RESUMEN
MOTIVATION: The full automation of digital neuronal reconstruction from light microscopic images has long been impeded by noisy neuronal images. Previous endeavors to improve image quality can hardly get a good compromise between robustness and computational efficiency. RESULTS: We present the image enhancement pipeline named Neuronal Image Enhancement through Noise Disentanglement (NIEND). Through extensive benchmarking on 863 mouse neuronal images with manually annotated gold standards, NIEND achieves remarkable improvements in image quality such as signal-background contrast (40-fold) and background uniformity (10-fold), compared to raw images. Furthermore, automatic reconstructions on NIEND-enhanced images have shown significant improvements compared to both raw images and images enhanced using other methods. Specifically, the average F1 score of NIEND-enhanced reconstructions is 0.88, surpassing the original 0.78 and the second-ranking method, which achieved 0.84. Up to 52% of reconstructions from NIEND-enhanced images outperform all other four methods in F1 scores. In addition, NIEND requires only 1.6 s on average for processing 256 × 256 × 256-sized images, and images after NIEND attain a substantial average compression rate of 1% by LZMA. NIEND improves image quality and neuron reconstruction, providing potential for significant advancements in automated neuron morphology reconstruction of petascale. AVAILABILITY AND IMPLEMENTATION: The study is conducted based on Vaa3D and Python 3.10. Vaa3D is available on GitHub (https://github.com/Vaa3D). The proposed NIEND method is implemented in Python, and hosted on GitHub along with the testing code and data (https://github.com/zzhmark/NIEND). The raw neuronal images of mouse brains can be found at the BICCN's Brain Image Library (BIL) (https://www.brainimagelibrary.org). The detailed list and associated meta information are summarized in Supplementary Table S3.
Asunto(s)
Compresión de Datos , Neuronas , Animales , Ratones , Tomografía Computarizada por Rayos X/métodos , Aumento de la Imagen , Encéfalo , Procesamiento de Imagen Asistido por Computador/métodosRESUMEN
OBJECTIVE: Mucosal T cells play a major role in inflammatory bowel disease (IBD). However, their immunometabolism during intestinal inflammation is poorly understood. Due to its impact on cellular metabolism and proinflammatory immune cell function, we here focus on the enzyme ATP citrate lyase (ACLY) in mucosal T cell immunometabolism and its relevance for IBD. DESIGN: ACLY expression and its immunometabolic impact on colitogenic T cell function were analysed in mucosal T cells from patients with IBD and in two experimental colitis models. RESULTS: ACLY was markedly expressed in colon tissue under steady-state conditions but was significantly downregulated in lamina propria mononuclear cells in experimental dextran sodium sulfate-induced colitis and in CD4+ and to a lesser extent in CD8+ T cells infiltrating the inflamed gut in patients with IBD. ACLY-deficient CD4+ T cells showed an impaired capacity to induce intestinal inflammation in a transfer colitis model as compared with wild-type T cells. Assessment of T cell immunometabolism revealed that ACLY deficiency dampened the production of IBD-relevant cytokines and impaired glycolytic ATP production but enriched metabolites involved in the biosynthesis of phospholipids and phosphatidylcholine. Interestingly, the short-chain fatty acid butyrate was identified as a potent suppressor of ACLY expression in T cells, while IL-36α and resolvin E1 induced ACLY levels. In a translational approach, in vivo administration of the butyrate prodrug tributyrin downregulated mucosal infiltration of ACLYhigh CD4+ T cells and ameliorated chronic colitis. CONCLUSION: ACLY controls mucosal T cell immunometabolism and experimental colitis. Therapeutic modulation of ACLY expression in T cells emerges as a novel strategy to promote the resolution of intestinal inflammation.
Asunto(s)
Colitis , Enfermedades Inflamatorias del Intestino , Linfocitos Intraepiteliales , Humanos , Animales , Linfocitos Intraepiteliales/metabolismo , ATP Citrato (pro-S)-Liasa/metabolismo , Linfocitos T CD8-positivos/metabolismo , Colitis/metabolismo , Inflamación/metabolismo , Butiratos , Mucosa Intestinal/metabolismo , Sulfato de Dextran , Modelos Animales de EnfermedadRESUMEN
BACKGROUND: Culex tritaeniorhynchus is widely distributed in China, from Hainan Island in the south to Heilongjiang in the north, covering tropical, subtropical, and temperate climate zones. Culex tritaeniorhynchus carries 19 types of arboviruses. It is the main vector of the Japanese encephalitis virus (JEV), posing a serious threat to human health. Understanding the effects of environmental factors on Culex tritaeniorhynchus can provide important insights into its population structure or isolation patterns, which is currently unclear. RESULTS: In total, 138 COI haplotypes were detected in the 552 amplified sequences, and the haplotype diversity (Hd) value increased from temperate (0.534) to tropical (0.979) regions. The haplotype phylogeny analysis revealed that the haplotypes were divided into two high-support evolutionary branches. Temperate populations were predominantly distributed in evolutionary branch II, showing some genetic isolation from tropical/subtropical populations and less gene flow between groups. The neutral test results of HNQH (Qionghai) and HNHK(Haikou) populations were negative (P < 0.05), indicating many low-frequency mutations in the populations and that the populations might be in the process of expansion. Moreover, Wolbachia infection was detected only in SDJN (Jining) (2.24%), and all Wolbachia genotypes belonged to supergroup B. To understand the influence of environmental factors on mosquito-borne viruses, we examined the prevalence of Culex tritaeniorhynchus infection in three ecological environments in Shandong Province. We discovered that the incidence of JEV infection was notably greater in Culex tritaeniorhynchus from lotus ponds compared to those from irrigation canal regions. In this study, the overall JEV infection rate was 15.27 per 1000, suggesting the current risk of Japanese encephalitis outbreaks in Shandong Province. CONCLUSIONS: Tropical and subtropical populations of Culex tritaeniorhynchus showed higher genetic diversity and those climatic conditions provide great advantages for the establishment and expansion of Culex tritaeniorhynchus. There are differences in JEV infection rates in wild populations of Culex tritaeniorhynchus under different ecological conditions. Our results suggest a complex interplay of genetic differentiation, population structure, and environmental factors in shaping the dynamics of Culex tritaeniorhynchus. The low prevalence of Wolbachia in wild populations may reflect the recent presence of Wolbachia invasion in Culex tritaeniorhynchus.
Asunto(s)
Culex , Haplotipos , Filogenia , Culex/genética , Culex/virología , Culex/microbiología , Animales , China , Clima , Variación Genética , Genética de Población , Wolbachia/genética , Mosquitos Vectores/genética , Mosquitos Vectores/virología , Mosquitos Vectores/microbiología , Complejo IV de Transporte de Electrones/genéticaRESUMEN
As emerging contaminants in the environment, antibiotic resistance genes (ARGs) have aroused a global health crisis and posed a serious threat to ecological safety and human health. Thus, efficient and accurate onsite detection of ARGs is crucial for environmental surveillance. Here, we presented a colorimetric-photoelectrochemical (PEC) dual-mode bioassay for simultaneous detection of multiple ARGs by smartly incorporating rolling circle amplification (RCA) into a stimuli-responsive DNA nanoassembly, using the tetracycline resistance genes tetA and tetC as models. The tailored DNA nanoassembly containing RCA amplicons hybridized with specific signal probes: CuO nanoflowers-anchored signal DNA1 and HgO nanoparticles-anchored signal DNA2, respectively. Upon exposure to an acidic stimulus, numerous Cu2+ and Hg2+ were released, serving as the reporting agent of colorimetric/PEC dual-mode assay. The released Cu2+ and Hg2+ induced localized surface plasmon resonance shifts in Au nanorods and triangular Ag nanoplates through an etching process, respectively, enabling visual analysis of ARGs with distinguishing color changes. Meanwhile, numerous Cu2+ and Hg2+ triggered the amplified PEC variations via reacting with the photoactive layers of CuS/CdS and ZnS, respectively. Thus, a rapid and ultrasensitive colorimetric/PEC dual-mode detection of multiple ARGs was achieved with the detection limit down to 17.2 aM. Furthermore, such dual-mode bioassay could discriminate single-base mismatch and successfully determine ARGs in E. coli plasmids and sludge samples, holding great promise for point-of-care genetic diagnostics.
RESUMEN
Lactate dehydrogenase B (LDHB) reversibly catalyzes the conversion of pyruvate to lactate or lactate to pyruvate and expressed in various malignancies. However, the role of LDHB in modulating immune responses against hepatocellular carcinoma (HCC) remains largely unknown. Here, we found that down-regulation of lactate dehydrogenase B (LDHB) was coupled with the promoter hypermethylation and knocking down the DNA methyltransferase 3A (DNMT 3A) restored LDHB expression levels in HCC cell lines. Bioinformatics analysis of the HCC cohort from The Cancer Genome Atlas revealed a significant positive correlation between LDHB expression and immune regulatory signaling pathways and immune cell infiltrations. Moreover, immune checkpoint inhibitors (ICIs) have shown considerable promise for HCC treatment and patients with higher LDHB expression responded better to ICIs. Finally, we found that overexpression of LDHB suppressed HCC growth in immunocompetent but not in immunodeficient mice, suggesting that the host immune system was involved in the LDHB-medicated tumor suppression. Our findings indicate that DNMT3A-mediated epigenetic silencing of LDHB may contribute to HCC progression through remodeling the tumor immune microenvironment, and LDHB may become a potential prognostic biomarker and therapeutic target for HCC immunotherapy.
Asunto(s)
Carcinoma Hepatocelular , ADN Metiltransferasa 3A , Epigénesis Genética , L-Lactato Deshidrogenasa , Neoplasias Hepáticas , Microambiente Tumoral , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/metabolismo , Microambiente Tumoral/inmunología , Humanos , Animales , Ratones , L-Lactato Deshidrogenasa/metabolismo , L-Lactato Deshidrogenasa/genética , ADN Metiltransferasa 3A/metabolismo , Regulación Neoplásica de la Expresión Génica , Metilación de ADN , Isoenzimas/genética , Isoenzimas/metabolismo , Línea Celular Tumoral , Silenciador del Gen , PronósticoRESUMEN
Chronic inflammation is an important pathogenetic factor that leads to the progression of Alzheimer's disease (AD), and specialized pro-resolving lipid mediators (SPMs) play critical role in regulating inflammatory responses during AD pathogenesis. Maresin1 (MaR1) is the latest discovered SPMs, and it is found that MaR1 improves AD cognitive impairment by regulating neurotrophic pathways to protect AD synapses and reduce Aß production, which made MaR1 as candidate agent for AD treatment. Unfortunately, the underlying mechanisms are still largely known. In this study, the AD mice and cellular models were subjected to MaR1 treatment, and we found that MaR1 reduced Aß production to ameliorate AD-related symptoms and increased the expression levels of ADAM10/17, sAPPα and sAPPß to exert its anti-inflammatory role. In addition, as it was determined by Western Blot analysis, we observed that MaR1 could affected the neuroprotective signal pathways. Specifically, MaR1 downregulated p57NTR and upregulated TrkA to activate the p75NTR/TrkA signal pathway, and it could increase the expression levels of p-PI3K and p-Akt, and downregulated p-mTOR to activate the PI3K/AKT/ERK/mTOR pathway. Finally, we verified the role of ADAM10/17 in regulating AD progression, and we found that silencing of ADAM10/17 inactivated the above neuroprotective signal pathways to aggravate AD pathogenesis. In conclusion, MaR1 is verified as potential therapeutic agent for AD by eliminating Aß production, upregulating ADAM10/17, sAPPα and sAPPß, and activating the neuroprotective p75NTR/TrkA pathway and the PI3K/AKT/ERK/mTOR pathway.
Asunto(s)
Proteína ADAM10 , Enfermedad de Alzheimer , Secretasas de la Proteína Precursora del Amiloide , Péptidos beta-Amiloides , Ácidos Docosahexaenoicos , Transducción de Señal , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/tratamiento farmacológico , Animales , Proteína ADAM10/metabolismo , Proteína ADAM10/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Transducción de Señal/efectos de los fármacos , Ácidos Docosahexaenoicos/farmacología , Ácidos Docosahexaenoicos/metabolismo , Péptidos beta-Amiloides/metabolismo , Ratones , Inflamación/metabolismo , Proyectos Piloto , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Humanos , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/genética , MasculinoRESUMEN
Erastin can induce ferroptosis in tumor cells as an effective small molecule inhibitor. However, its application is hampered by a lack of water solubility. This study investigated the effects of superparamagnetic iron oxide (SPIO)-erastin-polyethylene glycol (PEG) nanoparticles prepared by loading SPIO-PEG nanoparticles with erastin on ferroptosis. SPIO-erastin-PEG nanoparticles exhibited square and spherical shapes with good dispersibility. The zeta potential and hydrodynamic size of SPIO-erastin-PEG were measured as (-37.68 ± 2.706) mV and (45.75 ± 18.88) nm, respectively. On T2-weighted imaging, the nanosystem showed significant contrast enhancement compared to no-enhancement magnetic resonance imaging (MRI). SPIO-erastin-PEG induced ferroptosis by increasing reactive oxygen species and iron content and promoting the accumulation of lipid peroxides and the degradation of glutathione peroxidase 4. Pharmacokinetic experiments revealed a half-life of 1.25 ± 0.05 h for the SPIO-erastin-PEG solution in circulation. Moreover, significant antitumorigenic effects of SPIO-erastin-PEG have been demonstrated in 5-8F cells and mouse-bearing tumors. These results indicated that the synthesized SPIO-erastin-PEG nanoplatform could induce ferroptosis effects in vitro and in vivo while exhibiting favorable physical characteristics. This approach may provide a new strategy for theranostic nanoplatform for nasopharyngeal cancer.
Asunto(s)
Ferroptosis , Neoplasias Nasofaríngeas , Polietilenglicoles , Ferroptosis/efectos de los fármacos , Animales , Polietilenglicoles/química , Ratones , Humanos , Neoplasias Nasofaríngeas/tratamiento farmacológico , Línea Celular Tumoral , Imagen por Resonancia Magnética/métodos , Especies Reactivas de Oxígeno/metabolismo , Nanopartículas Magnéticas de Óxido de Hierro/química , Ratones Endogámicos BALB C , Ratones Desnudos , Ensayos Antitumor por Modelo de Xenoinjerto , Compuestos Férricos/química , Femenino , PiperazinasRESUMEN
BACKGROUND: Mutations in human ether-à-go-go-related gene (hERG) potassium channels are closely associated with long QT syndrome (LQTS). Previous studies have demonstrated that macrolide antibiotics increase the risk of cardiovascular diseases. To date, the mechanisms underlying acquired LQTS remain elusive. METHODS: A novel hERG mutation I1025N was identified in an azithromycin-treated patient with acquired long QT syndrome via Sanger sequencing. The mutant I1025N plasmid was transfected into HEK-293 cells, which were subsequently incubated with azithromycin. The effect of azithromycin and mutant I1025N on the hERG channel was evaluated via western blot, immunofluorescence, and electrophysiology techniques. RESULTS: The protein expression of the mature hERG protein was down-regulated, whereas that of the immature hERG protein was up-regulated in mutant I1025N HEK-293 cells. Azithromycin administration resulted in a negative effect on the maturation of the hERG protein. Additionally, the I1025N mutation exerted an inhibitory effect on hERG channel current. Moreover, azithromycin inhibited hERG channel current in a concentration-dependent manner. The I1025N mutation and azithromycin synergistically decreased hERG channel expression and hERG current. However, the I1025N mutation and azithromycin did not alter channel gating dynamics. CONCLUSIONS: These findings suggest that hERG gene mutations might be involved in the genetic susceptibility mechanism underlying acquired LQTS induced by azithromycin.
Asunto(s)
Azitromicina , Síndrome de QT Prolongado , Humanos , Azitromicina/efectos adversos , Células HEK293 , Antibacterianos/efectos adversos , Síndrome de QT Prolongado/inducido químicamente , Síndrome de QT Prolongado/genética , MutaciónRESUMEN
Primary Breast Angiosarcoma (PBA) is an exceptionally rare form of breast cancer, accounting for less than 0.05% of all breast cancers. It is characterized by a high level of malignancy, invasiveness, and has a prognosis that is typically poor. The lack of distinctive clinical features makes it prone to underdiagnosis and misdiagnosis. This study retrospectively examines a case utilizing multimodal ultrasound imaging techniques (including 2D ultrasound, contrast-enhanced ultrasound, and ultrasound elastography) for diagnosing PBA. Furthermore, the study reviews relevant literature to summarize the ultrasound characteristics of PBA, with the aim of improving understanding of this elusive condition.
Asunto(s)
Neoplasias de la Mama , Mama , Hemangiosarcoma , Imagen Multimodal , Ultrasonografía Mamaria , Humanos , Hemangiosarcoma/diagnóstico por imagen , Femenino , Neoplasias de la Mama/diagnóstico por imagen , Ultrasonografía Mamaria/métodos , Imagen Multimodal/métodos , Mama/diagnóstico por imagen , Medios de Contraste , Persona de Mediana Edad , Diagnóstico por Imagen de Elasticidad/métodos , Diagnóstico DiferencialRESUMEN
Cotton Verticillium wilt is mainly caused by the fungus Verticillium dahliae, which threatens the production of cotton. Its pathogen can survive in the soil for several years in the form of microsclerotia, making it a destructive soil-borne disease. The accurate, sensitive, and rapid detection of V. dahliae from complex soil samples is of great significance for the early warning and management of cotton Verticillium wilt. In this study, we combined the loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a technology to develop an accurate, sensitive, and rapid detection method for V. dahliae. Initially, LAMP primers and CRISPR RNA (crRNA) were designed based on a specific DNA sequence of V. dahliae, which was validated using several closely related Verticillium spp. The lower detection limit of the LAMP-CRISPR/Cas12a combined with the fluorescent visualization detection system is approximately ~10 fg/µL genomic DNA per reaction. When combined with crude DNA-extraction methods, it is possible to detect as few as two microsclerotia per gram of soil, with the total detection process taking less than 90 min. Furthermore, to improve the method's user and field friendliness, the field detection results were visualized using lateral flow strips (LFS). The LAMP-CRISPR/Cas12a-LFS system has a lower detection limit of ~1 fg/µL genomic DNA of the V. dahliae, and when combined with the field crude DNA-extraction method, it can detect as few as six microsclerotia per gram of soil, with the total detection process taking less than 2 h. In summary, this study expands the application of LAMP-CRISPR/Cas12a nucleic acid detection in V. dahliae and will contribute to the development of field-deployable diagnostic productions.
Asunto(s)
Sistemas CRISPR-Cas , Técnicas de Amplificación de Ácido Nucleico , Enfermedades de las Plantas , Microbiología del Suelo , Técnicas de Amplificación de Ácido Nucleico/métodos , Enfermedades de las Plantas/microbiología , Ascomicetos/genética , Ascomicetos/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Gossypium/microbiología , ADN de Hongos/genética , ADN de Hongos/aislamiento & purificación , Verticillium/genéticaRESUMEN
Balancing environmental protection and social-economic development in agricultural land use management is a dilemma for decision-makers. Based on the modelling of the impacts of land use changes on river water pollution by SWAT model, the tradeoff between tea plantation expansion and river water quality was detected. SWAT model performs well in simulating the non-point source (NPS) pollution in agricultural watershed. The results showed that the tea plantation area expanded dramatically from 44 km2 in 2000 to 169 km2 in 2020 at the high cost of forest land. Consequently, the mean contents of NO3--N and TN have significantly increased by 100% and 91% respectively in the past 20 years. And the NO3--N in river water accounted for over 80% of TN in the tea plantation area. The NO3--N and TN concentrations were positively related with the proportions of tea plantation area (Tea%) at different periods. The high pollution levels of NO3--N and TN are priority control targets for river water quality management. The results indicated that the proportion of tea plantation thresholds lead to abrupt changes in river water quality. When the Tea% exceeded 3.0% in 2000, the probability of N pollution increased sharply. Whereas in 2020, it is suggested that the Tea% should not exceeds 18% to avoid sudden deterioration of water quality. The critical interval value of the Tea% for sudden change in N pollution showed an obvious increase tendency. The accelerating of nutrient pollution in rivers reduced the sensitivity of water quality to tea plantation expansion. Our results can provide new insights and empirical evidence for balancing the tradeoff between agricultural development and river water quality protection by demonstrating the carrying capacity threshold of river water environment for the expansion tea plantation.
Asunto(s)
Agricultura , Ríos , Contaminación del Agua , Ríos/química , Contaminación del Agua/análisis , Calidad del Agua , Monitoreo del AmbienteRESUMEN
In the context of rapid global urbanization, the sustainable development of ecosystems should be considered. Accordingly, the Planetary Boundaries theory posits that reducing the amount of nitrogen and phosphorus pollutants entering bodies of water is necessary as excess levels may harm the aquatic environment and reduce in water quality. Thus, based on the long-term monitoring data of representative urban rivers in the Yangtze River Delta region, we evaluated the nitrogen and phosphorus pollution of water bodies in different urbanization stages and further quantified the effect of urban forests on water quality improvement. The results showed that, with the continuous progression of urbanization, the proportion of impervious surface area increased, along with the levels of nitrogen and phosphorus pollution in water bodies. The critical period of water quality deterioration in urban rivers occurred during the medium urbanization level when the proportion of impervious surface area reached 55-65 %, and the probability of an abrupt increase in total nitrogen (TN) and total phosphorus (TP) concentration exceeded 95 %. However, increasing the area of urban forests during this period reduced TN pollution by 36.64 % and TP pollution by 49.03 %. The results of this study support the expansion of urban forests during the medium urbanization stage to improve water quality. Furthermore, our results provide a reference and theoretical basis for urban forest construction as a key aspect of the sustainable development of the urban ecosystem in the Yangtze River Delta and similar regions around world.
Asunto(s)
Contaminantes Químicos del Agua , Calidad del Agua , Ecosistema , Urbanización , Mejoramiento de la Calidad , Bosques , Nitrógeno/análisis , Fósforo/análisis , China , Monitoreo del Ambiente/métodos , Contaminantes Químicos del Agua/análisisRESUMEN
OBJECTIVE: To explore the clinical manifestations and genetic basis for a Chinese pedigree affected with atypical Charcot-Marie-Tooth disease type 1 A (CMT1A). METHODS: A patient admitted to the Department of Neurology, Xijing Hospital Affiliated to Air Force Medical University in June 2022 was selected as the study subject. Clinical data of the patient was collected, and 17 family members from four generations of this pedigree were traced based on pes arcuatus and atypical clinical symptoms. Neuroultrasound and genetic testing were carried out on available family members. Whole exome sequencing and multiple ligation-dependent probe amplification assay were carried out for the proband and some of the affected members of the pedigree. RESULTS: The proband, a 15-year-old male, had presented with paroxystic limb pain with weakness, accompanied by pes cavus and hypertrophy of gastrocnemius muscles, without stork leg sign caused by muscles atrophy in the distal lower extremities. MRI has revealed no sign of fat infiltration in the muscles of both legs. Nerve conduction examination had indicated damages of the sensory and motor nerves of the limbs, mainly with demyelinating changes. Seven members of the pedigree had pes arcuatus, including 5 presenting with paroxysmal neuropathic pain and myasthenia in the limbs, whilst 2 were without any clinical symptoms. Neurosonography of the proband, his brother, father and aunt showed thickened peripheral nerves of the extremities with unclear bundle structure. Genetic analysis revealed a large repeat encompassing exons 1 to 5 of the PMP22 gene and flanking regions (chr17: 15133768_15502298) in some of the affected members, which was predicted to be pathogenic. CONCLUSION: The duplication of PMP22 gene was considered to be pathogenic for this CMT1A pedigree.
Asunto(s)
Enfermedad de Charcot-Marie-Tooth , Masculino , Humanos , Adolescente , Enfermedad de Charcot-Marie-Tooth/genética , Enfermedad de Charcot-Marie-Tooth/patología , Linaje , Proteínas de la Mielina/genética , Músculo Esquelético , China , Duplicación de GenRESUMEN
Reduced chemotactic migration of polymorphonuclear neutrophil (PMN) in sepsis patients leads to decreased bacterial clearance and accelerates the progression of sepsis disease. Quantification of PMN chemotaxis in sepsis patients can help characterize the immune health of sepsis patients. Microfluidic microarrays have been widely used for cell chemotaxis analysis because of the advantages of low reagent consumption, near-physiological environment, and visualization of the migration process. Currently, the study of PMN chemotaxis using microfluidic chips is mainly limited by the cumbersome cell separation operation and low throughput of microfluidic chips. In this paper, we first designed an inertial cell sorting chip to achieve label-free separation of the two major cell types by using the basic principle that leukocytes (mainly granulocytes, lymphocytes and monocytes) and erythrocytes move to different positions of the spiral microchannel when they move in the spiral microchannel under different strength of inertial force and Dean's resistance. Subsequently, in this paper, we designed a multi-channel cell migration chip and constructed a microfluidic PMN inertial label-free sorting and chemotaxis analysis platform. The inertial cell sorting chip separates leukocyte populations and then injects them into the multi-channel cell migration chip, which can complete the chemotaxis test of PMN to chemotactic peptide (fMLP) within 15 min. The remaining cells, such as monocytes with slow motility and lymphocytes that require pre-activation with proliferative culture, do not undergo significant chemotactic migration. The test results of sepsis patients ( n=6) and healthy volunteers ( n=3) recruited in this study showed that the chemotaxis index (CI) and migration velocity ( v) of PMN from sepsis patients were significantly weaker than those from healthy volunteers. In conclusion, the microfluidic PMN inertial label-free sorting and chemotaxis analysis platform constructed in this paper can be used as a new tool for cell label-free sorting and migration studies.
Asunto(s)
Quimiotaxis , Sepsis , Humanos , Neutrófilos/metabolismo , Microfluídica , Movimiento Celular , Sepsis/metabolismoRESUMEN
The pond wolf spider Pardosa pseudoannulata Bösenberg & Strand, 1906 (Araneae: Lycosidae) is an important predator of agricultural pests in southern, eastern and southeastern Asia. Here, we report the complete mitogenome of this spider reconstructed from Illumina sequencing data. The circular mitogenome length is 14,533 bp with the nucleotide composition A (33.3%), C (8.2%), G (15.2%), and T (43.3%). The P. pseudoannulata mitogenome comprises 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. Phylogenetic analyses of Lycosidae mitogenomes supported the monophyly of the subfamily Pardosinae and the two genera Pardosa and Alopecosa, and indicated the polyphyly of the subfamily Lycosinae and the paraphyly of its type genus Lycosa. In this study, P. pseudoannulata is the closest relative to P. pusiola. These results provide useful genetic information for future studies on the diversity, phylogeny, and evolution for wolf spiders.
RESUMEN
Compared with primary tumor sites, metastatic sites appear more resistant to treatments and respond differently to the treatment regimen. It may be due to the heterogeneity in the microenvironment between metastatic sites and primary tumors. Cancerassociated fibroblasts (CAFs) are widely present in the tumor stroma as key components of the tumor microenvironment. Primary tumor CAFs (pCAFs) and metastatic CAFs (mCAFs) are heterogeneous in terms of source, activation mode, markers and functional phenotypes. They can shape the tumor microenvironment according to organ, showing heterogeneity between primary tumors and metastases, which may affect the sensitivity of these sites to treatment. It was hypothesized that understanding the heterogeneity between pCAFs and mCAFs can provide a glimpse into the difference in treatment outcomes, providing new ideas for improving the rate of metastasis control in various cancers.
Asunto(s)
Fibroblastos Asociados al Cáncer , Neoplasias , Humanos , Fibroblastos Asociados al Cáncer/patología , Fibroblastos/patología , Neoplasias/tratamiento farmacológico , Neoplasias/patología , Resultado del Tratamiento , Microambiente TumoralRESUMEN
Imazethapyr is a widely used imidazolinone herbicide worldwide, and its potential adverse effects on non-target plants have raised concerns. Understanding the mechanisms of imazethapyr phytotoxicity is crucial for its agro-ecological risk assessment. Here, the comprehensive molecular responses and metabolic alterations of Arabidopsis in response to imazethapyr were investigated. Our results showed that root exposure to imazethapyr inhibited shoot growth, reduced chlorophyll contents, induced photoinhibition and decreased photosynthetic activity. By non-target metabolomic analysis, we identified 75 metabolites that were significantly changed after imazethapyr exposure, and they are mainly enriched in carbohydrate, lipid and amino acid metabolism. Transcriptomic analysis confirmed that imazethapyr significantly downregulated the genes involved in photosynthetic electron transport and the carbon cycle. In detail, 48 genes in the photosynthetic lightreaction and 11 genes in Calvin cycle were downregulated. Additionally, the downregulation of genes related to electron transport in mitochondria provides strong evidence for imazethapyr inhibiting photosynthetic carbon fixation and cellular energy metabolism as one of mechanisms of toxicity. These results revealed the molecular and metabolic basis of imazethapyr toxicity on non-target plants, contributing to environmental risk assessment and mitigate negative impact of imazethapyr residues in agricultural soils.
Asunto(s)
Arabidopsis , Herbicidas , Metabolómica , Transcriptoma , Herbicidas/toxicidad , Transcriptoma/efectos de los fármacos , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Fotosíntesis/efectos de los fármacos , Ácidos Nicotínicos/toxicidadRESUMEN
The influence of ferulic acid (FA) on rice starch was investigated by incorporating it at various concentrations (0, 2.5, 5, 7.5, and 10 %, w/w, on dry starch basis) and subjecting the resulting composites to hot-extrusion 3D printing (HE-3DP) process. This study examined the effects of FA addition and HE-3DP on the structural, rheological, and physicochemical properties as well as the printability and digestibility of rice starch. The results indicated that adding 0-5 % FA had no significant effect; however, as the amount of FA increased, the printed product edges became less defined, the product's overall stability decreased, and it collapsed. The addition of FA reduced the elasticity and viscosity, making it easier to extrude the composite gel from the nozzle. Moreover, the crystallinity and short-range ordered structure of the HE-3D printed rice starch gel decreased with the addition of FA, resulting in a decrease in the yield stress and an increase in fluidity. Furthermore, the addition of FA reduced the digestibility of the HE-3D-printed rice starch. The findings of this study may be useful for the development of healthier modified starch products by adding bioactive substances and employing the 3D printing technology.
Asunto(s)
Ácidos Cumáricos , Oryza , Impresión Tridimensional , Reología , Almidón , Almidón/química , Oryza/química , Ácidos Cumáricos/química , Viscosidad , Calor , Digestión/efectos de los fármacosRESUMEN
The wide application of plastics has led to the ubiquitous presence of nanoplastics and microplastics in terrestrial environments. However, few studies have focused on the mechanism underlying the effects of plastic particles on soil microbiomes and resistomes, especially the differences between nanoplastics and microplastics. This study investigated the microbiome and resistome in soil exposed to polystyrene microplastics (mPS) or nanoplastics (nPS) through 16S rRNA and shotgun metagenomic sequencing. Distinct microbial communities were observed between mPS and nPS exposure groups, and nPS exposure significantly changed the bacterial composition even at the lowest amended rate (0.01%, w/w). The abundance of antibiotic resistance genes (ARGs) in nPS exposure (1%) was 0.26 copies per cell, significantly higher than that in control (0.21 copies per cell) and mPS exposure groups (0.21 copies per cell). It was observed that nanoplastics, bacterial community, and mobile genetic elements (MGEs) directly affected the ARG abundance in nPS exposure groups, while in mPS exposure groups, only MGEs directly induced the change of ARGs. Streptomyces was the predominant host for multidrug in the control and mPS exposure, whereas the primary host was changed to Bacillus in nPS exposure. Additionally, exposure to nPS induced several bacterial hosts to exhibit possible multi-antibiotic resistance characteristics. Our results indicated that the effects of plastic particles on the soil microbial community were size-dependent, and nano-sized plastic particles exhibited more substantial impacts. Both microplastics and nanoplastics promoted ARG transfer and diversified their bacterial hosts. These findings bear implications for the regulation of plastic waste and ARGs.
RESUMEN
The aggravation of antibiotic resistance genes (ARGs) in the environment has posed a significant global health crisis. Accurate evaluation of ARGs levels in a facile manner is a pressing issue for environmental surveillance. Here, we demonstrate a unique dumbbell-shaped cascade nanozyme for visual/photoelectrochemical (PEC) dual-mode detection of ARGs. Gold nanoparticles (AuNPs) with tunable exposed facets are controllably anchored onto ZIF-8 dodecahedrons, exhibiting glucose oxidase (GOx)-like (ZIF-8@Au/G) and peroxidase (POD)-like (ZIF-8@Au/P) activities. Upon the occurrence of ARGs, an asymmetric cascade-amplified "dumbbell" configuration is spontaneously generated via target-induced DNA hybridization, comprising GOx-like ZIF-8@Au/G with capture DNA on one side and POD-like ZIF-8@Au/P with signal DNA on the opposite side. Such a cascade nano-system can efficiently oxidize colorless 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) into its green oxidation state and synergistically decompose H2O2, realizing colorimetric/PEC dual-mode ARGs detection with a detection limit of 0.112 nM. The applicability of the present bioassay is validated through measuring ARGs in real sludge samples. This work suggests the possibility to rationally design task-specific nanozymes and develop target-responsive nano-cascade assays for environmental monitoring.