ABSTRACT
Revealing the mechanisms underlying soil microbial community assembly is a fundamental objective in molecular ecology. However, despite increasing body of research on overall microbial community assembly mechanisms, our understanding of subcommunity assembly mechanisms for different prokaryotic and fungal taxa remains limited. Here, soils were collected from more than 100 sites across southwestern China. Based on amplicon high-throughput sequencing and iCAMP analysis, we determined the subcommunity assembly mechanisms for various microbial taxa. The results showed that dispersal limitation and homogenous selection were the primary drivers of soil microbial community assembly in this region. However, the subcommunity assembly mechanisms of different soil microbial taxa were highly variable. For instance, the contribution of homogenous selection to Crenarchaeota subcommunity assembly was 70%, but it was only around 10% for the subcommunity assembly of Actinomycetes, Gemmatimonadetes and Planctomycetes. The assembly of subcommunities including microbial taxa with higher occurrence frequencies, average relative abundance and network degrees, as well as wider niches tended to be more influenced by homogenizing dispersal and drift, but less affected by heterogeneous selection and dispersal limitation. The subcommunity assembly mechanisms also varied substantially among different functional guilds. Notably, the subcommunity assembly of diazotrophs, nitrifiers, saprotrophs and some pathogens were predominantly controlled by homogenous selection, while that of denitrifiers and fungal pathogens were mainly affected by stochastic processes such as drift. These findings provide novel insights into understanding soil microbial diversity maintenance mechanisms, and the analysis pipeline holds significant value for future research.
Subject(s)
Soil Microbiology , Soil , Bacteria/genetics , ChinaABSTRACT
Objectives: To determine the clinical significance of Ro52 protein/tripartite motif-containing 21 antibody and specific antinuclear antibody patterns using indirect immunofluorescence technique. METHODS: The retrospective study was conducted at the clinical laboratory of the First Affiliated Hospital of Chongqing Medical University, China, and comprised data from January 2017 to December 2021 of patients who underwent antinuclear antibody and anti-extractable nuclear antigen antibody detection. Inpatients with Ro52 antibody-positive status were taken as the cases, while anti-Ro52 negative patients with clear clinical diagnosis were taken as the controls. Data was analysed using SPSS 19. RESULTS: There were 1802 cases and 1211 controls. Positive Ro52 showed significantly greater frequency in patients with primary Sjogren's syndrome, systemic lupus erythematosus, inflammatory myositis, dry eyes and interstitial lung disease (p<0.05). Ro52 antibody showed high positive predictive value for primary Sjogren's syndrome 25(96.15%), systemic lupus erythematosus 259(91.20%), connective tissue disease-associated interstitial lung disease 45(86.67%) and inflammatory myositis 60(86.67%). Antinuclear antibody indirect immunofluorescence patterns most frequently detected were nuclear speckled 128(40.89%) and cytoplasmic speckled 126(40.26%) (p<0.05). Interstitial lung disease was associated with the presence of cytoplasmic speckled antinuclear antibody indirect immunofluorescence pattern 24(19.2%), while tumours 47(36.5%) and hepatitis B 26(20.3%) seemed to be more frequent with nuclear speckled pattern (p<0.05). The simultaneous reactivity extractable nuclear antigen antibodies most frequently detected were antinuclear antibody+Ro52+anti-Sjogren's syndrome A+ 558(33.96%). CONCLUSIONS: Ro52 antibody positivity was found to be associated with Sjogren's syndrome, systemic lupus erythematosus, inflammatory myositis, dry eye and interstitial lung disease. The antinuclear antibody immunofluorescence pattern of Ro52 positive was single and primarily granular cytoplasm type. Antinuclear antibody negative and Ro52 positive in the serum of patients also had certain significance in auxiliary disease diagnosis.
Subject(s)
Lung Diseases, Interstitial , Lupus Erythematosus, Systemic , Myositis , Sjogren's Syndrome , Humans , Antibodies, Antinuclear , Sjogren's Syndrome/diagnosis , Retrospective Studies , Fluorescent Antibody Technique, Indirect , Clinical Relevance , Ribonucleoproteins , Lupus Erythematosus, Systemic/diagnosis , Lung Diseases, Interstitial/diagnosisABSTRACT
The compound 3'-phosphoadenosine-5'-phosphosulfate (PAPS) serves as a sulfate group donor in the production of valuable sulfated compounds. However, elevated costs and low conversion efficiency limit the industrial applicability of PAPS. Here, we designed and constructed an efficient and controllable catalytic system for the conversion of adenosine triphosphate (ATP) (disodium salt) into PAPS without inhibition from by-products. In vitro and in vivo testing in Escherichia coli identified adenosine-5'-phosphosulfate kinase from Penicillium chrysogenum (PcAPSK) as the rate-limiting enzyme. Based on analysis of the catalytic steps and molecular dynamics simulations, a mechanism-guided "ADP expulsion" strategy was developed to generate an improved PcAPSK variant (L7), with a specific activity of 48.94 U·mg-1 and 73.27-fold higher catalytic efficiency (kcat/Km) that of the wild-type enzyme. The improvement was attained chiefly by reducing the ADP-binding affinity of PcAPSK, as well as by changing the enzyme's flexibility and lid structure to a more open conformation. By introducing PcAPSK L7 in an in vivo catalytic system, 73.59 mM (37.32 g·L-1 ) PAPS was produced from 150 mM ATP in 18.5 h using a 3-L bioreactor, and achieved titer is the highest reported to date and corresponds to a 98.13% conversion rate. Then, the PAPS catalytic system was combined with the chondroitin 4-sulfotransferase using a one-pot method. Finally, chondroitin sulfate was transformed from chondroitin at a conversion rate of 98.75%. This strategy has great potential for scale biosynthesis of PAPS and chondroitin sulfate.
Subject(s)
Adenosine Triphosphate/metabolism , Chondroitin Sulfates , Escherichia coli , Fungal Proteins , Penicillium chrysogenum/genetics , Phosphoadenosine Phosphosulfate , Phosphotransferases (Alcohol Group Acceptor) , Chondroitin Sulfates/biosynthesis , Chondroitin Sulfates/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Penicillium chrysogenum/enzymology , Phosphoadenosine Phosphosulfate/biosynthesis , Phosphoadenosine Phosphosulfate/genetics , Phosphotransferases (Alcohol Group Acceptor)/genetics , Phosphotransferases (Alcohol Group Acceptor)/metabolismABSTRACT
OBJECTIVE: To explore the role of diffusion kurtosis magnetic resonance (MR) imaging in the noninvasive identification of synovitis in hand arthritis. METHODS: A total of 30 patients with rheumatoid arthritis (RA) and 10 patients suspected of RA were enrolled in the prospective study. A 3.0-T MR imaging including the diffusion kurtosis MR imaging sequence (b = 0, 500, 1000, 1500, 2000 s·mm2) was performed. A total of 210 regions of interest were confirmed and diffusion kurtosis MR imaging parameters were generated. The suspected synovitis or effusion was scored on a scale of 0 (effusion) to 3 (mild, moderate, severe synovitis), according to RA-MR imaging scoring system. The performance of diffusion kurtosis MR imaging parameters (the apparent diffusion coefficient [ADC], diffusion coefficient [D], and kurtosis [K]) in distinguishing different synovitis scores was evaluated. RESULTS: There were significant differences in ADC, D, and K values among different synovitis scores (all P < 0.001). Synovitis scores were negatively correlated with the ADC and D values significantly (r = -0.725, -0.757, respectively, all P < 0.001), but positively correlated with the K values significantly (r = 0.429, P < 0.001). The area under the curve values of D, ADC, and K values were 0.884, 0.874, and 0.728 for differentiating score 1-3 from score 0, respectively. Diffusion coefficient and ADC had similar diagnostic performance, and both were higher than K in detecting synovitis. No significant difference was found between the ADC and D values in detecting synovitis. CONCLUSIONS: The diffusion kurtosis MR imaging may be feasible as a noninvasive method for the diagnosis and grading of synovitis in the hands of RA patients, and the D and ADC values showed similar diagnostic performance, both of which were higher than K values.
Subject(s)
Arthritis, Rheumatoid/diagnostic imaging , Diffusion Magnetic Resonance Imaging/methods , Image Interpretation, Computer-Assisted/methods , Synovitis/diagnostic imaging , Adult , Arthritis, Rheumatoid/complications , Female , Humans , Male , Middle Aged , Prospective Studies , Reproducibility of Results , Sensitivity and Specificity , Synovial Membrane/diagnostic imaging , Synovitis/complicationsABSTRACT
Two strains of a novel yeast species were isolated from traditional Chinese sauerkraut samples collected in Nanyang, Henan Province, central China. Phylogenetic analysis based on the concatenated sequences of the D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions showed that these strains belong to the Yarrowia clade, with seven clones of uncultured Yarrowia as their closest phylogenetic neighbours. They differed from their closest known species, Yarrowia divulgata CBS 11013T, by 3.2â% sequence divergence (14 substitutions and 2 gaps) in the D1/D2 domains and by 5.4â% sequence divergence (12 substitutions and 5 gaps) in the ITS regions. The two strains of novel species reproduced asexually, and no ascospores could be found. The name Yarrowia brassicae f.a., sp. nov. is proposed to accommodate these strains, with NYNU 17218T (=CICC 33263T=CBS 15225T) as the type strain.
Subject(s)
Fermented Foods/microbiology , Phylogeny , Yarrowia/classification , China , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Mycological Typing Techniques , Sequence Analysis, DNA , Spores, Fungal , Yarrowia/genetics , Yarrowia/isolation & purificationABSTRACT
Four yeast strains were isolated from rotting wood samples collected in the Baotianman Nature Reserve in Henan Province, Central China. On the basis of sequence analysis of the D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer regions, they were suggested to be two novel species of the genus Pichia. Pichia nanzhaoensis sp. nov. produces one to four spherical ascospores per ascus, and is most closely related to Candida pseudolambica. Pichia paraexigua f.a., sp. nov. is a sister taxa to Pichia exigua, but the formation of ascospores was not observed on various sporulation media. P. nanzhaoensis sp. nov. can weakly assimilate inulin, whereas P. paraexigua sp. nov. can weakly assimilate d-glucosamine. The type strain of Pichia nanzhaoensis is NYNU 178136T (=CICC 33279T=CBS 15346T) and the type strain of Pichia paraexigua is NYNU 178135T (=CICC 33278T=CBS 15237T).
Subject(s)
Phylogeny , Pichia/classification , Wood/microbiology , China , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Mycological Typing Techniques , Pichia/genetics , Pichia/isolation & purification , Sequence Analysis, DNA , Spores, FungalABSTRACT
Three strains representing a novel yeast species were isolated from rotten wood samples collected in Xishuangbanna Tropical Rainforest in Yunnan Province, PR China. Sequence analysis of the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) regions indicated that the novel species represented a member of the genus Deakozyma. The novel species was related most closely to the type strain of Deakozyma indianensis but they differed by 5.8â% sequence divergence (20 substitutions and 5 gaps) in the D1/D2 domain and by 12.2â% sequence divergence (27 substitutions and 15 gaps) in the ITS region. Interestingly, the novel yeast species was able to ferment glucose and sucrose in Durham tubes, a clear difference from its closest relative D. indianensis, the type species of the genus Deakozyma. The species name of Deakozyma yunnanensis sp. nov. is proposed to accommodate these strains, with NYNU 16742T (=CICC 33160T=CBS 14688T) designated the type strain. As the current description of the genus does not allow the inclusion of sugar fermenting species, the emendation of the diagnosis of the genus Deakozyma Kurtzman and Robnett is proposed.
Subject(s)
Phylogeny , Saccharomycetales/classification , Wood/microbiology , China , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Mycological Typing Techniques , Rainforest , Saccharomycetales/genetics , Saccharomycetales/isolation & purification , Sequence Analysis, DNA , Tropical ClimateABSTRACT
In this study, we designed a wearable patch-type ECG sensor (WPES), which is mainly composed of the electrodes, an ECG acquisition unit, a Bluetooth unit, and a power supplies module. By using BMD101 chip as the core of ECG acquisition, the WPES can communicate with smart phones through Bluetooth low energy (BLE). The WEPS can be well used in monitoring epileptic seizure prediction, based on the advantages of simplification of circuits, miniaturization, low-power consumption, light-weight, convenience of wearing, high-comfort etc.
Subject(s)
Seizures , Wearable Electronic Devices , Electric Power Supplies , Electrocardiography , Electrodes , Humans , Miniaturization , Monitoring, Physiologic , Smartphone , Wireless TechnologyABSTRACT
The benign and malignant status of solitary pulmonary nodules (SPNs) is a key determinant of treatment decisions. The main objective of this study was to validate the efficacy of machine learning (ML) models featured with dual-layer detector spectral computed tomography (DLCT) parameters in identifying the benign and malignant status of SPNs. 250 patients with pathologically confirmed SPN were included in this study. 8 quantitative and 16 derived parameters were obtained based on the regions of interest of the lesions on the patients' DLCT chest enhancement images. 6 ML models were constructed from 10 parameters selected after combining the patients' clinical parameters, including gender, age, and smoking history. The logistic regression model showed the best diagnostic performance with an area under the receiver operating characteristic curve (AUC) of 0.812, accuracy of 0.813, sensitivity of 0.750 and specificity of 0.791 on the test set. The results suggest that the ML models based on DLCT parameters are superior to the traditional CT parameter models in identifying the benign and malignant nature of SPNs, and have greater potential for application.
Subject(s)
Lung Neoplasms , Solitary Pulmonary Nodule , Humans , Solitary Pulmonary Nodule/diagnostic imaging , Solitary Pulmonary Nodule/pathology , Diagnosis, Differential , Tomography, X-Ray Computed/methods , ROC Curve , Lung Neoplasms/diagnostic imaging , Lung Neoplasms/pathologyABSTRACT
Background: The accurate assessment of lymph node metastasis (LNM) is crucial for the staging, treatment, and prognosis of lung cancer. In this study, we explored the potential value of dual-layer spectral detector computed tomography (SDCT) quantitative parameters in the prediction of LNM in non-small cell lung cancer (NSCLC). Methods: In total, 91 patients presenting with solid solitary pulmonary nodules (8 mm < diameter ≤30 mm) with pathologically confirmed NSCLC (57 without LNM, and 34 with LNM) were enrolled in the study. The patients' basic clinical data and the SDCT morphological features were analyzed using the chi-square test or Fisher's exact test. The Mann-Whitney U-test and independent sample t-test were used to analyze the differences in multiple SDCT quantitative parameters between the non-LNM and LNM groups. The diagnostic efficacy of the corresponding parameters in predicting LNM in NSCLC was evaluated by plotting the receiver operating characteristic (ROC) curves. A multivariate logistic regression analysis was conducted to determine the independent predictive factors of LNM in NSCLC. Interobserver agreement was assessed using intraclass correlation coefficients (ICCs) and Bland-Altman plots. Results: There were no significant differences between the non-LNM and LNM groups in terms of age, sex, and smoking history. Lesion size and vascular convergence sign differed significantly between the two groups (P<0.05), but there were no significant differences in the six tumor markers. The SDCT quantitative parameters [SAR40keV, SAR70keV, Δ40keV, Δ70keV, CER40keV, CER70keV, NEF40keV, NEF70keV, λ, normalized iodine concentration (NIC) and NZeff] were significantly higher in the non-LNM group than the LNM group (P<0.05). The ROC analysis showed that CER40keV, NIC, and CER70keV had higher diagnostic efficacy than other quantitative parameters in predicting LNM [areas under the curve (AUCs) =0.794, 0.791, and 0.783, respectively]. The multivariate logistic regression analysis showed that size, λ, and NIC were independent predictive factors of LNM. The combination of size, λ, and NIC had the highest diagnostic efficacy (AUC =0.892). The interobserver repeatability of the SDCT quantitative and derived quantitative parameters in the study was good (ICC: 0.801-0.935). Conclusions: The SDCT quantitative parameters combined with the clinical data have potential value in predicting LNM in NSCLC. The size + λ + NIC combined parameter model could further improve the prediction efficacy of LNM.
ABSTRACT
Compared with chemical synthesis and tissue extraction methods, microbial synthesis of glycosaminoglycans (GAGs) is attractive because of the advantages of eco-friendly processes, production safety, and sustainable development. However, boosting the efficiency of microbial cell factories, precisely regulating GAG molecular weights, and rationally controlling the sulfation degree of GAGs remain challenging. To address these issues, various strategies, including genetic, enzymatic, metabolic, and fermentation engineering, have been developed. In this review, we summarize the recent progress in the construction of efficient GAG-producing microbial cell factories, regulation of the molecular weight of GAGs, and modification of GAG chains. Moreover, future studies, remaining challenges, and potential solutions in this field are discussed.
Subject(s)
Glycosaminoglycans , Oligosaccharides , Glycosaminoglycans/chemistry , Glycosaminoglycans/metabolism , Oligosaccharides/chemistryABSTRACT
The experiment was conducted to investigate the effects of dietary leucine on growth, antioxidant capacity, immune response, and inflammation in juvenile yellow catfish. Five diets were formulated to contain five dietary leucine levels: 12.00 (control), 19.00, 26.00, 33.00, and 40.00 g kg-1. Each diet was randomly assigned to triplicate groups of 30 juvenile fish (5.02 ± 0.15 g) twice daily to apparent satiation for 56 days. Weight gain rate, specific growth rate, and activities of liver superoxide dismutase, glutathione peroxidase, and serum lysozyme, as well as immunoglobulin M content, significantly increased with increase in dietary leucine levels up to 26.00 g kg-1, but those values decreased significantly with a further increase in dietary leucine. On the contrary, the lowest malondialdehyde content was found in 26.00 and 33.00 g kg-1 leucine groups. The expression levels of IGF 1 and MYF 5 genes in muscle were significantly upregulated with increase in dietary leucine levels up to 26.00 g kg-1, but the expression of MSTN level showed the opposite trend. The lowest expression levels of IL 8 and TNFÉ genes in the liver were found in 26.00 g kg-1 leucine groups. The quadratic regression analysis on weight gain, specific growth rate, and feed conversion ratio against dietary leucine levels indicated that the optimal dietary leucine requirement was estimated to be 26.84-27.00 g kg-1of the dry diet.
ABSTRACT
Objective: This study aimed to investigate the role of spectral detector computed tomography (SDCT) quantitative parameters and their derived quantitative parameters combined with lesion morphological information in the differential diagnosis of solid SPNs. Methods: This retrospective study included basic clinical data and SDCT images of 132 patients with pathologically confirmed SPNs (102 and 30 patients in the malignant and benign groups, respectively). The morphological signs of SPNs were evaluated and the region of interest (ROI) was delineated from the lesion to extract and calculate the relevant SDCT quantitative parameters, and standardise the process. Differences in qualitative and quantitative parameters between the groups were statistically analysed. A receiver operating characteristic (ROC) curve was constructed to evaluate the efficacy of the corresponding parameters in the diagnosis of benign and malignant SPNs. Statistically significant clinical data, CT signs and SDCT quantitative parameters were analysed using multivariate logistic regression to determine the independent risk factors for predicting benign and malignant SPNs, and the best multi-parameter regression model was established. Inter-observer repeatability was assessed using the intraclass correlation coefficient (ICC) and Bland-Altman plots. Results: Malignant SPNs differed from benign SPNs in terms of size, lesion morphology, short spicule sign, and vascular enrichment sign (P< 0.05). The SDCT quantitative parameters and their derived quantitative parameters of malignant SPNs (SAR40keV, SAR70keV, Δ40keV, Δ70keV, CER40keV, CER70keV, NEF40keV, NEF70keV, λ, NIC, NZeff) were significantly higher than those of benign SPNs (P< 0.05). In the subgroup analysis, most parameters could distinguish between benign and adenocarcinoma groups (SAR40keV, SAR70keV, Δ40keV, Δ70keV, CER40keV, CER70keV, NEF40keV, NEF70keV, λ, NIC, and NZeff), and between benign and squamous cell carcinoma groups (SAR40keV, SAR70keV, Δ40keV, Δ70keV, NEF40keV, NEF70keV, λ, and NIC). However, there were no significant differences between the parameters in the adenocarcinoma and squamous cell carcinoma groups. ROC curve analysis indicated that NIC, NEF70keV, and NEF40keV had higher diagnostic efficacy for differentiating benign and malignant SPNs (area under the curve [AUC]:0.869, 0.854, and 0.853, respectively), and NIC was the highest. Multivariate logistic regression analysis showed that size (OR=1.138, 95% CI 1.022-1.267, P=0.019), Δ70keV (OR=1.060, 95% CI 1.002-1.122, P=0.043), and NIC (OR=7.758, 95% CI 1.966-30.612, P=0.003) were independent risk factors for the prediction of benign and malignant SPNs. ROC curve analysis showed that the AUC of size, Δ70keV, NIC, and a combination of the three for differential diagnosis of benign and malignant SPNs were 0.636, 0.846, 0.869, and 0.903, respectively. The AUC for the combined parameters was the largest, and the sensitivity, specificity, and accuracy were 88.2%, 83.3% and 86.4%, respectively. The SDCT quantitative parameters and their derived quantitative parameters in this study exhibited satisfactory inter-observer repeatability (ICC: 0.811-0.997). Conclusion: SDCT quantitative parameters and their derivatives can be helpful in the differential diagnosis of benign and malignant solid SPNs. The quantitative parameter, NIC, is superior to the other relevant quantitative parameters and when NIC is combined with lesion size and Δ70keV value for comprehensive diagnosis, the efficacy could be further improved.
ABSTRACT
Background: Conventional dynamic computed tomography (CT) has a low specificity for the distinction between benign and malignant solitary pulmonary nodules (SPNs), and spectral CT has been proposed as a potential alternative. We aimed to investigate the role of quantitative parameters based on full-volume spectral CT in the differential diagnosis of SPNs. Methods: This retrospective study included spectral CT images of 100 patients with pathologically confirmed SPNs (78 and 22 in the malignant and benign groups, respectively). All cases were confirmed by postoperative pathology, percutaneous biopsy, and bronchoscopic biopsy. Multiple quantitative parameters derived from spectral CT were extracted from whole-tumor volume and standardized. Differences in quantitative parameters between groups were statistically analyzed. Diagnostic efficiency was evaluated by generating a receiver operating characteristic (ROC) curve. Between-group differences were evaluated using an independent sample t-test or Mann-Whitney U test. Interobserver repeatability was assessed using intraclass correlation coefficients (ICCs) and Bland-Altman plots. Results: Spectral CT-derived quantitative parameters, except attenuation difference between the SPN in the 70 keV and arterial enhancement [ΔS-A(70 keV)], were significantly higher for malignant SPNs than for benign nodules (P<0.05). In the subgroup analysis, most parameters could distinguish between the benign and adenocarcinoma groups, and between the benign and squamous cell carcinoma groups (P<0.05). Only 1 parameter could differentiate the adenocarcinoma and squamous cell carcinoma groups (P=0.020). ROC curve analysis indicated that normalized arterial enhancement fraction in the 70 keV (NEF70 keV), normalized iodine concentration (NIC), and Δ70 keV had high diagnostic efficacy for differentiating SPNs between the benign and malignant SPNs [area under the curve (AUC): 0.867, 0.866, and 0.848, respectively] and between the benign and adenocarcinoma groups (AUC: 0.873, 0.872, and 0.874, respectively). The multiparameters derived from spectral CT exhibited satisfactory interobserver repeatability (ICC: 0.856-0.996). Conclusions: Our study suggests that quantitative parameters derived from whole-volume spectral CT may be useful to improve discrimination of SPNs.
ABSTRACT
Vibrio alginolyticus, one of the prevalently harmful Vibrio species found in the ocean, causes significant economic damage in the shrimp farming industry. Its flagellum serves as a crucial virulence factor in the invasion of host organisms. However, the processes of bacteria flagella recognition and activation of the downstream immune system in shrimp remain unclear. To enhance comprehension of this, a ΔflhG strain was created by in-frame deletion of the flhG gene in V. alginolyticus strain HN08155. Then we utilized the transcriptome analysis to examine the different immune responses in Litopenaeus vannamei hepatopancreas after being infected with the wild type and the mutant strains. The results showed that the ΔflhG strain, unlike the wild type, lost its ability to regulate flagella numbers negatively and displayed multiple flagella. When infected with the hyperflagella-type strain, the RNA-seq revealed the upregulation of several immune-related genes in the shrimp hepatopancreas. Notably, two C-type lectins (CTLs), namely galactose-specific lectin nattectin and macrophage mannose receptor 1, and the TNF receptor-associated factor (TRAF) 6 gene were upregulated significantly. These findings suggested that C-type lectins were potentially involved in flagella recognition in shrimp and the immune system was activated through the TRAF6 pathway after flagella detection by CTLs.
Subject(s)
Hepatopancreas , Vibrio alginolyticus , Animals , Vibrio alginolyticus/genetics , Immunity, Innate/genetics , Gene Expression Profiling , Flagella/genetics , Lectins, C-Type/geneticsABSTRACT
Tuft cells are a type of rare epithelial cells that have been recently found to utilize taste signal transduction pathways to detect and respond to various noxious stimuli and pathogens, including allergens, bacteria, protists and parasitic helminths. It is, however, not fully understood how many different types of pathogens they can sense or what exact molecular mechanisms they employ to initiate targeted responses. In this study, we found that an anaerobic pathobiont microbe, Ruminococcus gnavus (R. gnavus), can induce tuft cell proliferation in the proximal colon whereas the microbe's lysate can stimulate these proximal colonic tuft cells to release interleukin-25 (IL-25). Nullification of the Gng13 and Trpm5 genes that encode the G protein subunit Gγ13 and transient receptor potential ion channel Trpm5, respectively, or application of the Tas2r inhibitor allyl isothiocyanate (AITC), G protein Gßγ subunit inhibitor Gallein or the phospholipase Cß2 (PLCß2) inhibitor U73122 reduces R. gnavus-elicited tuft cell proliferation or IL-25 release or both. Furthermore, Gng13 conditional knockout or Trpm5 knockout diminishes the expression of gasdermins C2, C3 and C4, and concomitantly increases the activated forms of caspases 3, 8 and 9 as well as the number of TUNEL-positive apoptotic cells in the proximal colon. Together, our data suggest that taste signal transduction pathways are not only involved in the detection of R. gnavus infection, but also contribute to helping maintain gasdermin expression and prevent apoptotic cell death in the proximal colon, and these findings provide another strategy to combat R. gnavus infection and sheds light on new roles of taste signaling proteins along with gasdermins in protecting the integrity of the proximal colonic epithelium.
Subject(s)
Taste , Transient Receptor Potential Channels , Ruminococcus , Signal Transduction , ColonABSTRACT
Interstitial Cells of Cajal (ICC) plays a critical role in the peristaltic contractions of the gastrointestinal and urinary tract. The dysfunction and loss of ICC contributes to hypokinetic disease, such as gallstoneand ureteropelvic junction obstruction . In the present study, we identified the underlying driving molecular signals of oxidative stress and apoptosis in ICC. ICC was isolated from small intestine of Balb/c mice, and stimulated with tumor necrosis factor-alpha (TNF-α). MTT and flow cytometry were performed to assess cell viability, apoptosis, and the level of reactive oxygen species in ICC, respectively. The level of malondialdehyde, superoxide dismutase, and glutathione peroxidase in cells were measured to assess oxidative stress. The expression of inflammatory factors (interleukin, IL-1 and IL-6) and apoptosis-related proteins were detected by western blot. We observed that TNF-αinduced inflammation, oxidative stress and cell apoptosis in ICC. By using quantitative real-time PCR , we verified that the expression of long non-coding RNAMEG3 was elevated by TNF-α in ICC. Silencing MEG3 reversed inflammation, oxidative stress, and cell apoptosisin TNF-α-treated ICC. Subsequently, we confirmed that MEG3 sponged cytoprotective miR-21 to upregulate the expression of I-kappa-B-kinase beta (IKKB) and activate the nuclear factor kappa-B (NF-κB) pathway. Both miR-21 overexpression and IKKB knockdown reduced TNF-α-induced above symptoms in ICC. Taken together, we can conclude that MEG3 mediates inflammation, oxidative stress and apoptosis in TNF-α-treated ICC via the miR-21/IKKB-NF-κB axis. The study improves our understanding of the molecular mechanism of ICC reduction related diseases.