Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 137
Filtrar
1.
Plants (Basel) ; 13(6)2024 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-38592747

RESUMO

The species Senecio nutans Sch. Bip., commonly called "chachacoma", is widely used as a medicinal plant by the Andean communities of Northern Chile. Ethanolic extracts of S. nutans and the main compound, 4-hydroxy-3-(3-methyl-2-butenyl) acetophenone, have shown interesting biological activity. However, due to the high-altitude areas where this species is found, access to S. nutans is very limited. Due to the latter, in this work, we carried out micropropagation in vitro and ex vitro adaptation techniques as an alternative for the massive multiplication, conservation, and in vitro production of high-value metabolites from this plant. The micropropagation and ex vitro adaptation techniques were successfully employed, and UHPLC-DAD analysis revealed no significant changes in the phenolic profile, with acetophenone 4 being the most abundant metabolite, whose antioxidant and antibacterial activity was studied. Independently of the applied culture condition, the ethanolic extracts of S. nutans presented high activity against both Gram-positive and Gram-negative bacteria, demonstrating their antimicrobial capacity. This successful initiation of in vitro and ex vitro cultures provides a biotechnological approach for the conservation of S. nutans and ensures a reliable and consistent source of acetophenone 4 as a potential raw material for pharmacological applications.

2.
Nano Lett ; 24(3): 790-796, 2024 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-38189790

RESUMO

We experimentally and computationally investigate the magneto-conductance across the radial heterojunction of InAs-GaSb core-shell nanowires under a magnetic field, B, up to 30 T and at temperatures in the range 4.2-200 K. The observed double-peak negative differential conductance markedly blue-shifts with increasing B. The doublet accounts for spin-polarized currents through the Zeeman split channels of the InAs (GaSb) conduction (valence) band and exhibits strong anisotropy with respect to B orientation and marked temperature dependence. Envelope function approximation and a semiclassical (WKB) approach allow to compute the magnetic quantum states of InAs and GaSb sections of the nanowire and to estimate the B-dependent tunneling current across the broken-gap interface. Disentangling different magneto-transport channels and a thermally activated valence-to-valence band transport current, we extract the g-factor from the spin-up and spin-down dI/dV branch dispersion, revealing a giant, strongly anisotropic g-factor in excess of 60 (100) for the radial (tilted) field configurations.

3.
bioRxiv ; 2023 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-38045341

RESUMO

Here we report the discovery of MED6-189, a new analogue of the kalihinol family of isocyanoterpene (ICT) natural products. MED6-189 is effective against drug-sensitive and -resistant P. falciparum strains blocking both intraerythrocytic asexual replication and sexual differentiation. This compound was also effective against P. knowlesi and P. cynomolgi. In vivo efficacy studies using a humanized mouse model of malaria confirms strong efficacy of the compound in animals with no apparent hemolytic activity or apparent toxicity. Complementary chemical biology, molecular biology, genomics and cell biological analyses revealed that MED6-189 primarily targets the parasite apicoplast and acts by inhibiting lipid biogenesis and cellular trafficking. Genetic analyses in P. falciparum revealed that a mutation in PfSec13, which encodes a component of the parasite secretory machinery, reduced susceptibility to the drug. The high potency of MED6-189 in vitro and in vivo, its broad range of efficacy, excellent therapeutic profile, and unique mode of action make it an excellent addition to the antimalarial drug pipeline.

4.
J Chem Inf Model ; 63(21): 6642-6654, 2023 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-37909535

RESUMO

There is still growing interest in graphene interactions with proteins, both for its possible biological applications and due to concerns over detrimental effects at the cellular level. As with any process involving proteins, an understanding of amino acid composition is desirable. In this work, we systematically studied the adsorption process of amino acids onto pristine graphene via rigorous free-energy calculations. We characterized the free energy, potential energy, and entropy of the adsorption of all proteinogenic amino acids. The energetic components were further separated into pair interaction contributions. A linear correlation was found between the free energy and the solvent accessible surface area change during adsorption (ΔSASAads) over pristine graphene and uncharged amino acids. Free energies over pristine graphene were compared with adsorption onto graphene oxide, finding an almost complete loss of the favorability of amino acid adsorption onto graphene. Finally, the correlation with ΔSASAads was used to successfully predict the free energy of adsorption of several penta-l-peptides in different structural states and sequences. Due to the relative ease of calculating the ΔSASAads compared to free-energy calculations, it could prove to be a cost-effective predictor of the free energy of adsorption for proteins onto nonpolar surfaces.


Assuntos
Aminoácidos , Grafite , Aminoácidos/química , Entropia , Grafite/química , Adsorção , Solventes
5.
Opt Express ; 31(20): 31734-31748, 2023 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-37858991

RESUMO

A new procedure to measure the extinction coefficient k of film materials that are relatively transparent is presented. This procedure does not require the use of an optical-constant model or the knowledge of extra physical properties of the material, such as the specific heat capacity. It involves preparing a sample with two areas, at least one of them coated with the film, whereas the other may remain uncoated or may be coated with a different thickness of the same material. The differential transmittance between the two sample areas is shown to be proportional to k of the film material in the following measurement conditions: the incident light is p polarized and it impinges at the film material Brewster angle. The differential transmittance is obtained with a single measurement by making the light beam or the sample to oscillate with respect to one another and by using a lock-in amplifier; for normalization purposes, the transmittance in one of the sample areas is also measured. The proportionality factor between the normalized differential transmittance and k only involves the wavelength, the film thickness, and the Brewster angle. The knowledge of the film Brewster angle requires that the film refractive index (n) is measured beforehand; this can be performed with standard procedures, such as ellipsometry, since such techniques are efficient at measuring n of a transparent material, but are inefficient at measuring a small k. The procedure is exemplified with the calculation of k in the far ultraviolet of AlF3 films deposited by evaporation. The dependence of the uncertainty of k obtained with this procedure is analyzed in terms of the uncertainty of the film n, of wavelength, and of the degree of polarization of the incident beam. The selection of a substrate with similar n to the film material is also discussed. The uncertainties involved with the present procedure were analyzed for a specific example and an uncertainty of 2 × 10-5 in k calculation is considered feasible.

6.
ACS Infect Dis ; 9(10): 1821-1833, 2023 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-37722671

RESUMO

Each year, approximately 50,000 children under 5 die as a result of diarrhea caused by Cryptosporidium parvum, a protozoan parasite. There are currently no effective drugs or vaccines available to cure or prevent Cryptosporidium infection, and there are limited tools for identifying and validating targets for drug or vaccine development. We previously reported a high throughput screening (HTS) of a large compound library against Plasmodium N-myristoyltransferase (NMT), a validated drug target in multiple protozoan parasite species. To identify molecules that could be effective against Cryptosporidium, we counter-screened hits from the Plasmodium NMT HTS against Cryptosporidium NMT. We identified two potential hit compounds and validated them against CpNMT to determine if NMT might be an attractive drug target also for Cryptosporidium. We tested the compounds against Cryptosporidium using both cell-based and NMT enzymatic assays. We then determined the crystal structure of CpNMT bound to Myristoyl-Coenzyme A (MyrCoA) and structures of ternary complexes with MyrCoA and the hit compounds to identify the ligand binding modes. The binding site architectures display different conformational states in the presence of the two inhibitors and provide a basis for rational design of selective inhibitors.


Assuntos
Criptosporidiose , Cryptosporidium , Plasmodium , Criança , Humanos , Criptosporidiose/tratamento farmacológico , Desenvolvimento de Medicamentos
7.
mSphere ; 8(5): e0015423, 2023 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-37565760

RESUMO

Shigella flexneri is a facultative intracellular pathogen that causes shigellosis, a human diarrheal disease characterized by the destruction of the colonic epithelium. Novel antimicrobial compounds to treat infections are urgently needed due to the proliferation of bacterial antibiotic resistance and lack of new effective antimicrobials in the market. Our approach to find compounds that block the Shigella virulence pathway has three potential advantages: (i) resistance development should be minimized due to the lack of growth selection pressure, (ii) no resistance due to environmental antibiotic exposure should be developed since the virulence pathways are not activated outside of host infection, and (iii) the normal intestinal microbiota, which do not have the targeted virulence pathways, should be unharmed. We chose to utilize two phenotypic assays, inhibition of Shigella survival in macrophages and Shigella growth inhibition (minimum inhibitory concentration), to interrogate the 1.7 M compound screening collection subset of the GlaxoSmithKline drug discovery chemical library. A number of secondary assays on the hit compounds resulting from the primary screens were conducted, which, in combination with chemical, structural, and physical property analyses, narrowed the final hit list to 44 promising compounds for further drug discovery efforts. The rapid development of antibiotic resistance is a critical problem that has the potential of returning the world to a "pre-antibiotic" type of environment, where millions of people will die from previously treatable infections. One relatively newer approach to minimize the selection pressures for the development of resistance is to target virulence pathways. This is anticipated to eliminate any resistance selection pressure in environmental exposure to virulence-targeted antibiotics and will have the added benefit of not affecting the non-virulent microbiome. This paper describes the development and application of a simple, reproducible, and sensitive assay to interrogate an extensive chemical library in high-throughput screening format for activity against the survival of Shigella flexneri 2457T-nl in THP-1 macrophages. The ability to screen very large numbers of compounds in a reasonable time frame (~1.7 M compounds in ~8 months) distinguishes this assay as a powerful tool in further exploring new compounds with intracellular effect on S. flexneri or other pathogens with similar pathways of pathogenesis. The assay utilizes a luciferase reporter which is extremely rapid, simple, relatively inexpensive, and sensitive and possesses a broad linear range. The assay also utilized THP-1 cells that resemble primary monocytes and macrophages in morphology and differentiation properties. THP-1 cells have advantages over human primary monocytes or macrophages because they are highly plastic and their homogeneous genetic background minimizes the degree of variability in the cell phenotype (1). The intracellular and virulence-targeted selectivity of our methodology, determined via secondary screening, is an enormous advantage. Our main interest focuses on hits that are targeting virulence, and the most promising compounds with adequate physicochemical and drug metabolism and pharmacokinetic (DMPK) properties will be progressed to a suitable in vivo shigellosis model to evaluate the therapeutic potential of this approach. Additionally, compounds that act via a host-directed mechanism could be a promising source for further research given that it would allow a whole new, specific, and controlled approach to the treatment of diseases caused by some pathogenic bacteria.


Assuntos
Disenteria Bacilar , Shigella , Humanos , Shigella flexneri , Virulência/genética , Disenteria Bacilar/tratamento farmacológico , Bibliotecas de Moléculas Pequenas/farmacologia , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Macrófagos
8.
EMBO J ; 42(18): e113987, 2023 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-37577760

RESUMO

Dysregulation of the PI3K/AKT pathway is a common occurrence in high-grade serous ovarian carcinoma (HGSOC), with the loss of the tumour suppressor PTEN in HGSOC being associated with poor prognosis. The cellular mechanisms of how PTEN loss contributes to HGSOC are largely unknown. We here utilise time-lapse imaging of HGSOC spheroids coupled to a machine learning approach to classify the phenotype of PTEN loss. PTEN deficiency induces PI(3,4,5)P3 -rich and -dependent membrane protrusions into the extracellular matrix (ECM), resulting in a collective invasion phenotype. We identify the small GTPase ARF6 as a crucial vulnerability of HGSOC cells upon PTEN loss. Through a functional proteomic CRISPR screen of ARF6 interactors, we identify the ARF GTPase-activating protein (GAP) AGAP1 and the ECM receptor ß1-integrin (ITGB1) as key ARF6 interactors in HGSOC regulating PTEN loss-associated invasion. ARF6 functions to promote invasion by controlling the recycling of internalised, active ß1-integrin to maintain invasive activity into the ECM. The expression of the CYTH2-ARF6-AGAP1 complex in HGSOC patients is inversely associated with outcome, allowing the identification of patient groups with improved versus poor outcome. ARF6 may represent a therapeutic vulnerability in PTEN-depleted HGSOC.


Assuntos
Proteínas Monoméricas de Ligação ao GTP , Neoplasias Ovarianas , Humanos , Feminino , Integrinas/metabolismo , Proteômica , Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Proteínas Monoméricas de Ligação ao GTP/metabolismo , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo
9.
Epilepsy Behav ; 147: 109384, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37634373

RESUMO

PURPOSE: Urgent seizures are a medical emergency for which new therapies are still needed. This study evaluated the use of intravenous brivaracetam (IV-BRV) in an emergency setting in clinical practice. METHODS: BRIV-IV was a retrospective, multicenter, observational study. It included patients ≥18 years old who were diagnosed with urgent seizures (including status epilepticus (SE), acute repetitive seizures, and high-risk seizures) and who were treated with IV-BRV according to clinical practice in 14 hospital centers. Information was extracted from clinical charts and included in an electronic database. Primary effectiveness endpoints included the rate of IV-BRV responder patients, the rate of patients with a sustained response without seizure relapse in 12 h, and the time between IV-BRV administration and clinical response. Primary safety endpoints were comprised the percentage of patients with adverse events and those with adverse events leading to discontinuation. RESULTS: A total of 156 patients were included in this study. The mean age was 57.7 ± 21.5 years old with a prior diagnosis of epilepsy for 57.1% of patients. The most frequent etiologies were brain tumor-related (18.1%) and vascular (11.2%) epilepsy. SE was diagnosed in 55.3% of patients. The median time from urgent seizure onset to IV treatment administration was 60.0 min (range: 15.0-360.0), and the median time from IV treatment to IV-BRV was 90.0 min (range: 30.0-2400.0). Regarding dosage, the mean bolus infusion was 163.0 ± 73.0 mg and the mean daily dosage was 195.0 ± 87.0 mg. A total of 77.6% of patients responded to IV-BRV (66.3% with SE vs. 91% other urgent seizures) with a median response time of 30.0 min (range: 10.0-60.0). A sustained response was achieved in 62.8% of patients. However, adverse events were reported in 14.7%, which were predominantly somnolence and fatigue, with 4.5% leading to discontinuation. Eighty-six percent of patients were discharged with oral brivaracetam. CONCLUSION: IV-BRV in emergency settings was effective, and tolerability was good for most patients. However, a larger series is needed to confirm the outcomes.


Assuntos
Epilepsia , Estado Epiléptico , Adolescente , Adulto , Idoso , Humanos , Pessoa de Meia-Idade , Anticonvulsivantes/efeitos adversos , Quimioterapia Combinada , Epilepsia/tratamento farmacológico , Recidiva Local de Neoplasia , Pirrolidinonas/efeitos adversos , Estudos Retrospectivos , Convulsões/tratamento farmacológico , Convulsões/induzido quimicamente , Estado Epiléptico/tratamento farmacológico , Resultado do Tratamento
10.
Int J Mol Sci ; 24(14)2023 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-37511161

RESUMO

This study takes a step in understanding the physiological implications of the nanosecond pulsed electric field (nsPEF) by integrating molecular dynamics simulations and machine learning techniques. nsPEF, a state-of-the-art technology, uses high-voltage electric field pulses with a nanosecond duration to modulate cellular activity. This investigation reveals a relatively new and underexplored phenomenon: protein-mediated electroporation. Our research focused on the voltage-sensing domain (VSD) of the NaV1.5 sodium cardiac channel in response to nsPEF stimulation. We scrutinized the VSD structures that form pores and thereby contribute to the physical chemistry that governs the defibrillation effect of nsPEF. To do so, we conducted a comprehensive analysis involving the clustering of 142 replicas simulated for 50 ns under nsPEF stimuli. We subsequently pinpointed the representative structures of each cluster and computed the free energy between them. We find that the selected VSD of NaV1.5 forms pores under nsPEF stimulation, but in a way that significant differs from the traditional VSD opening. This study not only extends our understanding of nsPEF and its interaction with protein channels but also adds a new effect to further study.


Assuntos
Eletricidade , Eletroporação , Eletroporação/métodos , Terapia com Eletroporação , Coração
11.
Dalton Trans ; 52(25): 8620-8630, 2023 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-37284797

RESUMO

A series of new Me2Si-bridged cyclopentadiene/indene proligands {Me2Si(R2',5'2-R3',4'2-Cp)(R2,R4,R5,R6-Ind)H2} (1a-j) with various substitutions both on the indene and cyclopentadiene moieties was prepared. The corresponding C1-symmetric group 4 ansa-metallocene complexes (M = Zr, Hf), namely, {Me2Si(Me4Cp)(Ind)}ZrCl2 (2a-Zr), {Me2Si(Me4Cp)(2-Me,4-Ph-Ind)}MCl2 (2b-M), {Me2Si(Me4Cp)(2-Me,4-Ph,6-tBu-Ind)}ZrCl2 (2c-Zr), {Me2Si(Me4Cp)(2-Me,4-Ph,5-OMe,6-tBu-Ind)}MCl2 (2d-M), {Me2Si(Me4Cp)(2-R',4-(3',5'-tBu2,4'-OMe-C6H2),5-OMe,6-tBu-Ind)}ZrCl2, R' = Me (2e-Zr), R' = Et (2f-Zr), {Me2Si(2,5-Ph2-3,4-Me2-Cp)(2-Me,4-(3',5'-tBu2,4'-OMe-C6H2),5-OMe,6-tBu-Ind)}ZrCl2 (2g-Zr), {Me2Si(Me4Cp)(2-Me,4-(3',6'-tBu2-carbazol-4'-yl)-Ind)}ZrCl2 (2h-Zr), {Me2Si(2,5-Me2,3,4-iPr2-Cp)(2-Me,4-Ph-Ind)}ZrCl2 (2i-Zr), {Me2Si(2,5-Me2,3,4-iPr2-Cp)(2-Me,4-Ph,6-tBu-Ind)}ZrCl2 (2j-Zr) and {Me2Si(Me4Cp)(2-Me-4,5-[a]anthracene-Ind)}MCl2 (2k-Zr) were synthesized and characterized by NMR spectroscopy and mass spectrometry. The solid-state molecular structures of 2b-Zr, 2d-Zr, 2e-Zr, 2f-Zr, 2j-Zr and 2k-Zr were determined by X-ray crystallography. The zirconocene complexes, once activated with MAO in toluene solution, exhibited propylene polymerization activities at 60 °C up to 161 000 kg(PP) mol(Zr)-1 h-1, affording highly isotactic polypropylenes (iPP) with [m]4 up to 96.5% and Tm up to 157 °C. Also, metallocene complexes 2b-e-Zr were supported on SiO2-MAO and evaluated in slurry bulk propylene polymerization at 70 °C, producing iPPs with [m]4 = 91.7-96.6 mol% and low regiodefects (0.2-0.3 mol%) content, with productivities up to 636 000 kg(PP) mol(Zr)-1 h-1. DFT calculations allowed rationalizing a polymerization reaction mechanism occurring through "chain-stationary" enchainment with preference for 1,2-insertions.

12.
Opt Express ; 31(10): 15392-15408, 2023 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-37157642

RESUMO

Imaging at H Ly-α (121.6 nm), among other spectral lines in the short far UV (FUV), is of high interest for astrophysics, solar, and atmosphere physics, since this spectral line is ubiquitously present in space observations. However, the lack of efficient narrowband coatings has mostly prevented such observations. Present and future space observatories like GLIDE and the IR/O/UV NASA concept, among other applications, can benefit from the development of efficient narrowband coatings at Ly-α. The current state of the art of narrowband FUV coatings lacks performance and stability for coatings that peak at wavelengths shorter than ∼135 nm. We report highly reflective AlF3/LaF3 narrowband mirrors at Ly-α prepared by thermal evaporation, with, to our knowledge, the highest reflectance (over 80%) of a narrowband multilayer at such a short wavelength obtained so far. We also report a remarkable reflectance after several months of storage in different environments, including relative humidity levels above 50%. For astrophysics targets in which Ly-α may mask a close spectral line, such as in the search for biomarkers, we present the first coating in the short FUV for imaging at the OI doublet (130.4 and 135.6 nm), with the additional requirement of rejecting the intense Ly-α, which might mask the OI observations. Additionally, we present coatings with the symmetric design, aimed to observe at Ly-α, and reject the strong OI geocoronal emission, that could be of interest for atmosphere observations.

13.
Proc Natl Acad Sci U S A ; 120(22): e2303480120, 2023 05 30.
Artigo em Inglês | MEDLINE | ID: mdl-37216519

RESUMO

Metacaspases are part of an evolutionarily broad family of multifunctional cysteine proteases, involved in disease and normal development. As the structure-function relationship of metacaspases remains poorly understood, we solved the X-ray crystal structure of an Arabidopsis thaliana type II metacaspase (AtMCA-IIf) belonging to a particular subgroup not requiring calcium ions for activation. To study metacaspase activity in plants, we developed an in vitro chemical screen to identify small molecule metacaspase inhibitors and found several hits with a minimal thioxodihydropyrimidine-dione structure, of which some are specific AtMCA-IIf inhibitors. We provide mechanistic insight into the basis of inhibition by the TDP-containing compounds through molecular docking onto the AtMCA-IIf crystal structure. Finally, a TDP-containing compound (TDP6) effectively hampered lateral root emergence in vivo, probably through inhibition of metacaspases specifically expressed in the endodermal cells overlying developing lateral root primordia. In the future, the small compound inhibitors and crystal structure of AtMCA-IIf can be used to study metacaspases in other species, such as important human pathogens, including those causing neglected diseases.


Assuntos
Arabidopsis , Caspases , Humanos , Caspases/química , Simulação de Acoplamento Molecular , Apoptose , Proteínas de Ligação a DNA
14.
Free Radic Biol Med ; 200: 117-129, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36870374

RESUMO

Alternative splicing is a key posttranscriptional gene regulatory process, acting in diverse adaptive and basal plant processes. Splicing of precursor-messenger RNA (pre-mRNA) is catalyzed by a dynamic ribonucleoprotein complex, designated the spliceosome. In a suppressor screen, we identified a nonsense mutation in the Smith (Sm) antigen protein SME1 to alleviate photorespiratory H2O2-dependent cell death in catalase deficient plants. Similar attenuation of cell death was observed upon chemical inhibition of the spliceosome, suggesting pre-mRNA splicing inhibition to be responsible for the observed cell death alleviation. Furthermore, the sme1-2 mutants showed increased tolerance to the reactive oxygen species inducing herbicide methyl viologen. Both an mRNA-seq and shotgun proteomic analysis in sme1-2 mutants displayed a constitutive molecular stress response, together with extensive alterations in pre-mRNA splicing of transcripts encoding metabolic enzymes and RNA binding proteins, even under unstressed conditions. Using SME1 as a bait to identify protein interactors, we provide experimental evidence for almost 50 homologs of the mammalian spliceosome-associated protein to reside in the Arabidopsis thaliana spliceosome complexes and propose roles in pre-mRNA splicing for four uncharacterized plant proteins. Furthermore, as for sme1-2, a mutant in the Sm core assembly protein ICLN resulted in a decreased sensitivity to methyl viologen. Taken together, these data show that both a perturbed Sm core composition and assembly results in the activation of a defense response and in enhanced resilience to oxidative stress.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Animais , Arabidopsis/genética , Arabidopsis/metabolismo , Precursores de RNA/genética , Precursores de RNA/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Peróxido de Hidrogênio/farmacologia , Peróxido de Hidrogênio/metabolismo , Paraquat , Proteômica , Processamento Alternativo , Mutação , RNA Mensageiro/metabolismo , Estresse Oxidativo , Regulação da Expressão Gênica de Plantas , Mamíferos/metabolismo
15.
J Cell Biol ; 222(4)2023 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-36880595

RESUMO

ARF GTPases are central regulators of membrane trafficking that control local membrane identity and remodeling facilitating vesicle formation. Unraveling their function is complicated by the overlapping association of ARFs with guanine nucleotide exchange factors (GEFs), GTPase-activating proteins (GAPs), and numerous interactors. Through a functional genomic screen of three-dimensional (3D) prostate cancer cell behavior, we explore the contribution of ARF GTPases, GEFs, GAPs, and interactors to collective invasion. This revealed that ARF3 GTPase regulates the modality of invasion, acting as a switch between leader cell-led chains of invasion or collective sheet movement. Functionally, the ability of ARF3 to control invasion modality is dependent on association and subsequent control of turnover of N-cadherin. In vivo, ARF3 levels acted as a rheostat for metastasis from intraprostatic tumor transplants and ARF3/N-cadherin expression can be used to identify prostate cancer patients with metastatic, poor-outcome disease. Our analysis defines a unique function for the ARF3 GTPase in controlling how cells collectively organize during invasion and metastasis.


Assuntos
Fatores de Ribosilação do ADP , Proteínas Ativadoras de GTPase , Proteínas Monoméricas de Ligação ao GTP , Neoplasias da Próstata , Humanos , Masculino , Fatores de Ribosilação do ADP/genética , Caderinas/genética , Endocitose , Proteínas Ativadoras de GTPase/genética , Neoplasias da Próstata/genética
16.
Cell Death Differ ; 30(4): 1047-1058, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36755073

RESUMO

In plants, proteolysis is emerging as an important field of study due to a growing understanding of the critical involvement of proteases in plant cell death, disease and development. Because proteases irreversibly modify the structure and function of their target substrates, proteolytic activities are stringently regulated at multiple levels. Most proteases are produced as dormant isoforms and only activated in specific conditions such as altered ion fluxes or by post-translational modifications. Some of the regulatory mechanisms initiating and modulating proteolytic activities are restricted in time and space, thereby ensuring precision activity, and minimizing unwanted side effects. Currently, the activation mechanisms and the substrates of only a few plant proteases have been studied in detail. Most studies focus on the role of proteases in pathogen perception and subsequent modulation of the plant reactions, including the hypersensitive response (HR). Proteases are also required for the maturation of coexpressed peptide hormones that lead essential processes within the immune response and development. Here, we review the known mechanisms for the activation of plant proteases, including post-translational modifications, together with the effects of proteinaceous inhibitors.


Assuntos
Endopeptidases , Peptídeo Hidrolases , Peptídeo Hidrolases/metabolismo , Endopeptidases/metabolismo , Proteólise , Plantas/metabolismo , Processamento de Proteína Pós-Traducional
17.
Sci Adv ; 9(5): eabq1858, 2023 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-36735782

RESUMO

The glycocalyx component and sialomucin podocalyxin (PODXL) is required for normal tissue development by promoting apical membranes to form between cells, triggering lumen formation. Elevated PODXL expression is also associated with metastasis and poor clinical outcome in multiple tumor types. How PODXL presents this duality in effect remains unknown. We identify an unexpected function of PODXL as a decoy receptor for galectin-3 (GAL3), whereby the PODXL-GAL3 interaction releases GAL3 repression of integrin-based invasion. Differential cortical targeting of PODXL, regulated by ubiquitination, is the molecular mechanism controlling alternate fates. Both PODXL high and low surface levels occur in parallel subpopulations within cancer cells. Orthotopic intraprostatic xenograft of PODXL-manipulated cells or those with different surface levels of PODXL define that this axis controls metastasis in vivo. Clinically, interplay between PODXL-GAL3 stratifies prostate cancer patients with poor outcome. Our studies define the molecular mechanisms and context in which PODXL promotes invasion and metastasis.


Assuntos
Glicocálix , Sialoglicoproteínas , Masculino , Humanos , Glicocálix/metabolismo , Sialoglicoproteínas/metabolismo , Xenoenxertos , Transplante Heterólogo
18.
Cir. Urug ; 7(1): e307, 2023. ilus
Artigo em Espanhol | LILACS, UY-BNMED, BNUY | ID: biblio-1505953

RESUMO

Los lipomas colónicos son tumores benignos poco frecuentes, extremadamente raros a nivel rectal. A pesar de ello, ocupan el segundo lugar en frecuencia detrás de los pólipos adenomatosos. Aunque la mayoría de los lipomas colorrectales son asintomáticos y se descubren incidentalmente, en ocasiones pueden ser sintomáticos y determinar complicaciones agudas que requieren tratamiento quirúrgico de urgencia. Estas formas de presentación pueden confundirse con las del cáncer colorrectal, constituyendo un desafío diagnóstico y terapéutico. Presentamos el caso de una paciente que consultó en emergencia por una gran masa dolorosa de aparición aguda a nivel del ano, resultando ser un lipoma rectal prolapsado a través del canal anal.


Colonic lipomas are infrequent benign tumors, extremely rare at the rectum. Nevertheless, they follow in frequency polyp adenomas. Even though most colorectal lipomas are asymptomatic and incidental, they can occasionally be symptomatic and develop acute complications that require urgent surgical treatment. This form of presentation can be confounded with colorectal cancer, therefore impairing diagnosis and treatment. We present the case of a female patient who came to the emergency room with an acute painful mass through the anus, that resulted in a prolapsed rectal lipoma.


Os lipomas colônicos são tumores benignos pouco frequentes, extremamente raros no nível retal. Apesar disso, ocupam o segundo lugar em frequência atrás dos pólipos adenomatosos. Embora a maioria dos lipomas colorretais sejam assintomáticos e descobertos incidentalmente, às vezes eles podem ser sintomáticos e levar a complicações agudas que requerem tratamento cirúrgico de emergência. Essas formas de apresentação podem ser confundidas com as do câncer colorretal, constituindo um desafio diagnóstico e terapêutico. Apresentamos o caso de um paciente que consultou na sala de emergência por uma grande massa dolorosa de início agudo ao nível do ânus, que acabou por ser um lipoma retal prolapsado pelo canal anal.


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Canal Anal/patologia , Neoplasias Retais/diagnóstico , Prolapso Retal/diagnóstico , Lipoma/diagnóstico , Canal Anal/cirurgia , Neoplasias Retais/cirurgia , Prolapso Retal/cirurgia , Dor do Câncer , Lipoma/cirurgia
20.
Artigo em Inglês | MEDLINE | ID: mdl-36360629

RESUMO

Whole-body electromyostimulation (WB-EMS) training is effective in improving training adaptation. However, WB-EMS may have side effects and contraindications that can lead to excessive muscle damage and physiological impairment. This randomized crossover study aimed to analyze the acute effects of WB-EMS on muscle damage, autonomic modulation and performance during a single maximal strength session in physically active participants. Twenty healthy and physically active participants randomly performed three maximal strength training sessions (90% 1RM) consisting of bench presses and squat exercises, with a continuous stimulus, a coordinated stimulus with concentric and eccentric phases, and without WB-EMS. Data showed no significant differences between the trials for muscle damage (blood creatine kinase levels), lactate blood levels and performance after exercise. Likewise, the heart rate, blood oxygen saturation and the rate of perceived exertion were similar during exercise between trials. The heart rate variability analysis also showed a similar autonomic response among the trials. Training with WB-EMS seemed to be safe at the observed time intervals while offering a stimulus similar to regular training in physically active participants, regardless of the delivery of the electrical stimuli. More studies are needed to assess the effectiveness of WB-EMS in improving exercise adaptations during training programs.


Assuntos
Terapia por Estimulação Elétrica , Treinamento Resistido , Humanos , Músculo Esquelético/fisiologia , Estudos Cross-Over , Terapia por Exercício , Força Muscular/fisiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...