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K+ channel Kir7.1 expressed at the apical membrane of the retinal pigment epithelium (RPE) plays an essential role in retinal function. An isoleucine-to-threonine mutation at position 120 of the protein is responsible for blindness-causing vitreo-retinal dystrophy. We have studied the molecular mechanism of action of Kir7.1-I120T in vitro by heterologous expression and in vivo in CRISPR-generated knockin mice. Full-size Kir7.1-I120T reaches the plasma membrane but lacks any activity. Analysis of Kir7.1 and the I120T mutant in mixed transfection experiments, and that of tandem tetrameric constructs made by combining wild type (WT) and mutant protomers, leads us to conclude that they do not form heterotetramers in vitro. Homozygous I120T/I120T mice show cleft palate and tracheomalacia and do not survive beyond P0, whereas heterozygous WT/I120T develop normally. Membrane conductance of RPE cells isolated from WT/WT and heterozygous WT/I120T mice is dominated by Kir7.1 current. Using Rb+ as a charge carrier, we demonstrate that the Kir7.1 current of WT/I120T RPE cells corresponds to approximately 50% of that in cells from WT/WT animals, in direct proportion to WT gene dosage. This suggests a lack of compensatory effects or interference from the mutated allele product, an interpretation consistent with results obtained using WT/- hemizygous mouse. Electroretinography and behavioral tests also show normal vision in WT/I120T animals. The hypomorphic ion channel phenotype of heterozygous Kir7.1-I120T mutants is therefore compatible with normal development and retinal function. The lack of detrimental effect of this degree of functional deficit might explain the recessive nature of Kir7.1 mutations causing human eye disease.NEW & NOTEWORTHY Human retinal pigment epithelium K+ channel Kir7.1 is affected by generally recessive mutations leading to blindness. We investigate one such mutation, isoleucine-to-threonine at position 120, both in vitro and in vivo in knockin mice. The mutated channel is inactive and in heterozygosis gives a hypomorphic phenotype with normal retinal function. Mutant channels do not interfere with wild-type Kir7.1 channels which are expressed concomitantly without hindrance, providing an explanation for the recessive nature of the disease.
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Isoleucina , Retina , Camundongos , Humanos , Animais , Isoleucina/metabolismo , Retina/metabolismo , Cegueira/metabolismo , Mutação/genética , Treonina/metabolismoRESUMO
Electrospun nanofibrous membranes have garnered significant attention in antimicrobial applications, owing to their intricate three-dimensional network that confers an interconnected porous structure, high specific surface area, and tunable physicochemical properties, as well as their notable capacity for loading and sustained release of antimicrobial agents. Tailoring polymer or hybrid-based nanofibrous membranes with stimuli-responsive characteristics further enhances their versatility, enabling them to exhibit broad-spectrum or specific activity against diverse microorganisms. In this review, we elucidate the pivotal advancements achieved in the realm of stimuli-responsive antimicrobial electrospun nanofibers operating by light, temperature, pH, humidity, and electric field, among others. We provide a concise introduction to the strategies employed to design smart electrospun nanofibers with antimicrobial properties. The core section of our review spotlights recent progress in electrospun nanofiber-based systems triggered by single- and multi-stimuli. Within each stimulus category, we explore recent examples of nanofibers based on different polymers and antimicrobial agents. Finally, we delve into the constraints and future directions of stimuli-responsive nanofibrous materials, paving the way for their wider application spectrum and catalyzing progress toward industrial utilization.
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Alpinia zerumbet is a plant popularly used to treat hypertension and anxiety. Studies with Alpinia zerumbet demonstrate antihypertensive and vasodilator effects, among others. The objective of this study was to analyze the effect of essential oil of Alpinia zerumbet (EOAz) on cardiovascular and autonomic function in rats with isoproterenol-induced myocardial infarction. Male Wistar rats (n=32) were equally allocated into four groups: Control, ISO (150mg/kg, subcutaneous), EOAz (100mg/kg by gavage), ISO+EOAz. The rats were evaluated for cardiovascular and, autonomic parameters, electrocardiogram, and infarct size. EOAz was not able to reduce the electrocardiographic variations induced by ISO. Heart rate variability showed a decrease in sympathetic modulation on the heart in the groups treated with EOAz. The cardiopulmonary reflex induced by serotonin invoked a superior blood pressure variation at the 2 µg/kg dose in the EOAz treated groups, while the heart rate variation was significantly higher at the 16 µg/kg dose, when compared to other doses, in all groups, except EOAz+ISO. The sympathetic vagal index was higher in ISO group than in control. EOAz did not reduce the infarct size. We conclude that pretreatment with EOAz does not reverse the hemodynamic and electrocardiographic damage caused by isoproterenol but does reduce sympathetic modulation.
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Alpinia , Infarto do Miocárdio , Óleos Voláteis , Ratos , Animais , Óleos Voláteis/farmacologia , Óleos Voláteis/uso terapêutico , Isoproterenol , Ratos Wistar , Folhas de Planta , Infarto do Miocárdio/induzido quimicamente , Infarto do Miocárdio/tratamento farmacológicoRESUMO
Sea louse ectoparasitosis is a major threat to fish aquaculture. Avermectins such as ivermectin and emamectin have been effectively used against sea louse infestation, but the emergence of resistance has limited their use. A better understanding of the molecular targets of avermectins is essential to the development of novel treatment strategies or new, more effective drugs. Avermectins are known to act by inhibiting neurotransmission through allosteric activation of glutamate-gated chloride channels (GluCls). We have investigated the GluCl subunit present in Caligus rogercresseyi, a sea louse affecting aquaculture in the Southern hemisphere. We identify four new subunits, CrGluCl-B to CrGluCl-E, and characterise them functionally. CrGluCl-A (previously reported as CrGluClα), CrGluCl-B and CrGluCl-C all function as glutamate channel receptors with different sensitivities to the agonist, but in contrast to subunit -A and -C, CrGluCl-B is not activated by ivermectin but is rather antagonised by the drug. CrGluCl-D channel appears active in the absence of any stimulation by glutamate or ivermectin and CrGluCl-E does not exhibit any activity. Notably, the expression of CrGluCl-B with either -A or -C subunits gives rise to receptors unresponsive to ivermectin and showing altered response to glutamate, suggesting that coexpression has led to the preferential formation of heteromers to which the presence of CrGluCl-B confers the property of ivermectin-activation refractoriness. Furthermore, there was evidence for heteromer formation with novel properties only when coexpressing pairs E/C and D/B CrGluCl subtypes. Site-directed mutagenesis shows that three transmembrane domain residues contribute to the lack of activation by ivermectin, most crucially Gln 15' in M2, with mutation Q15'T (the residue present in ivermectin-activated subunits A and C) conferring ivermectin activation to CrGluCl-B. The differential response to avermectin of these Caligus rogercresseyi GluClsubunits, which are highly conserved in the Northern hemisphere sea louse Lepeophtheirus salmonis, could have an influence on the response of these parasites to treatment with macrocyclic lactones. They could serve as molecular markers to assess susceptibility to existing treatments and might be useful molecular targets in the search for novel antiparasitic drugs.
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Copépodes , Parasitos , Ftirápteros , Animais , Ivermectina/farmacologia , Ivermectina/metabolismo , Ftirápteros/metabolismo , Parasitos/metabolismo , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Ácido Glutâmico/farmacologiaRESUMO
Chemical structures of selective blockers of TASK channels contain aromatic groups and amide bonds. Using this rationale, we designed and synthesized a series of compounds based on 3-benzamidobenzoic acid. These compounds block TASK-1 channels by binding to the central cavity. The most active compound is 3-benzoylamino-N-(2-ethyl-phenyl)-benzamide or F3, blocking TASK-1 with an IC50 of 148 nM, showing a reduced inhibition of TASK-3 channels and not a significant effect on different K+ channels. We identified putative F3-binding sites in the TASK-1 channel by molecular modeling studies. Mutation of seven residues to A (I118A, L122A, F125A, Q126A, L232A, I235A, and L239A) markedly decreased the F3-induced inhibition of TASK-1 channels, consistent with the molecular modeling predictions. F3 blocks cell proliferation and viability in the MCF-7 cancer cell line but not in TASK-1 knockdown MCF-7 cells, indicating that it is acting in TASK-1 channels. These results indicated that TASK-1 is necessary to drive proliferation in the MCF-7 cancer cell line.
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Neoplasias , Humanos , Relação Estrutura-Atividade , Sítios de Ligação , Proliferação de Células , Modelos Moleculares , Células MCF-7RESUMO
In this research, a set of CuNiCrSiCoTi (H-0Nb), CuNiCrSiCoTiNb0.5 (H-0.5Nb) and CuNiCrSiCoTiNb1 (H-1Nb) high-entropy alloys (HEAs) were melted in a vacuum induction furnace. The effects of Nb additions on the microstructure, hardness, and wear behavior of these HEAs (compared with a CuBe commercial alloy) in the as-cast (AC) condition, and after solution (SHT) and aging (AT) heat treatments, were investigated using X-ray diffraction, optical microscopy, and electron microscopy. A ball-on-disc configuration tribometer was used to study wear behavior. XRD and SEM results showed that an increase in Nb additions and modification by heat treatment (HT) favored the formation of BCC and FCC crystal structures (CS), dendritic regions, and the precipitation of phases that promoted microstructure refinement during solidification. Increases in hardness of HEA systems were recorded after heat treatment and Nb additions. Maximum hardness values were recorded for the H-1Nb alloy with measured increases from 107.53 HRB (AC) to 112.98 HRB, and from 1104 HV to 1230 HV (aged for 60 min). However, the increase in hardness caused by Nb additions did not contribute to wear resistance response. This can be attributed to a high distribution of precipitated phases rich in high-hardness NiSiTi and CrSi. Finally, the H-0Nb alloy exhibited the best wear resistance behavior in the aged condition of 30 min, with a material loss of 0.92 mm3.
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OBJECTIVE: To summarize the existing evidence on the acute response of low-load (LL) resistance exercise (RE) with blood flow restriction (BFR) on hemodynamic parameters. DATA SOURCES: MEDLINE (via PubMed), EMBASE (via Scopus), SPORTDiscus, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, Web of Science and MedRxiv databases were searched from inception to February 2022. REVIEW METHODS: Cross-over trials investigating the acute effect of LLRE + BFR versus passive (no exercise) and active control methods (LLRE or HLRE) on heart rate (HR), systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure responses. RESULTS: The quality of the studies was assessed using the PEDro scale, risk of bias using the RoB 2.0 tool for cross-over trials and certainty of the evidence using the GRADE method. A total of 15 randomized cross-over studies with 466 participants were eligible for analyses. Our data showed that LLRE + BFR increases all hemodynamic parameters compared to passive control, but not compared to conventional resistance exercise. Subgroup analysis did not demonstrate any differences between LLRE + BFR and low- (LL) or high-load (HL) resistance exercise protocols. Studies including younger volunteers presented higher chronotropic responses (HR) than those with older volunteers. CONCLUSIONS: Despite causing notable hemodynamic responses compared to no exercise, the short-term LL resistance exercise with BFR modulates all hemodynamic parameters HR, SBP, DBP and MBP, similarly to a conventional resistance exercise protocol, whether at low or high-intensity. The chronotropic response is slightly higher in younger healthy individuals despite the similarity regarding pressure parameters.
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Treinamento Resistido , Estudos Cross-Over , Hemodinâmica , Humanos , Músculo Esquelético/irrigação sanguínea , Ensaios Clínicos Controlados Aleatórios como Assunto , Fluxo Sanguíneo Regional/fisiologia , Treinamento Resistido/métodosRESUMO
CONTEXT: During the 2016-2017 Zika virus outbreak in Puerto Rico, preventing unintended pregnancy was a primary strategy to reduce Zika-related adverse birth outcomes. The Zika Contraception Access Network (Z-CAN) was a short-term emergency response intervention that used contraception to prevent unintended pregnancy among women who chose to delay or avoid pregnancy. OBJECTIVE: This analysis reports on the identified policy and practice change strategies to increase access to or provision of contraceptive services in Puerto Rico between 2015 and 2018. METHODS: A policy review was conducted to document federal- and territorial-level programs with contraceptive coverage and payment policies in Puerto Rico and to identify policy and practice change. Semistructured interviews with key stakeholders in Puerto Rico were also conducted to understand perceptions of policy and practice change efforts following the Zika virus outbreak, including emergency response, local, and policy efforts to improve contraception access in Puerto Rico. RESULTS: Publicly available information on federal and territorial programs with policies that facilitate access, delivery, and utilization of contraceptive coverage and family planning services in Puerto Rico to support contraceptive access was documented; however, interview results indicated that the implementation of the policies was often limited by barriers and that policy and practice changes as the result of the Zika virus outbreak were short-term. CONCLUSION: Consideration of long-term policy and practice changes related to contraceptive access is warranted. Similar analyses can be used to identify policies, practices, and perceptions in other settings in which the goal is to increase access to contraception or reduce unintended pregnancy.
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Infecção por Zika virus , Zika virus , Anticoncepcionais , Serviços de Planejamento Familiar , Feminino , Humanos , Políticas , Gravidez , Porto Rico/epidemiologia , Infecção por Zika virus/epidemiologia , Infecção por Zika virus/prevenção & controleRESUMO
Natural pterocarpans and synthetic 5-carba-pterocarpans are isosteres in which the oxygen atom at position 5 in the pyran-ring of pterocarpans is replaced by a methylene group. These 5-carba-analogues were obtained in good yields through the palladium-catalyzed oxyarylation of alcoxy-1,2-dihydronaphthalens with o-iodophenols in PEG-400. They were evaluated on human cancer cell lineages derived respectively from prostate tumor (PC3, IC50 = 11.84 µmol L-1, SI > 12)) and acute myeloid leukemia (HL-60, IC50 = 8.81 µmol L-1, SI > 16), highly incident cancer types presenting resistance against traditional chemotherapeutics. Compound 6c (LQB-492) was the most potent (IC50 = 3.85 µmol L-1, SI > 37) in SF-295 cell lineage (glioblastoma). Such findings suggest that 5-carba-pterocarpan can potentially be new hit compounds for further development of novel antiproliferative agents.
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Antineoplásicos/farmacologia , Pterocarpanos/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Camundongos , Estrutura Molecular , Pterocarpanos/síntese química , Pterocarpanos/química , Relação Estrutura-AtividadeRESUMO
KEY POINTS: Kir7.1 K+ channel expressed in retinal pigment epithelium is mutated in inherited retinal degeneration diseases. We study Kir7.1 in heterologous expression to test the hypothesis that pathological R162 mutation to neutral amino acids results in loss of a crucial site that binds PI(4,5)P2 . Although R162W mutation inactivates Kir7.1, changes to smaller volume (e.g. Gln) amino acids are tolerated or even enhance function (Ala or Cys). Chemical modification of Kir7.1-R162C confirms that large residues of the size of Trp are incompatible with normal channel function even if positively charged. In addition to R162, K164 (and possibly K159) forms a binding site for the phosphoinositide and is essential for channel activity. R162 substitution with a large, neutral side chain like Trp exerts a dominant negative effect on Kir7.1 activity such that less than one fifth of the full activity is expected in a cell expressing the same amount of mutant and wild-type channels. ABSTRACT: Mutations in the Kir7.1 K+ channel, highly expressed in retinal pigment epithelium, have been linked to inherited retinal degeneration diseases. Examples are mutations changing Arg 162 to Trp in snowflake vitreoretinal degeneration (SVD) and Gln in retinitis pigmentosa. R162 is believed to be part of a site that binds PI(4,5)P2 and stabilises the open state. We have tested the hypothesis that R162 mutation to neutral amino acids will result in the loss of this crucial interaction to the detriment of channel function. Our findings indicate that although R612W mutation inactivates Kir7.1, changes to smaller volume (e.g. Gln) amino acids are tolerated or even enhance function (Ala or Cys). Cys chemical modification of Kir7.1-R162C confirms that large residues of the size of Trp are incompatible with normal channel function even if positively charged. Experiments titrating the levels of plasma membrane PI(4,5)P2 with voltage-dependent phosphatase DrVSP reveal that, in addition to R162, K164 (and possibly K159) forms a binding site for the phosphoinositide and ensures channel activity. Finally, the use of a concatemeric approach shows that substitution of R162 with a large, neutral side chain mimicking a Trp residue exerts a dominant negative effect on Kir7.1 activity such that less than one fifth of the full activity is expected in heterozygous cells carrying the SVD mutation. Our results suggest that if mutations in the human KCNJ13 gene resulting in the neutralisation of R162 and Kir7.1 malfunction led to retinal degeneration diseases, their severity might depend on the nature of the side chain of the replacing amino acid.
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Degeneração Retiniana , Membrana Celular , Humanos , Mutação , Fosfatidilinositóis , Degeneração Retiniana/genética , Epitélio Pigmentado da RetinaRESUMO
Aneurysms of the sinus of Valsalva are rare. Unruptured sinus of Valsalva aneurysm is usually asymptomatic and rarely presents as right ventricular outflow obstruction, myocardial infarction as a result of coronary artery compression, conduction disturbances, or endocarditis. They have only been reported as the presumed source of embolism in six cases. We report a patient with right sinus of Valsalva rupture to the right atrium and embolization of aneurysm contents to the pulmonary vasculature.
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Lubiprostone, a 20-carbon synthetic fatty acid used for the treatment of constipation, is thought to act through an action on Cl- channel ClC-2. Short chain fatty acids (SCFAs) are produced and absorbed in the distal intestine. We explore whether SCFAs affect ClC-2, re-examine a possible direct effect of lubiprostone on ClC-2, and use mice deficient in ClC-2 to stringently address the hypothesis that the epithelial effect of lubiprostone targets this anion channel. Patch-clamp whole cell recordings of ClC-2 expressed in mammalian cells are used to assay SCFA and lubiprostone effects. Using chamber measurements of ion current in mice deficient in ClC-2 or CFTR channels served to analyze the target of lubiprostone in the distal intestinal epithelium. Intracellular SCFAs had a dual action on ClC-2, partially inhibiting conduction but, importantly, facilitating the voltage activation of ClC-2. Intra- or extracellular lubiprostone had no effect on ClC-2 currents. Lubiprostone elicited a secretory current across colonic epithelia that was increased in mice deficient in ClC-2, consistent with the channel's proposed proabsorptive function, but absent from those deficient in CFTR. Whilst SCFAs might exert a physiological effect on ClC-2 as part of their known proabsorptive effect, ClC-2 plays no part in the lubiprostone intestinal effect that appears mediated by CFTR activation.
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Agonistas dos Canais de Cloreto/uso terapêutico , Canais de Cloreto/efeitos dos fármacos , Ácidos Graxos Voláteis/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Lubiprostona/uso terapêutico , Canais de Cloro CLC-2 , Agonistas dos Canais de Cloreto/farmacologia , Células HEK293 , Humanos , Lubiprostona/farmacologiaRESUMO
BACKGROUND: Invasive Africanized honey bees potentially compete with cavity-nesting birds in South America. However, the impacts of this competition and its conservation consequences to threatened species are poorly known. We quantified the presence of these bees and assessed their competition for cliff cavities used by nesting Lear's macaws Anodorhynchus leari, a globally endangered parrot endemic to the Caatinga biome of Brazil. We treated beehives with permethrin by shooting them with a crossbow bolt that distributed the compound upon impact. When feasible, we removed the comb and applied an insecticide (fipronil) to deter bee recolonization. We subsequently surveyed the macaw breeding population to verify whether our treatment allowed for nest recruitment in cavities previously occupied by bees. RESULTS: We recorded > 100 beehives in the nesting cliffs. Hives outnumbered macaw nests tenfold in two areas recently recolonized by macaws. Cavities occupied by bees were significantly higher than those occupied by macaws, suggesting that macaws may be forced to breed in lower cavities. None of the untreated cavities (n = 50) were occupied by nesting macaws, whereas 15% of treated cavities (n = 52) were occupied within 2 years post treatment. Treated cavities occupied by macaws were significantly higher than those not occupied. Hive management increased macaw breeding population by 71% of the macaw breeding population increase. CONCLUSION: Experimental hive treatments were effective in restoring nesting resources lost due to bee infestation. An intensive and continued eradication program is recommended to enhance macaw habitat restoration, facilitating its expansion into historical areas. © 2020 Society of Chemical Industry.
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Comportamento de Nidação , Papagaios , Animais , Abelhas , Brasil , Cruzamento , Densidade DemográficaRESUMO
BACKGROUND: Pharmaceutical nanotechnology represents an efficient alternative for the delivery of pharmacologically active plant-derived compounds, considering their protective capacity, oral bioavailability and drug vectorization capacity. In this context, butters obtained from plant seeds have emerged as promising products for the development of pharmacologically active nanostructures. They possess a complex lipid composition, allowing the formation of different emulsion systems with solid cores, since this mixture of different triglycerides is solid at room temperature and body temperature. Therefore, the systematic mapping around the technological development of nanostructures produced from plant-derived butters is potentially valuable for researchers interested in novel alternative formulations for pharmacological therapy, with potential industrial, economic, health and societal impacts. METHODS: Systematic review was carried out by the search of scientific papers and patents deposited in official databases concerning the development of nanostructured pharmaceutical products using plantderived butters as starting material. The publications obtained were subjected to sorting and analysis by applying the following inclusion/exclusion criteria. RESULTS: The Solid Lipid Nanoparticle (SLN) was the type of nanostructure produced in all the analyzed scientific papers, due to the physicochemical characteristics of the lipid constituents of plantderived butters. In this sense, 54% of the articles have reported the use of Cocoa Butter for the production of nanostructures; 28% for Shea Butter; 6% for Cupuacu Butter, 6% for Murumuru Butter and 6% for Bacuri Butter. DISCUSSION: In the technological prospection, only two patents exhibited SLN as an invention based on cocoa butter and on shea butter, respectively. The production methods employed have included: phase inversion temperature, microemulsion, hot high pressure homogenization, high shear homogenization and ultrasonication. CONCLUSION: In light of this prospective review, the encouragement of novel studies in lipids-based nanotechnology is evident, considering the small number of findings so far, in order to stimulate new research involving plant-derived butters from easily cultivated fruits in tropical regions, then stimulating the pharmaceutical development of new therapeutic alternatives using biocompatible and sustainable raw materials.
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Manteiga/análise , Portadores de Fármacos/química , Lipídeos/química , Nanopartículas/química , Plantas/química , Emulsões , Lipossomos , PublicaçõesRESUMO
The cancer multidrug resistance is involved in the failure of several treatments during cancer treatment. It is a phenomenon that has been receiving great attention in the last years due to the sheer amount of mechanisms discovered and involved in the process of resistance which hinders the effectiveness of many anti-cancer drugs. Among the mechanisms involved in the multidrug resistance, the participation of ATP-binding cassette (ABC) transporters is the main one. The ABC transporters are a group of plasma membrane and intracellular organelle proteins involved in the process of externalization of substrates from cells, which are expressed in cancer. They are involved in the clearance of intracellular metabolites as ions, hormones, lipids and other small molecules from the cell, affecting directly and indirectly drug absorption, distribution, metabolism and excretion. Other mechanisms responsible for resistance are the signaling pathways and the anti- and pro-apoptotic proteins involved in cell death by apoptosis. In this study we evaluated the influence of three nanosystem (Graphene Quantum Dots (GQDs), mesoporous silica (MSN) and poly-lactic nanoparticles (PLA)) in the main mechanism related to the cancer multidrug resistance such as the Multidrug Resistance Protein-1 and P-glycoprotein. We also evaluated this influence in a group of proteins involved in the apoptosis-related resistance including cIAP-1, XIAP, Bcl-2, BAK and Survivin proteins. Last, colonogenic and MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assays have also been performed. The results showed, regardless of the concentration used, GQDs, MSN and PLA were not cytotoxic to MDA-MB-231 cells and showed no impairment in the colony formation capacity. In addition, it has been observed that P-gp membrane expression was not significantly altered by any of the three nanomaterials. The results suggest that GQDs nanoparticles would be suitable for the delivery of other multidrug resistance protein 1 (MRP1) substrate drugs that bind to the transporter at the same binding pocket, while MSN can strongly inhibit doxorubicin efflux by MRP1. On the other hand, PLA showed moderate inhibition of doxorubicin efflux by MRP1 suggesting that this nanomaterial can also be useful to treat MDR (Multidrug resistance) due to MRP1 overexpression.
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Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Resistência a Múltiplos Medicamentos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos/administração & dosagem , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Imunofluorescência , Expressão Gênica , Grafite/química , Humanos , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Nanopartículas/química , Nanoestruturas/química , Nanomedicina TeranósticaRESUMO
Chaotic systems implemented by artificial neural networks are good candidates for data encryption. In this manner, this paper introduces the cryptographic application of the Hopfield and the Hindmarsh-Rose neurons. The contribution is focused on finding suitable coefficient values of the neurons to generate robust random binary sequences that can be used in image encryption. This task is performed by evaluating the bifurcation diagrams from which one chooses appropriate coefficient values of the mathematical models that produce high positive Lyapunov exponent and Kaplan-Yorke dimension values, which are computed using TISEAN. The randomness of both the Hopfield and the Hindmarsh-Rose neurons is evaluated from chaotic time series data by performing National Institute of Standard and Technology (NIST) tests. The implementation of both neurons is done using field-programmable gate arrays whose architectures are used to develop an encryption system for RGB images. The success of the encryption system is confirmed by performing correlation, histogram, variance, entropy, and Number of Pixel Change Rate (NPCR) tests.
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OBJECTIVES: To assess the diagnostic accuracy of signal loss on in-phase (IP) gradient-echo (GRE) images for differentiation between renal cell carcinomas (RCCs) and lipid-poor angiomyolipomas (lpAMLs). METHODS: We retrospectively searched our institutional database for histologically proven small RCCs (<5.0 cm) and AMLs without visible macroscopic fat (lpAMLs). Two experienced radiologists assessed MRIs qualitatively, to depict signal loss foci on IP GRE images. A third radiologist drew regions of interest (ROIs) on the same lesions, on IP and out-of-phase (OP) images to calculate the ratio of signal loss. Diagnostic accuracy parameters were calculated for both techniques and the inter-reader agreement for the qualitative analysis was evaluated using the κ test. RESULTS: 15 (38.4%) RCCs lost their signal on IP images, with a sensitivity of 38.5% (95% CI = 23.4-55.4), a specificity of 100% (71.1-100), a positive predictive value (PPV) of 100% (73.4-100), a negative predictive value (NPV) of 31.4% (26.3-37.0), and an overall accuracy of 52% (37.4-66.3%). In terms of the quantitative analysis, the signal intensity index (SII= [(SIIP - SIOP) / SIOP] x 100) for RCCs was -0.132 ± 0.05, while for AMLs it was -0.031 ± 0.02, p = 0.26. The AUC was 0.414 ± -0.09 (0.237-0.592). Using 19% of signal loss as the threshold, sensitivity was 16% and specificity was 100%. The κappa value for subjective analysis was 0.63. CONCLUSION: Signal loss in "IP" images, assessed subjectively, was highly specific for distinction between RCCs and lpAMLs, although with low sensitivity. The findings can be used to improve the preoperative diagnostic accuracy of MRI for renal masses. ADVANCES IN KNOWLEDGE: Signal loss on "IP" GRE images is a reliable sign for differentiation between RCC and lpAMLs.
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Angiomiolipoma/diagnóstico por imagem , Carcinoma de Células Renais/diagnóstico por imagem , Carcinoma de Células Pequenas/diagnóstico por imagem , Neoplasias Renais/diagnóstico por imagem , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Angiomiolipoma/patologia , Área Sob a Curva , Carcinoma de Células Renais/patologia , Carcinoma de Células Pequenas/patologia , Criança , Diagnóstico Diferencial , Feminino , Humanos , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto JovemRESUMO
Two-pore domain potassium (K2P) channels maintain the cell's background conductance by stabilizing the resting membrane potential. They assemble as dimers possessing four transmembrane helices in each subunit. K2P channels were crystallized in "up" and "down" states. The movements of the pore-lining transmembrane TM4 helix produce the aperture or closure of side fenestrations that connect the lipid membrane with the central cavity. When the TM4 helix is in the up-state, the fenestrations are closed, while they are open in the down-state. It is thought that the fenestration states are related to the activity of K2P channels and the opening of the channels preferentially occurs from the up-state. TASK-2, a member of the TALK subfamily of K2P channels, is opened by intracellular alkalization leading the deprotonation of the K245 residue at the end of the TM4 helix. This charge neutralization of K245 could be sensitive or coupled to the fenestration state. Here, we describe the relationship between the states of the intramembrane fenestrations and K245 residue in TASK-2 channel. By using molecular modeling and simulations, we show that the protonated state of K245 (K245+) favors the open fenestration state and, symmetrically, that the open fenestration state favors the protonated state of the lysine residue. We show that the channel can be completely blocked by Prozac, which is known to induce fenestration opening in TREK-2. K245 protonation and fenestration aperture have an additive effect on the conductance of the channel. The opening of the fenestrations with K245+ increases the entrance of lipids into the selectivity filter, blocking the channel. At the same time, the protonation of K245 introduces electrostatic potential energy barriers to ion entrance. We computed the free energy profiles of ion penetration into the channel in different fenestration and K245 protonation states, to show that the effects of the two transformations are summed up, leading to maximum channel blocking. Estimated rates of ion transport are in qualitative agreement with experimental results and support the hypothesis that the most important barrier for ion transport under K245+ and open fenestration conditions is the entrance of the ions into the channel.
Assuntos
Concentração de Íons de Hidrogênio , Canais de Potássio de Domínios Poros em Tandem/química , Canais de Potássio de Domínios Poros em Tandem/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Células HEK293 , Humanos , Ativação do Canal Iônico , Íons/química , Íons/metabolismo , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Ligação Proteica , Conformação Proteica , Relação Estrutura-AtividadeRESUMO
In 2018, the Mexican Caribbean coast received a massive influx of pelagic Sargassum spp. that accumulated and decayed on beaches producing organic decomposition products that made the water turbid and brown. Between May and September of the same year there were several reports of mass mortality of marine biota in this area. From these reports we estimate that organisms belonging to 78 faunal species died as result of this event, with demersal neritic fish and Crustacea being the most affected groups. The cause of mortality appears to be the combined effect of high ammonium and hydrogen sulfide concentrations, together with hypoxic conditions. If massive arrival of pelagic Sargassum spp. continues and algae is left to decay on the beach in large volumes then deterioration in water quality could affect coral reefs close to shore. Furthermore, barriers placed in lagoons to intercept the Sargassum spp. before it reaches the beach could impact reef fauna if the algae is left to die and sink on site.
Assuntos
Crustáceos , Peixes , Sargassum/fisiologia , Água do Mar/química , Compostos de Amônio/análise , Animais , Organismos Aquáticos , Região do Caribe , Sulfeto de Hidrogênio/análise , México , Mortalidade , Água do Mar/análise , Qualidade da ÁguaRESUMO
Inwardly rectifying K+ channel Kir7.1 is expressed in epithelia where it shares membrane localisation with the Na+/K+-pump. The ciliary body epithelium (CBE) of the eye is a determinant of intraocular pressure (IOP) through NaCl-driven fluid secretion of aqueous humour. In the present study we explored the presence Kir7.1 in this epithelium in the mouse and its possible functional role in the generation of IOP. Use heterozygous animals for total Kir7.1 knockout expressing ß-galactosidase under the control of Kir7.1 promoter, identified the expression of Kir7.1 in non-pigmented epithelial cells of CBE. Using conditional, floxed knockout Kir7.1 mice as negative controls, we found Kir7.1â¯at the basolateral membrane of the same CBE cell layer. This was confirmed using a knockin mouse expressing the Kir7.1 protein tagged with a haemagglutinin epitope. Measurements using the conditional knockout mouse show only a minor effect of Kir7.1 inactivation on steady-state IOP. Transient increases in IOP in response to general anaesthetics, or to water injection, are absent or markedly curtailed in Kir7.1-deficient mice. These results suggest a role for Kir7.1 in IOP regulation through a possible modulation of aqueous humour production by the CBE non-pigmented epithelial cells. The location of Kir7.1 in the CBE, together with the effect of its removal on dynamic changes in IOP, point to a possible role of the channel as a leak pathway preventing cellular overload of K+ during the secretion process. Kir7.1 could be used as a potential therapeutic target in pathological conditions leading to elevated intraocular pressure.