Microwave Depolymerization of Lignin via Dynamic Vapor Flow Reaction System: HCOOH as Pretreatment Solvent or Reforming Solvent Vapor.

Different forms of HCOOH in the depolymerization system play an important role in governing the monomeric products from lignin. We reported two strategies for the introduction of HCOOH to enrich the monophenols from kraft lignin by microwave-assisted depolymerization. The reaction of lignin models showed that HCOOH was in favor of the cleavage of C-O bonds (ß-O-4 typically) and partial C-C bonds (Cα-Cß). Subsequently, Microwave-assisted depolymerization of lignin with two strategies was conducted via a designed dynamic vapor flow reaction system. Strategy A with HCOOH as pretreatment solvent showed excellent monophenols enrichment with total mass yields of 193.71 mg/g (lignin basis). Strategy B using HCOOH as reforming solvent vapor significantly increased the monophenols selectivity. It presented unique reforming and upgrading performance by generating catechol (42.59 mg/g, lignin basis) and homovanillic acid (17.58 mg/g, lignin basis). This study provided potential strategies for the efficient conversion of kraft lignin into high-value platform chemicals.

Arabidopsis 3ß-Hydroxysteroid Dehydrogenases/C4-Decarboxylases Are Essential for the Pollen and Embryonic Development.

The biosynthesis of C27-29 sterols from their C30 precursor squalene involves C24-alkylation and the removal of three methyl groups, including two at the C4 position. The two C4 demethylation reactions require a bifunctional enzyme known as 3ß-hydroxysteroid dehydrogenase/C4-decarboxylase (3ßHSD/D), which removes an oxidized methyl (carboxylic) group at C4 while simultaneously catalyzing the 3ß-hydroxyl→3-keto oxidation. Its loss-of-function mutations cause ergosterol-dependent growth in yeast and congenital hemidysplasia with ichthyosiform erythroderma and limb defect (CHILD) syndrome in humans. Although plant 3ßHSD/D enzymes were well studied enzymatically, their developmental functions remain unknown. Here we employed a CRISPR/Cas9-based genome-editing approach to generate knockout mutants for two Arabidopsis 3ßHSD/D genes, HSD1 and HSD2, and discovered the male gametophytic lethality for the hsd1 hsd2 double mutation. Pollen-specific expression of HSD2 in the heterozygous hsd1 hsd2/+ mutant not only rescued the pollen lethality but also revealed the critical roles of the two HSD genes in embryogenesis. Our study thus demonstrated the essential functions of the two Arabidopsis 3ßHSD/D genes in male gametogenesis and embryogenesis.

Enhanced beetle antennae search algorithm for complex and unbiased optimization.

Beetle Antennae Search algorithm is a kind of intelligent optimization algorithms, which has the advantages of few parameters and simplicity. However, due to its inherent limitations, BAS has poor performance in complex optimization problems. The existing improvements of BAS are mainly based on the utilization of multiple beetles or combining BAS with other algorithms. The present study improves BAS from its origin and keeps the simplicity of the algorithm. First, an adaptive step size reduction method is used to increase the usability of the algorithm, which is based on an accurate factor and curvilinearly reduces the step size; second, the calculated information of fitness functions during each iteration are fully utilized with a contemporary optimal update strategy to promote the optimization processes; third, the theoretical analysis of the multi-directional sensing method is conducted and utilized to further improve the efficiency of the algorithm. Finally, the proposed Enhanced Beetle Antennae Search algorithm is compared with many other algorithms based on unbiased test functions. The test functions are unbiased when their solution space does not contain simple patterns, which may be used to facilitate the searching processes. As a result, EBAS outperformed BAS with at least 1 orders of magnitude difference. The performance of EBAS was even better than several state-of-the-art swarm-based algorithms, such as Slime Mold Algorithm and Grey Wolf Optimization, with similar running times. In addition, a WSN coverage optimization problem is tested to demonstrate the applicability of EBAS on real-world optimizations.

CircRNA Lrp6 promotes cementoblast differentiation via miR-145a-5p/Zeb2 axis.

BACKGROUND AND OBJECTIVE: Cementum is a part of the periodontium and anchors periodontal ligaments to the alveolar bone. Cementoblasts are responsible for the cementum formation via matrix deposition and subsequently mineralization. Thus, exploring novel mechanisms underlying the function of cementoblast contributes to the treatment of cementum damage. Recently, circRNA Lrp6 (circLRP6) has been of interest due to its active role in cell differentiation, but its potential role in cementoblast differentiation remains unclear. Herein, we attempted to elucidate the role of circLRP6 in cementoblast differentiation and clarify any associated mechanisms. MATERIAL AND METHODS: The mRNA expressions of circLRP6, miR-145a-5p, zinc finger E-box binding homeobox 2 (Zeb2), runt-related transcription factor 2 (Runx2), osteopontin (Opn), and bone sialoprotein (Bsp) were evaluated by qRT-PCR. The protein levels of Zeb2 were measured by Western blot. Bioinformatic analysis and dual-luciferase reporter assays were used to test the potential binding targets of miR-145a-5p. The differentiation potentials of the cementoblasts were assessed by Alkaline phosphatase (ALP) staining, ALP activity assay, Alizarin red S (ARS) staining, and quantification. RESULTS: In this study, circLRP6 was significantly upregulated in cementoblast differentiation. Furthermore, circLRP6 knockdown inhibited ALP levels, reduced calcium nodule formation and the expression of Runx2, Opn, and Bsp. Mechanically, bioinformatic analysis and dual-luciferase reporter assays confirmed miR-145a-5p was a potential binding target of circLRP6. miR-145a-5p can negatively regulate cementoblast differentiation. Subsequently, bioinformatic analysis and dual-luciferase reporter assays confirmed Zeb2 was a potential miR-145a-5p target. miR-145a-5p overexpression resulted in a downregulation of Zeb2. Furthermore, Zeb2 inhibition partially reversed the effect of circLRP6 during cementoblast differentiation. CONCLUSION: Taken together, circLRP6 appears to modulate cementoblast differentiation by antagonizing the function of miR-145a-5p, thereby increasing Zeb2. This study serves as a stepping stone for the potential development of an approach to promote cementum formation.

Plastid genome sequencing, identification of nuclear SNP markers, and quality assessment of medicinal rhizomatous herb Polygonatum odoratum (Asparagaceae) cultivars.

Polygonatum odoratum (Mill.) Druce (Asparagaceae, Asparagales) is a widely cultivated medicinal herb in China. However, this useful herb is understudied despite being known as a medicinal resource with top grade medical and edible properties since long. In this study, P. odoratum and four cultivars were investigated. The variations in morphological characteristics and vegetative phases of each cultivar were observed. For genetic aspect, the plastid genome of P. odoratum varies in length from 154,569 bp to 155,491 bp, containing a large single-copy region of 83,486-84,459 bp, a small single-copy region of 18,292-18,471 bp, and two inverted repeats of 26,302-26,370 bp. A total of 131 genes were predicted, including 85 protein-coding, 38 tRNA, and eight rRNA genes. Genome comparisons revealed a slight variation in the sequence across the five accessions, but two highly variable regions (trnC-petN and rpl32-trnL) were detected when comparing the four different cultivars. For the RAD-seq markers, a total of 33.64 Gb of clean data, with an average value of 1.08 Gb per sample, were analyzed for the presence of single nucleotide polymorphisms (SNPs). Well-resolved phylogenies of the P. odoratum cultivars are constructed; the nonmonophyletic relationship in the plastome-based phylogenetic trees, yet monophyletic form in the RAD-based linkage map suggested possibility of hybrid cultivar for P. odoratum "Dazhu" (GDDZ), which was further supported by morphological observations. Quality assessment based on the standards of the Chinese Pharmacopoeia on Polygonati Odorati Rhizoma (POR) on the four cultivars used in this study recorded that PORs from P. odoratum "Zhongzhu" (GDZZ) met the minimum criteria for the acceptance as raw material for medicinal drug production. This study has provided insights on the morphological variations, genetic background, and medicinal qualities of P. odoratum cultivars that could be explored for future genetic improvement as well as breeding programs of P. odoratum for POR production.

miR-146a-5p attenuates IL-1ß-induced IL-6 and IL-1ß expression in a cementoblast-derived cell line.

OBJECTIVE: miR-146a is widely induced during the immune response. However, little is known about the biogenesis, function and mechanism of miR-146a in cementoblasts during the pathogenesis of periodontitis. This study aimed to investigate the effects of miR-146a in murine cementoblast-derived OCCM-30 cells following IL-1ß stimulation. MATERIALS AND METHODS: OCCM-30 cells were cultured and exposed to IL-1ß. IL-6, IL-1ß and TNF-α, and miR-146a-5p expression was assessed by qRT-PCR. Mimics/inhibitors were transiently transfected into cells to determine the function of miR-146a-5p. Signalling pathways including p38 MAPK, ERK1/2 and NF-κB were studied by using specific inhibitors. The indicated proteins were measured by Western blot analysis and ELISA. RESULTS: In IL-1ß-stimulated OCCM-30 cells, the expression levels of miR-146a-5p along with IL-6 and IL-1ß increased in a time-dependent manner. The ERK1/2, p38 MAPK and NF-κB pathway were activated upon IL-1ß stimulation. Blocking the NF-κB pathway decreased IL-6, IL-1ß and miR-146a-5p expression. The overexpression of miR-146a-5p reduced IL-6 and IL-1ß expression, while the inhibition of miR-146a-5p increased IL-6 and IL-1ß expression in IL-1ß-treated OCCM-30 cells. miR-146a-5p attenuated IL-6 and IL-1ß expression via the IRAK1/TRAF6 pathway. CONCLUSION: This study suggested that miR-146a-5p attenuates IL-1ß-induced inflammatory factors in cementoblast-derived cell line.

Irisin promotes cementoblast differentiation via p38 MAPK pathway.

OBJECTIVE: Irisin is a newly identified exercise-induced myokine which can affect glucose metabolism and cortical bone mass and strength. However, the influence of irisin on cementoblasts remains largely unknown. MATERIAL AND METHODS: An immortalized mouse cementoblast cell line OCCM-30 was used in this study. Cementoblast differentiation markers and PGC-1α in cells cultured with mineral induction medium were evaluated by qRT-PCR. Cementoblast mineralization was evaluated by alizarin red staining. Differentiation markers and the activity of p38 MAPK pathway under irisin stimulation were assessed by qRT-PCR or Western blot analysis. p38 MAPK pathway inhibitor SB203580 or p38 siRNA was used to further identify the regulatory mechanism. Cell proliferation treated with irisin was examined by CCK-8 method. RESULTS: The expression of Runx2, osterix, ALP, and PGC-1α was up-regulated consistently under mineral induction. The formation of mineralized nodules was increased by irisin. Runx2, osterix, ALP, and osteocalcin were obviously up-regulated under irisin stimulation as well as the activity of p38 MAPK pathway. When pretreated with SB203580 or p38 siRNA before irisin stimulation, the irisin-induced differentiation was distinctly suppressed. OCCM-30 cell proliferation was enhanced when treated with high-dose irisin for long time. CONCLUSION: Irisin can promote the differentiation of cementoblasts via p38 MAPK pathway.

Detoxification Strategies for Zearalenone Using Microorganisms: A Review.

Zearalenone (ZEA) is a mycotoxin produced by Fusarium fungi that is commonly found in cereal crops. ZEA has an estrogen-like effect which affects the reproductive function of animals. It also damages the liver and kidneys and reduces immune function which leads to cytotoxicity and immunotoxicity. At present, the detoxification of mycotoxins is mainly accomplished using biological methods. Microbial-based methods involve zearalenone conversion or adsorption, but not all transformation products are nontoxic. In this paper, the non-pathogenic microorganisms which have been found to detoxify ZEA in recent years are summarized. Then, two mechanisms by which ZEA can be detoxified (adsorption and biotransformation) are discussed in more detail. The compounds produced by the subsequent degradation of ZEA and the heterogeneous expression of ZEA-degrading enzymes are also analyzed. The development trends in the use of probiotics as a ZEA detoxification strategy are also evaluated. The overall purpose of this paper is to provide a reliable reference strategy for the biological detoxification of ZEA.

Bacillus velezensis A2 fermentation exerts a protective effect on renal injury induced by Zearalenone in mice.

Zearalenone (ZEN) is an estrogen-like mycotoxin occurring in food and feeds, and it can cause oxidative damage and apoptosis in the testis, liver, and kidney. A current concern for researchers is how to reduce the harm it causes to humans and animals. In this study, our aim was to isolate and identify a novel and efficient ZEN-detoxifying strain of bacteria, and we aimed to assess the protective effect of the isolated strain on kidney damage caused by ZEN in mice. Our results indicated that a strain of Bacillus velezensis (B. velezensis), named A2, could completely degrade ZEN (7.45 µg/mL) after three days of incubation at 37 °C in the Luria-Bertani (LB) medium. This fermentation broth of the B. velezensis A2 strain was given to mice. The histopathological analysis indicated that the fermentation broth from the B. velezensis A2 strain reduced the degree of renal injury that is induced by ZEN. Furthermore, it greatly reduced the increase in serum levels of creatinine (CRE), uric acid (UA), and urea nitrogen (BUN) caused by ZEN. In addition, B. velezensis A2 strain also significantly inhibited the increase of malonaldehyde (MDA) content, and reversed the decreases of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) activities caused by ZEN. Studies have shown that ZEN is involved in the regulation of mRNA and protein levels of genes involved in the ER stress-induced apoptotic pathway, such as heavy chain binding protein (BIP), C-/-EBP homologous protein (CHOP), cysteine Aspartate-specific protease-12 (Caspase-12), c-Jun N-terminal kinase (JNK), and BCL2-related X protein (Bcl-2 and Bax). However, when mice were administered the fermentation broth of the B. velezensis A2 strain, it significantly reversed the expressions of these genes in their kidney tissue. In conclusion, our results indicate that the newly identified strain of B. velezensis A2, has a protective effect from renal injury induced by ZEN in mice. This strain has a potential application in the detoxification of ZEN in feed and protects animals from ZEN poisoning.

Transcriptome analysis to identify the Ras and Rap1 signal pathway genes involved in the response of TM3 Leydig cells exposed to zearalenone.

The mechanism of action of zearalenone (ZEA) in inducing germ cell tumors is unclear, and little is known about the change in the transcriptome of germ cells after ZEA exposure. To explore the molecular basis of the ZEA oncogene, we examined the median lethal concentration (50 µmol/L) and pro-apoptotic effect of ZEA on TM3 Leydig cells by MTT and TUNEL assay. Subsequently, we investigated the genetic changes in the transcriptome of TM3 Leydig cells exposed to 50 µmol/L ZEA. The transcriptome sequencing results show that 772 genes are significantly down-regulated, while 204 genes are significantly up-regulated. Gene ontology (GO) enrichment analysis shows that ZEA has a major effect on the connective function, cell composition, cell cycle, and energy metabolism of the TM3 Leydig cells. Using the results of the GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, we select the Ras and Rap1 signaling pathways that are closely related to the occurrence of cancer. The differentially expressed genes visualized in the KEGG pathway were selected for RT-qPCR differential gene expression validation. The results show that the gene expression results are consistent with the transcriptome sequencing results. This study thus provides a theoretical molecular basis for the mechanism of ZEA carcinogenesis.

Combined Use of C. butyricum Sx-01 and L. salivarius C-1-3 Improves Intestinal Health and Reduces the Amount of Lipids in Serum via Modulation of Gut Microbiota in Mice.

The study was conducted to investigate whether combined use of C. butyricum Sx-01 and L. salivarius C-1-3 could improve the intestinal health and reduce the lipid levels in sera of mice and whether these benefits were related to regulating the intestinal microflora. Eighty Kunming male mice were divided into four groups with five replicates per group and four mice per replicate. Mice in the control group were administrated with 0.2 mL normal saline; mice in three experimental groups were daily orally administrated with 4 × 108 cfu of L. salivarius, 4 × 108 cfu of C. butyricum, and a combination thereof (2 × 108 cfu of L. salivarius, and 2 × 108 cfu of C. butyricum), respectively. The experiment lasted for 14 days. The results showed that the average daily feed intake (ADFI) and feed/gain (F/G) ratio of growing mice underwent no significant changes (p > 0.05); however, the average daily gain (ADG) tended to increase over short periods of time. The activities of SOD and GSH-Px in serum in the combination group were significantly increased (p < 0.05); The triglyceride, and total cholesterol, contents in serum in the combined treatment group were significantly decreased (p < 0.05); The total volatile fatty acids and butyric acid in faecal matter of mice in the experimental groups were all significantly increased at 14 days (p < 0.05); The length of villi, and the mucosal thickness of colon and caecum (p < 0.05) were significantly improved; The relative abundance of some bacteria with antioxidant capacity or decomposing cholesterol capacity or butyrate producing capacity was increased, while the relative abundance of some pathogenic bacteria was decreased in the colon. Furthermore, our results showed that the beneficial effects of the combined use of the two strains was higher than that of single use. Overall, the results demonstrated that the combined use of C. butyricum Sx-01 and L. salivarius C-1-3 can significantly improve intestinal health and reduce the amount of lipids in sera of mice. The reason for these effects might be that besides their own probiotic effects, combined use of the two strains could regulate the intestinal microflora.

Proanthocyanidins Protect Epithelial Cells from Zearalenone-Induced Apoptosis via Inhibition of Endoplasmic Reticulum Stress-Induced Apoptosis Pathways in Mouse Small Intestines.

This study evaluated the protective effect of proanthocyanidins (PCs) on reducing apoptosis in the mouse intestinal epithelial cell model MODE-K exposed to zearalenone (ZEA) through inhibition of the endoplasmic reticulum stress (ERS)-induced apoptosis pathway. Our results showed that PCs could reduce the rate of apoptosis in MODE-K cells exposed to ZEA (p < 0.01). PCs significantly increased the ZEA-induced antioxidant protective effects on the enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and on the content of GSH. PCs also significantly decreased the ZEA-induced increase in the content of malondialdehyde (MDA). The analysis indicated that ZEA increased both mRNA and protein expression levels of C/EBP homologous protein (CHOP), GRP78, c-Jun N-terminal kinase (JNK), and cysteinyl aspartate specific proteinase 12 (caspase-12) (p < 0.05), which are related to the ERS-induced apoptosis pathway. ZEA decreased levels of the pro-apoptotic related protein Bcl-2 (p < 0.05) and increased the anti-apoptotic related protein Bax (p < 0.05). Co-treatment with PCs was also shown to significantly reverse the expression levels of these proteins in MODE-K cells. The results demonstrated that PCs could protect MODE-K cells from oxidative stress and apoptosis induced by ZEA. The underlying mechanism may be that PCs can alleviate apoptosis in mouse intestinal epithelial cells by inhibition of the ERS-induced apoptosis pathway.

Miro1-mediated mitochondrial dysfunction under high nutrient stress is linked to NOD-like receptor 3 (NLRP3)-dependent inflammatory responses in rat pancreatic beta cells.

Type 2 Diabetes (T2D) is associated with a state of low-grade inflammation that leads to insulin resistance under sustained high-fat and glucose (HFG) stress. Mitochondria from pancreatic beta cells play an essential role by metabolizing nutrients and generating signals required for both triggering and amplifying pathways of insulin secretion responding to HFG. However, the underlying pathway linking mitochondrial function to initiate and integrate inflammatory responses within the pancreatic beta cells under HFG stress remains poorly defined. Here, we demonstrated that HFG induced Ca(2+)-mediated deleterious effects on mitochondrial rho GTPase 1 (Miro1), a protein allowing mitochondria to move along microtubules to regulate mitochondria dynamics. This redistribution of Miro1 by HFG led to aggravation of proinflammatory responses in rat islets due to damaged mitochondria-producing reactive oxygen species (ROS). In addition, HFG-induced Ca(2+)-mediated increased expression of mitochondrial dynamin-like protein (DLP1) was assembled on the outer membrane of mitochondria to initiate fission events. Higher expression of DLP1 induced mitochondria fragmentation as expected but was not essential for ROS-induced proinflammatory responses, while Miro1-mediated mitochondrial dysfunction induced proinflammatory responses under HFG stress. Combined, we proposed in this study that HFG stress caused mtROS release mainly through Miro1-mediated effects on mitochondria in pancreatic beta cells triggering the NLRP3-dependent proinflammatory responses and, subsequently, damaged insulin secretion.