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1.
J Nutr Biochem ; : 109637, 2024 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-38574828

RESUMO

Adequate dietary L-tryptophan (Trp) governs intestinal homeostasis in piglets. However, the defensive role of Trp in the diet against enterotoxigenic E. coli F4 (K88) in pigs is still poorly understood. Here, sixty (6.15 ± 1.52 kg, 24-day-old, Duroc × Landrace × Yorkshire) weaned piglets were used for an E. coli F4 attack test in a 2 × 2 factorial design. The growth (ADG, ADFI, GH), immune factors (IL-10, IgA, IgG, IgM), Trp metabolite 5-HT, intestinal morphology (jejunal and colonic VH), mRNA expression of ß-defensins (jejunal BD-127, BD-119, ileal BD-1, BD-127), and abundance of beneficial microorganisms in the colon (Prevotella 9, Lactobacillus, Phascolarctobacterium, Faecalibacterium) were higher in the piglets in the HT (High Trp) and HTK (High Trp, K88) groups than in the LT (Low Trp) and LTK (Low Trp, K88) groups (P < 0.05), while FCR, diarrhea rate, diarrhea index, serum Trp, Kyn, IDO, D-LA, ET, and abundance of harmful microorganisms in the colon (Spirochaetes, Fusobacteria, Prevotella, Christensenellaceae R7) were lower in the HT and HTK groups than in the LT and LTK groups (P < 0.05). High Trp reduced the expression of virulence genes (K88 and LT) after E. coli F4 attack (P < 0.05). The IL-6, TNF-α was lower in the HTK group than in the LT, LTK group (P < 0.05). In short, a diet containing 0.35% Trp protected piglets from enterotoxigenic E. coli F4 (K88) via Trp metabolism promoting BD expression in the intestinal mucosa, which improved growth and intestinal health.

2.
Microbiome ; 12(1): 73, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38605412

RESUMO

BACKGROUND: The utilization of mulberry branch fiber (MF), the largest by-product of the sericulture industry, is an important issue. Supplementation with MF as a dietary fiber for poultry may serve as a useful application. However, little is known about the effects of MF on liver lipid metabolism and egg yolk fatty acid composition of laying hens and their underlying mechanisms. In this study, we performed a multi-omics investigation to explore the variations in liver lipid metabolism, egg yolk fatty acid composition, gut microbiota, and the associations among them induced by dietary MF in laying hens. RESULTS: Dietary MF had no harmful effects on the laying performance or egg quality in laying hens. The enzyme activities associated with lipid metabolism in the liver were altered by the addition of 5% MF, resulting in reduced liver fat accumulation. Furthermore, dietary 5% MF induced the variation in the fatty acid profiles of egg yolk, and increased the polyunsaturated fatty acid (PUFA) content. We observed a significant reduction in the diversity of both gut bacteria and changes in their compositions after the addition of MF. Dietary MF significantly increased the abundance of genes involved in fatty acid biodegradation, and short-chain fatty acids biosynthesis in the gut microbiota of laying hens. The significant correlations were observed between the liver lipid metabolism enzyme activities of hepatic lipase, lipoprotein lipase, and total esterase with gut microbiota, including negative correlations with gut microbiota diversity, and multiple correlations with gut bacteria and viruses. Moreover, various correlations between the contents of PUFAs and monounsaturated fatty acids in egg yolk with the gut microbiota were obtained. Based on partial-least-squares path modeling integrated with the multi-omics datasets, we deduced the direct effects of liver enzyme activities and gut bacterial compositions on liver fat content and the roles of liver enzyme activities and gut bacterial diversity on egg yolk fatty acid composition. CONCLUSIONS: The results indicate that dietary MF is beneficial to laying hens as it reduces the liver fat and improves egg yolk fatty acid composition through the enterohepatic axis. Video Abstract.


Assuntos
Ácidos Graxos , Morus , Animais , Feminino , Ácidos Graxos/metabolismo , Gema de Ovo/metabolismo , Morus/metabolismo , Metabolismo dos Lipídeos , Galinhas/metabolismo , Dieta , Ácidos Graxos Insaturados/metabolismo , Ração Animal/análise , Suplementos Nutricionais
3.
Front Med (Lausanne) ; 11: 1359962, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38638935

RESUMO

Background: Few studies have focused on the clinical characteristics and intestinal flora of Tibetan patients with irritable bowel syndrome (IBS). The study aimed to compare the difference of between Tibetan and Han patients with IBS. Methods: Patients who met inclusion and exclusion criteria were divided into the Tibet and Han groups. A simplified Gastrointestinal Symptom Rating Scale (GSRS)-based questionnaire was used to assess the IBS severity. Fecal samples from all subjects were collected for the analysis of gut microbiota using 16sRNA Illumina sequencing. Results: No significant difference was found in the total symptom scores between two groups. However, Tibetans with IBS are more prone to bloating than Hans (17.41% vs 9.09%, p < 0.001). A profit shift in the gut microbiota was shown between the two groups. The ratio of Firmicutes/Bacteroidetes was significantly lower in the Tibet group than in the Han group (2.954 ± 0.78 vs 8.23 ± 2.04, p = 0.004). In the Tibet group, the level of the genus Blautia decreased significantly compared to the Han group, and there was a significant negative correlation between the level of Blautia and the bloating scores (Pearson r = -0.33, p = 0.025). Conclusion: The characteristics of Tibetan patients differ from those of Han patients with IBS, not only in terms of the clinical symptoms, but also in the characteristics of intestinal flora. Tibetans with IBS are more prone to bloating, which might be due to the different gut microbiota. The genus Blautia may play a role in this mechanism.

4.
Food Chem ; 450: 139380, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38640535

RESUMO

Pyrimethanil (PYR) is a fungicide that is harmful to consumers when present in foods at concentrations greater than maximum permitted residue levels. High-performance immunoprobes and dual-readout strategy may be useful for constructing sensitive lateral flow immunoassay (LFIA). Herein, the prepared litchi-like Au-Ag bimetallic nanospheres (LBNPs) exhibited high mass extinction coefficients and fluorescence quenching constants. Benefiting from LBNPs and dual-readout mode, the limits of detection of LBNPs-CM-LFIA and LBNPs-FQ-LFIA for PYR were 0.957 and 0.713 ng mL-1, which were 2.54- and 3.41-fold lower than that of gold nanoparticles-based LFIA, respectively. The limits of quantitation of LBNPs-CM-LFIA and LBNPs-FQ-LFIA were 3.740 and 1.672 ng mL-1, respectively. LBNPs-LFIA was applied to detect PYR in cucumber and grape samples with satisfactory recovery (90%-111%). LBNPs-LFIA showed good agreement with LC-MS/MS for the detection of PYR in the samples. Accordingly, this sensitive and accurate dual-readout LFIA based on LBNPs can be effectively applied for food safety.

5.
J Agric Food Chem ; 72(15): 8684-8692, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38564621

RESUMO

Polyhydroxyalkanoates (PHAs) are promising alternatives to petroleum-based plastics, owing to their biodegradability and superior material properties. Here, the controllable biosynthesis of scl-co-mcl PHA containing 3-hydroxybutyrate (3HB) and mcl 3-hydroxyalkanoates was achieved in Pseudomonas chlororaphis HT66. First, key genes involved in fatty acid ß-oxidation, the de novo fatty acid biosynthesis pathway, and the phaC1-phaZ-phaC2 operon were deleted to develop a chassis strain. Subsequently, an acetoacetyl-CoA reductase gene phaB and a PHA synthase gene phaC with broad substrate specificity were heterologously expressed for producing and polymerizing the 3HB monomer with mcl 3-hydroxyalkanoates under the assistance of native ß-ketothiolase gene phaA. Furthermore, the monomer composition of scl-co-mcl PHA was regulated by adjusting the amount of glucose and dodecanoic acid supplemented. Notably, the cell dry weight and scl-co-mcl PHA content reached 14.2 g/L and 60.1 wt %, respectively, when the engineered strain HT11Δ::phaCB was cultured in King's B medium containing 5 g/L glucose and 5 g/L dodecanoic acid. These results demonstrated that P. chlororaphis can be a platform for producing scl-co-mcl PHA and has the potential for industrial application.


Assuntos
Poli-Hidroxialcanoatos , Pseudomonas chlororaphis , Ácido 3-Hidroxibutírico , Pseudomonas chlororaphis/genética , Pseudomonas chlororaphis/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Glucose/metabolismo
6.
Work ; 2024 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-38517833

RESUMO

BACKGROUND: Pushing is a typical workplace activity that may result in musculoskeletal disorders (MSDs). A convenient method of assessing the risk of MSDs is to determine the association between the force applied and the subjective rating of force exertion. OBJECTIVES: This study aimed to determine the relationship between the subjective rating of force exertion and the force applied to both a fixed and a movable handle. It also aimed to confirm that the participants could apply a higher one-handed push force at shoulder height than at elbow height. METHODS: A one-handed pushing experiment was conducted. In the push-to-scale session, the participants pushed a handle fixed on a rig on four levels on the CR-10 scale. The push force was measured. In the force-rating session, the participants rated their force exertions when pushing a movable handle with metal blocks suspended on the two sides. The subjective ratings on the CR-10 were recorded. RESULTS: The one-handed push force at shoulder height was significantly higher than that at elbow height. More effort was needed to push on a movable handle than a fixed one. Two regression models were established to describe the relationship between the push force and the subjective ratings of pushing for each of the fixed and movable handles, respectively. The correlation coefficient results showed that these models are adequate to describe the relationship between the one-handed push force and the ratings of the CR-10 scale. CONCLUSION: Participants applied a higher one-handed push force at shoulder height than at elbow height was confirmed. Participants needed more efforts to push the movable handle under the same CR-10 level than the fixed handle. The predictive models developed in this study may be used to estimate the push force for one-handed pushing tasks when assessing the risk of MSDs.

7.
Carbohydr Polym ; 333: 121987, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38494238

RESUMO

Limited bone regeneration, uncontrollable degradation rate, mismatched defect zone and poor operability have plagued the reconstruction of irregular bone defect by tissue-engineered materials. A combination of biomimetic scaffolds with hydroxyapatite has gained great popularity in promoting bone regeneration. Therefore, we designed an injectable, photocurable and in-situ curing hydrogel by methacrylic anhydride -modified carboxymethyl cellulose (CMC-MA) loading with spherical hydroxyapatite (HA) to highly simulate the natural bony matrix and match any shape of damaged tissue. The prepared carboxymethyl cellulose-methacrylate/ hydroxyapatite(CMC-MA/HA) composite presented good rheological behavior, swelling ratio and mechanical property under light illumination. Meanwhile, this composite hydrogel promoted effectively proliferation, supported adhesion and upregulated the osteogenic-related genes expression of MC3T3-E1 cells in vitro, as well as the activity of the osteogenic critical protein, Integrin α1, ß1, Myosin 9, Myosin 10, BMP-2 and Smad 1 in Integrin/BMP-2 signal pathway. Together, the composite hydrogels realized promotion of bone regeneration, deformity improvement, and the enhanced new bone strength in skull defect. It also displayed a good histocompatibility and stability of subcutaneous implantation in vivo. Overall, this study laid the groundwork for future research into developing a novel biomaterial and a minimally invasive therapeutic strategies for reconstructing bone defects and contour deficiencies.


Assuntos
Durapatita , Tecidos Suporte , Carboximetilcelulose Sódica , Crânio , Hidrogéis/farmacologia , Miosinas
8.
Heliyon ; 10(5): e27196, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38486782

RESUMO

Various preclinical and a limited number of clinical studies of CAR-NK cells have shown promising results: efficient elimination of target cells without side effects similar to CAR-T therapy. However, the homing and infiltration abilities of CAR-NK cells are poor due to the inhibitory tumor microenvironment. From the perspective of clinical treatment strategies, combined with the biological and tumor microenvironment characteristics of NK cells, CAR-NK combination therapy strategies with anti-PD-1/PD-L1, radiotherapy and chemotherapy, kinase inhibitors, proteasome inhibitors, STING agonist, oncolytic virus, photothermal therapy, can greatly promote the proliferation, migration and cytotoxicity of the NK cells. In this review, we will summarize the targets selection, structure constructions and combinational therapies of CAR-NK cells for tumors to provide feasible combination strategies for overcoming the inhibitory tumor microenvironment and improving the efficacy of CAR-NK cells.

9.
Anal Chem ; 96(13): 5205-5214, 2024 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-38481140

RESUMO

Pathogenic diseases that trigger food safety remain a noteworthy concern due to substantial public health, economic, and social burdens worldwide. It is vital for developing an integrated diagnosis and treatment strategy for bacteria, which could achieve quick detection of pathogenic bacteria and the inhibition of multidrug-resistant bacteria. Herein, we reported an organic molecule (M-3) possessed strong light capture capacity, emerging a low energy gap and ΔEST. Subsequently, M-3 was integrated into a nanostructured system (BTBNPs) with excellent ROS generation, light absorption capability, and photothermal performance. Reactive oxygen species (ROS) generated by BTBNPs were mainly free radicals from a type I mechanism, and the high photothermal conversion efficiency of BTBNPs was 41.26%. Benefiting from these advantages of BTBNPs, BTBNPs could achieve a ∼99% antibacterial effect for Escherichia coli O157:H7 with 20 µM dosage and 5 min of irradiation. Furthermore, the limit of detection (LoD) of the proposed BTBNPs-LFIA (colorimetric and photothermal modalities) for detecting E. coli O157:H7 was 4105 and 419 CFU mL-1, respectively. Overall, this work is expected to provide a new and sophisticated perspective for integrated diagnosis and treatment systems regarding pathogenic bacteria.


Assuntos
Escherichia coli O157 , Nanopartículas Multifuncionais , Microbiologia de Alimentos , Espécies Reativas de Oxigênio , Limite de Detecção
10.
Bioresour Technol ; 399: 130561, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38460558

RESUMO

During the wastewater treatment and resource recovery process by attached microalgae, the chemical oxygen demand (COD) can cause biotic contamination in algal culture systems, which can be mitigated by adding an appropriate dosage of antibiotics. The transport of COD and additive antibiotic (chloramphenicol, CAP) in algal biofilms and their influence on algal physiology were studied. The results showed that COD (60 mg/L) affected key metabolic pathways, such as photosystem II and oxidative phosphorylation, improved biofilm autotrophic and heterotrophic metabolic intensities, increased nutrient demand, and promoted biomass accumulation by 55.9 %, which was the most suitable COD concentration for attached microalgae. CAP (5-10 mg/L) effectively stimulated photosynthetic pigment accumulation and nutrient utilization in pelagic microalgal cells. In conclusion, controlling the COD concentration (approximately 60 mg/L) in the medium and adding the appropriate CAP concentration (5-10 mg/L) are conducive to improving attached microalgal biomass production and resource recovery potential from wastewater.


Assuntos
Microalgas , Microalgas/metabolismo , Cloranfenicol/metabolismo , Análise da Demanda Biológica de Oxigênio , Águas Residuárias , Biofilmes , Biomassa , Nitrogênio/metabolismo
11.
New Phytol ; 242(3): 1257-1274, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38481385

RESUMO

Plant pathogenic fungi elaborate numerous detoxification strategies to suppress host reactive oxygen species (ROS), but their coordination is not well-understood. Here, we show that Sirt5-mediated protein desuccinylation in Magnaporthe oryzae is central to host ROS detoxification. SIRT5 encodes a desuccinylase important for virulence via adaptation to host oxidative stress. Quantitative proteomics analysis identified a large number of succinylated proteins targeted by Sirt5, most of which were mitochondrial proteins involved in oxidative phosphorylation, TCA cycle, and fatty acid oxidation. Deletion of SIRT5 resulted in hypersuccinylation of detoxification-related enzymes, and significant reduction in NADPH : NADP+ and GSH : GSSG ratios, disrupting redox balance and impeding invasive growth. Sirt5 desuccinylated thioredoxin Trx2 and glutathione peroxidase Hyr1 to activate their enzyme activity, likely by affecting proper folding. Altogether, this work demonstrates the importance of Sirt5-mediated desuccinylation in controlling fungal process required for detoxifying host ROS during M. oryzae infection.


Assuntos
Ascomicetos , Magnaporthe , Oryza , Espécies Reativas de Oxigênio/metabolismo , Lisina/metabolismo , Estresse Oxidativo , Ascomicetos/metabolismo , Proteínas Fúngicas/metabolismo , Oryza/metabolismo , Doenças das Plantas/microbiologia
12.
Liver Int ; 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38436551

RESUMO

BACKGROUND AND AIMS: Accurate preoperative prediction of microvascular invasion (MVI) and recurrence-free survival (RFS) is vital for personalised hepatocellular carcinoma (HCC) management. We developed a multitask deep learning model to predict MVI and RFS using preoperative MRI scans. METHODS: Utilising a retrospective dataset of 725 HCC patients from seven institutions, we developed and validated a multitask deep learning model focused on predicting MVI and RFS. The model employs a transformer architecture to extract critical features from preoperative MRI scans. It was trained on a set of 234 patients and internally validated on a set of 58 patients. External validation was performed using three independent sets (n = 212, 111, 110). RESULTS: The multitask deep learning model yielded high MVI prediction accuracy, with AUC values of 0.918 for the training set and 0.800 for the internal test set. In external test sets, AUC values were 0.837, 0.815 and 0.800. Radiologists' sensitivity and inter-rater agreement for MVI prediction improved significantly when integrated with the model. For RFS, the model achieved C-index values of 0.763 in the training set and ranged between 0.628 and 0.728 in external test sets. Notably, PA-TACE improved RFS only in patients predicted to have high MVI risk and low survival scores (p < .001). CONCLUSIONS: Our deep learning model allows accurate MVI and survival prediction in HCC patients. Prospective studies are warranted to assess the clinical utility of this model in guiding personalised treatment in conjunction with clinical criteria.

13.
Ecotoxicol Environ Saf ; 273: 116169, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38447518

RESUMO

Parkinson's disease (PD) is among the most prevalent neurodegenerative diseases, and approximately one third of patients with PD are estimated to have depression. Paraquat (PQ) exposure is an important environmental risk factor for PD. In this study, we established a mouse model of PQ-induced PD with depression to comprehensively investigate cellular heterogeneity and the mechanisms underlying the progression of depression in the context of PD. We utilized single-cell RNA-seq (scRNA-seq) to acquire the transcriptomic atlas of individual cells from model mice and characterize the gene expression profiles in each differentially expressed cell type. We identified a specific glutamatergic neuron cluster responsible for the development of heterogeneous depression-associated changes and established a comprehensive gene expression atlas. Furthermore, functional enrichment and cell trajectory analyses revealed that the mechanisms underlying the progression of PD with depression were associated with specific glutamatergic neurons. Together, our findings provide a valuable resource for deciphering the cellular heterogeneity of PD with depression. The suggested connection between intrinsic transcriptional states of neurons and the progression of depression can provide insight into potential biomarkers and specific targets for anti-depression treatment in patients with PD. SYNOPSIS: Our results obtained using model mice confirm the core effects of PQ exposure on glutamatergic neurons and their potential role in the development of PD with depression.


Assuntos
Paraquat , Doença de Parkinson , Humanos , Animais , Camundongos , Paraquat/toxicidade , Doença de Parkinson/genética , Depressão/induzido quimicamente , Depressão/genética , Perfilação da Expressão Gênica , RNA
14.
Environ Pollut ; 349: 123875, 2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38548152

RESUMO

With the evidence emerging that abnormal expression of long noncoding RNAs (lncRNAs) are involved in onset of Parkinson's disease (PD), the role of NR_030777 contributing to this disease is of great interest. We recently found that a novel lncRNA "NR_030777" demonstrates protective effects on PQ-induced neurodegeneration. However, the underlying molecular mechanisms of NR_030777 in the regulation of mitochondrial fission and mitophagy involved in PQ-induced neuronal damage remain to be explored. NR_030777 brain conditional overexpressing mice as well as in vitro primary neuronal cells from cerebral cortex and Neuro2a cells were adopted. Immunofluorescence, Immunohistochemistry, qRT-PCR and Western blotting were used to evaluate the expression levels of RNA and proteins. RNA immunoprecipitation and RNA pulldown experiment were used to evaluate the interaction of NR_030777 with its target proteins. NR_030777 and mitophagy were increased, and tyrosine hydroxylase (TH) levels recovered after NR_030777 overexpression upon PQ treatment. The overexpression and knockdown of NR_030777 unveiled that NR_030777 positively regulated mitophagy such as the upregulation of LC3B-II:I, ATG12-ATG5, p62 and NBR1. Moreover, the application of mdivi-1, a DRP-1 inhibitor, in combination with NR_030777 genetic modified cells unveiled that NR_030777 promoted DRP1-mediated mitochondrial fission and mitophagy. Furthermore, NR_030777 were directly bound to CDK1 to increase p-DRP1 levels at the Ser616 site, leading to mitochondrial fission and mitophagy. On the other hand, NR_030777 acted directly on ATG12 within the ATG12-ATG5 complex in the 800-1400 nt region to modulate the membrane formation. Accordingly, NR_030777 deficiency in neuron cells compromised cell mitophagy. Finally, the above findings were confirmed using NR_030777-overexpressing mice. NR_030777 exerted a protective effect on PQ-exposed mice by enhancing mitophagy. Our data provide the first scientific evidence for the precise invention of PQ-induced PD. Our findings further propose a breakthrough for understanding the regulatory relationship between NR_030777, CDK1, ATG12 and mitophagy in PQ-induced PD.

15.
Huan Jing Ke Xue ; 45(3): 1803-1811, 2024 Mar 08.
Artigo em Chinês | MEDLINE | ID: mdl-38471891

RESUMO

To investigate the effect of chelating agents on plant uptake of heavy metals, castor (Ricinus communis L.) was used as the test plant. Soil culture and pot experiments were conducted to study the effects of different concentrations of ethylenediamine disuccinic acid (EDDS) on the forms of Cu and Cd in soil and their absorption and transport by castor. The results showed that the application of EDDS significantly increased the content of available Cu and Cd. After 15 days of cultivation, the available Cu and Cd concentrations in the soil increased by 43.01%-103.55% and 51.78%-69.43%, respectively. EDDS promoted the conversion of reducible Cu to weak acid extractable and increased the mobility of Cu. Meanwhile, the application of EDDS promoted the absorption, transport, and enrichment of Cu in castor. Under the application of 2.5 mmol·kg-1 EDDS and 5.0 mmol·kg-1 EDDS, the Cu concentrations in the shoots were 4.88 times and 16.65 times higher than that of the control (P< 0.05), and the Cu concentrations in the roots were 2.89 times and 3.60 times higher than that of the control (P< 0.05), respectively. The Cu transport coefficient significantly increased by 72.73% and 381.82% when treated with EDDS 2.5 and EDDS 5.0. Simultaneously, the phytoextraction of Cu in shoots, roots, and their sum were 14.08, 2.16, and 4.70 times higher than that of the control (P<0.05), respectively, when treated with EDDS 5.0. Furthermore, EDDS significantly increased the Cd concentrations in castor. When treated with EDDS 2.5 the shoots and roots increased by 15.15% and 57.42%, respectively, and the phytoextraction of total Cd significantly increased by 13.44%. Generally, the EDDS treatment could increase the available Cu and Cd in soil, promote the uptake of Cu and Cd, and improve the phytoremediation efficiency of castor. Among them, the addition of 5.0 mmol·kg-1 EDDS had the best effect for Cu, whereas the addition of 2.5 mmol kg-1 EDDS had a higher increase in the phytoextraction of Cd.


Assuntos
Metais Pesados , Poluentes do Solo , Cádmio/análise , Solo , Poluentes do Solo/análise , Metais Pesados/análise , Etilenodiaminas , Quelantes/farmacologia , Biodegradação Ambiental , Succinatos/farmacologia
16.
Int J Biol Sci ; 20(5): 1578-1601, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38481806

RESUMO

Background: The application of chimeric antigen receptor (CAR) NK cells in solid tumors is hindered by lack of tumor-specific targets and inefficient CAR-NK cell efficacy. Claudin-6 (CLDN6) has been reported to be overexpressed in ovarian cancer and may be an attractive target for CAR-NK cells immunotherapy. However, the feasibility of using anti-CLDN6 CAR-NK cells to treat ovarian cancer remains to be explored. Methods: CLDN6 expression in primary human ovarian cancer, normal tissues and cell lines were detected by immunohistochemistry and western blot. Two types of third-generation CAR NK-92MI cells targeting CLDN6, CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) and CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB) were constructed by lentivirus transfection, sorted by flow cytometry and verified by western blot and qPCR. OVCAR-3, SK-OV-3, A2780, Hey and PC-3 cells expressing the GFP and luciferase genes were transduced. Subcutaneous and intraperitoneal tumor models were established via NSG mice. The ability of CLDN6-CAR NK cells to kill CLDN6-positive ovarian cancer cells were evaluated in vitro and in vivo by live cell imaging and bioluminescence imaging. Results: Both CLDN6-CAR1 and CLDN6-CAR2 NK-92MI cells could specifically killed CLDN6-positive ovarian cancer cells (OVCAR-3, SK-OV-3, A2780 and Hey), rather than CLDN6 negative cell (PC-3), in vitro. CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) exhibited stronger cytotoxicity than CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB). Furthermore, CLDN6-CAR1 NK cells could effectively eliminate ovarian cancer cells in subcutaneous and intraperitoneal tumor models. More importantly, CAR-NK cells combined with immune checkpoint inhibitors, anti-PD-L1, could synergistically enhance the antitumor efficacy of CLDN6-targeted CAR-NK cells. Conclusions: These results indicate that CLDN6-CAR NK cells possess strong antitumor activity and represent a promising immunotherapeutic modality for ovarian cancer.


Assuntos
Claudinas , Neoplasias Ovarianas , Receptores de Antígenos Quiméricos , Humanos , Animais , Camundongos , Feminino , Receptores de Antígenos Quiméricos/genética , Neoplasias Ovarianas/terapia , Neoplasias Ovarianas/metabolismo , Linhagem Celular Tumoral , Apoptose , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Antígenos CD28/metabolismo , Células Matadoras Naturais , Imunoterapia/métodos , Imunoterapia Adotiva/métodos
18.
Sci Total Environ ; 919: 170784, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38340834

RESUMO

Reclaimed water with nitrogen, phosphorus, and other contaminants may trigger algal blooms during its ecological utilization in replenishing rivers or lakes. However, the effect of reclaimed water on algal growth rates is not well understood. In this study, the growth potentials of algae in terms of Cyanophyta, Chlorophyta, and Bacillariophyta, as well as mixed algae in both regular culture medium and reclaimed water produced from treatment plants in Beijing with similar N and P concentrations, were compared to evaluate whether reclaimed water could facilitate algal growth. In addition, reclaimed water was also sterilized to verify the impact of bacteria's presence on algal growth. The results indicated that most algae grew faster in reclaimed water, among which the growth rate of Microcystis aeruginosa even increased by 5.5 fold. The growth of mixed algae in reclaimed water was not enhanced due to the strong adaptive ability of the community structure. Residual bacteria in the reclaimed water were found to be important contributors to algal growth. This work provided theoretical support for the safe and efficient utilization of reclaimed water.


Assuntos
Cianobactérias , Microcystis , Pequim , Água , Eutrofização , Fósforo/análise , China
19.
Proc Natl Acad Sci U S A ; 121(8): e2309504121, 2024 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-38346190

RESUMO

Graph neural networks (GNNs) excel in modeling relational data such as biological, social, and transportation networks, but the underpinnings of their success are not well understood. Traditional complexity measures from statistical learning theory fail to account for observed phenomena like the double descent or the impact of relational semantics on generalization error. Motivated by experimental observations of "transductive" double descent in key networks and datasets, we use analytical tools from statistical physics and random matrix theory to precisely characterize generalization in simple graph convolution networks on the contextual stochastic block model. Our results illuminate the nuances of learning on homophilic versus heterophilic data and predict double descent whose existence in GNNs has been questioned by recent work. We show how risk is shaped by the interplay between the graph noise, feature noise, and the number of training labels. Our findings apply beyond stylized models, capturing qualitative trends in real-world GNNs and datasets. As a case in point, we use our analytic insights to improve performance of state-of-the-art graph convolution networks on heterophilic datasets.

20.
ACS Synth Biol ; 13(3): 913-920, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38377538

RESUMO

The advancement of metabolic engineering and synthetic biology has promoted in-depth research on the nonmodel microbial metabolism, and the potential of nonmodel organisms in industrial biotechnology is becoming increasingly evident. The nonmodel organism Pseudomonas chlororaphis is a safe plant growth promoting bacterium for the production of phenazine compounds; however, its application is seriously hindered due to the lack of an effective gene expression precise regulation toolkit. In this study, we constructed a library of 108 promoter-5'-UTR (PUTR) and characterized them through fluorescent protein detection. Then, 6 PUTRs with stable low, intermediate, and high intensities were further characterized by report genes lacZ encoding ß-galactosidase from Escherichia coli K12 and phzO encoding PCA monooxygenase from P. chlororaphis GP72 and thus developed as a static gene expression regulation system. Furthermore, the stable and high-intensity expressed PMOK_RS0128085UTR was fused with the LacO operator to construct an IPTG-induced plasmid, and a self-induced plasmid was constructed employing the high-intensity PMOK_RS0116635UTR regulated by cell density, resulting in a dynamic gene expression regulation system. In summary, this study established two sets of static and dynamic regulatory systems for P. chlororaphis, providing an effective toolkit for fine-tuning gene expression and reprograming the metabolism flux.


Assuntos
Pseudomonas chlororaphis , Pseudomonas chlororaphis/genética , Pseudomonas chlororaphis/metabolismo , Engenharia Metabólica/métodos , Regulação Bacteriana da Expressão Gênica/genética , Regiões Promotoras Genéticas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo
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