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1.
Integr Comp Biol ; 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38992237

RESUMO

As global temperatures continue to rise, accurate predicted species distribution models will be important for forecasting the movement of range-shifting species. These predictions rely on measurements of organismal thermal tolerance, which can be measured using classical threshold concepts such as Arrhenius Break Temperatures and Critical Thermal Temperatures, or through ecologically relevant measurements-such as the temperature at which reproduction and growth occur. Many species, including invasive species, exhibit thermal plasticity, so these thresholds may change based on ambient temperature, life stage, and measurement techniques. Here, we review thermal thresholds for 15 invertebrate species invasive to the Gulf of Maine. The high degree of variability within a species and between applied conceptual frameworks suggests that modeling the future distribution of these species in all ecosystems, but especially in the rapidly warming Northwest Atlantic and Gulf of Maine, will be challenging. While each of these measurement techniques are valid, we suggest contextualization and integration of threshold measurements for accurate modeling.

2.
bioRxiv ; 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38853846

RESUMO

Microsporidia are single-celled intracellular parasites that cause opportunistic diseases in humans. Encephalitozoon intestinalis is a prevalent human-infecting species that invades the small intestine. Dissemination to other organ systems is also observed, and is potentially facilitated by macrophages. The macrophage response to infection and the developmental trajectory of the parasite are not well studied. Here we use single cell RNA sequencing to investigate transcriptional changes in both the host and parasite during infection. While a small population of infected macrophages mount a response, most remain transcriptionally unchanged, suggesting that the majority of parasites may avoid host detection. The parasite transcriptome reveals large transcriptional changes throughout the life cycle, providing a blueprint for parasite development. The stealthy microsporidian lifestyle likely allows these parasites to harness macrophages for replication and dissemination. Together, our data provide insights into the host response in primary human macrophages and the E. intestinalis developmental program.

3.
Cell ; 187(14): 3726-3740.e43, 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38861993

RESUMO

Many growth factors and cytokines signal by binding to the extracellular domains of their receptors and driving association and transphosphorylation of the receptor intracellular tyrosine kinase domains, initiating downstream signaling cascades. To enable systematic exploration of how receptor valency and geometry affect signaling outcomes, we designed cyclic homo-oligomers with up to 8 subunits using repeat protein building blocks that can be modularly extended. By incorporating a de novo-designed fibroblast growth factor receptor (FGFR)-binding module into these scaffolds, we generated a series of synthetic signaling ligands that exhibit potent valency- and geometry-dependent Ca2+ release and mitogen-activated protein kinase (MAPK) pathway activation. The high specificity of the designed agonists reveals distinct roles for two FGFR splice variants in driving arterial endothelium and perivascular cell fates during early vascular development. Our designed modular assemblies should be broadly useful for unraveling the complexities of signaling in key developmental transitions and for developing future therapeutic applications.


Assuntos
Diferenciação Celular , Fatores de Crescimento de Fibroblastos , Receptores de Fatores de Crescimento de Fibroblastos , Transdução de Sinais , Animais , Humanos , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Camundongos , Ligantes , Cálcio/metabolismo , Sistema de Sinalização das MAP Quinases
4.
Nat Commun ; 15(1): 4695, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38824138

RESUMO

Which isoforms of apolipoprotein E (apoE) we inherit determine our risk of developing late-onset Alzheimer's Disease (AD), but the mechanism underlying this link is poorly understood. In particular, the relevance of direct interactions between apoE and amyloid-ß (Aß) remains controversial. Here, single-molecule imaging shows that all isoforms of apoE associate with Aß in the early stages of aggregation and then fall away as fibrillation happens. ApoE-Aß co-aggregates account for ~50% of the mass of diffusible Aß aggregates detected in the frontal cortices of homozygotes with the higher-risk APOE4 gene. We show how dynamic interactions between apoE and Aß tune disease-related functions of Aß aggregates throughout the course of aggregation. Our results connect inherited APOE genotype with the risk of developing AD by demonstrating how, in an isoform- and lipidation-specific way, apoE modulates the aggregation, clearance and toxicity of Aß. Selectively removing non-lipidated apoE4-Aß co-aggregates enhances clearance of toxic Aß by glial cells, and reduces secretion of inflammatory markers and membrane damage, demonstrating a clear path to AD therapeutics.


Assuntos
Doença de Alzheimer , Peptídeos beta-Amiloides , Apolipoproteína E4 , Apolipoproteínas E , Doença de Alzheimer/metabolismo , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Humanos , Apolipoproteínas E/metabolismo , Apolipoproteínas E/genética , Animais , Apolipoproteína E4/metabolismo , Apolipoproteína E4/genética , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/genética , Camundongos , Feminino , Agregados Proteicos , Masculino , Agregação Patológica de Proteínas/metabolismo , Camundongos Transgênicos , Neuroglia/metabolismo
5.
bioRxiv ; 2024 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-38585835

RESUMO

In Mycobacterium tuberculosis proteins that are post-translationally modified with Pup, a prokaryotic ubiquitin-like protein, can be degraded by proteasomes. While pupylation is reversible, mechanisms regulating substrate specificity have not been identified. Here, we identify the first depupylation regulators: CoaX, a pseudokinase, and pantothenate, an essential, central metabolite. In a Δ coaX mutant, pantothenate synthesis enzymes were more abundant, including PanB, a substrate of the Pup-proteasome system. Media supplementation with pantothenate decreased PanB levels in a coaX and Pup-proteasome-dependent manner. In vitro , CoaX accelerated depupylation of Pup∼PanB, while addition of pantothenate inhibited this reaction. Collectively, we propose CoaX contributes to proteasomal degradation of PanB by modulating depupylation of Pup∼PanB in response to pantothenate levels. One Sentence Summary: A pseudo-pantothenate kinase regulates proteasomal degradation of a pantothenate synthesis enzyme in M. tuberculosis .

6.
Sci Total Environ ; 930: 172526, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38636866

RESUMO

Pesticide contamination poses a significant threat to non-target wildlife, including amphibians, many of which are already highly threatened. This study assessed the extent of pesticide exposure in dead frogs collected during a mass mortality event across eastern New South Wales, Australia between July 2021 and March 2022. Liver tissue from 77 individual frogs of six species were analysed for >600 legacy and contemporary pesticides, including rodenticides. More than a third (36 %) of the liver samples contained at least one of the following pesticides: brodifacoum, dieldrin, DDE, heptachlor/heptachlor epoxide, fipronil sulfone, and 2-methyl-4-chlorophenoxyacetic acid (MCPA). Brodifacoum, a second-generation anticoagulant rodenticide, was found in four of the six frog species analysed: the eastern banjo frog (Limnodynastes dumerilii), cane toad (Rhinella marina), green tree frog (Litoria caerulea) and Peron's tree frog (Litoria peronii). This is the first report of anticoagulant rodenticide detected in wild amphibians, raising concerns about potential impacts on frogs and extending the list of taxa shown to accumulate rodenticides. Dieldrin, a banned legacy pesticide, was also detected in two species: striped marsh frog (Limnodynastes peronii) and green tree frog (Litoria caerulea). The toxicological effects of these pesticides on frogs are difficult to infer due to limited comparable studies; however, due to the low frequency of detection the presence of these pesticides was not considered a major contributing factor to the mass mortality event. Additional research is needed to investigate the effects of pesticide exposure on amphibians, particularly regarding the impacts of second-generation anticoagulant rodenticides. There is also need for continued monitoring and improved conservation management strategies for the mitigation of the potential threat of pesticide exposure and accumulation in amphibian populations.


Assuntos
Anticoagulantes , Anuros , Monitoramento Ambiental , Praguicidas , Rodenticidas , Animais , Rodenticidas/análise , Anticoagulantes/análise , New South Wales , Austrália
7.
Heliyon ; 10(5): e26805, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38455564

RESUMO

Over the years, the release of potential radiological components around the oil exploration environment has increased with potential health implications.Yet; the mechanism and health associated assessment have remained fuzzy to most experimental scientists. The current study determines the activity concentration of radionuclides in sediments and the corresponding health risk assessments from the hydrocarbon exploration environment of the southern coastal area of Delta State, Nigeria. A Sodium-iodide NaI(Tl) detector, with a well-calibrated multichannel analyzer (MCA) to ensure efficiency and energy was utilized. A total of seventy-five sediment samples (Five sediment samples each per community) were collected from the southern coastal area of Delta State, Nigeria. The mean activity concentrations of 40K, 238U, and 232Th of the sediment samples were 3361.48 ± 194.26 Bqkg-1, 40.11 ± 16.17 Bqkg-1, and 45.73 ± 19.27 Bqkg-1 respectively. The obtained mean values exceeded the world standard limit of 400 Bqkg-1, 35 Bqkg-1, and 30 Bqkg-1 respectively. Also, the computed mean radiological health hazard risk of radium equivalent activity (Raeq), representative level index (Iyr), external hazard index (Hex), internal hazard index (Hin), absorbed gamma dose rate (D), annual effective dose equivalent outdoor and indoor (AEDE) and lifetime cancer risk (ELCR) values are 363.94 ± 32.37 Bkgl-1, 2.9657 Bkgl-1, 0.9839, 1.0919, 175.82 nGyh-1, 2.1556 mSvyr-1, 0.8625 mSvyr-1, and 7.5447 mSvyr-1 respectively. The values were found to be slightly higher than the world standard limit. Therefore, the residents that are using the sediments of the southern coastal area for the construction of buildings as well as dwelling in houses built with such sediments are exposed to these radiological materials. This may pose a radiological health risk concern. The obtained results will serve as radiation and radiological baseline data for sediments of the southern coastal area of Delta State, Nigeria.

8.
Sci Total Environ ; 926: 171743, 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38494020

RESUMO

Per- and poly-fluoroalkyl substances (PFAS) pose a threat to organisms and ecosystems due to their persistent nature. Ecotoxicology endpoints used in regulatory guidelines may not reflect multiple, low-level but persistent stressors. This study examines the biological effects of PFAS on Eastern short-necked turtles in Queensland, Australia. In this study, blood samples were collected and analysed for PFAS, hormone levels, and functional omics endpoints. High levels of PFAS were found in turtles at the impacted site, with PFOS being the dominant constituent. The PFAS profiles of males and females differed, with males having higher PFAS concentrations. Hormone concentrations differed between impacted and reference sites in male turtles, with elevated testosterone and corticosterone indicative of stress. Further, energy utilisation, nucleotide synthesis, nitrogen metabolism, and amino acid synthesis were altered in both male and female turtles from PFAS-impacted sites. Both sexes show similar metabolic responses to environmental stressors from the PFAS-contaminated site, which may adversely affect their reproductive fitness. Purine metabolism, caffeine metabolism, and ferroptosis pathway changes in turtles can cause gout, cell death, and overall health problems. Further, the study showed that prolonged exposure to elevated PFAS levels in the wild could compromise turtle reproductive fitness by disrupting reproductive steroids and metabolic pathways.


Assuntos
Ácidos Alcanossulfônicos , Poluentes Ambientais , Fluorocarbonos , Tartarugas , Animais , Masculino , Feminino , Ecossistema , Aptidão Genética , Água Doce , Hormônios , Fluorocarbonos/toxicidade
9.
Int J Mol Sci ; 25(5)2024 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-38473716

RESUMO

Despite the well-known relevance of polyamines to many forms of life, little is known about how polyamines regulate osteogenesis and skeletal homeostasis. Here, we report a series of in vitro studies conducted with human-bone-marrow-derived pluripotent stromal cells (MSCs). First, we show that during osteogenic differentiation, mRNA levels of most polyamine-associated enzymes are relatively constant, except for the catabolic enzyme spermidine/spermine N1-acetyltransferase 1 (SAT1), which is strongly increased at both mRNA and protein levels. As a result, the intracellular spermidine to spermine ratio is significantly reduced during the early stages of osteoblastogenesis. Supplementation of cells with exogenous spermidine or spermine decreases matrix mineralization in a dose-dependent manner. Employing N-cyclohexyl-1,3-propanediamine (CDAP) to chemically inhibit spermine synthase (SMS), the enzyme catalyzing conversion of spermidine into spermine, also suppresses mineralization. Intriguingly, this reduced mineralization is rescued with DFMO, an inhibitor of the upstream polyamine enzyme ornithine decarboxylase (ODC1). Similarly, high concentrations of CDAP cause cytoplasmic vacuolization and alter mitochondrial function, which are also reversible with the addition of DFMO. Altogether, these studies suggest that excess polyamines, especially spermidine, negatively affect hydroxyapatite synthesis of primary MSCs, whereas inhibition of polyamine synthesis with DFMO rescues most, but not all of these defects. These findings are relevant for patients with Snyder-Robinson syndrome (SRS), as the presenting skeletal defects-associated with SMS deficiency-could potentially be ameliorated by treatment with DFMO.


Assuntos
Células-Tronco Mesenquimais , Espermidina , Humanos , Espermidina/metabolismo , Espermina/metabolismo , Espermina Sintase/genética , Ornitina Descarboxilase/metabolismo , Osteogênese , Poliaminas/metabolismo , Células-Tronco Mesenquimais/metabolismo , RNA Mensageiro
10.
Arch Environ Contam Toxicol ; 86(2): 178-186, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38383776

RESUMO

There is growing recognition of the threat posed to wildlife by pollutants. Waterbirds are robust bioindicators of ecosystem health, and metal toxicity is a threat to these species in waterways worldwide. Urban waterbirds are likely to be at the highest risk of heavy metal exposure, but this issue has not been widely explored in Australia. Our aim was to estimate contemporary heavy metal exposure in a sedentary urban waterbird population: black swans (Cygnus atratus) inhabiting an inner-city wetland in one of Australia's largest cities, Melbourne. To investigate the physiological implications of legacy heavy metal exposure in these birds, we quantified blood biochemistry profiles and examined their relationships with metal concentrations in feathers. We caught 15 swans in 2021 and took feather samples to measure the concentration of eight heavy metals (chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), lead (Pb), and mercury (Hg)), and blood samples to measure the concentration of 13 plasma analytes. Multivariate regression analysis revealed few associations between heavy metals and biochemistry markers, and no differences between sexes or age classes. This study presents a baseline dataset of these contaminants and blood biochemical profiles of swans at this wetland that can be used for future monitoring and is an important step toward a better understanding of the threat posed by heavy metals to Australian urban waterbirds.


Assuntos
Mercúrio , Metais Pesados , Animais , Humanos , Ecossistema , População Urbana , Austrália , Metais Pesados/análise , Zinco/análise , Mercúrio/análise , Monitoramento Ambiental
11.
Elife ; 122024 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-38381133

RESUMO

Microsporidia are eukaryotic, obligate intracellular parasites that infect a wide range of hosts, leading to health and economic burdens worldwide. Microsporidia use an unusual invasion organelle called the polar tube (PT), which is ejected from a dormant spore at ultra-fast speeds, to infect host cells. The mechanics of PT ejection are impressive. Anncaliia algerae microsporidia spores (3-4 µm in size) shoot out a 100-nm-wide PT at a speed of 300 µm/s, creating a shear rate of 3000 s-1. The infectious cargo, which contains two nuclei, is shot through this narrow tube for a distance of ∼60-140 µm (Jaroenlak et al, 2020) and into the host cell. Considering the large hydraulic resistance in an extremely thin tube and the low-Reynolds-number nature of the process, it is not known how microsporidia can achieve this ultrafast event. In this study, we use Serial Block-Face Scanning Electron Microscopy to capture 3-dimensional snapshots of A. algerae spores in different states of the PT ejection process. Grounded in these data, we propose a theoretical framework starting with a systematic exploration of possible topological connectivity amongst organelles, and assess the energy requirements of the resulting models. We perform PT firing experiments in media of varying viscosity, and use the results to rank our proposed hypotheses based on their predicted energy requirement. We also present a possible mechanism for cargo translocation, and quantitatively compare our predictions to experimental observations. Our study provides a comprehensive biophysical analysis of the energy dissipation of microsporidian infection process and demonstrates the extreme limits of cellular hydraulics.


Assuntos
Anatomia Regional , Núcleo Celular , Biofísica
12.
Heliyon ; 10(2): e24344, 2024 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-38298659

RESUMO

The continued increase in the global population has resulted in increased water demand for domestic, agricultural, and industrial purposes. These activities have led to the generation of high volumes of wastewater, which has an impact on water quality. Consequently, more practical solutions are needed to improve the current wastewater treatment systems. The use of improved ceramic membranes for wastewater treatment holds significant prospects for advancement in water treatment and sanitation. Hence, different studies have employed ceramic membranes in wastewater treatment and the search for low-cost and environmentally friendly starting materials has continued to engender research interests. This review focuses on the application of coal fly ash in membrane technology for wastewater treatment. The processes of membrane fabrication and the various limitations of the material. Several factors that influence the properties and performance of coal fly ash ceramic membranes in wastewater treatment are also presented. Some possible solutions to the limitations are also proposed, while cost analysis of coal fly ash-based membranes is explored to evaluate its potential for large-scale applications.

13.
Int Orthop ; 48(1): 261-265, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37938321

RESUMO

PURPOSE: Intramedullary nail is the gold standard in the management of long bone diaphyseal fractures of tibia and femur. The jig of these nails has corresponding extraction device whose pitch for nail coupling come in various sizes. This unlike plate and screws may be difficult to predict preoperatively and may pose a problem during removal. Difficulties in removal may arise due to the proliferation of nail brands especially in developing countries. The study aims to identify the incidence of extraction device mismatch among orthopaedic surgeons in Nigeria as well as the indications and complications associated with intramedullary nail removal. METHODS: A two-page questionnaire was administered to 87 orthopaedic surgeons attending the Annual General Meeting of the Nigerian Medical Association. The attitudes of the participants towards intramedullary nail were assessed. RESULTS: All participants agree to asymptomatic removal. Patients wish was the leading indication for asymptomatic removal among the participants. Sixty-one percent of the surgeons have had the need to remove a nail different from the brand in the hospital their practice. The commonest indication for symptomatic removal was infections. Forty-seven percent of the participant encountered nail extraction-device mismatch. CONCLUSIONS: The incidence of extraction device mismatch may portend a public health issue. There may be need for patient who had intra medullary nail insertion to be told their brand. We advocate for standardization of extraction device pitch for intramedullary nail.


Assuntos
Pinos Ortopédicos , Fixação Intramedular de Fraturas , Humanos , Pinos Ortopédicos/efeitos adversos , Fixação Intramedular de Fraturas/efeitos adversos , Países em Desenvolvimento , Fixação Interna de Fraturas , Tíbia , Remoção de Dispositivo/efeitos adversos
14.
Rev Sci Instrum ; 94(8)2023 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38065140

RESUMO

This report details the analyses and related uncertainties in measuring longitudinal-stress-density paths in indirect laser-driven ramp equation-of-state (EOS) experiments [Smith et al., Nat. Astron. 2(6), 452-458 (2018); Smith et al., Nature 511(7509), 330-333 (2014); Fratanduono et al., Science 372(6546), 1063-1068 (2021); and Fratanduono et al., Phys. Rev. Lett. 124(1), 015701 (2020)]. Experiments were conducted at the National Ignition Facility (NIF) located at the Lawrence Livermore National Laboratory. The NIF can deliver up to 2 MJ of laser energy over 30 ns and provide the necessary laser power and control to ramp compress materials to TPa pressures (1 TPa = 10 × 106 atmospheres). These data provide low-temperature solid-state EOS data relevant to the extreme conditions found in the deep interiors of giant planets. In these experiments, multi-stepped samples with thicknesses in the range of 40-120 µm experience an initial shock compression followed by a time-dependent ramp compression to peak pressure. Interface velocity measurements from each thickness combine to place a constraint on the Lagrangian sound speed as a function of particle velocity, which in turn allows for the determination of a continuous stress-density path to high levels of compressibility. In this report, we present a detailed description of the experimental techniques and measurement uncertainties and describe how these uncertainties combine to place a final uncertainty in both stress and density. We address the effects of time-dependent deformation and the sensitivity of ramp EOS techniques to the onset of phase transformations.

15.
Heliyon ; 9(11): e21423, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38027928

RESUMO

Fabrication of heterojunction and surface defective engineering, through the formation of oxygen vacancies, are among the various photocatalytic enhancement techniques. A combination of these techniques has the prospect of enhancing photocatalytic activities through improved light absorption capabilities and charge separation process of the photocatalysts. In this study, a heterojunction of black titanium oxide-zinc oxide (BTiO2-ZnO) nanocomposite was synthesized using the conventional sol-gel approach, coupled with aluminum foil-assisted NaBH4 reduction. The structure, morphology, surface properties, and optical characteristics of the synthesized material were studied using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), UV-vis absorption spectra, scanning electron microscope (SEM), Energy-dispersive X-ray spectroscopy (EDS), and transmission electron microscope (TEM). The XRD confirmed the successful formation of BTiO2-ZnO heterostructure, while SEM revealed the structural morphology as pseudo-spherical with slight agglomeration. BTiO2-ZnO was found to be more efficient than BTiO2 and BZnO for the removal of tetracycline with degradation efficiencies of 63, 58, and 56 % respectively. The effects of process parameters such as the amount of photocatalyst, pollutant's concentration, and the initial solution pH on photocatalytic degradation study were systematically explored. The results confirm that the formation of the heterostructure from BTiO2 and BZnO could offer a facile route to improving the catalytic degradation of tetracycline. Therefore, this study offers a novel perspective on the design of efficient metal oxide photocatalyst systems that rely on the integration of defect engineering and heterojunction for the removal of organic contaminants.

16.
Nat Commun ; 14(1): 7662, 2023 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-37996434

RESUMO

Microsporidia are an early-diverging group of fungal pathogens with a wide host range. Several microsporidian species cause opportunistic infections in humans that can be fatal. As obligate intracellular parasites with highly reduced genomes, microsporidia are dependent on host metabolites for successful replication and development. Our knowledge of microsporidian intracellular development remains rudimentary, and our understanding of the intracellular niche occupied by microsporidia has relied on 2D TEM images and light microscopy. Here, we use serial block-face scanning electron microscopy (SBF-SEM) to capture 3D snapshots of the human-infecting species, Encephalitozoon intestinalis, within host cells. We track E. intestinalis development through its life cycle, which allows us to propose a model for how its infection organelle, the polar tube, is assembled de novo in developing spores. 3D reconstructions of parasite-infected cells provide insights into the physical interactions between host cell organelles and parasitophorous vacuoles, which contain the developing parasites. The host cell mitochondrial network is substantially remodeled during E. intestinalis infection, leading to mitochondrial fragmentation. SBF-SEM analysis shows changes in mitochondrial morphology in infected cells, and live-cell imaging provides insights into mitochondrial dynamics during infection. Our data provide insights into parasite development, polar tube assembly, and microsporidia-induced host mitochondria remodeling.


Assuntos
Encephalitozoon , Microsporídios , Parasitos , Animais , Humanos , Imageamento Tridimensional
17.
bioRxiv ; 2023 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-37873249

RESUMO

The outer membrane of Gram-negative bacteria provides a formidable barrier, essential for both pathogenesis and antimicrobial resistance. Biogenesis of the outer membrane requires the transport of phospholipids across the cell envelope. Recently, YhdP was implicated as a major protagonist in the transport of phospholipids from the inner membrane to the outer membrane however the molecular mechanism of YhdP mediated transport remains elusive. Here, utilising AlphaFold, we observe YhdP to form an elongated assembly of 60 ß strands that curve to form a continuous hydrophobic groove. This architecture is consistent with our negative stain electron microscopy data which reveals YhdP to be approximately 250 Å in length and thus sufficient to span the bacterial cell envelope. Furthermore, molecular dynamics simulations and in vivo bacterial growth assays indicate essential helical regions at the N- and C-termini of YhdP, that may embed into the inner and outer membranes respectively, reinforcing its envelope spanning nature. Our in vivo crosslinking data reveal phosphate-containing substrates captured along the length of the YhdP groove, providing direct evidence that YhdP transports phospholipids. This finding is congruent with our molecular dynamics simulations which demonstrate the propensity for inner membrane lipids to spontaneously enter the groove of YhdP. Collectively, our results support a model in which YhdP bridges the cell envelope, providing a hydrophobic environment for the transport of phospholipids to the outer membrane.

18.
J Biol Chem ; 299(12): 105321, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37802313

RESUMO

Staphylococcus aureus (S. aureus) is a serious global pathogen that causes a diverse range of invasive diseases. S. aureus utilizes a family of pore-forming toxins, known as bi-component leukocidins, to evade the host immune response and promote infection. Among these is LukAB (leukocidin A/leukocidin B), a toxin that assembles into an octameric ß-barrel pore in the target cell membrane, resulting in host cell death. The established cellular receptor for LukAB is CD11b of the Mac-1 complex. Here, we show that hydrogen voltage-gated channel 1 is also required for the cytotoxicity of all major LukAB variants. We demonstrate that while each receptor is sufficient to recruit LukAB to the plasma membrane, both receptors are required for maximal lytic activity. Why LukAB requires two receptors, and how each of these receptors contributes to pore-formation remains unknown. To begin to resolve this, we performed an alanine scanning mutagenesis screen to identify mutations that allow LukAB to maintain cytotoxicity without CD11b. We discovered 30 mutations primarily localized in the stem domains of LukA and LukB that enable LukAB to exhibit full cytotoxicity in the absence of CD11b. Using crosslinking, electron microscopy, and hydroxyl radical protein footprinting, we show these mutations increase the solvent accessibility of the stem domain, priming LukAB for oligomerization. Together, our data support a model in which CD11b binding unlatches the membrane penetrating stem domains of LukAB, and this change in flexibility promotes toxin oligomerization.


Assuntos
Proteínas de Bactérias , Leucocidinas , Staphylococcus aureus , Toxinas Biológicas , Humanos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Membrana Celular/metabolismo , Leucocidinas/genética , Leucocidinas/metabolismo , Leucocidinas/toxicidade , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Staphylococcus aureus/patogenicidade , Toxinas Biológicas/metabolismo , Mutação , Ligação Proteica/genética , Domínios Proteicos , Linhagem Celular , Células CHO , Cricetulus , Animais
19.
iScience ; 26(10): 107935, 2023 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-37841587

RESUMO

The halide lead perovskite single crystals (HLPSCs) have great potential in gamma-ray detection with high attenuation coefficient, strong defects tolerance, and large mobility-lifetime product. However, mobile halide ions would migrate under high external bias, which would both weaken the gamma-ray response and cause additional noise. Here, we report the gamma-ray PIN photodiodes made of cascade HLPSCs including both ion-formed and electron-hole-formed electrical junctions that could suppress the ions migration and improve the charges collection. Our photodiodes based on cascade HLPSCs (MAPbBr3/MAPbBr2.5Cl0.5/MAPbCl3) show a wide halide-ion-formed depletion layer of ∼52 µm. The built-in potential along the wide ionic-formed junction ensures a high mobility-lifetime product of 1.1 × 10-2 cm2V-1. As a result, our gamma-ray PIN photodiodes exhibit compelling response to 241Am, 137Cs, and 60Co; the energy resolution can reach 9.4%@59.5keV and 5.9%@662keV, respectively. This work provides a new path toward constructing high-performance gamma-ray detectors based on HLPSCs.

20.
PLoS One ; 18(9): e0290707, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37756252

RESUMO

Most vertebrate offspring must transition from the relative security of parental care (nutrition and protection) to independent foraging. Offspring face many challenges during this critical period, particularly in species where parental care ends at weaning, such as the grey seal (Halichoerus grypus). We studied the development of movement behaviour in naïve grey seal pups from their first trips to sea to about five months of age. Twenty-five (12 males and 13 females) newly-weaned pups were fitted with satellite-linked GPS tags on Sable Island, Nova Scotia, Canada in January 2016. The influence of fixed effects (pup size, sex, week) and the random effect of pup identity on trip characteristics were examined. Movement behaviour was analyzed using a move persistence mixed-effects model. Habitat use was highly variable among individuals and covered much of the geographic distribution of the population. Unlike older juveniles, subadults, and adults in this population, most naïve pups used multiple haulout sites to begin and end trips. There was little evidence of area-restricted search behaviour during trips, suggesting that naïve pups were using an opportunistic foraging tactic that may result in more variable foraging success than that of older, experienced animals. Naïve pups made longer trips with longer haulout durations between them than observed for older greys seals. Males and females differed in some trip characteristics, but sex effects were small over the first few months of life. Offspring size at weaning was not a useful predictor of trip characteristics. Move persistence of grey seal pups was initially high and then decreased over time as individuals gained experience. Both intrinsic and extrinsic factors were influential on the movements of grey seal pups. Greater body length at weaning, longer duration spent on shore after weaning, shallower water column depth, and farther distance from shore were all associated with lower move persistence. Female grey seal pups had lower move persistence than males. Overall, the movements of naïve grey seal pups during the first few months of life were characterized by extensive exploration, but move persistence decreased over time suggesting they may be using an exploration-refinement foraging tactic.


Assuntos
Transtorno do Espectro Autista , Mustelidae , Focas Verdadeiras , Animais , Feminino , Masculino , Estatura , Ecossistema , Nova Escócia
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