the accounting methodologies used in LCA for abiotic resource depletion still have some shortcomings and need to be improved. In this article a new thermodynamic approach is presented for the evaluation of the depletion of nonfuel minerals. The method is based on quantifying the exergy costs required to replace the extracted minerals with current available technologies, from a completely degraded state in what we term 'Thanatia' to the conditions currently found in nature. Thanatia is an estimated reference model of a commercial end of the planet, where all resources have been extracted and dispersed, and all fossil fuels have been burned. Mineral deposits constitute an exergy bonus that nature gives us for free by providing minerals in a concentrated state and not dispersed in the crust. The exergy replacement costs provide a measure of the bonus lost through extraction. This approach allows performing an LCA by including a new stage in the analysis: namely the grave to cradle path. The methodology is explained through the case study of nickel depletion. [ABSTRACT FROM AUTHOR]

Abstract: This paper makes an inventory of the natural capital on earth in terms of exergy, which includes not only renewable and non-renewable energy resources, but also non-fuel minerals. The exergy method is very suitable for the accounting of our natural capital because all kinds of resources can be assessed with a single property. For the case of minerals, exergy allows to unify properties tonnage and grade. Furthermore, the exergy replacement costs of minerals includes additional information of the state of technology. The aggregation capacity of the exergy and exergy replacement cost indicators increases the analysis potential of the results. This way, the non-fuel mineral''s wealth can be compared to that of fuel minerals or even to other natural resources. The results of this study reveal that the real scarcity problems that humankind are facing are not based on the lack of energy sources, but on the lack of minerals. [Copyright &y& Elsevier]

A conventional passenger car demands almost 50 different types of metals, along other raw materials. Some of these metals, such as tantalum, indium, niobium or rare earths elements, are considered critical by the European Commission and many other institutions. Additionally, their functional recycling is practically absent. The transition to fully electric vehicles will require more electrical and electronic devices, motors and batteries, that will need an increasing amount of critical metals. A methodology has been developed to identify strategic elements for the automobile sector. This approach defines a variable called Strategic Metal Index (SMI) which is calculated for each metal. This index is the result of combining the following parameters: (1) Automobile sector demand with respect to world production; (2) known resources compared to total cumulative demand and (3) Supply risk. The index has been applied to 50 metals used by different types of vehicle powertrains. The assessment covers metal demand from 2018 to 2050 according to vehicle sales projections. Using this approach, the most strategic elements for the automobile manufacturing sector are Ni, Li and Co (used in batteries), Nd and Dy (permanent magnets), Tb (lighting and fuel injectors), Sb, Bi and and B (steel alloys, paintings), Au and Ag (electronics), In (screens) and Te (steel alloys, electronics). The search for substitutes, implementation of eco-design measures and the increase of the functional recyclability of these elements should be strongly encouraged in the sector. [ABSTRACT FROM AUTHOR]

• Around 50 metals are contained in both the ICEV and the BEV. • The BEV contains 50% more metal than the ICEV in terms of mass. • The BEV represents between 2.5 and 4 times the metal thermodynamic rarity of the ICEV. • The metals' thermodynamic rarity of the BEV decreases with the evolution of the battery. • Identification of the most exergy intensive parts within the BEV. The transition to full electromobility must be carefully evaluated, as large amounts of strategic metals will be required, for which there is presently little to no recovery or recycling (e.g. gold, silver, tantalum or cobalt). In this study, we perform a comprehensive metal assessment of two passenger cars (conventional and battery electric models) in terms of mass and thermodynamic rarity. Thermodynamic rarity is based on the property of exergy and is defined as "the amount of exergy resources needed to obtain a mineral commodity from average crustal concentration using the best available technology" (measured in kJ). Thus, the thermodynamic rarity approach assigns a greater exergetic value to scarce (understood as having a relative low average crustal concentration) and difficult-to-extract minerals. Of the 60 metals analyzed, almost 50 metals have been identified within the studied cars, representing 800 (conventional) and 1,200 kg (battery electric), showcasing the fact that a car constitutes a "road mine". Furthermore, given that the technology behind battery electric cars is in development, three generations of Li-ion batteries were analyzed to study the effect on resource use of a metal changing composition over time. Albeit the battery modules of the three generations present a similar mass content (approximately 70 kgs), the thermodynamic rarity decreases from 275 to 100 Gigajoules, due to the reduced proportion of cobalt, which is by far the most exergetic metal within the battery. Additionally, with the thermodynamic rarity approach, the most exergy intensive parts within a battery electric car have been identified – the high-voltage battery modules, the electric drive, the power module, the charger, the electrical air conditioning compressor and the electromechanical brake servo – providing an indicator facilitating proactive mid- to long-term ecodesign measures and recycling strategies. [ABSTRACT FROM AUTHOR]

• A lyophilisation-based method to obtain rat brain tissue powder was developed. • Different biomolecule specialist single-extractions can be carried out in powder aliquots. • Lyophilisation results in no RNA/protein losses relative to only flash-freezing. • The method allows coordinated RNA and protein expression analyses in ischemic stroke. • It is sample saving and contributes to the reduction principle in animal research. Optimisation of tissue processing procedures in preclinical studies reduces the number of animals used and allows integrated multilevel study in the same sample. Multiple extraction of different biomolecules from the same sample has several limitations. Using brain samples from rats subjected to ischemic stroke, we combined lyophilisation of flash-frozen tissue, mechanical pulverisation and cryopreservation in a method to optimise tissue handling and preservation for independent RNA or protein single-extract methods, and subsequent RT-qPCR or Western blot analyses. Lyophilisation resulted in 70% tissue weight loss. RNA (OD 260/280 ∼1.8) and protein yields were similar in non-ischemic and ischemic brain samples, subjected to either flash freezing (FF) or flash freezing followed by lyophilisation (FF + Lyo). RNA transcription of reference genes (Actb and Rn18s), expression of housekeeping proteins (β-actin and α-tubulin), and mRNA overexpression of stroke-regulated genes (Nos2 , Mmp9 and Tnfa) was similar in FF and FF + Lyo samples. Contrary to high heat stress of baking method in a drying oven, lyophilisation maintains the integrity of dried samples for subsequent extractions and analyses. Sample lyophilisation allows different manual representative extractions/analyses from the same rat, it is much cheaper than using commercial kits, and shows higher yields that multiple manual or kit-based extractions. The lyophilisation-based method for different biomolecule single-extractions from tissue powder aliquots, representing the same rat brain sample, is sample saving, contributes to the reduction principle in animal research, and allows coordinated analysis for accurate correlations between the transcriptome and proteome in stroke and other neuroscience research. [ABSTRACT FROM AUTHOR]

Because neuroprotection in stroke should be revisited in the era of recanalisation, the present study analysed the potential neuroprotective effect of the selective oestrogen receptor modulator, bazedoxifene acetate (BZA), in an animal model of diabetic ischaemic stroke that mimics thrombectomy combined with adjuvant administration of a putative neuroprotectant. Four weeks after induction of diabetes (40 mg kg‐1 streptozotocin, i.p.), male Wistar rats were subjected to transient middle cerebral artery occlusion (intraluminal thread technique, 60 minutes) and assigned to one of three groups treated with either: vehicle, BZA (3 mg kg‐1 day‐1, i.p.) or 17β‐oestradiol (E2) (100 μg kg‐1 day‐1, i.p.). At 24 hours post‐ischaemia‐reperfusion, brain damage (neurofunctional score, infarct size and apoptosis), expression of oestrogen receptors (ER)α, ERβ and G protein‐coupled oestrogen receptor), and activity of the mitogen‐activated protein kinase/extracellular signal‐regulated kinase (MAPK/ERK)1/2 and phosphoinositide 3‐kinase/Akt pathways were analysed. At 24 hours after the ischaemic insult, both BZA‐ and E2‐treated animals showed lower brain damage in terms of improved neurofunctional condition, decreased infarct size and decreased apoptotic cell death. Ischaemia‐reperfusion induced a significant decrease in ERα and ERβ expression without affecting that of G protein‐coupled oestrogen receptor, whereas BZA and E2 reversed such a decrease. The ischaemic insult up‐regulated the activity of both the MAPK/ERK1/2 and phosphoinositide 3‐kinase/Akt pathways; BZA and E2 attenuated the increased activity of the ERK1/2 pathway, without affecting that of the Akt pathway. The results of the present study lend further support to the consideration of BZA as an effective and safer alternative overcoming the drawbacks of E2 with respect to improving diabetic ischaemic stroke outcome after successful reperfusion. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H 2 S) is a potential endothelium-derived hyperpolarizing factor (EDHF) and adventitium- or adipocyte-derived relaxing factor (ADRF) which vasorelaxant action is mediated by potassium channels. H 2 S could also play an important role in the pathophysiology of diabetic cardiovascular complications. The present study has investigated the influence of alloxan-induced diabetes on the role of potassium channels mediating the relaxant response of the rabbit carotid artery to NaHS, a donor of H 2 S. NaHS (10−8-3 × 10−5 M) relaxed phenylephrine-precontracted carotid arteries, with higher potency in diabetic than in control rabbits. The selective blockers of potassium channels charybdotoxin, 4-amynopiridine and glibenclamide significantly inhibited the relaxant action of NaHS in diabetic rabbits, but not in control rabbits. When compared to control rabbits, carotid arteries from diabetic rabbits showed significantly reduced expression of big conductance Ca+2-activated potassium channels (BK Ca), significantly enhanced expression of intermediate conductance Ca+2-activated potassium channels (IK Ca) and not significant different expression of voltage-sensitive potassium channels (K V) and ATP-sensitive potassium channels (K ATP). These results suggest that an enhanced role of IK Ca , K V and K ATP potassium channels could be involved in the increased sensitivity of the rabbit carotid artery to H 2 S in diabetes. [ABSTRACT FROM AUTHOR]

Despite the overwhelming advances in the understanding of the pathogenesis of stroke, a devastating disease affecting millions of people worldwide, currently there are only a limited number of effective treatments available. Preclinical and clinical studies show that stroke is a sexually dimorphic disorder, affecting males and females differently. Strong experimental evidence indicates that estrogen may play a role in this difference and that exogenous 17β-estradiol (E2) is neuroprotective against stroke in both male and female rodents. However, the molecular mechanisms by which E2 intervenes in ischemia-induced cell death, revealing these sex differences, remain unclear. The present study was aimed to determine, in female rats, the molecular mechanisms of two well-known pro-survival signaling pathways, MAPK/ERK1/2 and PI3K/Akt, that mediate E2 neuroprotection in response to acute ischemic stroke. E2 pretreatment reduced brain damage and attenuated apoptotic cell death in ovariectomized female rats after an ischemic insult. Moreover, E2 decreased phosphorylation of ERK1/2 and prevented ischemia/reperfusion-induced dephosphorylation of both Akt and the pro-apoptotic protein, BAD. However, MAPK/ERK1/2 inhibitor PD98059, but not the PI3K inhibitor LY294002, attenuated E2 neuroprotection. Thus, these results suggested that E2 pretreatment in ovariectomized female rats modulates MAPK/ERK1/2 and activates Akt independently of PI3K to promote cerebroprotection in ischemic stroke. A better understanding of the mechanisms and the influence of E2 in the female sex paves the way for the design of future successful hormone replacement therapies. [ABSTRACT FROM AUTHOR]

In recent years, heightened environmental concerns linked to agriculture have surged, with soil degradation standing out as a global issue. However, prevailing sustainability assessment methodologies in agriculture often overlook soil systems due to their intricate nature. This study aims to develop a methodology for evaluating soil degradation in agricultural practices using exergy regeneration costs. These costs determine the exergy required to restore soil fertility to pre-harvest levels. The methodology covers key soil factors like nutrients, organic matter, and prevalent issues like salinity, acidification, and erosion. For each of these factors, exergy regeneration costs are determined based on the energy needed to execute an optimal process for reverting the soil to its original or ideal state. The methodology has been applied to data from agricultural trials, showing that the calculated soil replacement cost is significantly higher compared to one of the most energy-demanding processes in agriculture, the use of urea. This demonstrates that agricultural soil degradation needs to be quantified for a correct evaluation of agricultural practices and their sustainability. [Display omitted] • Assessment of soil fertility using exergy as a unifying tool. • Quantification of the degradation of soil in agricultural practices using the exergy regeneration cost. • Analysis of the impact of agricultural practices on the soil ecosystem. [ABSTRACT FROM AUTHOR]

Along with its role in regulating blood pressure and fluid homeostasis, the natriuretic peptide system could be also part of an endogenous protective mechanism against brain damage. We aimed to assess the possibility that exogenous atrial natriuretic peptide (ANP) could protect against acute ischemic stroke, as well as the molecular mechanisms involved. Three groups of rats subjected to transient middle cerebral artery occlusion (tMCAO, intraluminal filament technique, 60 min) received intracerebroventricular vehicle, low-dose ANP (0.5 nmol) or high-dose ANP (2.5 nmol), at 30 min reperfusion. Neurofunctional condition, and brain infarct and edema volumes were measured at 24 h after tMCAO. Apoptotic cell death and expression of natriuretic peptide receptors (NPR-A and NPR-C), K + channels (K ATP , K V and BK Ca ), and PI3K/Akt and MAPK/ERK1/2 signaling pathways were analyzed. Significant improvement in neurofunctional status, associated to reduction in infarct and edema volumes, was shown in the high-dose ANP group. As to the molecular mechanisms analyzed, high-dose ANP: 1) reduced caspase-3-mediated apoptosis; 2) did not modify the expression of NPR-A and NPR-C, which had been downregulated by the ischemic insult; 3) induced a significant reversion of ischemia-downregulated K ATP channel expression; and 4) induced a significant reversion of ischemia-upregulated pERK2/ERK2 expression ratio. In conclusion, ANP exerts a significant protective role in terms of both improvement of neurofunctional status and reduction in infarct volume. Modulation of ANP on some molecular mechanisms involved in ischemia-induced apoptotic cell death (K ATP channels and MAPK/ERK1/2 signaling pathway) could account, at least in part, for its beneficial effect. Therefore, ANP should be considered as a potential adjunctive neuroprotective agent improving stroke outcome after successful reperfusion interventions. [ABSTRACT FROM AUTHOR]

Atrial natriuretic peptide (ANP) is a vasodilator with significant regional differences and controversial effects in the cerebral circulation, a vascular bed particularly prone to diabetes-induced complications. The present study has investigated how alloxan-induced diabetes modifies the mechanisms involved in the response of the rabbit basilar artery to ANP. ANP (10 –12 –10 −7 M) relaxed precontracted basilar arteries, with higher potency in diabetic than in control rabbits. In arteries from both groups of animals, endothelium removal reduced ANP-induced relaxations. Inhibition of NO-synthesis attenuated ANP-induced relaxation but this attenuation was lower in diabetic than in control rabbits. In control rabbits, indomethacin displaced to the left the concentration-response curve to ANP, without significantly modifying the E max value. In diabetic rabbits, indomethacin significantly enhanced arterial relaxations to ANP. In KCl-depolarised arteries, relaxation to ANP was almost abolished both in control and in diabetic rabbits. Iberiotoxin inhibited relaxations to ANP in both groups of rabbits. Glibenclamide and 4-aminopyridine inhibited the ANP-induced relaxations more in diabetic than in control rabbits. Basilar arteries from diabetic rabbits showed decreased natriuretic peptide receptor C expression and no changes in natriuretic peptide receptor A, large conductance calcium-activated K + channels (BK Ca ), ATP-sensitive K + channels (K ATP ) and voltage-sensitive K + channels (K V ) expression. These results suggest that diabetes enhances the sensitivity of the rabbit basilar artery to ANP by mechanisms that at least include reduced expression of natriuretic peptide receptor C, and enhanced activity of K ATP and K V channels. Furthermore, diabetes reduces endothelial NO and prostacyclin which mediate arterial relaxation to ANP. [ABSTRACT FROM AUTHOR]

As the knowledge on the estrogenic system in the brain grows, the possibilities to modulate it in order to afford further neuroprotection in brain damaging disorders so do it. We have previously demonstrated the ability of the selective estrogen receptor modulator, bazedoxifene (BZA), to reduce experimental ischemic brain damage. The present study has been designed to gain insight into the molecular mechanisms involved in such a neuroprotective action by investigating: 1) stroke-induced apoptotic cell death; 2) expression of estrogen receptors (ER) ERα, ERβ and the G-protein coupled estrogen receptor (GPER); and 3) modulation of MAPK/ERK1/2 and PI3K/Akt signaling pathways. For comparison, a parallel study was done with 17β-estradiol (E2)-treated animals. Male Wistar rats subject to transient right middle cerebral artery occlusion (tMCAO, intraluminal thread technique, 60 min), were distributed in vehicle-, BZA- (20.7 ± 2.1 ng/mL in plasma) and E2- (45.6 ± 7.8 pg/mL in plasma) treated groups. At 24 h from the onset of tMCAO, RT-PCR, Western blot and histochemical analysis were performed on brain tissue samples. Ischemia-reperfusion per se increased apoptosis as assessed by both caspase-3 activity and TUNEL-positive cell counts, which were reversed by both BZA and E2. ERα and ERβ expression, but not that of GPER, was reduced by the ischemic insult. BZA and E2 had different effects: while BZA increased both ERα and ERβ expression, E2 increased ERα expression but did not change that of ERβ. Both MAPK/ERK1/2 and PI3K/Akt pathways were stimulated under ischemic conditions. While BZA strongly reduced the increased p-ERK1/2 levels, E2 did not. Neither BZA nor E2 modified ischemia-induced increase in p-Akt levels. These results show that modulation of ERα and ERβ expression, as well as of the ERK1/2 signaling pathway accounts, at least in part, for the inhibitory effect of BZA on the stroke-induced apoptotic cell death. This lends mechanistic support to the consideration of BZA as a potential neuroprotective drug in acute ischemic stroke treatment. [ABSTRACT FROM AUTHOR]

• Soil fertility evaluation using exergy as a unifying tool. • Establishment of a reference copiously fertile soil to perform exergy evaluation of soils. • Inorganic and organic soil components determining is degradation. Soil degradation, affecting around 38% of the world's cropland, threatens the global food supply. Due to the soil's complexity, the measure of soil degradation that involves the loss of soil fertility due to crop system management processes represents an unsolved problem. Exergy is a property with the potential to be used in soil fertility and/or degradation analysis. A methodology to determine the exergy value fenced in a fertile soil due to its inorganic and organic components is established in this study and will be applied to evaluate soil fertility, degradation, and quality. As a first step, the exergy of perfect topsoil with optimum characteristics called "OptSOIL" is determined. The "OptSOIL" is established by agronomic expertise and will allow establishing a general theoretical reference suitable to execute exergy assessments of soils and compare the degradation grade of any soil concerning the best possible. Consequently, we introduce a perfect fertile planetary crust made of "OptNUT" and "OptSOM" invariant and independent of the different local textures, but not independent of their water content and aeration. We call this imaginary crust -copiously fertile- Pristinia as opposed to Thanatia, a dead state referring to abiotic resources. Thus, any real agricultural soil will be an intermediate soil between Pristinia and Thanatia. This idea might serve to quantitatively diagnose an assessment of all the concepts by which soil is degraded. The methodology has been validated through laboratory agronomic tests for different soils, concluding that exergy is a rigorous indicator to measure topsoil fertility. [ABSTRACT FROM AUTHOR]

Fungal pathogens pose a threat to human health and food security. Few antifungals are available and resistance to all has been reported. Novel strategies to control plant and human pathogens as well as food spoilers are urgently required. Environmental yeasts provide a functionally diverse, yet underexploited potential for fungal control based on their natural competition via the secretion of proteins and other small molecules such as iron chelators, volatile organic compounds or biosurfactants. However, there is a lack of standardized workflows to systematically access application-relevant yeast-based compounds and understand their molecular functioning. Towards this goal, we developed a workflow to identify and characterize yeast isolates that are active against spoilage yeasts and relevant human and plant pathogens, herein focusing on discovering yeasts that secrete antifungal proteins. The workflow includes the classification of the secreted molecules and cross-comparison of their antifungal capacity using an independent synthetic calibrant. Our workflow delivered a collection of 681 yeasts of which 212 isolates (31 %) displayed antagonism against at least one target strain. While 57.5 % of the active yeasts showed iron-depended antagonism, likely due to pulcherrimin-like iron chelators, 31.7 % secreted antifungal proteins. Those yeast candidates clustered within twelve OTUs, showed narrow and broad target spectra, and several showed a broad pH and temperature activity profile. Given the tools for yeast biotechnology and protein engineering available, our collection can serve as a rich starting point for genetic and molecular characterization of the various antifungal phenotypes, their mode of action and their future exploitation., Competing Interests: Declaration of Competing Interest The authors declare no competing interest., (Copyright © 2025 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Etiological classification of ischemic stroke is fundamental for secondary prevention, but frequently results in undetermined cause. We aimed to develop a Deep Learning (DL)-based model for automatic etiological classification of ischemic stroke using digital images of thrombi retrieved by mechanical thrombectomy., Methods: Patients with large vessel occlusion stroke subjected to mechanical thrombectomy between April 2016 and January 2023 at La Fe University and Polytechnic Hospital in Valencia were included. Thrombus digital images were obtained and clinical characteristics, including TOAST etiological classification as reference standard, were retrieved. Statistical analysis was performed to compare clinical characteristics between atherothrombotic and cardioembolic strokes. A DL method was designed based on two deep neural networks for: (1) image segmentation and (2) image classification including clinical characteristics. The metrics used were DICE coefficient for the segmentation network, and accuracy, precision, sensitivity, specificity and area under the curve (AUC) for the predictions of the classification network., Results: A total of 166 patients (mean age 69 [SD, 13], 67 female) were included. TOAST classification was: 31 atherothrombotic, 87 cardioembolic, and 48 cryptogenic. The segmentation network achieved an average DICE coefficient of 0.96 [SD, 0.13]. The optimal fused imaging and clinical classification network had a 0.968 accuracy [95% CI, 0.935-0.994], and AUC of 0.947 [95% CI, 0.870-1]. Cryptogenic thrombi were classified as cardioembolic (96%) or atherothrombotic (4%)., Conclusion: Two convolutional neural networks perform the automatic segmentation of thrombus images and, combined with selected clinical characteristics, their accurate and precise classification into atherothrombotic or cardioembolic etiology in patients with acute ischemic stroke., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Lucero-Garófano, Aliena-Valero, Vielba-Gómez, Escudero-Martínez, Morales-Caba, Aparici-Robles, Tarruella Hernández, Fortea, Tembl, Salom and Manjón.)

Oligo pools are array-synthesized, user-defined mixtures of single-stranded oligonucleotides that can be used as a source of synthetic DNA for library cloning. While currently offering the most affordable source of synthetic DNA, oligo pools also come with limitations such as a maximum synthesis length (approximately 350 bases), a higher error rate compared to alternative synthesis methods, and the presence of truncated molecules in the pool due to incomplete synthesis. Here, we provide users with a comprehensive protocol that details how oligo pools can be used in combination with Golden Gate cloning to create user-defined protein mutant libraries, as well as single-guide RNA libraries for CRISPR applications. Our methods are optimized to work within the Yeast Toolkit Golden Gate scheme, but are in principle compatible with any other Golden Gate-based modular cloning toolkit and extendable to other restriction enzyme-based cloning methods beyond Golden Gate. Our methods yield high-quality, affordable, in-house variant libraries., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

ApTOLL, a TLR4 modulator aptamer, has demonstrated cerebroprotective effects in a permanent ischemic stroke mouse model, as well as safety and efficacy in early phase clinical trials. We carried out reverse translation research according to STAIR recommendations to further characterize the effects and mechanisms of ApTOLL after transient ischemic stroke in rats and to better inform the design of pivotal clinical trials. Adult male rats subjected to transient middle cerebral artery occlusion were treated either with ApTOLL or the vehicle intravenously at different doses and time-points. ApTOLL was compared with TAK-242 (a TLR4 inhibitor). Female rats were also studied. After neurofunctional evaluation, brains were removed for infarct/edema volume, hemorrhagic transformation, and histologic determinations. Peripheral leukocyte populations were assessed via flow cytometry. ApTOLL showed U-shaped dose-dependent cerebroprotective effects. The maximum effective dose (0.45 mg/kg) was cerebroprotective when given both before reperfusion and up to 12 h after reperfusion and reduced the hemorrhagic risk. Similar effects occurred in female rats. Both research and clinical ApTOLL batches induced slightly superior cerebroprotection when compared with TAK-242. Finally, ApTOLL modulated circulating leukocyte levels, reached the brain ischemic tissue to bind resident and infiltrated cell types, and reduced the neutrophil density. These results show the cerebroprotective effects of ApTOLL in ischemic stroke by reducing the infarct/edema volume, neurofunctional impairment, and hemorrhagic risk, as well as the peripheral and local immune response. They provide information about ApTOLL dose effects and its therapeutic time window and target population, as well as its mode of action, which should be considered in the design of pivotal clinical trials.

Mesenchymal stem cell therapy after stroke is a promising option investigated in animal models and clinical trials. The intravenous route is commonly used in clinical settings guaranteeing an adequate safety profile although low yields of engraftment. In this report, rats subjected to ischemic stroke were injected with adipose-derived stem cells (ADSCs) labeled with superparamagnetic iron oxide nanoparticles (SPIONs) applying an external magnetic field in the skull to retain the cells. Although most published studies demonstrate viability of ADSCs, only a few have used ultrastructural techniques. In our study, the application of a local magnetic force resulted in a tendency for higher yields of SPION-ADSCs targeting the brain. However, grafted cells displayed morphological signs of death, one day after administration, and correlative microscopy showed active microglia and astrocytes associated in the process of scavenging. Thus, we conclude that, although successfully targeted within the brain, SPION-ADSCs viability was rapidly compromised., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Despite the promising neuroprotective effects of uric acid (UA) in acute ischemic stroke, the seemingly pleiotropic underlying mechanisms are not completely understood. Recent evidence points to transcription factors as UA targets. To gain insight into the UA mechanism of action, we investigated its effects on pertinent biomarkers for the most relevant features of ischemic stroke pathophysiology: (1) oxidative stress (antioxidant enzyme mRNAs and MDA), (2) neuroinflammation (cytokine and Socs3 mRNAs, STAT3, NF-κB p65, and reactive microglia), (3) brain swelling (Vegfa, Mmp9, and Timp1 mRNAs), and (4) apoptotic cell death (Bcl-2, Bax, caspase-3, and TUNEL-positive cells). Adult male Wistar rats underwent intraluminal filament transient middle cerebral artery occlusion (tMCAO) and received UA (16 mg/kg) or vehicle (Locke's buffer) i.v. at 20 min reperfusion. The outcome measures were neurofunctional deficit, infarct, and edema. UA treatment reduced cortical infarct and brain edema, as well as neurofunctional impairment. In brain cortex, increased UA: (1) reduced tMCAO-induced increases in Vegfa and Mmp9/Timp1 ratio expressions; (2) induced Sod2 and Cat expressions and reduced MDA levels; (3) induced Il6 expression, upregulated STAT3 and NF-κB p65 phosphorylation, induced Socs3 expression, and inhibited microglia activation; and (4) ameliorated the Bax/Bcl-2 ratio and induced a reduction in caspase-3 cleavage as well as in TUNEL-positive cell counts. In conclusion, the mechanism for morphological and functional neuroprotection by UA in ischemic stroke is multifaceted, since it is associated to activation of the IL-6/STAT3 pathway, attenuation of edematogenic VEGF-A/MMP-9 signaling, and modulation of relevant mediators of oxidative stress, neuroinflammation, and apoptotic cell death.

Progression on therapy in non-small cell lung carcinoma (NSCLC) is often evaluated radiographically, however, image-based evaluation of said therapies may not distinguish disease progression due to intrinsic tumor drug resistance or inefficient tumor penetration of the drugs. Here we report that the inhibition of mutated EGFR promotes the secretion of a potent vasoconstrictor, endothelin-1 (EDN1), which continues to increase as the cells become resistant with a mesenchymal phenotype. As EDN1 and its receptor (EDNR) is linked to cancer progression, EDNR-antagonists have been evaluated in several clinical trials with disappointing results. These trials were based on a hypothesis that the EDN1-EDNR axis activates the MAPK-ERK signaling pathway that is vital to the cancer cell survival; the trials were not designed to evaluate the impact of tumor-derived EDN1 in modifying tumor microenvironment or contributing to drug resistance. Ectopic overexpression of EDN1 in cells with mutated EGFR resulted in poor drug delivery and retarded growth in vivo but not in vitro . Intratumoral injection of recombinant EDN significantly reduced blood flow and subsequent gefitinib accumulation in xenografted EGFR -mutant tumors. Furthermore, depletion of EDN1 or the use of endothelin receptor inhibitors bosentan and ambrisentan improved drug penetration into tumors and restored blood flow in tumor-associated vasculature. Correlatively, these results describe a simplistic endogenous yet previously unrealized resistance mechanism inherent to a subset of EGFR -mutant NSCLC to attenuate tyrosine kinase inhibitor delivery to the tumors by limiting drug-carrying blood flow and the drug concentration in tumors. SIGNIFICANCE: EDNR antagonists can be repurposed to improve drug delivery in VEGFA-secreting tumors, which normally respond to TKI treatment by secreting EDN1, promoting vasoconstriction, and limiting blood and drug delivery., (©2020 American Association for Cancer Research.)

Preclinical and clinical studies support a promising, albeit not definitive, neuroprotective effect of emergent uric acid (UA) administration in ischemic stroke. We assessed the effects of UA in an ischemic stroke model relevant to the current treatment paradigm of mechanical thrombectomy within the STAIR/RIGOR recommendations. A cohort of male and female Wistar rats was subjected to ischemic stroke with mechanical recanalization under physiological monitoring. The effects of transient middle cerebral artery occlusion (tMCAO) with adjunctive UA (IV, 16 mg/kg) or vehicle treatment were assessed at 24 h and 7 days. Outcomes included neurofunctional impairment, brain infarct (TTC staining, MRI imaging and cresyl violet staining) and edema. At 24 h after tMCAO, neurofunctional scores and brain infarct were significantly reduced in rats subjected to UA treatment compared to vehicle, with a selective effect of UA on cortical infarct. No differential effect of UA between male and female rats was evidenced, as no significant interaction of sex with stroke outcomes was found. Rats achieving higher reperfusion levels after tMCAO showed superior reduction of neurofunctional impairment, cortical infarct and edema by UA. After a 7-day follow-up, male rats subjected to UA treatment still showed reductions in neurofunctional impairment and infarct size, compared to vehicle treatment. In conclusion, UA treatment immediately after transient ischemia results in a sex-independent, maintained reduction of brain damage and neurological impairment, better manifested in hyperperfusion conditions. This synergistic effect of UA with mechanical recanalization supports additional clinical testing of UA as an adjunctive treatment to mechanical thrombectomy., (Copyright © 2018 IBRO. Published by Elsevier Ltd. All rights reserved.)

Diabetic nephropathy is associated with increased risk of cardiovascular disease. B-type natriuretic peptide (BNP) plays an important role in cardiovascular pathophysiology and therapeutics. The aim of the present study was to investigate the influence of experimental diabetes on the mechanisms that regulate the relaxant response of the rabbit renal artery to BNP. Arterial relaxations to BNP were enhanced in diabetic rabbits. Indomethacin enhanced BNP-induced relaxation in control rabbits but showed no effect in diabetic rabbits. BNP-induced release of thromboxane A 2 or prostacyclin was not different in both groups of animals. Iberiotoxin had no effect on relaxations to BNP in both groups of animals. Charybdotoxin displaced to the right the concentration-response curve to BNP in both group of animals, and inhibited BNP-induced relaxation only in diabetic rabbits. Glibenclamide did not modify the BNP-induced relaxations in control rabbits, but inhibited it in diabetic rabbits. These results suggest that diabetes induces hypereactivity of the rabbit renal artery to BNP by mechanisms that at least include (1) a reduced vasoconstrictor influence of arachidonic acid metabolites via cyclooxygenase 2, which is not related with changes in thromboxane A 2 and prostacyclin release from the arterial wall and (2) a selectively increased modulatory activity of K ATP and endothelial IK Ca channels.

Background: Nowadays, severe deficiency of vitamin D is not a common finding in most developed countries. However, the prevalence of vitamin D insufficiency is relatively high and it can contribute to the descent of bone mass in osteoporosis risk populations. The objective of our study was to evaluate the prevalence of vitamin D insufficiency in postmenopausal women (PMW), patients with inflammatory bowel disease (IBD) and corticosteroid-dependent asthmatic patients (CAP) and to analyze its relationship with bone mineral density (BMD) and calciotropic hormones., Patients and Method: We studied 299 patients (PMW: 161; IBD: 61; CAP: 77). In all cases, serum levels of PTH and 25OHD were determined and the BMD (DXA, Hologic QDR1000) in lumbar spine (LS) and femoral neck (FN) was measured., Results: Vitamin D insufficiency (25OHD < 15 ng/ml) was observed in 39.1% patients with PMW, 70.7% patients with IBD and 44.2% patients with CAP. 25OHD concentrations were lower in EII patients (p = 0.003) and PTH concentrations were higher in MPM (p < 0.001). We found a negative correlation between PTH and 25OHD in the overall group and this correlation persisted after considering each group separately. After adjusting for remaining variables, 25OHD was found to be significantly associated with BMD at lumbar spine and/or femoral neck in the three groups., Conclusions: In populations at risk of osteoporosis, there is a high prevalence of vitamin D insufficiency. This insufficiency has a significant effect on bone integrity.