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Introduction: Manganese (Mn) plays a pivotal role in plant growth and development. Aside aiding in plant growth and development, Mn as heavy metal (HM) can be toxic in soil when applied in excess. Morus alba is an economically significant plant, capable of adapting to a range of environmental conditions and possessing the potential for phytoremediation of contaminated soil by HMs. The mechanism by which M. alba tolerates Mn stresses remains obscure. Methods: In this study, Mn concentrations comprising sufficiency (0.15 mM), higher regimes (1.5 mM and 3 mM), and deficiency (0 mM and 0.03 mM), were applied to M. alba in pot treatment for 21 days to understand M. alba Mn tolerance. Mn stress effects on the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), intercellular CO2 concentration (Ci), chlorophyll content, plant morphological traits, enzymatic and non-enzymatic parameters were analyzed as well as metabolome signatures via non-targeted LC-MS technique. Results: Mn deficiency and toxicity decrease plant biomass, Pn, Ci, Gs, Tr, and chlorophyll content. Mn stresses induced a decline in the activities of catalase (CAT) and superoxide dismutase (SOD), while peroxidase (POD) activity, and leaf Mn content, increased. Soluble sugars, soluble proteins, malondialdehyde (MDA) and proline exhibited an elevation in Mn deficiency and toxicity concentrations. Metabolomic analysis indicates that Mn concentrations induced 1031 differentially expressed metabolites (DEMs), particularly amino acids, lipids, carbohydrates, benzene and derivatives and secondary metabolites. The DEMs are significantly enriched in alpha-linolenic acid metabolism, biosynthesis of unsaturated fatty acids, galactose metabolism, pantothenate and CoA biosynthesis, pentose phosphate pathway, carbon metabolism, etc. Discussion and conclusion: The upregulation of Galactinol, Myo-inositol, Jasmonic acid, L-aspartic acid, Coproporphyrin I, Trigonelline, Pantothenol, and Pantothenate and their significance in the metabolic pathways makes them Mn stress tolerance metabolites in M. alba. Our findings reveal the fundamental understanding of DEMs in M. alba's response to Mn nutrition and the metabolic mechanisms involved, which may hold potential significance for the advancement of M. alba genetic improvement initiatives and phytoremediation programs.
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There is growing interest in the potential health benefits of probiotics for both humans and animals. The study aimed to investigate the effects of feeding the canine-derived probiotic Pediococcus acidilactici GLP06 to adult beagles by analysing the microbiome and metabolome. Twenty-four healthy adult beagles were randomly assigned to four groups. The CK group received a standard diet, while the three probiotic groups, the LG group (2 × 108 CFU/day/dog), MG group (2 × 109 CFU/day/dog), and HG group (2 × 1010 CFU/day/dog), received the standard diet supplemented with varying amounts of probiotics. The results show that, compared to the CK group, total antioxidant capacity was significantly increased in the MG and HG groups (p < 0.05), and superoxide dismutase and catalase were significantly increased in the HG group (p < 0.05). Compared to the CK group, malondialdehyde and blood urea nitrogen content were significantly decreased in the MG and HG groups (p < 0.05). Additionally, secretory immunoglobulin A activity was significantly increased in the HG group compared to the CK and LG groups (p < 0.05), and immunoglobulin G activity was significantly increased in the HG group compared to the CK, LG, and MG groups (p < 0.05). In addition, compared with the CK group, the abundance of Faecalitalea and Collinsella increased in the LG group, and the relative abundance of Tyzzerella and Parasutterella increased in the MG group. The α diversity and the relative abundances of beneficial bacteria (Faecalibacterium, Lachnospiraceae_NK4A1316, and Ruminococcaceae_UCG-005) were higher in the HG group than in the CK group. Furthermore, acetic acid content was significantly increased in the HG group compared to the CK, LG, and MG groups (p < 0.05). Butyric acid, isobutyric acid, and the total SCFA content were significantly increased in the HG group compared to the CK group (p < 0.05). Moreover, metabolome analysis revealed 111 upregulated and 171 downregulated metabolites in the HG group. In conclusion, this study presents evidence that supplementing with P. acidilactici GLP06 can have a positive impact on antioxidant activity, immunoproteins, SCFAs, and gut microbiota in adult beagles. These findings highlight the potential of probiotics as a dietary intervention to enhance gut health and overall wellbeing in companion animals.
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Tropical and subtropical forests play a crucial role in global carbon (C) pools, and their responses to warming can significantly impact C-climate feedback and predictions of future global warming. Despite earth system models projecting reductions in land C storage with warming, the magnitude of this response varies greatly between models, particularly in tropical and subtropical regions. Here, we conducted a field ecosystem-level warming experiment in a subtropical forest in southern China, by translocating mesocosms (ecosystem composed of soils and plants) across 600 m elevation gradients with temperature gradients of 2.1°C (moderate warming), to explore the response of ecosystem C dynamics of the subtropical forest to continuous 6-year warming. Compared with the control, the ecosystem C stock decreased by 3.8% under the first year of 2.1°C warming; but increased by 13.4% by the sixth year of 2.1°C warming. The increased ecosystem C stock by the sixth year of warming was mainly attributed to a combination of sustained increased plant C stock due to the maintenance of a high plant growth rate and unchanged soil C stock. The unchanged soil C stock was driven by compensating and offsetting thermal adaptation of soil microorganisms (unresponsive soil respiration and enzyme activity, and more stable microbial community), increased plant C input, and inhibitory C loss (decreased C leaching and inhibited temperature sensitivity of soil respiration) from soil drying. These results suggest that the humid subtropical forest C pool would not necessarily diminish consistently under future long-term warming. We highlight that differential and asynchronous responses of plant and soil C processes over relatively long-term periods should be considered when predicting the effects of climate warming on ecosystem C dynamics of subtropical forests.
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Sequestro de Carbono , Ecossistema , Mudança Climática , Florestas , Carbono , SoloRESUMO
The present study describes the preparation of woven silk fabric (WSF) and poly(ethylene glycol) diacrylate (PEGDA) hydrogel composite reinforced artificial heart valve (SPAHV). Interestingly, the longitudinal and latitudinal elastic modulus of the SPAHV composite can achieve at 54.08 ± 3.29 MPa and 23.96 ± 2.18 MPa, respectively, while its volume/mass swelling ratio and water permeability was 1.9 %/2.8 % and 3 mL/(cm2âmin), respectively, revealing remarkable anisotropic mechanical properties, low water swelling property and water permeability. The in vitro & in vivo biocompatibility and anti-calcification ability of SPAHV were further examined using L929 mouse fibroblasts and Sprague Dawley (SD) male rat model under 8 weeks of subcutaneous implantation. The expression of pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 was determined by immunohistochemical staining, as well as the H&E staining and alizarin red staining were accessed. The results showed that the composites possess better biocompatibility, resistance to degradation and anti-calcification ability compared to the control group (p < 0.05). Thus, the SPAHV composite with robust mechanical properties and biocompatibility has potential application for artificial heart valves.
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Coração Artificial , Hidrogéis , Ratos , Camundongos , Animais , Masculino , Hidrogéis/farmacologia , Hidrogéis/química , Ratos Sprague-Dawley , Polietilenoglicóis/química , Citocinas , Água , SedaRESUMO
BACKGROUND: In order to study the effect of adsorption of surfactant at the two interfacial layers on emulsion stability, the kinetically stable water-in-oil-in-water (W/O/W) emulsion carriers were prepared using polyglycerol polyricinoleate (PGPR) and gum arabic (GA) as emulsifiers. The relationship between the adsorption of the surfactant and the stability mechanism of the emulsions was elucidated. RESULTS: When the contents of PGPR and GA were low, the interfaces between oil and the inner and outer water phases, respectively, could not be completely covered. However, when the concentration of PGPR was higher than 60 g kg-1 , the excess PGPR was adsorbed on the interface between the oil phase and the outer water phase. When the concentration of GA reached 80 g kg-1 , more GA was adsorbed to the oil-in-water interface. Moreover, the presence of PGPR on the interface could reduce the adsorption capacity of GA. Two types of kinetically stable emulsions were obtained by optimizing the interface composition (60 g kg-1 GA/80 g kg-1 PGPR and 60 g kg-1 PGPR/80 g kg-1 GA). The kinetically stable W/O/W emulsions prepared in this study were successfully used to encapsulate a hydrophilic vitamin (vitamin B12) with an encapsulation efficiency (EE) of 80% and release efficiency (RE) of 95%. The interfacial adsorption GA can accelerate the hydrolysis of fat. CONCLUSION: Overall, this study provides a new strategy for the preparation of W/O/W emulsions, which might be beneficial for application in food, cosmetic, chemical, and pharmaceutical industries. © 2023 Society of Chemical Industry.
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Goma Arábica , Tensoativos , Emulsões/química , Goma Arábica/química , Tensoativos/química , Água/químicaRESUMO
Recent developments in quantum light-matter coupled systems and quantum transducers have highlighted the need for cryogenic optical measurements. In this study, we present a packaged fiber-optic coupler with a coupling efficiency of over 50% for telecom wavelength light down to the mK temperature range. Besides the high coupling efficiency, our method enables sensitive photonic device measurements that are immune to mechanical vibrations present in cryogenic setups.
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Probiotics are available from various sources, including the gastrointestinal tract of healthy animals. In this study, Pediococcus acidilactici was isolated for the first time from Felis catus and evaluated for its functionality. The findings revealed that P. acidilactici CLP03 exhibited inhibitory properties against pathogenic bacteria (E. coli, Salmonella, S. aureus, P. aeruginosa, and L. monocytogenes). Then, survival of strains exposed to pH 2.5, 0.3% bile salts, 0.5% bile salts, and gastrointestinal fluids was 63.97%, 98.84%, 87.95%, and 52.45%, respectively. Also, P. acidilactici CLP03 demonstrated high hydrophobicity (69.63-82.03%) and self-aggregation (73.51-81.44%), negative for hemolytic, and was susceptible to clindamycin. Finally, the scavenging rates of DPPH, ABTS, and O2- were 53.55%, 54.81%, and 85.13%, respectively, which demonstrated that the strain CLP03 has good oxidation resistance. All these characteristics contribute to the survival, colonization, and functionality of the strain in the gastrointestinal tract, indicating their excellent probiotic potential. On the other hand, animal experiments (KM mice, randomly assigned to four groups) showed that the gavage of CLP03 had no toxic effects on mice, increased the serum SOD content, and decreased the MDA and BUN contents, which revealed gavage of CLP03 significantly increased the antioxidant capacity of mice in vivo. In addition, complete genome annotation showed that P. acidilactici CLP03 had 1976 CDS genes, and the numbers of CRISPR, gene islands, and phages were 8, 3, and 6, respectively. In conclusion, P. acidilactici CLP03 could be a candidate functional cat probiotic to enhance animal health and welfare.
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Based on a computational approach that can accurately describe their geometric structures and electronic spectra, we have theoretically studied the nonlinear optical (NLO) properties of H-capped carbon chains, H-(CîC)n-H (n = 3-15), for the first time. Special attention was paid to the size dependence of the molecular (hyper)polarizability of these species through the nonlinear fitting of the data, which formed two power-law formulas of αiso(∞) = -0.206 + 0.264n1.498 and γâ(∞) = -0.624 + 0.006n3.368 and was thoroughly discussed at the electronic structure level by in-depth wavefunction analyses. The fundamental gap (ΔE) between vertical ionization energy (VIE) and vertical electron affinity (VEA) is found to be related to the molecular (hyper)polarizability. The calculated (hyper)polarizability of the carbon chains H-(CîC)n-H (n = 3-15) is more sensitive to the density functional theory (DFT) applied than to the basis set selected. The results are expected to provide theoretical guidance for the property prediction of arbitrarily long carbon chains not yet synthesized.
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Osteoporosis, a systemic bone disease defined by decreased bone mass and deterioration of bone microarchitecture, is becoming a global concern. Nodakenin (NK) is a furanocoumarin-like compound isolated from the traditional Chinese medicine Radix Angelicae biseratae (RAB). NK has been reported to have various pharmacological activities, but osteoporosis has not been reported to be affected by NK. In this study, we used network pharmacology, molecular docking and molecular dynamics simulation techniques to identify potential targets and pathways of NK in osteoporosis. We found that NK treatment significantly promoted osteogenic differentiation of BMSCs while activating the PI3K/AKT/mTOR signalling pathway by measuring alkaline phosphatase activity and the expression of various osteogenic markers. In contrast, LY294002, an inhibitor of PI3K, reversed these changes and inhibited the osteogenic differentiation-enabling effect of NK. Meanwhile, prevent the Akt and NFκB signalling pathways by down-regulating c-Src and TRAF6 thereby effectively inhibiting RANKL-induced osteoclastogenesis. In addition, oral administration of NK to mice significantly elevated bone mass and ameliorated ovariectomized (OVX)-mediated bone microarchitectural disorders. In conclusion, these data suggest that NK attenuates OVX-induced bone loss by enhancing osteogenesis and inhibiting osteoclastogenesis.
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Osteogênese , Osteoporose , Feminino , Camundongos , Animais , Humanos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Simulação de Acoplamento Molecular , Osteoporose/tratamento farmacológico , Osteoporose/etiologia , Osteoporose/metabolismo , Ovariectomia/efeitos adversos , Osteoclastos , Diferenciação Celular , Ligante RANK/farmacologiaRESUMO
Ca-biochar is an efficient material for As(III)-containing acid mine drainage (AMD) decontamination, while it is challenging to fabricate Ca-biochar with oyster shell waste as the Ca source due to its complex structure. Herein, a mechanochemical method was proposed to activate oyster shell waste and wood waste for Ca-biochar design and production, and its efficacy and relevant mechanisms for AMD detoxification were evaluated. The smaller size Ca-biochar produced by the medium-speed ball milling showed a higher As(III) removal (74.0 %) compared to high-speed ball milling (60.9 %), attributed to the formation of finer Ca(OH)2 while avoiding particle aggregation, which could release more Ca (89.0 mg/g) and alkalinity for the co-precipitation of As. Meanwhile, wood-based biochar substrate served as a platform for co-precipitation, and its surface functionality supported the oxidative immobilization of As. This study presents a promising route for upcycling food and wood waste to produce Ca-biochar for AMD decontamination.
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Arsênio , Cálcio , Descontaminação , Carvão Vegetal/química , AdsorçãoRESUMO
[This corrects the article DOI: 10.3389/fmicb.2023.1179953.].
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BACKGROUND: Fat significantly affects the properties of ice cream. Prior studies have investigated the correlation between fat crystallization, fat destabilization, and ice cream quality. However, the role of fatty acid composition, the similarity between fat and emulsifier in these characteristics, and their impact on final product quality remains unclear. RESULTS: To investigate the influence of the fatty acid composition of fats, as well as their similarity to glycerol monostearate (GMS), on fat crystallization and destabilization during the aging and freezing stages, ice creams were formulated using a combination of two types of fats (coconut oil and palm olein) in five different ratios. In oil phases, decreased saturation of fatty acids (from 93.38% to 46.69%) and increased similarity to GMS (from 11.96% to 46.01%) caused a reduction in the maximum solid fat content. Moreover, the rise in unsaturated long-chain fatty acids (from 34.61% to 99.57%) and similarity to GMS enhanced the formation of rare and coarse fat crystals, leading to a sparse crystalline network. This, in turn, reduced the crystallization rate and the stiffness of the fat in emulsions. Assuming consistent overrun across all ice creams, the enhanced interactions between fat globules in ice cream improved its hardness, melting properties, and shrinkage. CONCLUSION: The crystalline properties of fat in emulsions were influenced by oil phases, impacting fat destabilization and ultimately enhancing the quality of ice cream. The present study offers valuable insights for the optimization of fat and monoglyceride fatty acid ester selection, with the potential to improve ice cream quality. © 2023 Society of Chemical Industry.
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Ácidos Graxos , Sorvetes , Cristalização , Sorvetes/análise , Glicerol , Gorduras/química , Emulsões/químicaRESUMO
The beneficial effects of lactic acid bacteria are well known and recognized as functional foods that are health benefits for companion animals. This study, for the first time, reports the probiotic properties, safety, and whole-genome sequence of Pediococcus acidilactici GLP06 isolated from feces of beagles. In this study, candidate probiotic bacteria P. acidilactici GLP02 and GLP06 were morphologically characterized and tested for their antimicrobial capacity, tolerance to different conditions (low pH, bile salts, an artificial gastrointestinal model, and high temperature), antibiotic sensitivity, hemolytic activity, cell surface hydrophobicity, autoaggregation activity, and adhesion to Caco-2 cells. P. acidilactici GLP06 showed better probiotic potential. Therefore, P. acidilactici GLP06 was evaluated for in vivo safety in mice and whole-genome sequencing. The results showed, that the supplemented MG06 group (1010 cfu/mL), GLP06 was not only nontoxic to mice, but also promoted the development of the immune system, improved resistance to oxidative stress, and increased the diversity of intestinal microorganisms and the abundance of Lactobacillus. Whole-genome sequencing showed that P. acidilactici GLP06 was 2,014,515 bp and contained 1,976 coding sequences, accounting for 86.12% of the genome, with no drug resistance genes and eight CRISPR sequences. In conclusion, the newly isolated canine-derived P. acidilactici GLP06 had good probiotic potential, was nontoxic to mice and promoted the development of immune organs, improved the biodiversity of the intestinal flora, and had no risk of drug-resistant gene transfer, indicating that P. acidilactici GLP06 can be used as a potential probiotic for the prevention and treatment of gastrointestinal diseases in companion animals.
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Ovalbumin (OVA)/sodium carboxymethylcellulose (CMC) colloidal particles were prepared with different compactness and morphologies by regulating the interaction between proteins and polysaccharides during heating. Electrostatic interactions between the amine groups of OVA (-NH3+) and carboxyl groups of CMC (-COO-) enhanced complex formation. The protein conformation change benefited the hydrophobic interaction between the particles. Proteins in colloidal particles were unfolded/folded under thermal induction to form aggregates having more ß-sheet structures. When the OVA/CMC ratio was 1:2, the initially loosely connected OVA/CMC aggregation changed into a uniform sphere between 25 and 90 °C. The mass ratio of OVA to CMC within the final colloidal particle (90 °C) was about 1:1.4. The OVA/CMC particle stability was maintained with hydrogen bonding, hydrophobicity, and disulfide bond. When OVA levels were predominant, OVA and CMC developed an approximately hollow sphere. Moreover, the final colloidal particle composition showed the OVA-to-CMC ratio as 3:1 (w/w). OVA bound into colloidal particle pores to increase compactness. Moreover, OVA and CMC bound to the colloidal particle while the particle shrank, thereby increasing the compactness of colloidal particles. There was a significant decrease in ABTSâ¢+ scavenging activity of curcumin compared with that of the particles with a ratio of 1:2. Thus, the rational adjustment of the structure of colloidal particles could effectively enhance their functional characteristics, providing a new way for the controlled release of the active ingredients.
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Carboximetilcelulose Sódica , Ovalbumina/química , Carboximetilcelulose Sódica/químicaRESUMO
Herein, the electronic structure and bonding character of the stable cyclo[18]carbon (C18 ) precursor, C18 Br6 , are thoroughly characterized by molecular orbital (MO), density of states (DOS), bond order (BO), and interaction region indicator (IRI) analyses. The delocalization characters of out-of-plane and in-plane π-electrons (labeled as πout - and πin -electrons, respectively) in bonding regions were examined using localized orbital locator (LOL) and electron localization function (ELF). The aromaticity was investigated, studying the molecular magnetic response to external magnetic field by computing the magnetically induced current density (Jind ), iso-chemical shielding surface (ICSS), anisotropy of the induced current density (AICD), and the induced magnetic field (Bind ). All these analyses indicate that C18 Br6 is a globally aromatic species with lower aromaticity than C18 , and the blocking of in-plane π-conjugation (labeled as πin -conjugation) by the presence of -Br substituents in it is the underlying cause for the weakening of molecular aromaticity.
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The photophysical property and optical nonlinearity of an electronic push-pull mesoionic compound, 2-(4-trifluoromethophenyl)-3-methyl-4-(4-methoxyphenyl)-1,3-thiazole-5-thiolate, were theoretically investigated with a reliable computing strategy. The essence of the optical properties were then explored through a variety of wavefunction analysis methods, including the natural transition orbital analysis, hole-electron analysis, (hyper)polarizability density analysis, decomposition of the (hyper)polarizability contribution, and (hyper)polarizability tensor analysis, at the level of electronic structure. The influences of the electric field and solvation on the absorption spectrum and (hyper)polarizability of the molecule are highlighted and clarified. The results are expected to provide guidance for people to understand the effects of external field wavelength and solvation on the optical properties of mesoscopic molecules.
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Conformação Molecular , HumanosRESUMO
Background: The burden of depression symptoms has increased among individuals infected with SARS-CoV-2 during COVID-19 pandemic. However, the prevalence and associated factors of depressive symptoms among individuals infected with SARS-CoV-2 remain uncertain after optimizing the COVID-19 response in China. Methods: An online cross-sectional survey was conducted among the public from January 6 to 30, 2023, using a convenience sampling method. Sociodemographic and COVID-19 pandemic-related factors were collected. The depression symptoms were assessed using the Patient Health Questionnaire-9 (PHQ-9). Logistic regression analysis was performed to explore the associated factors with depressive symptoms. Results: A total of 2,726 participants completed the survey. The prevalence of depression symptoms was 35.3%. About 58% of the participants reported experiencing insufficient drug supply. More than 40% of participants reported that they had missed healthcare appointments or delayed treatment. One-third of participants responded experiencing a shortage of healthcare staff and a long waiting time during medical treatment. Logistic regression analysis revealed several factors that were associated with depression symptoms, including sleep difficulties (OR, 2.84; 95% CI, 2.34-3.44), chronic diseases (OR, 2.15; 95% CI, 1.64-2.82), inpatient treatment for COVID-19 (OR, 3.24; 95% CI, 2.19-4.77), with COVID-19 symptoms more than 13 days (OR, 1.30, 95% CI 1.04-1.63), re-infection with SARS-CoV-2 (OR, 1.52; 95% CI, 1.07-2.15), and the increased in demand for healthcare services (OR, 1.32; 95% CI, 1.08-1.61). Conclusion: This study reveals a moderate prevalence of depression symptoms among individuals infected with SARS-CoV-2. The findings underscore the importance of continued focus on depressive symptoms among vulnerable individuals, including those with sleeping difficulties, chronic diseases, and inpatient treatment for COVID-19. It is necessary to provide mental health services and psychological interventions for these vulnerable groups during the COVID-19 epidemic.
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COVID-19 , Humanos , COVID-19/epidemiologia , Autorrelato , SARS-CoV-2 , Depressão/epidemiologia , Estudos Transversais , Pandemias , Prevalência , China/epidemiologia , Doença CrônicaRESUMO
Drought stress has been a key environmental factor affecting plant growth and development. The plant genome is capable of producing long noncoding RNAs (lncRNAs). To better understand white mulberry (Morus alba L.) drought response mechanism, we conducted a comparative transcriptome study comparing two treatments: drought-stressed (EG) and well-watered (CK) plants. A total of 674 differentially expressed lncRNAs (DElncRNAs) were identified. In addition, 782 differentially expressed messenger RNAs (DEmRNAs) were identified. We conducted Gene Ontology (GO) and KEGG enrichment analyses focusing on the differential lncRNAs cis-target genes. The target genes of the DElncRNAs were most significantly involved in the biosynthesis of secondary metabolites. Gene regulatory networks of the target genes involving DElncRNAs-mRNAs-DEmRNAs and DElncRNA-miRNA-DEmRNA were constructed. In the DElncRNAs-DEmRNAs network, 30 DEmRNAs involved in the biosynthesis of secondary metabolites are collocated with 46 DElncRNAs. The interaction between DElncRNAs and candidate genes was identified using LncTar. In summary, quantitative real-time polymerase chain reaction (qRT-PCR) validated nine candidate genes and seven target lncRNAs including those identified by LncTar. We predicted that the DElncRNAs-DEmRNAs might recruit microRNAs (miRNAs) to interact with gene regulatory networks under the drought stress response in mulberry. The findings will contribute to our understanding of the regulatory functions of lncRNAs under drought stress and will shed new light on the mulberry-drought stress interactions.
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The incidence of Candida infections in intensive care units (ICU) has significantly increased in recent years, and these infections have become one of the most serious complications threatening the lives of ICU patients. The proportion of non-Candida albicans infections, such as Candida krusei and Candida glabrata infections, which are resistant to fluconazole, is increasing each year. Early identification of the strains causing Candida infections is important for the timely implementation of targeted treatments to save patients' lives. However, the current methods of direct microscopy, culture, and histopathology, as well as other diagnostic methods, have many shortcomings, such as their low sensitivity and long assay times; therefore, they cannot meet the needs for early clinical diagnosis. Recombinant polymerase amplification (RPA) is a promising isothermal amplification technique that can be performed without sophisticated instruments and equipment, and is suitable for use in resource-poor areas. RPA combined with lateral flow strips (LFS) can be used to rapidly amplify and visualize target genes within 20 min. In this study, RPA-LFS was used to amplify the internal transcribed spacer 2 (ITS2) region of C. krusei. The primer-probe design was optimized by introduction of base mismatches (probe modification of five bases) to obtain a specific and sensitive primer-probe combination for the detection of clinical specimens. Thirty-five common clinical pathogens were tested with RPA-LFS to determine the specificity of the detection system. The RPA-LFS system specifically detected C. krusei without cross-reaction with other fungi or bacteria. A gradient dilution of the template was tested to explore the lower limit of detection and sensitivity of the assay. The sensitivity was 10 CFU/50 µL per reaction, without interference from genomic DNA of other species. The RPA-LFS and qPCR assays were performed on 189 clinical specimens to evaluate the detection performance of the RPA-LFS system. Seventy-six specimens were identified as C. krusei, indicating a detection rate of 40.2%. The results were consistent with those of qPCR and conventional culture methods. The RPA-LFS system established in our study provides a reliable molecular diagnostic method for the detection of C. krusei, thus meeting the urgent need for rapid, specific, sensitive, and portable clinical field testing.
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Candidíase , Recombinases , Candidíase/diagnóstico , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , Nucleotidiltransferases , Pichia , Sensibilidade e EspecificidadeRESUMO
The back-propagating action potential (bpAP) is crucial for neuronal signal integration and synaptic plasticity in dendritic trees. Its properties (velocity and amplitude) can be affected by dendritic morphology. Due to limited spatial resolution, it has been difficult to explore the specific propagation process of bpAPs along dendrites and examine the influence of dendritic morphology, such as the dendrite diameter and branching pattern, using patch-clamp recording. By taking advantage of Optopatch, an all-optical electrophysiological method, we made detailed recordings of the real-time propagation of bpAPs in dendritic trees. We found that the velocity of bpAPs was not uniform in a single dendrite, and the bpAP velocity differed among distinct dendrites of the same neuron. The velocity of a bpAP was positively correlated with the diameter of the dendrite on which it propagated. In addition, when bpAPs passed through a dendritic branch point, their velocity decreased significantly. Similar to velocity, the amplitude of bpAPs was also positively correlated with dendritic diameter, and the attenuation patterns of bpAPs differed among different dendrites. Simulation results from neuron models with different dendritic morphology corresponded well with the experimental results. These findings indicate that the dendritic diameter and branching pattern significantly influence the properties of bpAPs. The diversity among the bpAPs recorded in different neurons was mainly due to differences in dendritic morphology. These results may inspire the construction of neuronal models to predict the propagation of bpAPs in dendrites with enormous variation in morphology, to further illuminate the role of bpAPs in neuronal communication.