[Purification and characterization of endoglucanase Egn21 from Fusarium sp. Q7-31T].

Objective: The objective of this research was to study plant cell wall degradation enzymes from Fusarium sp. Q7-31T. Methods: Strain was cultured in liquid medium with 1% (W/V) peptone as nitrogen source, 0.5% (W/V) oat straw as carbon source, 120 r/min shaking at 20 °C for 3 days. The endoglucanase Egn21 was purified by using Sephacry S-100 chromatography and DEAE-sepharose ion-exchange column chromatography. Then the enzymatic properties and MADIL-TOF-TOF identification were analyzed. Results: The molecular weight and isoelectric point (pI) of Egn21 was 44.25 kDa and 4.91, respectively. Egn21 had optimal activity with carboxymethyl cellulose at 40 °C and pH 6.0, stable at 45 °C and pH between 5.0 and 8.0, inhibited by Fe2+, Ca2+, K+, Na+, Mn2+ and inactivated by Hg2+, whereas Co2+, Zn2+ and Mg2+ had no effect. Conclusion: The enzymatic properties and MADIL-TOF-TOF results suggested that Egn21 belongs to GH5 family.

[Purification, identification and characterization of an endoglucanase Egn20 from Fusarium sp. Q7-31T].

OBJECTIVE: The endoglucanase from Fusarium sp. Q7-31T was isolated, purified, identified and characterized to provide data for enzyme system of Fusarium sp. . [Methods] Strain was cultured in liquid fermentation with oat straw as carbon source, the endoglucanase was purified by using Sephacry S-100 chromatography and DEAE-sepharose ion-exchange column chromatography and the enzymatic properties were studied. The protein was identified using MADIL-TOF-TOF. RESULTS: An endoglucanase was purified and named Egn20. The molecular weight was 55.37 kDa and isoelectric point (pI) was 7.44. Egn20 had optimal activity with carboxymethyl cellulose at 40 degrees C and pH 6.0, stabilized at 45 degrees C and pH 5.0 - 7.0, activated by Fe2+, inhibited by Na+, Ca2+, Mg2+, Zn2+, K+ and inactivated by Hg2+. The enzymatic properties and MADIL-TOF-TOF results suggested that Egn20 belongs to GH7 family. CONCLUSION: Our results may provide important data for the study of Fusarium sp. enzyme system.

Expression and correlation analysis of silent information regulator 1 (SIRT1), sterol regulatory element-binding protein-1 (SREBP1), and pyroptosis factor in gestational diabetes mellitus.

BACKGROUND AND AIM: Globally, the prevalence of gestational diabetes mellitus (GDM) is rising each year, yet its pathophysiology is still unclear. To shed new light on the pathogenesis of gestational diabetes mellitus and perhaps uncover new therapeutic targets, this study looked at the expression levels and correlations of SIRT1, SREBP1, and pyroptosis factors like NLRP3, Caspase-1, IL-1, and IL-18 in patients with GDM. METHODS: This study involved a comparative analysis between two groups. The GDM group consisted of 50 GDM patients and the control group included 50 pregnant women with normal pregnancies. Detailed case data were collected for all participants. We utilized real-time quantitative PCR and Western Blot techniques to assess the expression levels of SIRT1 and SREBP1 in placental tissues from both groups. Additionally, we employed an enzyme-linked immunosorbent assay to measure the serum levels of SIRT1, SREBP1, and pyroptosis factors, namely NLRP3, Caspase-1, IL-1ß, and IL-18, in the patients of both groups. Subsequently, we analyzed the correlations between these factors and clinical. RESULTS: The results showed that there were significantly lower expression levels of SIRT1 in both GDM group placental tissue and serum compared to the control group (p < 0.01). In contrast, the expression of SREBP1 was significantly higher in the GDM group than in the control group (p < 0.05). Additionally, the serum levels of NLRP3, Caspase-1, IL-1ß, and IL-18 were significantly elevated in the GDM group compared to the control group (p < 0.01). The expression of SIRT1 exhibited negative correlations with the expression of FPG, OGTT-1h, FINS, HOMA-IR, SREBP1, IL-1ß, and IL-18. However, there was no significant correlation between SIRT1 expression and OGTT-2h, NLRP3, or Caspase-1. On the other hand, the expression of SREBP1 was positively correlated with the expression of IL-1ß, Caspase-1, and IL-18, but has no apparent correlation with NLRP3. CONCLUSIONS: Low SIRT1 levels and high SREBP1 levels in placental tissue and serum, coupled with elevated levels of pyroptosis factors NLRP3, Caspase-1, IL-1ß, and IL-18 in serum, may be linked to the development of gestational diabetes mellitus. Furthermore, these three factors appear to correlate with each other in the pathogenesis of GDM, offering potential directions for future research and therapeutic strategies.

Relationship between serum NLRP3 along with its effector molecules and pregnancy outcomes in women with hyperglycemia.

OBJECTIVE: The study aims to investigate the levels of serum NLRP3 along with its effector molecules (Caspase-1, IL-1ß, and IL-18) in the mid-pregnancy in pregnant women with hyperglycemia, and explore the relationship between NLRP3, along with its effector molecules (Caspase-1, IL-1ß, and IL-18) and insulin resistance, as well as pregnancy outcomes. METHODS: The levels of serum NLRP3 along with its effector molecules (Caspase-1, IL-1ß, and IL-18) in three groups of pregnant women with gestational diabetes mellitus (GDM), pregestational diabetes mellitus (PGDM) and normal glucose tolerance (NGT) were measured in mid-pregnancy, and their relationship with insulin resistance and pregnancy outcomes was analyzed. The ROC curve was also used to evaluate the predictive value of serum NLRP3 inflammasome and its effector molecules for pregnancy outcomes. RESULTS: There were no statistical differences in the general clinical data of the three groups, and the concentrations of serum NLRP3 along with its effector molecules were higher in the GDM and PGDM groups than in the NGT group, and NLRP3 along with its effector molecules were positively correlated with fasting blood glucose, fasting insulin, and insulin resistance index in both groups (r > 0, p < .05). The incidence of preterm delivery, hypertensive disorders of pregnancy, premature rupture of membranes, neonatal hypoglycemia and macrosomia was significantly higher in both groups than in the NGT group (p < .05). The value of the combined serum NLRP3 and its effector molecules in mid-pregnancy to predict adverse pregnancy outcomes was highest, and the AUCs for the combined prediction of late hypertensive disorders of pregnancy, premature rupture of membranes, preterm delivery, neonatal hypoglycemia and macrosomia were 0.84 (95% CI 0.79-0.88, p < .001), 0.81 (95% CI 0.75-0.85, p < .001), 0.76 (95% CI 0.70-0.81, p < .001), 0.76 (95% CI 0.70-0.81, p < .001) and 0.72 (95% CI 0.63-0.81, p < .001), respectively. CONCLUSIONS: Increased serum NLRP3 along with its effector molecules in pregnant women with hyperglycemia are associated with the levels of insulin resistance and the subsequent development of adverse pregnancy outcomes.

Bi-terminal fusion of intrinsically-disordered mussel foot protein fragments boosts mechanical strength for protein fibers.

Microbially-synthesized protein-based materials are attractive replacements for petroleum-derived synthetic polymers. However, the high molecular weight, high repetitiveness, and highly-biased amino acid composition of high-performance protein-based materials have restricted their production and widespread use. Here we present a general strategy for enhancing both strength and toughness of low-molecular-weight protein-based materials by fusing intrinsically-disordered mussel foot protein fragments to their termini, thereby promoting end-to-end protein-protein interactions. We demonstrate that fibers of a ~60 kDa bi-terminally fused amyloid-silk protein exhibit ultimate tensile strength up to 481 ± 31 MPa and toughness of 179 ± 39 MJ*m-3, while achieving a high titer of 8.0 ± 0.70 g/L by bioreactor production. We show that bi-terminal fusion of Mfp5 fragments significantly enhances the alignment of ß-nanocrystals, and intermolecular interactions are promoted by cation-π and π-π interactions between terminal fragments. Our approach highlights the advantage of self-interacting intrinsically-disordered proteins in enhancing material mechanical properties and can be applied to a wide range of protein-based materials.

OsbZIP60-mediated unfolded protein response regulates grain chalkiness in rice.

Grain chalkiness, an undesirable trait caused by complex factors, has great negative impacts on the quality and economic value of rice. However, little is known about the regulatory mechanism of grain chalkiness, particularly the effect of endoplasmic reticulum (ER) stress. Here, a genome-wide association study (GWAS) reveals that the transcription factor OsbZIP60 is a vital regulator of rice grain chalkiness. Genetic analysis demonstrates that knockout of OsbZIP60 results in extremely high grain chalkiness and aberrant structure of storage substances. Notably, the expression of unfolded protein response (UPR) genes, such as OsbZIP50, OsBiP1, OsBiP2 and OsBiP3, is up-regulated in the endosperm cells of osbzip60, and overexpression of all these UPR genes causes various degrees of chalkiness. Furthermore, OsbZIP60 is found to activate the expression of key genes related to grain chalkiness, such as GPA3, FSE1, FLO7, Chalk5, OsNF-YB1, and OsPK2, whose expression is significantly suppressed in osbzip60 and overexpression lines of OsbZIP50, OsBiP1, OsBiP2, and OsBiP3. Our study provides novel insights into the function of OsbZIP60 and the role of the UPR pathway in the formation of grain chalkiness in rice.

Analogy-Detail Networks for Object Recognition.

The human visual system can recognize object categories accurately and efficiently and is robust to complex textures and noises. To mimic the analogy-detail dual-pathway human visual cognitive mechanism revealed in recent cognitive science studies, in this article, we propose a novel convolutional neural network (CNN) architecture named analogy-detail networks (ADNets) for accurate object recognition. ADNets disentangle the visual information and process them separately using two pathways: the analogy pathway extracts coarse and global features representing the gist (i.e., shape and topology) of the object, while the detail pathway extracts fine and local features representing the details (i.e., texture and edges) for determining object categories. We modularize the architecture and encapsulate the two pathways into the analogy-detail block as the CNN building block to construct ADNets. For implementation, we propose a general principle that transmutes typical CNN structures into the ADNet architecture and applies the transmutation on representative baseline CNNs. Extensive experiments on CIFAR10, CIFAR100, street view house numbers, and ImageNet data sets demonstrate that ADNets significantly reduce the test error rates of the baseline CNNs by up to 5.76% and outperform other state-of-the-art architectures. Comprehensive analysis and visualizations further demonstrate that ADNets are interpretable and have a better shape-texture tradeoff for recognizing the objects with complex textures.

Microbial production of megadalton titin yields fibers with advantageous mechanical properties.

Manmade high-performance polymers are typically non-biodegradable and derived from petroleum feedstock through energy intensive processes involving toxic solvents and byproducts. While engineered microbes have been used for renewable production of many small molecules, direct microbial synthesis of high-performance polymeric materials remains a major challenge. Here we engineer microbial production of megadalton muscle titin polymers yielding high-performance fibers that not only recapture highly desirable properties of natural titin (i.e., high damping capacity and mechanical recovery) but also exhibit high strength, toughness, and damping energy - outperforming many synthetic and natural polymers. Structural analyses and molecular modeling suggest these properties derive from unique inter-chain crystallization of folded immunoglobulin-like domains that resists inter-chain slippage while permitting intra-chain unfolding. These fibers have potential applications in areas from biomedicine to textiles, and the developed approach, coupled with the structure-function insights, promises to accelerate further innovation in microbial production of high-performance materials.

Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases.

In-field molecular diagnosis of plant diseases via nucleic acid amplification is currently limited by cumbersome protocols for extracting and isolating pathogenic DNA from plant tissues. To address this challenge, a rapid plant DNA extraction method was developed using a disposable polymeric microneedle (MN) patch. By applying MN patches on plant leaves, amplification-assay-ready DNA can be extracted within a minute from different plant species. MN-extracted DNA was used for direct polymerase chain reaction amplification of plant plastid DNA without purification. Furthermore, using this patch device, extraction of plant pathogen DNA ( Phytophthora infestans) from both laboratory-inoculated and field-infected leaf samples was performed for detection of late blight disease in tomato. MN extraction achieved 100% detection rate of late blight infections for samples after 3 days of inoculation when compared to the conventional gold standard cetyltrimethylammonium bromide (CTAB)-based DNA extraction method and 100% detection rate for all blind field samples tested. This simple, cell-lysis-free, and purification-free DNA extraction method could be a transformative approach to facilitate rapid sample preparation for molecular diagnosis of various plant diseases directly in the field.

Assessment of heavy metals contamination in sediments from three adjacent regions of the Yellow River using metal chemical fractions and multivariate analysis techniques.

Metal chemical fractions obtained by optimized BCR three-stage extraction procedure and multivariate analysis techniques were exploited for assessing 7 heavy metals (Cr, Pb, Cd, Co, Cu, Zn and Ni) in sediments from Gansu province, Ningxia and Inner Mongolia Autonomous Regions of the Yellow River in Northern China. The results indicated that higher susceptibility and bioavailability of Cr and Cd with a strong anthropogenic source were due to their higher availability in the exchangeable fraction. A portion of Pb, Cd, Co, Zn, and Ni in reducible fraction may be due to the fact that they can form stable complexes with Fe and Mn oxides. Substantial amount of Pb, Co, Ni and Cu was observed as oxidizable fraction because of their strong affinity to the organic matters so that they can complex with humic substances in sediments. The high geo-accumulation indexes (I(geo)) for Cr and Cd showed their higher environmental risk to the aquatic biota. Principal component analysis (PCA) revealed that high toxic Cr and Cd in polluted sites (Cd in S10, S11 and Cr in S13) may be contributed to anthropogenic sources, it was consistent with the results of dual hierarchical clustering analysis (DHCA), which could give more details about contributing sources.

ZEB1-activated Notch1 promotes circulating tumor cell migration and invasion in lung squamous cell carcinoma

Background Lung squamous cell carcinoma (LUSC) is recognized as the major subtypes of non-small cell lung cancer (NSCLC). Circulating tumor cells (CTCs) are critical players in tumor metastasis. A molecular profiling of CTCs has previously identified notch receptor 1 (Notch1) as an important mediator in NSCLC. Therefore, we investigate Notch1 roles in LUSC and its related mechanisms. Methods The serum levels of Notch1 were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The CTCs isolated from blood samples were characterized via an immunofluorescence method. Cell motion was determined using Transwell chambers. The regulatory relationship between Notch1 and zinc finger E-box-binding homeobox 1 (ZEB1) was verified by chromatin immunoprecipitation (ChIP) and luciferase reporter assays. The protein levels were detected by western blotting. Results Higher Notch1 expression in patients with LUSC than that in normal controls was observed. Notch1 knockdown inhibited cell motion and epithelial–mesenchymal transition (EMT). ZEB1 transcriptionally activated Notch1. ZEB1 upregulation exacerbated the malignant phenotypes of CTCs. Conclusion ZEB1-activated Notch1 promotes malignant phenotypes of CTCs in LUSC and indicates poor prognosis (AU)