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1.
Front Mol Biosci ; 11: 1399225, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38962283

RESUMO

Periostin is a matricellular protein encoded by the POSTN gene that is alternatively spliced to produce ten different periostin isoforms with molecular weights ranging from 78 to 91 kDa. It is known to promote fibrillogenesis, organize the extracellular matrix, and bind integrin-receptors to induce cell signaling. As well as being a key component of the wound healing process, it is also known to participate in the pathogenesis of different diseases including atopic dermatitis, asthma, and cancer. In both health and disease, the functions of the different periostin isoforms are largely unknown. The ability to precisely determine the isoform profile of a given human sample is fundamental for characterizing their functional significance. Identification of periostin isoforms is most often carried out at the transcriptional level using RT-PCR based approaches, but due to high sequence homogeneity, identification on the protein level has always been challenging. Top-down proteomics, where whole proteins are measured by mass spectrometry, offers a fast and reliable method for isoform identification. Here we present a fully developed top-down mass spectrometry assay for the characterization of periostin splice isoforms at the protein level.

2.
Cogn Emot ; : 1-12, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38953391

RESUMO

Previous research on emotion-induced blindness (EIB) argues emotional distractors capture attention in a bottom-up manner due to their physical and emotional salience. However, recent research has shown it is controversial whether EIB will be modulated by top-down factors. The present study further investigated whether the magnitude of EIB would be modulated by top-down factors, specifically the emotional relevance between tasks and distractors. Participants were divided into two groups having the same targets except for different task instructions. The orientation judgment group was asked to judge the orientation of the target (an emotionally irrelevant task), and the emotion judgment group was required to judge the emotional valence of the target (an emotionally relevant task). It was found the emotional relevance between tasks and distractors has no modulation on the magnitudes of EIB in two groups when targets and distractors are from different categories (Experiment 1), but a modulation when they are from the same category (Experiment 2). Consequently, we contend top-down task relevance modulates the EIB effect and distractors' priority is regulated by the emotional relevance between tasks and distractors. The current study holds attentional capture by stimulus-driven is unconditional in EIB, while attentional capture by goal-driven requires certain conditions.

3.
Proc Biol Sci ; 291(2026): 20240868, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38955327

RESUMO

Biotic interactions play a critical role in shaping patterns of global biodiversity. While several macroecological studies provide evidence for stronger predation in tropical regions compared with higher latitudes, results are variable even within the tropics, and the drivers of this variability are not well understood. We conducted two complementary standardized experiments on communities of sessile marine invertebrate prey and their associated predators to test for spatial and seasonal differences in predation across the tropical Atlantic and Pacific coastlines of Panama. We further tested the prediction that higher predator diversity contributes to stronger impacts of predation, using both direct observations of predators and data from extensive reef surveys. Our results revealed substantially higher predation rates and stronger effects of predators on prey in the Pacific than in the Atlantic, demonstrating striking variation within tropical regions. While regional predator diversity was high in the Atlantic, functional diversity at local scales was markedly low. Peak predation strength in the Pacific occurred during the wet, non-upwelling season when ocean temperatures were warmer and predator communities were more functionally diverse. Our results highlight the importance of regional biotic and abiotic drivers that shape interaction strength and the maintenance of tropical communities, which are experiencing rapid environmental change.


Assuntos
Cadeia Alimentar , Comportamento Predatório , Estações do Ano , Clima Tropical , Animais , Biodiversidade , Panamá , Oceano Atlântico , Oceano Pacífico , Invertebrados/fisiologia
4.
Schizophr Bull ; 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38982879

RESUMO

BACKGROUND: Various neurocognitive models explore perceptual distortions and hallucinations in schizophrenia and the general population. A variant of predictive coding account suggests that strong priors, like cognitive expectancy, may influence perception. This study examines if stronger cognitive expectancies result in more auditory false percepts in clinical and healthy control groups, investigates group differences, and explores the association between false percepts and hallucinations. STUDY DESIGN: Patients diagnosed with schizophrenia with current auditory hallucinations (n = 51) and without hallucinations (n = 66) and healthy controls (n = 51) underwent the False Perception Task under various expectancy conditions. All groups were examined for the presence and severity of hallucinations or hallucinatory-like experiences. STUDY RESULTS: We observed a main effect of condition across all groups, ie, the stronger the cognitive expectancy, the greater the ratio of auditory false percepts. However, there was no group effect for the ratio of auditory false percepts. Despite modest pairwise correlations in the hallucinating group, the ratio of auditory false percepts was not predicted by levels of hallucinations and hallucinatory-like experiences in a linear mixed model. CONCLUSIONS: The current study demonstrates that strong priors in the form of cognitive expectancies affect perception and play a role in perceptual disturbances. There is also a tentative possibility that overreliance on strong priors may be associated with hallucinations in currently hallucinating subjects. Possible, avoidable confounding factors are discussed in detail.

5.
J Proteome Res ; 23(7): 2315-2322, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38913967

RESUMO

Native top-down mass spectrometry (nTDMS) allows characterization of protein structure and noncovalent interactions with simultaneous sequence mapping and proteoform characterization. The majority of nTDMS studies utilize purified recombinant proteins, with significant challenges hindering application to endogenous systems. To perform native top-down proteomics (nTDP), where endogenous proteins from complex biological systems are analyzed by nTDMS, it is essential to separate proteins under nondenaturing conditions. However, it remains difficult to achieve high resolution with MS-compatible online chromatography while preserving protein tertiary structure and noncovalent interactions. Herein, we report the use of online mixed-bed ion exchange chromatography (IEC) to enable separation of endogenous proteins from complex mixtures under nondenaturing conditions, preserving noncovalent interactions for nTDP analysis. We have successfully detected large proteins (>146 kDa) and identified endogenous metal-binding and oligomeric protein complexes in human heart tissue lysate. The use of a mixed-bed stationary phase allowed retention and elution of proteins over a wide range of isoelectric points without altering the sample or mobile phase pH. Overall, our method provides a simple online IEC-MS platform that can effectively separate proteins from complex mixtures under nondenaturing conditions and preserve higher-order structure for nTDP applications.


Assuntos
Proteômica , Cromatografia por Troca Iônica/métodos , Humanos , Proteômica/métodos , Miocárdio/química , Espectrometria de Massas/métodos , Misturas Complexas/química , Proteínas/química , Proteínas/análise , Proteínas/isolamento & purificação
6.
Animals (Basel) ; 14(12)2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38929393

RESUMO

Poultry managers can better understand the state of poultry through poultry behavior analysis. As one of the key steps in behavior analysis, the accurate estimation of poultry posture is the focus of this research. This study mainly analyzes a top-down pose estimation method of multiple chickens. Therefore, we propose the "multi-chicken pose" (MCP), a pose estimation system for multiple chickens through deep learning. Firstly, we find the position of each chicken from the image via the chicken detector; then, an estimate of the pose of each chicken is made using a pose estimation network, which is based on transfer learning. On this basis, the pixel error (PE), root mean square error (RMSE), and image quantity distribution of key points are analyzed according to the improved chicken keypoint similarity (CKS). The experimental results show that the algorithm scores in different evaluation metrics are a mean average precision (mAP) of 0.652, a mean average recall (mAR) of 0.742, a percentage of correct keypoints (PCKs) of 0.789, and an RMSE of 17.30 pixels. To the best of our knowledge, this is the first time that transfer learning has been used for the pose estimation of multiple chickens as objects. The method can provide a new path for future poultry behavior analysis.

7.
Ecology ; 105(7): e4334, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38887829

RESUMO

Ecological theory predicts that kelp forests structured by trophic cascades should experience recovery and persistence of their foundation species when herbivores become rare. Yet, climate change may be altering the outcomes of top-down forcing in kelp forests, especially those located in regions that have rapidly warmed in recent decades, such as the Gulf of Maine. Here, using data collected annually from 30+ sites spanning >350 km of coastline, we explored the dynamics of Maine's kelp forests in the ~20 years after a fishery-induced elimination of sea urchin herbivores. Although forests (Saccharina latissima and Laminaria digitata) had broadly returned to Maine in the late 20th century, we found that forests in northeast Maine have since experienced slow but significant declines in kelp, and forest persistence in the northeast was juxtaposed by a rapid, widespread collapse in the southwest. Forests collapsed in the southwest apparently because ocean warming has-directly and indirectly-made this area inhospitable to kelp. Indeed, when modeling drivers of change using causal techniques from econometrics, we discovered that unusually high summer seawater temperatures the year prior, unusually high spring seawater temperatures, and high sea urchin densities each negatively impacted kelp abundance. Furthermore, the relative power and absolute impact of these drivers varied geographically. Our findings reveal that ocean warming is redefining the outcomes of top-down forcing in this system, whereby herbivore removal no longer predictably leads to a sustained dominance of foundational kelps but instead has led to a waning dominance (northeast) or the rise of a novel phase state defined by "turf" algae (southwest). Such findings indicate that limiting climate change and managing for low herbivore abundances will be essential for preventing further loss of the vast forests that still exist in northeast Maine. They also more broadly highlight that climate change is "rewriting the rules" of nature, and thus that ecological theory and practice must be revised to account for shifting species and processes.


Assuntos
Pesqueiros , Cadeia Alimentar , Kelp , Animais , Kelp/fisiologia , Mudança Climática , Maine , Oceanos e Mares , Ouriços-do-Mar/fisiologia , New England , Fatores de Tempo , Herbivoria
8.
Brain Sci ; 14(6)2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38928609

RESUMO

While recent advancements have been made towards a better understanding of the involvement of the prefrontal cortex (PFC) in the context of cognitive control, the exact mechanism is still not fully understood. Successful behavior requires the correct detection of goal-relevant cues and resisting irrelevant distractions. Frontal parietal networks have been implicated as important for maintaining cognitive control in the face of distraction. The present study investigated the role of gamma-band power in distraction resistance and frontoparietal networks, as its increase is linked to cholinergic activity. We examined changes in gamma activity and their relationship to frontoparietal top-down modulation for distractor challenges and to bottom-up distractor processing. Healthy young adults were tested using a modified version of the distractor condition sustained attention task (dSAT) while wearing an EEG. The modified distractor was designed so that oscillatory activities could be entrained to it, and the strength of entrainment was used to assess the degree of distraction. Increased top-down control during the distractor challenge increased gamma power in the left parietal regions rather than the right prefrontal regions predicted from rodent studies. Specifically, left parietal gamma power increased in response to distraction where the amount of this increase was negatively correlated with the neural activity reflecting bottom-up distractor processing in the visual area. Variability in gamma power in right prefrontal regions was associated with increased response time variability during distraction. This may suggest that the right prefrontal region may contribute to the signaling needed for top-down control rather than its implementation.

9.
Curr Biol ; 34(13): 2801-2811.e9, 2024 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-38834064

RESUMO

Consolidation of initially encoded hippocampal representations in the neocortex through reactivation is crucial for long-term memory formation and is facilitated by the coordination of hippocampal sharp-wave ripples (SWRs) with cortical slow and spindle oscillations during non-REM sleep. Recent evidence suggests that high-frequency cortical ripples can also coordinate with hippocampal SWRs in support of consolidation; however, the contribution of cortical ripples to reactivation remains unclear. We used high-density, continuous recordings in the hippocampus (area CA1) and prefrontal cortex (PFC) over the course of spatial learning and show that independent PFC ripples dissociated from SWRs are prevalent in NREM sleep and predominantly suppress hippocampal activity. PFC ripples paradoxically mediate top-down suppression of hippocampal reactivation rather than coordination, and this suppression is stronger for assemblies that are reactivated during coordinated CA1-PFC ripples for consolidation of recent experiences. Further, we show non-canonical, serial coordination of independent cortical ripples with slow and spindle oscillations, which are known signatures of memory consolidation. These results establish a role for prefrontal cortical ripples in top-down regulation of behaviorally relevant hippocampal representations during consolidation.


Assuntos
Consolidação da Memória , Córtex Pré-Frontal , Córtex Pré-Frontal/fisiologia , Consolidação da Memória/fisiologia , Animais , Masculino , Hipocampo/fisiologia , Sono/fisiologia , Aprendizagem Espacial/fisiologia , Região CA1 Hipocampal/fisiologia
10.
Proteomics ; : e2400074, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38899939

RESUMO

Extracellular vesicles (EVs) are anucleate particles enclosed by a lipid bilayer that are released from cells via exocytosis or direct budding from the plasma membrane. They contain an array of important molecular cargo such as proteins, nucleic acids, and lipids, and can transfer these cargoes to recipient cells as a means of intercellular communication. One of the overarching paradigms in the field of EV research is that EV cargo should reflect the biological state of the cell of origin. The true relationship or extent of this correlation is confounded by many factors, including the numerous ways one can isolate or enrich EVs, overlap in the biophysical properties of different classes of EVs, and analytical limitations. This presents a challenge to research aimed at detecting low-abundant EV-encapsulated nucleic acids or proteins in biofluids for biomarker research and underpins technical obstacles in the confident assessment of the proteomic landscape of EVs that may be affected by sample-type specific or disease-associated proteoforms. Improving our understanding of EV biogenesis, cargo loading, and developments in top-down proteomics may guide us towards advanced approaches for selective EV and molecular cargo enrichment, which could aid EV diagnostics and therapeutics research.

11.
ACS Nano ; 18(24): 15779-15789, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38833666

RESUMO

The property of being stubborn and degradation resistant makes nanoplastic (NP) pollution a long-standing remaining challenge. Here, we apply a designed top-down strategy to leverage the natural hierarchical structure of waste crayfish shells with exposed functional groups for efficient NP capture. The crayfish shell-based organic skeleton with improved flexibility, strength (14.37 to 60.13 MPa), and toughness (24.61 to 278.98 MJ m-3) was prepared by purposefully removing the inorganic components of crayfish shells through a simple two-step acid-alkali treatment. Due to the activated functional groups (e.g., -NH2, -CONH-, and -OH) and ordered architectures with macropores and nanofibers, this porous crayfish shell exhibited effective removal capability of NPs (72.92 mg g-1) by physical interception and hydrogen bond/electrostatic interactions. Moreover, the sustainability and stability of this porous crayfish shell were demonstrated by the maintained high-capture performance after five cycles. Finally, we provided a postprocessing approach that could convert both porous crayfish shell and NPs into a tough flat sheet. Thus, our feasible top-down engineering strategy combined with promising posttreatment is a powerful contender for a recycling approach with broad application scenarios and clear economic advantages for simultaneously addressing both waste biomass and NP pollutants.


Assuntos
Exoesqueleto , Astacoidea , Animais , Adsorção , Porosidade , Exoesqueleto/química , Microplásticos/química , Tamanho da Partícula , Propriedades de Superfície
12.
Drug Metab Pharmacokinet ; 56: 101011, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38833901

RESUMO

Physiologically-based pharmacokinetic (PBPK) models and quantitative systems pharmacology (QSP) models have contributed to drug development strategies. The parameters of these models are commonly estimated by capturing observed values using the nonlinear least-squares method. Software packages for PBPK and QSP modeling provide a range of parameter estimation algorithms. To choose the most appropriate method, modelers need to understand the basic concept of each approach. This review provides a general introduction to the key points of parameter estimation with a focus on the PBPK and QSP models, and the respective parameter estimation algorithms. The latter part assesses the performance of five parameter estimation algorithms - the quasi-Newton method, Nelder-Mead method, genetic algorithm, particle swarm optimization, and Cluster Gauss-Newton method - using three examples of PBPK and QSP modeling. The assessment revealed that some parameter estimation results were significantly influenced by the initial values. Moreover, the choice of algorithms demonstrating good estimation results heavily depends on factors such as model structure and the parameters to be estimated. To obtain credible parameter estimation results, it is advisable to conduct multiple rounds of parameter estimation under different conditions, employing various estimation algorithms.


Assuntos
Algoritmos , Modelos Biológicos , Farmacocinética , Humanos , Animais , Software
13.
Cell Rep ; 43(7): 114371, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38923458

RESUMO

High-dimensional brain activity is often organized into lower-dimensional neural manifolds. However, the neural manifolds of the visual cortex remain understudied. Here, we study large-scale multi-electrode electrophysiological recordings of macaque (Macaca mulatta) areas V1, V4, and DP with a high spatiotemporal resolution. We find that the population activity of V1 contains two separate neural manifolds, which correlate strongly with eye closure (eyes open/closed) and have distinct dimensionalities. Moreover, we find strong top-down signals from V4 to V1, particularly to the foveal region of V1, which are significantly stronger during the eyes-open periods. Finally, in silico simulations of a balanced spiking neuron network qualitatively reproduce the experimental findings. Taken together, our analyses and simulations suggest that top-down signals modulate the population activity of V1. We postulate that the top-down modulation during the eyes-open periods prepares V1 for fast and efficient visual responses, resulting in a type of visual stand-by state.

14.
Int J Biol Macromol ; 275(Pt 1): 133340, 2024 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-38925195

RESUMO

As the traditional aerogel has defects such as poor mechanical properties, complicated preparation process, high energy consumption and non-renewable, wood aerogel as a new generation of aerogel shows unique advantages. With a natural cellulose framework, wood aerogel is a novel nano-porous material exhibiting exceptional properties such as light weight, high porosity, large specific surface area, and low thermal conductivity. Furthermore, its adaptability to further functionalization enables versatile applications across diverse fields. Driven by the imperative for sustainable development, wood aerogel as a renewable and eco-friendly material, has garnered significant attention from researchers. This review introduces preparation methods of wood aerogel based on the top-down strategy and analyzes the factors influencing their key properties intending to obtain wood aerogels with desirable properties. Avenues for realizing its functionality are also explored, and research progress across various domains are surveyed, including oil-water separation, conductivity and energy storage, as well as photothermal conversion. Finally, potential challenges associated with wood aerogel exploitation and utilization are addressed, alongside discussions on future prospects and research directions. The results emphasize the broad research value and future prospects of wood aerogels, which are poised to drive high-value utilization of wood and foster the development of green multifunctional aerogels.

15.
J Anim Ecol ; 2024 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-38790092

RESUMO

Marine heatwaves (MHWs) are episodes of anomalous warming in the ocean that can last from a few days to years. MHWs have different characteristics in terms of intensity, duration and frequency and generate thermal stress in marine ecosystems. In reef ecosystems, they are one of the main causes of the decreased presence and abundance of corals, invertebrates and fish. The deleterious capacity of thermal stress often depends on biotic factors, such as the trophic control of predators on prey. Despite the evidence of thermal stress and biotic factors affecting individual species, the combined effects of both stressors on entire reef ecosystems are much less studied. Here, using a food web modelling approach, we estimated the rate of change in species' biomass due to different MHW characteristics. Specifically, we modelled the mechanistic link between species' consumption rate and seawater temperature (thermal stressor), simulating species' biomass dynamics for different MHW characteristics under different trophic control assumptions (top-down, mixed trophic control and bottom-up). We find that total reef ecosystem biomass declined by 10% ± 5% under MHWs with severe intensity and a top-down control assumption. The bottom-up control assumption moderates the total ecosystem biomass reduction by 5% ± 5%. Irrespective of the MHW characteristics and the trophic control assumption, the most substantial biomass changes occur among top, mesopredators and corals (5% to 20% ± 10%). We show that reef ecosystems where predators exert top-down control on prey are prone to suffer species abundance declines under strong MHW events. We identify food web trophic control as a crucial driver that modulates the impacts of MHWs. Overall, our results provide a unified understanding of the interplay between abiotic stressors and biotic factors in reef ecosystems under extreme thermal events, offering insights into present baselines and future ecological states for reef ecosystems.

16.
Molecules ; 29(9)2024 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-38731523

RESUMO

This study reports an innovative approach for producing nanoplastics (NP) from various types of domestic waste plastics without the use of chemicals. The plastic materials used included water bottles, styrofoam plates, milk bottles, centrifuge tubes, to-go food boxes, and plastic bags, comprising polyethylene terephthalate (PET), polystyrene (PS), polypropylene (PP), high-density polyethylene (HDPE), and Poly (Ethylene-co-Methacrylic Acid) (PEMA). The chemical composition of these plastics was confirmed using Raman and FTIR spectroscopy, and they were found to have irregular shapes. The resulting NP particles ranged from 50 to 400 nm in size and demonstrated relative stability when suspended in water. To assess their impact, the study investigated the effects of these NP particulates on cell viability and the expression of genes involved in inflammation and oxidative stress using a macrophage cell line. The findings revealed that all types of NP reduced cell viability in a concentration-dependent manner. Notably, PS, HDPE, and PP induced significant reductions in cell viability at lower concentrations, compared to PEMA and PET. Moreover, exposure to NP led to differential alterations in the expression of inflammatory genes in the macrophage cell line. Overall, this study presents a viable method for producing NP from waste materials that closely resemble real-world NP. Furthermore, the toxicity studies demonstrated distinct cellular responses based on the composition of the NP, shedding light on the potential environmental and health impacts of these particles.


Assuntos
Sobrevivência Celular , Macrófagos , Microplásticos , Sobrevivência Celular/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Animais , Camundongos , Nanopartículas/química , Plásticos/química , Células RAW 264.7 , Expressão Gênica/efeitos dos fármacos , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Resíduos/análise , Tamanho da Partícula
17.
Bio Protoc ; 14(9): e4981, 2024 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-38737506

RESUMO

Ribosomes are an archetypal ribonucleoprotein assembly. Due to ribosomal evolution and function, r-proteins share specific physicochemical similarities, making the riboproteome particularly suited for tailored proteome profiling methods. Moreover, the structural proteome of ribonucleoprotein assemblies reflects context-dependent functional features. Thus, characterizing the state of riboproteomes provides insights to uncover the context-dependent functionality of r-protein rearrangements, as they relate to what has been termed the ribosomal code, a concept that parallels that of the histone code, in which chromatin rearrangements influence gene expression. Compared to high-resolution ribosomal structures, omics methods lag when it comes to offering customized solutions to close the knowledge gap between structure and function that currently exists in riboproteomes. Purifying the riboproteome and subsequent shot-gun proteomics typically involves protein denaturation and digestion with proteases. The results are relative abundances of r-proteins at the ribosome population level. We have previously shown that, to gain insight into the stoichiometry of individual proteins, it is necessary to measure by proteomics bound r-proteins and normalize their intensities by the sum of r-protein abundances per ribosomal complex, i.e., 40S or 60S subunits. These calculations ensure that individual r-protein stoichiometries represent the fraction of each family/paralog relative to the complex, effectively revealing which r-proteins become substoichiometric in specific physiological scenarios. Here, we present an optimized method to profile the riboproteome of any organism as well as the synthesis rates of r-proteins determined by stable isotope-assisted mass spectrometry. Our method purifies the r-proteins in a reversibly denatured state, which offers the possibility for combined top-down and bottom-up proteomics. Our method offers a milder native denaturation of the r-proteome via a chaotropic GuHCl solution as compared with previous studies that use irreversible denaturation under highly acidic conditions to dissociate rRNA and r-proteins. As such, our method is better suited to conserve post-translational modifications (PTMs). Subsequently, our method carefully considers the amino acid composition of r-proteins to select an appropriate protease for digestion. We avoid non-specific protease cleavage by increasing the pH of our standardized r-proteome dilutions that enter the digestion pipeline and by using a digestion buffer that ensures an optimal pH for a reliable protease digestion process. Finally, we provide the R package ProtSynthesis to study the fractional synthesis rates of r-proteins. The package uses physiological parameters as input to determine peptide or protein fractional synthesis rates. Once the physiological parameters are measured, our equations allow a fair comparison between treatments that alter the biological equilibrium state of the system under study. Our equations correct peptide enrichment using enrichments in soluble amino acids, growth rates, and total protein accumulation. As a means of validation, our pipeline fails to find "false" enrichments in non-labeled samples while also filtering out proteins with multiple unique peptides that have different enrichment values, which are rare in our datasets. These two aspects reflect the accuracy of our tool. Our method offers the possibility of elucidating individual r-protein family/paralog abundances, PTM status, fractional synthesis rates, and dynamic assembly into ribosomal complexes if top-down and bottom-up proteomic approaches are used concomitantly, taking one step further into mapping the native and dynamic status of the r-proteome onto high-resolution ribosome structures. In addition, our method can be used to study the proteomes of all macromolecular assemblies that can be purified, although purification is the limiting step, and the efficacy and accuracy of the proteases may be limited depending on the digestion requirements. Key features • Efficient purification of the ribosomal proteome: streamlined procedure for the specific purification of the ribosomal proteome or complex Ome. • Accurate calculation of fractional synthesis rates: robust method for calculating fractional protein synthesis rates in macromolecular complexes under different physiological steady states. • Holistic ribosome methodology focused on plants: comprehensive approach that provides insights into the ribosomes and translational control of plants, demonstrated using cold acclimation [1]. • Tailored strategies for stable isotope labeling in plants: methodology focusing on materials and labeling considerations specific to free and proteinogenic amino acid analysis [2].

18.
Cell Rep ; 43(5): 114212, 2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38743567

RESUMO

Diverse types of inhibitory interneurons (INs) impart computational power and flexibility to neocortical circuits. Whereas markers for different IN types in cortical layers 2-6 (L2-L6) have been instrumental for generating a wealth of functional insights, only the recent identification of a selective marker (neuron-derived neurotrophic factor [NDNF]) has opened comparable opportunities for INs in L1 (L1INs). However, at present we know very little about the connectivity of NDNF L1INs with other IN types, their input-output conversion, and the existence of potential NDNF L1IN subtypes. Here, we report pervasive inhibition of L2/3 INs (including parvalbumin INs and vasoactive intestinal peptide INs) by NDNF L1INs. Intersectional genetics revealed similar physiology and connectivity in the NDNF L1IN subpopulation co-expressing neuropeptide Y. Finally, NDNF L1INs prominently and selectively engage in persistent firing, a physiological hallmark disconnecting their output from the current input. Collectively, our work therefore identifies NDNF L1INs as specialized master regulators of superficial neocortex according to their pervasive top-down afferents.


Assuntos
Interneurônios , Interneurônios/metabolismo , Animais , Camundongos , Neuropeptídeo Y/metabolismo , Neocórtex/metabolismo , Neocórtex/citologia , Neocórtex/fisiologia , Peptídeo Intestinal Vasoativo/metabolismo , Masculino , Parvalbuminas/metabolismo
19.
Neuroimage ; 294: 120640, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38719154

RESUMO

Attentional control, guided by top-down processes, enables selective focus on pertinent information, while habituation, influenced by bottom-up factors and prior experiences, shapes cognitive responses by emphasizing stimulus relevance. These two fundamental processes collaborate to regulate cognitive behavior, with the prefrontal cortex and its subregions playing a pivotal role. Nevertheless, the intricate neural mechanisms underlying the interaction between attentional control and habituation are still a subject of ongoing exploration. To our knowledge, there is a dearth of comprehensive studies on the functional connectivity between subsystems within the prefrontal cortex during attentional control processes in both primates and humans. Utilizing stereo-electroencephalogram (SEEG) recordings during the Stroop task, we observed top-down dominance effects and corresponding connectivity patterns among the orbitofrontal cortex (OFC), the middle frontal gyrus (MFG), and the inferior frontal gyrus (IFG) during heightened attentional control. These findings highlighting the involvement of OFC in habituation through top-down attention. Our study unveils unique connectivity profiles, shedding light on the neural interplay between top-down and bottom-up attentional control processes, shaping goal-directed attention.


Assuntos
Atenção , Eletroencefalografia , Habituação Psicofisiológica , Córtex Pré-Frontal , Humanos , Córtex Pré-Frontal/fisiologia , Córtex Pré-Frontal/diagnóstico por imagem , Atenção/fisiologia , Masculino , Feminino , Eletroencefalografia/métodos , Habituação Psicofisiológica/fisiologia , Adulto , Adulto Jovem , Teste de Stroop
20.
Hear Res ; 447: 109023, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38733710

RESUMO

Limited auditory input, whether caused by hearing loss or by electrical stimulation through a cochlear implant (CI), can be compensated by the remaining senses. Specifically for CI users, previous studies reported not only improved visual skills, but also altered cortical processing of unisensory visual and auditory stimuli. However, in multisensory scenarios, it is still unclear how auditory deprivation (before implantation) and electrical hearing experience (after implantation) affect cortical audiovisual speech processing. Here, we present a prospective longitudinal electroencephalography (EEG) study which systematically examined the deprivation- and CI-induced alterations of cortical processing of audiovisual words by comparing event-related potentials (ERPs) in postlingually deafened CI users before and after implantation (five weeks and six months of CI use). A group of matched normal-hearing (NH) listeners served as controls. The participants performed a word-identification task with congruent and incongruent audiovisual words, focusing their attention on either the visual (lip movement) or the auditory speech signal. This allowed us to study the (top-down) attention effect on the (bottom-up) sensory cortical processing of audiovisual speech. When compared to the NH listeners, the CI candidates (before implantation) and the CI users (after implantation) exhibited enhanced lipreading abilities and an altered cortical response at the N1 latency range (90-150 ms) that was characterized by a decreased theta oscillation power (4-8 Hz) and a smaller amplitude in the auditory cortex. After implantation, however, the auditory-cortex response gradually increased and developed a stronger intra-modal connectivity. Nevertheless, task efficiency and activation in the visual cortex was significantly modulated in both groups by focusing attention on the visual as compared to the auditory speech signal, with the NH listeners additionally showing an attention-dependent decrease in beta oscillation power (13-30 Hz). In sum, these results suggest remarkable deprivation effects on audiovisual speech processing in the auditory cortex, which partially reverse after implantation. Although even experienced CI users still show distinct audiovisual speech processing compared to NH listeners, pronounced effects of (top-down) direction of attention on (bottom-up) audiovisual processing can be observed in both groups. However, NH listeners but not CI users appear to show enhanced allocation of cognitive resources in visually as compared to auditory attended audiovisual speech conditions, which supports our behavioural observations of poorer lipreading abilities and reduced visual influence on audition in NH listeners as compared to CI users.


Assuntos
Estimulação Acústica , Atenção , Implante Coclear , Implantes Cocleares , Surdez , Eletroencefalografia , Pessoas com Deficiência Auditiva , Estimulação Luminosa , Percepção da Fala , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Implante Coclear/instrumentação , Adulto , Estudos Prospectivos , Estudos Longitudinais , Pessoas com Deficiência Auditiva/psicologia , Pessoas com Deficiência Auditiva/reabilitação , Surdez/fisiopatologia , Surdez/reabilitação , Surdez/psicologia , Estudos de Casos e Controles , Idoso , Percepção Visual , Leitura Labial , Fatores de Tempo , Audição , Potenciais Evocados Auditivos , Córtex Auditivo/fisiopatologia , Potenciais Evocados
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