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Quick differentiation of current circulating variants and the emerging recombinant variants of SARS-CoV-2 is essential to monitor their transmissions. However, the widely applied gene sequencing method is time-consuming and costly especially when facing recombinant variants, because a large part or whole genome sequencing is required. Allele-specific reverse transcriptase real time RT-PCR (RT-qPCR) represents a quick and cost-effective method for SNP (single nucleotide polymorphism) genotyping and has been successfully applied for SARS-CoV-2 variant screening. In the present study, we developed a panel of 5 multiplex allele-specific RT-qPCR assays targeting 20 key mutations for quick differentiation of the Omicron subvariants (BA.1 to BA.5 and their descendants) and the recombinant variants (XBB.1 and XBB.1.5). Two parallel multiplex RT-qPCR reactions were designed to separately target the prototype allele and the mutated allele of each mutation in the allele-specific RT-qPCR assay. Optimal annealing temperatures, primer and probe dosage, and time for annealing/extension for each reaction were determined by multi-factor and multi-level orthogonal test. The variation of Cp (crossing point) values (ΔCp) between the two multiplex RT-qPCR reactions was applied to determine if a mutation occurs or not. SARS-CoV-2 subvariants and related recombinant variants were differentiated by their unique mutation patterns. The developed multiplex allele-specific RT-qPCR assays exhibited excellent analytical sensitivities (with limits of detection (LoDs) of 1.47-18.52 copies per reaction), wide linear detection ranges (109-100 copies per reaction), good amplification efficiencies (88.25 to 110.68%), excellent reproducibility (coefficient of variations (CVs) < 5% in both intra-assay and inter-assay tests), and good clinical performances (99.5-100% consistencies with Sanger sequencing). The developed multiplex allele-specific RT-qPCR assays in the present study provide an alternative tool for quick differentiation of the SARS-CoV-2 Omicron subvariants and their recombinant variants. KEY POINTS: ⢠A panel of five multiplex allele-specific RT-qPCR assays for quick differentiation of 11 SARS-CoV-2 Omicron subvariants (BA.1, BA.2, BA.4, BA.5, and their descendants) and 2 recombinant variants (XBB.1 and XBB.1.5). ⢠The developed assays exhibited good analytical sensitivities and reproducibility, wide linear detection ranges, and good clinical performances, providing an alternative tool for quick differentiation of the SARS-CoV-2 Omicron subvariants and their recombinant variants.
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COVID-19 , Humanos , Alelos , COVID-19/diagnóstico , Reprodutibilidade dos Testes , SARS-CoV-2/genéticaRESUMO
The neuropeptide orexin is involved in motor circuit function. However, its modulation on neuronal activities of motor structures, integrating orexin's diverse downstream molecular cascades, remains elusive. By combining whole-cell patch-clamp recordings and neuropharmacological methods, we revealed that both non-selective cationic conductance (NSCC) and endocannabinoids (eCBs) are recruited by orexin signalling on reticulospinal neurones in the caudal pontine reticular nucleus (PnC). The orexin-NSCC cascade provides a depolarizing force that proportionally enhances the firing-responsive gain of these neurones. Meanwhile, the orexin-eCB cascade selectively attenuates excitatory synaptic strength in these neurones by activating presynaptic cannabinoid receptor type 1. This cascade restrains the firing response of the PnC reticulospinal neurones to excitatory inputs. Intriguingly, non-linear or linear interactions between orexin postsynaptic excitation and presynaptic inhibition can influence the firing responses of PnC reticulospinal neurones in different directions. When presynaptic inhibition is in the lead, non-linear interactions can prominently downregulate or even gate the firing response. Conversely, linear interactions occur to promote the firing response, and these linear interactions can be considered a proportional reduction in the contribution of depolarization to firing by presynaptic inhibition. Through the dynamic employment of these interactions, adaptive modulation may be achieved by orexin to restrain or even gate the firing output of the PnC to weak/irrelevant input signals and facilitate those to salient signals. KEY POINTS: This study investigated the effects of orexin on the firing activity of PnC reticulospinal neurones, a key element of central motor control. We found that orexin recruited both the non-selective cationic conductances (NSCCs) and endocannabinoid (eCB)-cannabinoid receptor type 1 (CB1R) system to pontine reticular nucleus (PnC) reticulospinal neurones. The orexin-NSCC cascade exerts a postsynaptic excitation that enhances the firing response, whereas the orexin-eCB-CB1R cascade selectively attenuates excitatory synaptic strength that restrains the firing response. The postsynaptic and presynaptic actions of orexins occur in an overlapping time window and interact to dynamically modulate firings in PnC reticulospinal neurones. Non-linear interactions occur when presynaptic inhibition of orexin is in the lead, and these interactions can prominently downregulate or even gate firing responses in PnC reticulospinal neurones. Linear interactions occur when postsynaptic excitation of orexin is in the lead, and these interactions can promote the firing response. These linear interactions can be considered a proportional reduction of the contribution of depolarization to firing by presynaptic inhibition.
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Endocanabinoides , Neuropeptídeos , Orexinas/farmacologia , Endocanabinoides/farmacologia , Neurônios/fisiologia , Receptores de CanabinoidesRESUMO
BACKGROUND: There is a lack of a comprehensive evaluation for pediatric clinical practice guidelines (CPGs) published in recent years. Here, we assessed the quality of pediatric CPGs, considering factors that might affect their quality. The aim of the study is to promote a more coherent development and application of CPGs. METHODS: Pediatric CPGs published in PubMed, MedLive, Guidelines International Network, National Institute for Health and Care Excellence, and World Health Organization between 2017 and 2019 were searched and collected. Paired researchers conducted screening, data extraction, and quality assessment using the Appraisal of Guidelines for Research and Evaluation II (AGREE II). Linear regression analysis determined the factors affecting CPGs' quality. RESULTS: The study included a total of 216 CPGs, which achieved a mean score of 4.26 out of 7 points (60.86%) in the AGREE II assessment. Only 6.48% of the CPGs reached the "recommend" level. The remaining 69.91% should have been modified before recommendation, while the other 23.61% did not reach the recommended level at all. The overall quality of recent pediatric CPGs was higher than previously, and the proportion of CPGs with low-quality decreased over time. However, there were still too few CPGs that reached a high-quality level. The "applicability" and "rigor of development" domains had generally low scores. CPGs formulated by developing countries or regions, those that are not under an organizations or groups responsibility, and those that used non-evidence-based methods were found to be associated with poorer quality in different domains as independent or combinational factors. CONCLUSIONS: The quality of pediatric CPGs still needs to be improved. Specifically, a quality control before applying new CPGs should be essential to ensure their quality and applicability.
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Organização Mundial da Saúde , Criança , HumanosRESUMO
BACKGROUND: Vasomotor regulation of dermal blood vessels, which are critical in the function of the skin in thermoregulatory control, involves both neural and non-neural mechanisms. Whereas the role of sympathetic nerves in regulating vasomotor activities is comprehensively studied and well recognized, that of sensory nerves is underappreciated. Studies in rodents have shown that severance of the sciatic nerve leads to vasodilatation in the foot, but whether sympathetic or sensory nerve fibers or both are responsible for the neurogenic vasodilatation remains unknown. RESULTS: In adult Sprague-Dawley rats, vasodilatation after transection of the sciatic nerve gradually diminished to normal within 3-4 days. The neurotmesis-induced neurogenic vasodilatation was not detectable when the sciatic nerve was chronically deafferentated by selective resection of the dorsal root ganglia (DRGs) that supply the nerve. Specific activation of C-afferents by intra-neural injection of capsaicin resulted in neurogenic vasodilatation to a magnitude comparable to that by neurotmesis, and transection of the sciatic nerve pre-injected with capsaicin did not induce further vasodilatation. CONCLUSIONS: Our results collectively indicate that vasodilatation after traumatic nerve injury in rats is predominantly mediated by C-fiber afferents.
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Capsaicina/farmacologia , Nervo Isquiático/lesões , Neuropatia Ciática/fisiopatologia , Pele/inervação , Vasodilatação/fisiologia , Animais , Feminino , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/fisiopatologia , Fibras Nervosas/efeitos dos fármacos , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Neuropatia Ciática/tratamento farmacológico , Vasodilatação/efeitos dos fármacosRESUMO
An amendment to this paper has been published and can be accessed via the original article.
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Salinity (NaCl) was used in waste activated sludge (WAS) anaerobic fermentation system which had been presented to greatly enhance the extracellular polymeric substance (EPS) production including protein and polysaccharide and short-chain fatty acids (SCFAs). Salinity enhanced soluble protein and polysaccharide (SB-EPS) release which was 4.04 times (protein) and 1.83 times (polysaccharide) compared to 0 g/L NaCl level. More important, salinity restrained the coenzyme 420 activity (F420), but increased the hydrolase activity. Abundant hydrolysis of substrate and highly active hydrolase led to abundant SCFA production. Pearson correlation coefficient showed that the protein became the main reaction substrate for SCFA generation.
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Reatores Biológicos , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Salinidade , Esgotos/microbiologia , AnaerobioseRESUMO
Zinc oxide nanoparticles (ZnO NPs) production and usage might lead to a large discharge of ZnO NPs into the natural environment, raising concerns of pollution and ecological security. The effects of ZnO NPs on waste activated sludge hydrolytic acidification and microbial communities were studied in semi-continuous fermentation systems. The fermentation performance of eight ZnO NPs concentrations including ZnO NPs normal [0.01, 0.1, 1 and 10 mg/g mixed liquor suspended solids (MLSS)] and ZnO NPs shock (10, 1000, 1000 and 10,000 mg/g MLSS) were discussed, and their biodegradability was also analyzed. The experimental results showed that proteins, polysaccharides and short-chain fatty acids were enhanced by ZnO NPs, particularly by ZnO NPs shock. Low ZnO NPs concentrations inhibited coenzyme 420 (F420) and dehydrogenase activities but enhanced α-glucosidase and protease activities. Illumina MiSeq sequencing revealed that ZnO NPs addition enriched Azospira, Ottowia and Hyphomicrobium but not Anaerolineaceae.
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Fermentação , Microbiota/efeitos dos fármacos , Nanopartículas/toxicidade , Esgotos/microbiologia , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/toxicidade , Óxido de Zinco/toxicidade , Anaerobiose , Biodegradação Ambiental , Hidrólise , Nanopartículas/análise , Esgotos/química , Poluentes Químicos da Água/análise , Óxido de Zinco/análiseRESUMO
BACKGROUND: The Center for Epidemiological Studies Depression Scale for Children (CES-DC) is a widely used scale for screening depressive symptoms in children and adolescents. This study aims to uncover the optimal factor structure of the DES-DC and presents an alternate conceptualization of adolescent depression by estimating bifactor models and several competing models using a sample of Chinese adolescents. METHODS: The participants were 533 adolescents (49.7% boys, 49.7% girls, 3 participants did not report) between 12 and 18 years of age attending public secondary schools in the middle part of mainland China. Data were collected in classrooms using a questionnaire survey. A structural equation modeling approach was used to estimate and compare a series of competing models for the DES-DC. RESULTS: A Bifactor exploratory structural equation model (Bi-ESEM) with the best model fit was retained for representing the current data. Tests of measurement invariance demonstrated strict measurement equivalence across gender and age. No gender and age differences have been found in the general depression factor. Findings provided evidence for the composite reliability and construct validity of DES-DC. Depressive symptoms positively related to the Big Five trait neuroticism, negative emotions, loneliness, social anxious behaviors, and disruptive behaviors during school and negatively related to agreeableness, conscientiousness, extraversion, physical health status, school connectedness, and academic self-efficacy. CONCLUSIONS: This study provides support for the one general factor construct of the CES-DC and the continuum concept structure of adolescent depression. Moreover, this research offers empirical evidence for comparing depression symptoms among adolescent populations with diverse genders and age groups. Additionally, the findings replicate and expand upon the implications of depressive symptoms on adolescents' traits, well-being, social-relational adjustment, and academic adaptation.
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Acne vulgaris caused by antibiotic-resistant Cutibacterium acnes (C. acnes) infection is difficult to treat conventionally. Phages have been suggested as a potential solution, but research on the mechanism of phage treatment is inadequate. This research investigates the underlying molecular mechanisms of phage φPaP11-13 attenuating C. acnes-induced inflammation in rat models. We found that rats infected with C. acnes had higher average ear thickness, greater enrichment of inflammatory cells as shown by hematoxylin-eosin (HE) staining, and fewer TUNEL (TdT-mediated dUTP Nick-End Labeling)-positive keratinocytes visualized by IF staining. Moreover, an increase of IGF-1 and IGF-1 receptor (IGF-1r) was detected using the immunohistochemical (IHC) staining method, Western blot (WB), and quantitative real-time PCR (qRT-PCR) when infected with C. acnes, which was decreased after the application of phage φPaP11-13. By applying the IGF-1 antibody, it was demonstrated that the severity of C. acnes-induced inflammation was relevant to the expression of IGF-1. Through WB and qRT-PCR, activation of the PI3K/Akt pathway and a down-regulation of the BAD-mediated apoptosis pathway were discovered after C. acnes infection. Subsequently, it was shown that the activation of the PI3K/Akt pathway against BAD-mediated apoptosis pathway was alleviated after applying phage φPaP11-13. Furthermore, applying the IGF-1r inhibitor, Pan-PI3K inhibitor, and Akt inhibitor reversed the changing trends of BAD induced by C. acnes and phage φPaP11-13. This study demonstrates that one of the critical mechanisms underlying the attenuation of acne vulgaris by phage φPaP11-13 is lysing C. acnes and regulating keratinocyte apoptosis via the PI3K/Akt signaling pathway.IMPORTANCECutibacterium acnes infection-induced acne vulgaris may cause severe physical and psychological prognosis. However, the overuse of antibiotics develops drug resistance, bringing challenges in treating Cutibacterium acnes. Bacteriophages are currently proven effective in MDR (multiple drug-resistant) Cutibacterium acnes, but there is a significant lack of understanding of phage therapy. This study demonstrated a novel way of curing acne vulgaris by using phages through promoting cell death of excessive keratinocytes in acne lesions by lysing Cutibacterium acnes. However, the regulation of this cell cycle has not been proven to be directly mediated by phages. The hint of ternary relation among "phage-bacteria-host" inspires huge interest in future phage therapy studies.
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Acne Vulgar , Bacteriófagos , Animais , Ratos , Fator de Crescimento Insulin-Like I/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Queratinócitos/metabolismo , Queratinócitos/patologia , Acne Vulgar/microbiologia , Propionibacterium acnes/metabolismo , Inflamação/metabolismo , ApoptoseRESUMO
Effective interpersonal emotion regulation (IER) strategies have been found to be meaningful predictors for positive psychological functioning. The Difficulties in Interpersonal Regulation of Emotions Scale (DIRE) is a measure developed to assess maladaptive IER strategies. This study aimed to examine the psychometric properties of the Chinese version of DIRE using two college student samples (Sample 1: n = 296; Sample 2: n = 419). The two-factor structure of DIRE (venting and excessive reassurance-seeking) was confirmed through an exploratory structure equation modeling approach. Our results demonstrated that the Chinese version of DIRE exhibits a similar factor structure (in both samples) as the original DIRE. Measurement invariance across gender and samples was also achieved. Latent mean analyses demonstrated that females more frequently reported excessive reassurance-seeking (in both samples) and venting (in Sample 1) than males. Furthermore, venting and excessive reassurance-seeking were significantly related to intrapersonal emotion regulation and well-being indicators. Although in Chinese culture DIRE performs somewhat differently from the original DIRE, the current findings suggest that DIRE is a reliable and valid scale with which to measure the IER strategies in Chinese culture and the use of this measure in clinical practice may allow for an accurate assessment of emotion regulation deficits in clients from other diverse cultures.
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Starch chain-length distributions play a key role in regulating the processing and digestion characteristics of proso millet starch. Waxy proso millet starch has higher endothermic enthalpy (13.06-16.73 J/g) owing to its higher relative crystallinity (27.83%-32.04%), while nonwaxy proso millet starch has lower peak viscosity (1.0630-1.1930 Paâs) and stronger viscoelasticity owing to its higher amylose content (21.72%-24.34%). Non-waxy proso millet starch exhibited two different digestion phases and its resistant starch content (18.37%-20.80%) was higher than waxy proso millet starch. Correlation analysis showed proso millet starch with longer amylopectin B1 chains and more amylopectin B2 chains exhibited excellent thermal ability and retrograde resistance, whereas proso millet starch with shorter and more amylose medium/long-chains not only reduced the digestion rate and increased the resistant starch content but also exhibited stronger viscoelasticity and excellent retrogradation properties. These results could provide more insights into efficient utilization of proso millet starch.
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Digestão , Amido , Amido/química , Amido/metabolismo , Viscosidade , Amilose/química , Amilose/análise , Amilose/metabolismo , Amilopectina/química , Amilopectina/metabolismoRESUMO
This study aimed to reveal the underlying mechanisms of the differences in viscoelasticity and digestibility between mung bean starch (MBS) and proso millet starch (PMS) from the viewpoint of starch fine molecular structure. The contents of amylopectin B2 chains (14.94-15.09 %), amylopectin B3 chains (14.48-15.07 %) and amylose long chains (183.55-198.84) in MBS were significantly higher than PMS (10.45-10.76 %, 12.48-14.07 % and 70.59-88.03, respectively). MBS with higher amylose content (AC, 28.45-31.80 %) not only exhibited a lower weight-average molar mass (91,750.65-128,120.44 kDa) and R1047/1022 (1.1520-1.1904), but also was significantly lower than PMS in relative crystallinity (15.22-23.18 %, p < 0.05). MBS displayed a higher storage modulus (G') and loss modulus (G'') than PMS. Although only MBS-1 showed two distinct and discontinuous phases, MBS exhibited a higher resistant starch (RS) content than PMS (31.63-39.23 %), with MBS-3 having the highest RS content (56.15 %). Correlation analysis suggested that the amylopectin chain length distributions and AC played an important role in affecting the crystal structure, viscoelastic properties and in vitro starch digestibility of MBS and PMS. These results will provide a theoretical and scientific basis for the development of starch science and industrial production of low glycemic index starchy food.
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Amilopectina , Amilose , Panicum , Amido , Vigna , Amilopectina/análise , Amilose/análise , Vigna/química , Amido/química , Panicum/química , Pepsina A/metabolismo , Difração de Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Peso Molecular , CinéticaRESUMO
BACKGROUND: Hepatocellular carcinoma (HCC) is a highly aggressive liver cancer with significant morbidity and mortality rates. AXIN1 is one of the top-mutated genes in HCC, but the mechanism by which AXIN1 mutations contribute to HCC development remains unclear. METHODS: In this study, we utilized CRISPR/Cas9 genome editing to repair AXIN1-truncated mutations in five HCC cell lines. RESULTS: For each cell line we successfully obtained 2-4 correctly repaired clones, which all show reduced ß-catenin signaling accompanied with reduced cell viability and colony formation. Although exposure of repaired clones to Wnt3A-conditioned medium restored ß-catenin signaling, it did not or only partially recover their growth characteristics, indicating the involvement of additional mechanisms. Through RNA-sequencing analysis, we explored the gene expression patterns associated with repaired AXIN1 clones. Except for some highly-responsive ß-catenin target genes, no consistent alteration in gene/pathway expression was observed. This observation also applies to the Notch and YAP/TAZ-Hippo signaling pathways, which have been associated with AXIN1-mutant HCCs previously. The AXIN1-repaired clones also cannot confirm a recent observation that AXIN1 is directly linked to YAP/TAZ protein stability and signaling. CONCLUSIONS: Our study provides insights into the effects of repairing AXIN1 mutations on ß-catenin signaling, cell viability, and colony formation in HCC cell lines. However, further investigations are necessary to understand the complex mechanisms underlying HCC development associated with AXIN1 mutations.
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Proteína Axina , Sistemas CRISPR-Cas , Carcinoma Hepatocelular , Neoplasias Hepáticas , Mutação , beta Catenina , Proteína Axina/genética , Proteína Axina/metabolismo , Humanos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/metabolismo , Linhagem Celular Tumoral , beta Catenina/metabolismo , beta Catenina/genética , Regulação Neoplásica da Expressão Gênica , Edição de Genes , Transdução de Sinais/genéticaRESUMO
Intrapersonal emotion dysregulation has been found to be a transdiagnostic predictor in the development of almost all affective disorders. Interpersonal resources are also involved in achieving people's emotion regulation goals. The Interpersonal Regulation Questionnaire (IRQ) has been developed to assess the tendency and efficacy of people using external resources to help manage their emotions. Under the restrictions of the COVID-19 pandemic, the role of interpersonal emotion regulation in individuals' adjustment and well-being remains unclear. This study aimed to investigate the optimal factor structure of the IRQ in Chinese culture using an exploratory structural equation modeling approach and to examine the associations between interpersonal emotion regulation, tested by the IRQ, and young people's intrapersonal emotion dysregulation and social and emotional well-being. The sample consisted of 556 college students aged from 17 to 31 from Mainland China. Factor analyses suggested that the four-factor structure was the optimal model for the current data. Females reported a higher tendency to use external resources to regulate their negative emotions and higher efficacy in regulating negative emotions. The Chinese version of the IRQ (C-IRQ) presented adequate psychometric properties and would be a useful tool for measuring interpersonal emotion regulation behaviors.
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Purpose: In recent years, research on interpersonal emotion regulation has garnered increasing attention. Extrinsic emotion regulation (EER) or the regulation of others' emotions is associated with individual personal relationships and mental health outcomes. However, investigations into why people engage in extrinsic emotion regulation are relatively rare. The current study aimed to identify the underlying factor structure of the motives of extrinsic emotion regulation, based on the Extrinsic Emotion Regulation Motives Scale (EERMS), to examine how these factors related to people's moral identity and social relationships during the COVID-19 pandemic. Participants and Methods: The participants of this study were 464 adults (73.1% females) from China who completed an online survey based on EERMS and other related measures. An exploratory factor analysis (EFA) was utilized to examine the underlying structure of EER motives followed by a bifactor EFA (bi-EFA). Results: Analyses revealed that the bi-EFA model, with an overarching factor (other-oriented empathetic concern) and three specific factors (self-orientation, reciprocation, and obligation), was best applied to the data. These factors were differently associated with social well-being indicators (moral self-identity, peer relationship quality, and perceived social support). Conclusion: This preliminary study supports the use of a hierarchical perspective to understand the different dimensions of motives for extrinsic emotion regulation.
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Background: The triple transgenic mouse model of Alzheimer's disease (3×Tg-AD) has gained popularity in Alzheimer's research owing to the progressive development of both amyloid-ß and tau pathologies in its brain. Prior handling-habituation, a necessary preparation procedure that reduces anxiety and stress in rodents, was seldom described in the literature involving these mice and needs to be addressed. Objective: We sought to determine whether 3×Tg-AD mice differ from B6;129 genetic control mice in terms of tameness and prior habituation to handling. Methods: We devised hand-staying and hand-boarding assays to evaluate tameness in 3×Tg-AD and B6;129 genetic control mice at 2.5, 7, and 11.5 months of age, representing cognitively pre-symptomatic, early symptomatic and advanced symptomatic stages of the disease, respectively. We monitored the progress of handling-habituation across 8-15 daily handling sessions and assessed the animal behaviors in elevated plus maze. Results: We found that 3×Tg-AD mice were markedly tamer than age-matched control mice at the baseline. Whereas it took 2-3 days for 3×Tg-AD mice to reach the criteria for full tameness, it took an average of 7-9 days for young genetic control mice to do so. Prior handling-habituation enhanced risk assessment and coping strategy in mice in elevated plus maze. Completely handling-habituated mice exhibited comparable anxiety indices in the maze regardless of genotype and age. Conclusion: These findings collectively point to inherently heightened tameness and accelerated handling-habituation in 3×Tg-AD mice on a B6;129 genetic background. These traits should be carefully considered when behaviors are compared between 3×Tg-AD and the genetic control mice.
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Quick differentiation of the circulating variants and the emerging recombinant variants of SARS-CoV-2 is essential to monitor their transmission. However, the widely used gene sequencing method is time-consuming and costly when facing the viral recombinant variants, because partial or whole genome sequencing is required. Allele-specific real time RT-PCR (qRT-PCR) represents a quick and cost-effective method in SNP genotyping and has been successfully applied for SARS-CoV-2 variant screening. In the present study, we developed a panel of 3 multiplex allele-specific qRT-PCR assays targeting 12 key differential mutations for quick differentiation of SARS-CoV-2 recombinant variants (XD and XE) and Omicron subvariants (BA.1 and BA.2). Two parallel multiplex qRT-PCR reactions were designed to separately target the protype allele and the mutated allele of the four mutations in each allele-specific qRT-PCR assay. The variation of Cp values (ΔCp) between the two multiplex qRT-PCR reactions was applied for mutation determination. The developed multiplex allele-specific qRT-PCR assays exhibited outstanding analytical sensitivities (with limits of detection [LoDs] of 2.97-27.43 copies per reaction), wide linear detection ranges (107-100 copies per reaction), good amplification efficiencies (82% to 95%), good reproducibility (Coefficient of Variations (CVs) < 5% in both intra-assay and inter-assay tests) and clinical performances (99.5%-100% consistency with Sanger sequencing). The developed multiplex allele-specific qRT-PCR assays in this study provide an alternative tool for quick differentiation of SARS-CoV-2 recombinant variants (XD and XE) and Omicron subvariants (BA.1 and BA.2).
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COVID-19 , SARS-CoV-2 , Alelos , COVID-19/diagnóstico , Teste para COVID-19 , Humanos , RNA Viral/genética , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , SARS-CoV-2/genéticaRESUMO
BACKGROUND: Women have a two-fold higher risk than men to Alzheimer's disease (AD) at midlife. Larger brain tau burden was consistently shown in older women than age-matched men. The biological basis for this gender disparity remains elusive. OBJECTIVE: We sought to know whether tau expression and phosphorylation physiologically differ between males and females. METHODS: We used western blots and immunohistochemistry to compare the levels of total tau and phosphorylated tau in the hippocampus and entorhinal cortex (EC) between sexes in Wistar rats at 40 days, and 8 and 20 months of age. RESULTS: We detected no statistically significant difference in total tau, 3R-tau, and 4R-tau between sexes. However, female rats exhibited lower levels of tau unphosphorylated at the Tau-1 site at 40 days of age. At 8 months of age, females showed higher levels of tau phosphorylated at Ser190, Ser387, and Ser395 (Ser199, Ser396, and Ser404 of human tau, respectively) than males in EC. At 20 months of age, both brain regions of female rats consistently showed higher levels than males of tau phosphorylated at Ser253, Ser387, PHF-1 (Ser387/395), and Ser413 sites, which correspond to Ser262, Ser396, Ser396/404, and Ser422 of human tau, respectively. CONCLUSION: Rats of both sexes have comparable levels of total tau, 3R-tau, and 4R-tau, whereas females exhibit higher levels of tau phosphorylated at multiple sites that are implicated in AD tau pathology, indicating a sexual dimorphism of tau phosphorylation that may potentially underlie the disparity in brain tau burden and risk for AD between sexes.
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Doença de Alzheimer , Córtex Entorrinal/metabolismo , Hipocampo/metabolismo , Proteínas tau/metabolismo , Fatores Etários , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Animais , Causalidade , Feminino , Regulação da Expressão Gênica , Imuno-Histoquímica , Masculino , Fosforilação , Isoformas de Proteínas , Ratos , Ratos Wistar , Fatores SexuaisRESUMO
The fatigue energy consumption of independent gestures can be obtained by calculating the power spectrum of surface electromyography (sEMG) signals. The existing research studies focus on the fatigue of independent gestures, while the research studies on integrated gestures are few. However, the actual gesture operation mode is usually integrated by multiple independent gestures, so the fatigue degree of integrated gestures can be predicted by training neural network of independent gestures. Three natural gestures including browsing information, playing games, and typing are divided into nine independent gestures in this paper, and the predicted model is established and trained by calculating the energy consumption of independent gestures. The artificial neural networks (ANNs) including backpropagation (BP) neural network, recurrent neural network (RNN), and long short-term memory (LSTM) are used to predict the fatigue of gesture. The support vector machine (SVM) is used to assist verification. Mean square error (MSE), root mean square error (RMSE), and mean absolute error (MAE) are utilized to evaluate the optimal prediction model. Furthermore, the different datasets of the processed sEMG signal and its decomposed wavelet coefficients are trained, respectively, and the changes of error functions of them are compared. The experimental results show that LSTM model is more suitable for gesture fatigue prediction. The processed sEMG signals are appropriate for using as the training set the fatigue degree of one-handed gesture. It is better to use wavelet decomposition coefficients as datasets to predict the high-dimensional sEMG signals of two-handed gestures. The experimental results can be applied to predict the fatigue degree of complex human-machine interactive gestures, help to avoid unreasonable gestures, and improve the user's interactive experience.
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Algoritmos , Gestos , Eletromiografia , Fadiga , Mãos , Humanos , Redes Neurais de ComputaçãoRESUMO
Ziziphus Jujuba cv. Pozao has been consumed as a traditional fruit with regional characteristics in China for a long time; however, fewer studies on polysaccharides from Ziziphus Jujuba cv. Pozao (JP) have been documented. This study aimed to evaluate the effect of oral administration of JP on cyclophosphamide-induced ICR mice for 28 days. The results showed that oral administration of JP could significantly improve the lymphocyte proliferation in the spleen and decrease the proportion of CD3+ and CD4+ and the ratio of CD4+/CD8+ in cyclophosphamide-induced mice in a dose-dependent manner. JP treatment also increased the levels of IL-2, IL-4, IL-10, IFN-γ, and TNF-α in serum and the intestine, and the improvement effects were proportional to the dose of JP. Similarly, JP significantly increased the levels of IgA and SIgA, as well as the expressions of Claudin-1 and Occludin in the intestine. Particularly, the expressions of Claudin-1 and Occludin were the best in the M-JP group. Furthermore, JP positively regulated the gut microbiota as indicated by the enriched microbiota diversity. At the phylum level, the relative abundance of Firmicutes was significantly decreased by JP, while that of Bacteroidetes was increased by JP treatment. More importantly, the ratio of Firmicutes/Bacteroidetes was significantly increased. And a high dose of JP is the most effective. At the genus level, the abundances of the Bacteroidales-S24-7-group, Lachnospiraceae, Alloprevotella, Alistipes and Bacteroides were increased by JP treatment. These results provided evidence for the regulating effect of JP on the peripheral immunity and intestinal barrier function in cyclophosphamide-induced hypoimmune mice.