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This study investigated the effects of different storage times (0-24 months) and temperatures (-20 °C, 4 °C, 25 °C, and 35 °C) on the volatile compounds and color of vanilla beans from Taiwan using ISO 5565-1:1999 as the commercial quality indicators. The vanillin level was highest at -20 °C for 6 months (p < 0.05) and no color-perceived differences (ΔE < 1). At 35 °C, lipid metabolism, enzymatic oxidation, and Maillard reaction products, such as 2-acetylpyrrole and 5-hydroxymethylfurfural, were detected after 18 months of storage, leading to significant color-perceived differences (ΔE > 5) and changes in the odor profile, which did not meet ISO standards. To sum up, as the storage temperature and duration increased, oxidative metabolites increased. Furthermore, storage at -20 °C or 4 °C is stable, but considering vanillin quality, the storage temperature of -20 °C can extend the storage period by 12 months.
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BACKGROUND: Hemodialysis patients exhibit a reduced response to vaccination and have different vaccine dose regimens. Vaccines induce antibodies and affect the inflammatory balance through antibody glycosylation and effector functions. Therefore, we aimed to analyze the antibody glycosylation profiles in hemodialysis patients who were vaccinated against severe acute respiratory syndrome coronavirus 2, infected with the virus, or both, and compare them with those of dialysis patients in a control group. METHODS: Plasma samples from 112 hemodialysis patients were assigned to four groups: control, infected, vaccinated, and post-vaccine-infected. Paired plasma samples from 47 people with vaccination (vaccinees) were analyzed before and after the booster dose. The same analytical approach was applied to the four groups for a cross-sectional comparison. RESULTS: Our study found that both vaccination and infection groups showed decreased fucosylation of IgG1, which is associated with a proinflammatory biosignature. However, vaccination also leads to increased galactosylation and bisection of IgG antibodies, which are associated with anti-inflammatory effects and the additional regulation of immune responses. In contrast, infection led to an additional decrease in the fucosylation of IgG2 and IgA, demonstrating a more intense proinflammatory biosignature than vaccination. CONCLUSIONS: Our findings emphasize the proinflammatory biosignature of afucosylation in both vaccination and infection groups. Additionally, we uncovered further regulated profiles related to galactosylation in vaccinees. These findings suggest that antibody investigation for vaccination or infection should not solely focus on neutralization but should also consider effector function-related glycosylation profiling. This comprehensive information can be valuable for fine-tuning vaccine development in the future.
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This study delves into the application of Brain-Computer Interfaces (BCIs), focusing on exploiting Steady-State Visual Evoked Potentials (SSVEPs) as communication tools for individuals facing mobility impairments. SSVEP-BCI systems can swiftly transmit substantial volumes of information, rendering them suitable for diverse applications. However, the efficacy of SSVEP responses can be influenced by variables such as the frequency and color of visual stimuli. Through experiments involving participants equipped with electrodes on the brain's visual cortex, visual stimuli were administered at 4, 17, 25, and 40Hz, using white, red, yellow, green, and blue light sources. The results reveal that white and green stimuli evoke higher SSVEP responses at lower frequencies, with color's impact diminishing at higher frequencies. At low light intensity (1W), white and green stimuli elicit significantly higher SSVEP responses, while at high intensity (3W), responses across colors tend to equalize. Notably, due to seizure risks, red and blue lights should be used cautiously, with white and green lights preferred for SSVEP-BCI systems. This underscores the critical consideration of color and frequency in the design of effective and safe SSVEP-BCI systems, necessitating further research to optimize designs for clinical applications.
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Interfaces Cérebro-Computador , Eletroencefalografia , Potenciais Evocados Visuais , Estimulação Luminosa , Humanos , Potenciais Evocados Visuais/fisiologia , Masculino , Adulto , Estimulação Luminosa/métodos , Feminino , Adulto Jovem , Córtex Visual/fisiologiaRESUMO
The emergence of brain-computer interface (BCI) technology provides enormous potential for human medical and daily applications. Therefore, allowing users to tolerate the visual response of SSVEP for a long time has always been an important issue in the SSVEP-BCI system. We recruited three subjects and conducted visual experiments in groups using different frequencies (17 and 25Hz) and 60Hz light. After recording the physiological signal, use FFT to perform a time-frequency analysis on the physiological signal to check whether there is any difference in the signal-to-noise ratio and amplitude of the 60Hz light source compared with a single low-frequency signal source. The results show that combining a 60Hz light source with low-frequency LEDs can reduce participants' eye discomfort while achieving effective light stimulation control. At the same time, there was no significant difference in signal-to-noise ratio and amplitude between the groups. This also means that 60Hz can make vision more continuous and improve the subject's experience and comfort. At the same time, it does not affect the performance of the original SSVEP-induced response. This study highlights the importance of considering technical aspects and user comfort when designing SSVEP-BCI systems to increase the usability of SSVEP systems for long-term flash viewing.
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Interfaces Cérebro-Computador , Eletroencefalografia , Potenciais Evocados Visuais , Estimulação Luminosa , Humanos , Estimulação Luminosa/métodos , Masculino , Potenciais Evocados Visuais/fisiologia , Adulto , Feminino , Adulto Jovem , Percepção Visual/fisiologia , Razão Sinal-RuídoRESUMO
Antibody glycosylation plays a crucial role in the humoral immune response by regulating effector functions and influencing the binding affinity to immune cell receptors. Previous studies have focused mainly on the immunoglobulin G (IgG) isotype owing to the analytical challenges associated with other isotypes. Thus, the development of a sensitive and accurate analytical platform is necessary to characterize antibody glycosylation across multiple isotypes. In this study, we have developed an analytical workflow using antibody-light-chain affinity beads to purify IgG, IgA, and IgM from 16 µL of human plasma. Dual enzymes, trypsin and Glu-C, were used during on-bead digestion to obtain enzymatic glycopeptides and protein-specific surrogate peptides. Ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry was used in order to determine the sensitivity and specificity. Our platform targets 95 glycopeptides across the IgG, IgA, and IgM isotypes, as well as eight surrogate peptides representing total IgG, four IgG classes, two IgA classes, and IgM. Four stable isotope-labeled internal standards were added after antibody purification to calibrate the preparation and instrumental bias during analysis. Calibration curves constructed using serially diluted plasma samples showed good curve fitting (R2 > 0.959). The intrabatch and interbatch precision for all the targets had relative standard deviation of less than 29.6%. This method was applied to 19 human plasma samples, and the glycosylation percentages were calculated, which were comparable to those reported in the literature. The developed method is sensitive and accurate for Ig glycosylation profiling. It can be used in clinical investigations, particularly for detailed humoral immune profiling.
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Glicopeptídeos , Imunoglobulina G , Humanos , Glicosilação , Imunoglobulina G/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas , Glicopeptídeos/metabolismo , Digestão , Imunoglobulina A , Imunoglobulina MRESUMO
OBJECTIVE: To identify the feasibility and efficiency of deep convolutional neural networks (DCNNs) in the detection of ankle fractures and to explore ensemble strategies that applied multiple projections of radiographs.Ankle radiographs (AXRs) are the primary tool used to diagnose ankle fractures. Applying DCNN algorithms on AXRs can potentially improve the diagnostic accuracy and efficiency of detecting ankle fractures. METHODS: A DCNN was trained using a trauma image registry, including 3102 AXRs. We separately trained the DCNN on anteroposterior (AP) and lateral (Lat) AXRs. Different ensemble methods, such as "sum-up," "severance-OR," and "severance-Both," were evaluated to incorporate the results of the model using different projections of view. RESULTS: The AP/Lat model's individual sensitivity, specificity, positive-predictive value, accuracy, and F1 score were 79%/84%, 90%/86%, 88%/86%, 83%/85%, and 0.816/0.850, respectively. Furthermore, the area under the receiver operating characteristic curve (AUROC) of the AP/Lat model was 0.890/0.894 (95% CI: 0.826-0.954/0.831-0.953). The sum-up method generated balanced results by applying both models and obtained an AUROC of 0.917 (95% CI: 0.863-0.972) with 87% accuracy. The severance-OR method resulted in a better sensitivity of 90%, and the severance-Both method obtained a high specificity of 94%. CONCLUSION: Ankle fracture in the AXR could be identified by the trained DCNN algorithm. The selection of ensemble methods can depend on the clinical situation which might help clinicians detect ankle fractures efficiently without interrupting the current clinical pathway. ADVANCES IN KNOWLEDGE: This study demonstrated different ensemble strategies of AI algorithms on multiple view AXRs to optimize the performance in various clinical needs.
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Fraturas do Tornozelo , Aprendizado Profundo , Humanos , Fraturas do Tornozelo/diagnóstico por imagem , Tornozelo , Algoritmos , Redes Neurais de ComputaçãoRESUMO
To establish the analytic conditions for examining the aroma quality of vanilla pods, we compared different extraction methods and identified a suitable option. We utilized headspace solid-phase microextraction (HS-SPME), steam distillation (SD), simultaneous steam distillation (SDE) and alcoholic extraction combined with gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) to identify volatile components of vanilla pods. A total of 84 volatile compounds were identified in this experiment, of which SDE could identify the most volatile compounds, with a total of 51 species, followed by HS-SPME, with a total of 28 species. Ten volatile compounds were identified by extraction with a minimum of 35% alcohol. HS-SPME extraction provided the highest total aroma peak areas, and the peak areas of aldehydes, furans, alcohols, monoterpenes and phenols compounds were several times higher than those of the other extraction methods. The results showed that the two technologies, SDE and HS-SPME, could be used together to facilitate analysis of vanilla pod aroma.
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Vanilla , Compostos Orgânicos Voláteis , Cromatografia Gasosa-Espectrometria de Massas/métodos , Odorantes/análise , Microextração em Fase Sólida/métodos , Vapor/análise , Compostos Orgânicos Voláteis/análiseRESUMO
Alpha/beta hydrolase domain-containing protein 5 (ABHD5) is a highly conserved protein that regulates various lipid metabolic pathways via interactions with members of the perilipin (PLIN) and Patatin-like phospholipase domain-containing protein (PNPLA) protein families. Loss of function mutations in ABHD5 result in Chanarin-Dorfman Syndrome (CDS), characterized by ectopic lipid accumulation in numerous cell types and severe ichthyosis. Recent data demonstrates that ABHD5 is the target of synthetic and endogenous ligands that might be therapeutic beneficial for treating metabolic diseases and cancers. However, the structural basis of ABHD5 functional activities, such as protein-protein interactions and ligand binding is presently unknown. To address this gap, we constructed theoretical structural models of ABHD5 by comparative modeling and topological shape analysis to assess the spatial patterns of ABHD5 conformations computed in protein dynamics. We identified functionally important residues on ABHD5 surface for lipolysis activation by PNPLA2, lipid droplet targeting and PLIN-binding. We validated the computational model by examining the effects of mutating key residues in ABHD5 on an array of functional assays. Our integrated computational and experimental findings provide new insights into the structural basis of the diverse functions of ABHD5 as well as pathological mutations that result in CDS.
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1-Acilglicerol-3-Fosfato O-Aciltransferase/química , 1-Acilglicerol-3-Fosfato O-Aciltransferase/metabolismo , Biologia Computacional/métodos , Lipase/metabolismo , Gotículas Lipídicas/metabolismo , Mutação , 1-Acilglicerol-3-Fosfato O-Aciltransferase/genética , Humanos , Ligantes , Gotículas Lipídicas/química , Conformação ProteicaRESUMO
Vanilla (Vanilla planifolia) is a precious natural flavoring that is commonly used throughout the world. In the past, all vanilla used in Taiwan was imported; however, recent breakthroughs in cultivation and processing technology have allowed Taiwan to produce its own supply of vanilla. In this study, headspace solid-phase microextraction (HS-SPME) combined with GC-FID and GC-MS was used to analyze the volatile components of vanilla from different origins produced in Taiwan under different cultivation and processing conditions. The results of our study revealed that when comparing different harvest maturities, the composition diversity and total volatile content were both higher when the pods were matured for more than 38 weeks. When comparing different killing conditions, we observed that the highest vanillin percentage was present after vanilla pods were killed three times in 65 °C treatments for 1 min each. From the experiment examining the addition of different strains, the PCA results revealed that the volatiles of vanilla that was processed with Dekkera bruxellensis and Bacillus subtilis was clearly distinguished from which obtained by processing with the other strains. Vanilla processed with B. subtilis contained 2-ethyl-1-hexanol, and this was not detected in other vanillas. Finally, when comparing the vanillin percentage from seven different regions in Taiwan, vanilla percentage from Taitung and Taoyuan Longtan were the highest.
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Vanilla/química , Vanilla/metabolismo , Compostos Orgânicos Voláteis/química , Agricultura/métodos , Benzaldeídos/química , Benzaldeídos/isolamento & purificação , Cromatografia Gasosa/métodos , Aromatizantes/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Extratos Vegetais/análise , Microextração em Fase Sólida/métodos , Taiwan , Compostos Orgânicos Voláteis/análiseRESUMO
SARS-CoV-2 infects humans through the binding of viral S-protein (spike protein) to human angiotensin I converting enzyme 2 (ACE2). The structure of the ACE2-S-protein complex has been deciphered and we focused on the 27 ACE2 residues that bind to S-protein. From human sequence databases, we identified nine ACE2 variants at ACE2-S-protein binding sites. We used both experimental assays and protein structure analysis to evaluate the effect of each variant on the binding affinity of ACE2 to S-protein. We found one variant causing complete binding disruption, two and three variants, respectively, strongly and mildly reducing the binding affinity, and two variants strongly enhancing the binding affinity. We then collected the ACE2 gene sequences from 57 nonhuman primates. Among the 6 apes and 20 Old World monkeys (OWMs) studied, we found no new variants. In contrast, all 11 New World monkeys (NWMs) studied share four variants each causing a strong reduction in binding affinity, the Philippine tarsier also possesses three such variants, and 18 of the 19 prosimian species studied share one variant causing a strong reduction in binding affinity. Moreover, one OWM and three prosimian variants increased binding affinity by >50%. Based on these findings, we proposed that the common ancestor of primates was strongly resistant to and that of NWMs was completely resistant to SARS-CoV-2 and so is the Philippine tarsier, whereas apes and OWMs, like most humans, are susceptible. This study increases our understanding of the differences in susceptibility to SARS-CoV-2 infection among primates.
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COVID-19 , Resistência à Doença/genética , Peptidil Dipeptidase A , SARS-CoV-2 , Animais , COVID-19/genética , COVID-19/imunologia , Chlorocebus aethiops , Humanos , Macaca mulatta , Peptidil Dipeptidase A/genética , Peptidil Dipeptidase A/imunologia , SARS-CoV-2/genética , SARS-CoV-2/imunologiaRESUMO
Hepatitis C virus (HCV) NS3 protein possesses protease and helicase activities and is considered an oncoprotein in virus-derived hepatocellular carcinoma. The NS3-associated oncogenesis has been studied but not fully understood. In this study, we have identified novel interactions of the NS3 protein with DNA repair factors, Werner syndrome protein (WRN) and Ku70, in both an HCV subgenomic replicon system and Huh7 cells expressing NS3. HCV NS3 protein inhibits WRN-mediated DNA repair and reduces the repair efficiency of nonhomologous end joining. It interferes with Ku70 recruitment to the double-strand break sites and alters the nuclear distribution of WRN-Ku repair complex. In addition, WRN is a substrate of the NS3/4A protease; the level of WRN protein is regulated by both the proteasome degradation pathway and HCV NS3/4A protease activity. The dual role of HCV NS3 and NS3/4A proteins in regulating the function and expression level of the WRN protein intensifies the effect of impairment on DNA repair. This may lead to an accumulation of DNA mutations and genome instability and, eventually, tumor development.IMPORTANCE HCV infection is a worldwide problem of public health and a major contributor to hepatocellular carcinoma. The single-stranded RNA virus with RNA-dependent RNA polymerase experiences a high error rate and develops strategies to escape the immune system and hepatocarcinogenesis. Studies have revealed the involvement of HCV proteins in the impairment of DNA repair. The present study aimed to further elucidate mechanisms by which the viral NS3 protein impairs the repair of DNA damage. Our results clearly indicate that HCV NS3/4A protease targets WRN for degradation, and, at the same time, diminishes the repair efficiency of nonhomologous end joining by interfering with the recruitment of Ku protein to the DNA double-strand break sites. The study describes a novel mechanism by which the NS3 protein influences DNA repair and provides new insight into the molecular mechanism of HCV pathogenesis.
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Reparo do DNA por Junção de Extremidades , Hepacivirus/genética , Hepacivirus/metabolismo , Proteínas não Estruturais Virais/metabolismo , Helicase da Síndrome de Werner/metabolismo , Linhagem Celular , DNA/metabolismo , Quebras de DNA de Cadeia Dupla , Dano ao DNA , Reparo do DNA/fisiologia , Proteínas de Ligação a DNA/metabolismo , Exodesoxirribonucleases/metabolismo , Células HEK293 , Hepatite C Crônica/genética , Humanos , Autoantígeno Ku/genética , RNA Polimerase Dependente de RNA/genética , Proteínas não Estruturais Virais/genética , Helicase da Síndrome de Werner/fisiologiaRESUMO
Even with increasing evidence for roles of glycolytic enzymes in controlling cancerous characteristics, the best target of candidate metabolic enzymes for lessening malignancy remains under debate. Pyruvate is a main glycolytic metabolite that could be mainly converted into either lactate by Lactate Dehydrogenase A (LDHA) or acetyl-CoA by Pyruvate Dehydrogenase E1 component α subunit (PDHA1) catalytic complex. In tumor cells, accumulating lactate is produced whereas the conversion of pyruvate into mitochondrial acetyl-CoA is less active compared with their normal counterparts. This reciprocal molecular association makes pyruvate metabolism a potential choice of anti-cancer target. Cellular and molecular changes were herein assayed in Head and Neck Squamous Cell Carcinoma (HNSCC) cells in response to LDHA and PDHA1 loss in vitro, in vivo and in clinic. By using various human cancer databases and clinical samples, LDHA and PDHA1 levels exhibit reversed prognostic roles. In vitro analysis demonstrated that decreased cell growth and motility accompanied by an increased sensitivity to chemotherapeutic agents was found in cells with LDHA loss whereas PDHA1-silencing exhibited opposite phenotypes. At the molecular level, it was found that oncogenic Protein kinase B (PKB/Akt) and Extracellular signal-regulated kinase (ERK) singling pathways contribute to pyruvate metabolism mediated HNSCC cell growth. Furthermore, LDHA/PDHA1 changes in HNSCC cells resulted in a broad metabolic reprogramming while intracellular molecules including polyunsaturated fatty acids and nitrogen metabolism related metabolites underlie the malignant changes. Collectively, our findings reveal the significance of pyruvate metabolic fates in modulating HNSCC tumorigenesis and highlight the impact of metabolic plasticity in HNSCC cells.
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Carcinogênese/genética , L-Lactato Desidrogenase/genética , Piruvato Desidrogenase (Lipoamida)/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Animais , Linhagem Celular Tumoral , Proliferação de Células/genética , Glicólise/genética , Xenoenxertos , Humanos , Ácido Láctico/metabolismo , Camundongos , Mitocôndrias/genética , Ácido Pirúvico/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologiaRESUMO
With the rapid progress of cancer genome studies, many missense mutations in populations of somatic cells of different cancer types and at different stages have been identified. However, it is challenging to understand the implications of these cancer-related variants. We have developed a computational method that integrates structural, topographical, and evolutionary information for assessments of biochemical effects and the extent of deleteriousness of the cancer-related variants. We have mapped somatic missense mutations from the Catalogue of Somatic Mutations In Cancer (COSMIC) to 3D structures in the Protein Data Bank (PDB). Our results show that a large portion of these missense mutations is located on protein surface pockets, which often serve as a structural and functional unit of cancer variants. We provide detailed analysis of several examples and assessment on the importance of these variants, including prediction of previously unreported cancer-variants, along with independent evidence from the literature. Furthermore, we show our predictions can inform on the functional roles and the mechanism of predicted cancer variants.
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The aim of this study was to investigate changes in task-related brain oscillations and corticocortical connections in patients with mild cognitive impairment (MCI) and those with normal aging using cross-mutual information (CMI) analysis. We hypothesized that task-related brain oscillations and corticocortical connections were affected by age- and disease-related changes, which could be reflected in the CMI analysis. Electroencephalogram (EEG) recordings were measured in 16 MCI patients, 15 healthy age-matched controls, and 16 healthy younger individuals. The frequencies and interhemispheric CMI data were estimated in all groups. The specific EEG rhythms measured were delta (δ), theta (θ), alpha (α), beta (ß), and gamma (γ) bands. Significant differences in δ, θ, α, and ß bands were observed between the younger and elderly groups. However, only the θ band was significantly different between the elderly and MCI groups. Moreover, this study used EEG recordings to investigate age- and disease-related changes in the corticocortical connections of the brain. This study proves that the θ-band frequency of the connection between the parietal and occipital lobes for the age- and disease-related changes can be depicted using the CMI analysis.
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An important question in healthcare for older patients is whether age-related changes in cortical reorganization can be measured with advancing age. This study investigated the factors behind such age-related changes, using time-frequency analysis of event-related potentials (ERPs). We hypothesized that brain rhythms was affected by age-related changes, which could be reflected in the ERP indices. An oddball task was conducted in two experimental groups, namely young participants (N=15; mean age 23.7±2.8 years) and older participants (N=15; mean age 70.1±7.9 years). Two types of stimuli were used: the target (1 kHz frequency) and standard (2 kHz frequency). We scrutinized three ERP indices: event-related spectral power (ERPSP), inter-trial phase-locking (ITPL), and event-related cross-phase coherence (ERPCOH). Both groups performed equally well for correct response rate. However, the results revealed a statistically significant age difference for inter-trial comparison. Compared with the young, the older participants showed the following age-related changes: (a) power activity decreased; however, an increase was found only in the late (P3, 280-450 ms) theta (4-7 Hz) component over the bilateral frontal and temporo-frontal areas; (b) low phase-locking in the early (N1, 80-140 ms) theta band over the parietal/frontal (right) regions appeared; (c) the functional connections decreased in the alpha (7-13 Hz) and beta (13-30 Hz) bands, but no difference emerged in the theta band between the two groups. These results indicate that age-related changes in task-specific brain activity for a normal aging population can be depicted using the three ERP indices.