Carbohydrate Metabolism Regulated by Antisense vicR RNA in Cariogenicity

2019 ◽  
Vol 99 (2) ◽  
pp. 204-213 ◽  
Author(s):  
L. Lei ◽  
B. Zhang ◽  
M. Mao ◽  
H. Chen ◽  
S. Wu ◽  
...  
Keyword(s):  
Carbohydrate Metabolism ◽  
Messenger Rna ◽  
High Performance ◽  
Response Regulator ◽  
Gel Permeation Chromatography ◽  
Monosaccharide Composition ◽  
Composition Analysis ◽  
Rnase Iii ◽  
Bacterial Gene

Streptococcus mutans is a major cariogenic pathogen that resides in multispecies oral microbial biofilms. The VicRK 2-component system is crucial for bacterial adaptation, virulence, and biofilm organization and contains a global and vital response regulator, VicR. Notably, we identified an antisense vicR RNA (AS vicR) associated with an adjacent RNase III–encoding ( rnc) gene that was relevant to microRNA-size small RNAs (msRNAs). Here, we report that ASvicR overexpression significantly impeded bacterial growth, biofilm exopolysaccharide synthesis, and cariogenicity in vivo. Transcriptome analysis revealed that the AS vicR RNA mainly regulated carbohydrate metabolism. In particular, overproducing AS vicR demonstrated a reduction in galactose and glucose metabolism by monosaccharide composition analysis. The results of high-performance gel permeation chromatography revealed that the water-insoluble glucans isolated from AS vicR presented much lower molecular weights. Furthermore, direct evidence showed that total RNAs were disrupted by rnc-encoded RNase III. With the coexpression of T4 RNA ligase, putative msRNA1657, which is an rnc-related messenger RNA, was verified to bind to the 5′-UTR regions of the vicR gene. Furthermore, AS vicR regulation revealed a sponge regulatory-mediated network for msRNA associated with adjacent RNase III–encoding genes. There was an increase in AS vicR transcript levels in clinical S. mutans strains from caries-free children, while the expression of AS vicR was decreased in early childhood caries patients; this outcome may be explored as a potential strategy contributing to the management of dental caries. Taken together, our findings suggest an important role of AS vicR-mediated sponge regulation in S. mutans, indicating the characterization of lactose metabolism by a vital response regulator in cariogenicity. These findings have a number of implications and have reshaped our understanding of bacterial gene regulation from its transcriptional conception to the key roles of regulatory RNAs.

scholarly journals Alkaline Extraction, Structural Characterization, and Bioactivities of (1→6)-β-d-Glucan from Lentinus edodes

Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1610 ◽  
Author(s):  
Jia Li ◽  
Chao Cai ◽  
Mengmeng Zheng ◽  
Jiejie Hao ◽  
Ya Wang ◽  
...  
Keyword(s):  
High Performance ◽  
Antioxidant Activities ◽  
Gel Permeation Chromatography ◽  
Monosaccharide Composition ◽  
Biological Properties ◽  
Orthogonal Experimental Design ◽  
Lentinus Edodes ◽  
Anion Exchange Column ◽  
Strong Anion Exchange ◽  
Fast Flow

The purpose of this study is to develop a robust approach to obtain β glucans from Lentinus edodes and to characterize their structural and biological properties for sustainable utilization. The alkali extraction was optimized with an orthogonal experimental design, and a concise process for obtaining specific targeting polysaccharides from Lentinus edodes was developed in this study. After purification with a Q-Sepharose Fast Flow strong anion-exchange column, the monosaccharide composition, a methylation analysis, and NMR spectroscopy were employed for their structural characterizations. LeP-N2 was found to be composed of (1→6)-β-d-glucans with minor β-(1→3) glucosidic side chains. Atomic force microscopy (AFM) and high-performance gel permeation chromatography–refractive index–multi-angle laser light scattering (HPGPC-RI-MALLS) also revealed LeP-N2 exhibiting a compact unit in aqueous solution. This (1→6)-β-d-glucan was tested for antioxidant activities with IC50 at 157 μg/mL. Moreover, RAW 264.7 macrophage activation indicated that the release of nitric oxide (NO) and reactive oxygen species (ROS) was markedly increased with no cytotoxicity at a dose of 100 μg/mL. These findings suggest that the (1→6)-β-d-glucans obtained from Lentinus edodes could serve as potential agents in the fields of functional foods or medicine.

scholarly journals Physicochemical Characterization and Immunomodulatory Activity of a Novel Acid Polysaccharide from Solanum muricatum

Polymers ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1972 ◽  
Author(s):  
Heng Yue ◽  
Qianqian Xu ◽  
Xianheng Li ◽  
Jeevithan Elango ◽  
Wenhui Wu ◽  
...  
Keyword(s):  
High Performance ◽  
Average Molecular Weight ◽  
Thermal Studies ◽  
Physicochemical Characterization ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Immunomodulatory Activity ◽  
Composition Analysis ◽  
Solanum Muricatum ◽  
Acid Polysaccharide

To investigate the structure and immunomodulatory activity of polysaccharide from Solanum muricatum, a novel acid polysaccharide named SMP-3a was purified from Solanum muricatum pulp through DEAE-52 cellulose column and Sephadex G-200 chromatography. Monosaccharide composition analysis showed that SMP-3a was mainly composed of rhamnose, arabinose, galactose, and galacturonic acid with the molar ratio of 1.09:2.64:1.54:1. The average molecular weight was found to be 227 kDa by high performance gel permeation chromatography (HPGPC). Thermal studies revealed the SMP-3a was a thermally stable polymer. Based on the results of methylation and NMR analysis, the backbone chain of SMP-3a was composed of →2)-α-l-Rhap-(1→, →4)-α-d-GalpA-(1→ and →4)-α-d-Galp-(1→. The side chain was consisted of α-l-Araf-(1→ and →5)-α-l-Araf-(1→. Immunomodulatory assay indicated that SMP-3a could significantly promote the proliferation of macrophages and stimulate the secretion of cytokines, including TNF-α, IL-1β, and IL-6. Our results suggested that SMP-3a could be used as a novel potential immunomodulatory agent in functional food.

scholarly journals Antioxidant and Hepatoprotective Activities of Polysaccharides from Spent Mushroom Substrates (Laetiporus sulphureus) in Acute Alcohol-Induced Mice

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Huajie Zhao ◽  
Yufei Lan ◽  
Hui Liu ◽  
Yongfa Zhu ◽  
Weiru Liu ◽  
...  
Keyword(s):  
Functional Foods ◽  
Reducing Power ◽  
Monosaccharide Composition ◽  
Vital Role ◽  
Hot Water ◽  
Composition Analysis ◽  
Laetiporus Sulphureus ◽  
Exploitation And Utilization ◽  
Natural Drugs

In order to contribute to the exploitation and utilization of spent mushroom substrates (SMS) of Laetiporus sulphureus, hot-water-extractable polysaccharides (H-SMPS) and enzymatic-extractable polysaccharides (E-SMPS) were successfully isolated from SMS of L. sulphureus. Both H-SMPS and E-SMPS were found to have high reducing power and potential scavenging activities against hydroxyl, DPPH, and superoxide anion radicals. In vivo assays showed that the administration of H-SMPS and E-SMPS has potential hepatoprotective effects against alcohol-induced alcoholic liver disease (ALD), possibly brought about by improving liver function, increasing antioxidant status, and reducing lipid peroxidation. Furthermore, monosaccharide composition analysis showed that fucose may play a vital role in guaranteeing stronger hepatoprotection. These results may provide references for the exploitation of the SMS of L. sulphureus as a source of H-SMPS and E-SMPS, which in turn can be used as functional foods or natural drugs for the prevention of ALD and other liver diseases.

scholarly journals Characterization of bovine κ-casein fractions and the kinetics of chymosin-induced macropeptide release from carbohydrate-free and carbohydrate-containing fractions determined by high-performance gel-permeation chromatography

1986 ◽  
Vol 240 (1) ◽  
pp. 87-97 ◽  
Author(s):  
H J Vreeman ◽  
S Visser ◽  
C J Slangen ◽  
J A Van Riel
Keyword(s):  
High Performance ◽  
Substrate Inhibition ◽  
Deae Cellulose ◽  
Free Fraction ◽  
Gel Permeation Chromatography ◽  
Reaction Products ◽  
Milk Clotting ◽  
Milk Clotting Enzyme ◽  
Gel Permeation

Bovine kappa-casein was fractionated at pH 8.0 on DEAE-Sepharose with an NaCl gradient, followed by DEAE-cellulose chromatography using a decreasing pH gradient from pH 6.0 to 4.5. At least ten components could be identified, each differing in N-acetylneuraminic acid (NeuAc) and/or phosphorus content. Two components appeared to be multiply-phosphorylated, but did not contain NeuAc. The possible significance of this finding in relation to the mode of phosphorylation and glycosylation in vivo is discussed. A carbohydrate-free fraction as well as two NeuAc-containing fractions were compared in their substrate behaviour towards the action of the milk-clotting enzyme chymosin at pH 6.6 and 30 degrees C. To this end the trichloroacetic acid-soluble reaction products were analysed by high-performance gel-permeation chromatography. In order of increasing carbohydrate content the kcat. values found ranged from 40 to 25 s-1 and the Km values from 9 to 3 microM; the overall substrate properties of these components as reflected by the kinetic parameter kcat./Km ranged from 5 to 8 microM-1 X S-1. Irreversible polymerization of the carbohydrate-free fraction brought about a more-than-2-fold increase in Km, the kcat. value remaining virtually constant. The kcat./Km found for the cleavage of whole kappa-casein at pH 6.6 was of the same magnitude as the kcat./Km found for the polymerized carbohydrate-free fraction (i.e. about 3 microM-1 X S-1). No indication of substrate inhibition was found for the carbohydrate-free fraction.

scholarly journals Mapping and Identification of the Major Cell Wall-Associated Components of Mycobacterium leprae

1998 ◽  
Vol 66 (6) ◽  
pp. 2625-2631 ◽  
Author(s):  
Maria Angela M. Marques ◽  
Sadhana Chitale ◽  
Patrick J. Brennan ◽  
Maria Cristina V. Pessolani
Keyword(s):  
Cell Wall ◽  
Response Regulator ◽  
Cell Envelope ◽  
Elongation Factor ◽  
Mycobacterium Leprae ◽  
Monosaccharide Composition ◽  
Mycolic Acid ◽  
Diaminopimelic Acid ◽  
Two Dimensional Gel Electrophoresis

ABSTRACT Mycobacterium leprae, an obligate intracellular pathogen, can be derived only from host tissue and thus affords the opportunity to study in vivo-expressed products responsible for the particular pathogenesis of leprosy. Despite considerable progress in the characterization of the proteins and secondary gene products ofM. leprae, there is little information on the nature of the proteins associated with the cell envelope. M. leprae has been fractionated into its major subcellular components, cell wall, cytoplasmic membrane, and soluble cytosol. A number of biochemical markers, including diaminopimelic acid content, monosaccharide composition, mycolic acid, and glycolipid distribution, were applied to their characterization, and two-dimensional gel electrophoresis was used to map the component proteins. A total of 391 major proteins spots were resolved, and 8 proteins were identified based on their reactivity to a panel of monoclonal antibodies and/or relative pI size. Microsequencing of six protein spots present in the cell wall fraction allowed identification of new proteins, including the protein elongation factor EF-Tu and a homolog for the Mycobacterium tuberculosis MtrA response regulator. These results, together with previous studies, contribute to the progressive knowledge of the composition of the in vivo-expressed proteins of M. leprae.

scholarly journals Homologous Over-Expression of Chain Length Determination Protein EpsC Increases the Molecular Weight of Exopolysaccharide in Streptococcus thermophilus 05-34

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhengyuan Zhai ◽  
Shuxin Xie ◽  
Hongxing Zhang ◽  
Huaxi Yi ◽  
Yanling Hao
Keyword(s):  
Molecular Weight ◽  
Chain Length ◽  
High Performance ◽  
Gel Permeation Chromatography ◽  
Streptococcus Thermophilus ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Gas Chromatography Mass Spectrometry ◽  
Over Expression ◽  
Length Determination

In Streptococcus thermophilus, EpsC is a polysaccharide co-polymerase which is involved in determining the chain length of EPS synthesized by the Wzx/Wzy-dependent pathway. Our previous study found that there was a positive correlation between transcription level of epsC and molecular weight of EPS in S. thermophilus 05-34. To further investigate the effects of EpsC on EPS biosynthesis, this gene was over-expressed in S. thermophilus 05-34 in this study. Reverse transcription qPCR and Western blotting confirmed the successful transcription and translation of epsC in 05-34, respectively. The yield of EPS was not affected by the over-expression of EpsC. Gas chromatography-mass spectrometry (GC-MS) showed that the monosaccharide composition was still composed of galactose and glucose in a molar ratio of 1.0:0.8, whereas high performance gel permeation chromatography (HPGPC) indicated that the molecular weight of EPS was increased from 4.62 × 105 Da to 9.17 × 105 Da by the over-expression of EpsC. In addition, S. thermophilus 05epsC which could produce higher molecular weight EPS improved the viscoelasticity and water-holding capacity of yogurt, but significantly reduced the level of syneresis in yogurt. In summary, these results indicated that homologous over-expression of EpsC in S. thermophilus could increase the molecular weight of EPS and improve the microrheological or physical properties of yogurt.

scholarly journals The Effect of Hemicellulose and Lignin on Properties of Polysaccharides in Lentinus edodes and Their Antioxidant Evaluation

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1834 ◽  
Author(s):  
Feifei Wu ◽  
Xin Jia ◽  
Lijun Yin ◽  
Yongqiang Cheng ◽  
Yuxin Miao ◽  
...  
Keyword(s):  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Monosaccharide Composition ◽  
Control Group ◽  
Composition Analysis ◽  
Lentinus Edodes ◽  
Polysaccharide Biosynthesis ◽  
C Elegans

Lentinus edodes, whose polysaccharides possess diverse bioactivities, commonly grows on hardwood sawdust composed of hemicellulose, lignin and cellulose. In this study the effect of hemicellulose and lignin on the growth of mycelia, as well as the physicochemical properties of polysaccharides from L. edodes mycelia (LEPs) were investigated. The antioxidant properties of LEPs were evaluated through radical scavenging assays in vitro and through the Caenorhabditis elegans model in vivo. The results showed that hemicellulose at a concentration of 4% increased the yield of the mycelia biomass to twice that of the control group. Meanwhile, when cultured with 4.0% hemicellulose, the polysaccharide content of the mycelia was raised by 112.2%. In addition, the appropriate concentration of lignin could stimulate mycelia growth and polysaccharide biosynthesis in L. edodes. Monosaccharide composition analysis showed that a higher content of xylose was found when mycelia were cultured with higher concentrations of hemicellulose. The molecular structure, including the molecular weight distribution and configuration type, was affected by hemicellulose and lignin. Antioxidant assays indicated that LEPs supplemented with hemicellulose and/or lignin possessed higher radical scavenging abilities in vitro and exhibited a thermal resistance effect on C. elegans, implying that the antioxidant effect is potent in vivo. In summary, the addition of hemicellulose and lignin improved the biosynthesis and bioactivity of LEPs.

scholarly journals Reduced 11β-Hydroxysteroid Dehydrogenase Activity in the Remaining Kidney Following Nephrectomy*

Endocrinology ◽  
1998 ◽  
Vol 139 (4) ◽  
pp. 1533-1539 ◽  
Author(s):  
Geneviève Escher ◽  
Bruno Vogt ◽  
Thomas Beck ◽  
Daniel Guntern ◽  
Brigitte M. Frey ◽  
...  
Keyword(s):  
Messenger Rna ◽  
High Performance ◽  
Reductase Activity ◽  
Kidney Tissue ◽  
Internal Standard ◽  
Hydroxysteroid Dehydrogenase ◽  
Mineralocorticoid Receptors ◽  
Reduction Activity ◽  
Convoluted Tubules

Abstract Intracellular access of steroids to gluco- and mineralocorticoid receptors is regulated by reduced 11β-hydroxysteroid dehydrogenase (OHSD) 1 and 2. These enzymes convert active 11β-OH-steroids into inactive 11-keto-steroids. The purpose of the present study was to establish whether the 11β-OHSD1 and 11β-OHSD2 are modulated in the remnant kidney 24 h or 14 days after uninephrectomy (UNX) in rats. Overall, 11β-OHSD activity was analyzed by measuring the ratio of the exogenous 11β-OH-steroid prednisolone to its 11-keto metabolite prednisone in vivo in kidney tissue using high performance liquid chromatography. To determine which isoenzyme accounts for the changed activity 24 h after UNX, the oxidation and reduction attributable to 11β-OHSD1 and oxidation to 11β-OHSD2 were analyzed in total renal extracts and in isolated glomeruli, proximal convoluted tubules (PCT), cortical ascending limbs, and cortical convoluted tubules (CCT). The messenger RNA content of 11β-OHSD1 and 11β-OHSD2 was measured by RT-PCR in renal tissues and single segments, using glyceraldehyde-3-phosphate-dehydrogenase as an internal standard. Protein amounts of 11β-OHSD1 and 11β-OHSD2 were assessed by Western blot. The prednisolone/prednisone ratio increased 24 h after UNX in 9 out of 10 animals (P ≤ 0.0011), and was unchanged 14 days after UNX. 11β-OHSD1 oxidation (P ≤ 0.032) and reduction activity (P ≤ 0.002) declined 24 h after UNX in total extracts. 11β-OHSD1 oxidase activity was more than 3 times higher in PCT than in glomeruli, cortical ascending limbs, and CCT, and declined by 50% after UNX (P ≤ 0.001). The reductase activity did not change following UNX in PCT. 11β-OHSD2 activity was 5–15 times higher in CCT than in the other segments, and decreased significantly after UNX (P ≤ 0.008). UNX did not affect messenger RNA and protein levels of both enzymes in total renal extracts. In conclusion, 11β-OHSD1 and 11β-OHSD2 are predominantly expressed in PCT and CCT, respectively, and their corresponding oxidative activities decline after UNX. Thus, the access of 11β-glucocorticosteroids to gluco- and mineralocorticoid receptors in the remaining kidney is facilitated after UNX.

scholarly journals Content and composition analysis of polysaccharides from Blaps rynchopetera and its macrophage phagocytic activity

2021 ◽  
Vol 19 (1) ◽  
pp. 338-346
Author(s):  
Ying Wang ◽  
Yin-He Yang ◽  
Qing Luo ◽  
Yuan Hu ◽  
Qian Lu ◽  
...  
Keyword(s):  
Phagocytic Activity ◽  
High Performance ◽  
Synergistic Effects ◽  
Folk Medicine ◽  
Total Content ◽  
Monosaccharide Composition ◽  
Peritoneal Macrophages ◽  
Composition Analysis ◽  
Phagocytic Ability ◽  
Mouse Peritoneal Macrophages

Abstract Blaps rynchopetera Fairmaire has a long history of use as a folk medicine in China for treating fever, cough, gastritis, boils, and tumors. In the present study, the content analyses, monosaccharide composition analyses, and the macrophage phagocytic activity of rynchopetera polysaccharides (RPS) were reported. B. rhynchoptera is rich in polysaccharides (content value 3.97%). Through PMP (1-phenyl-3-methyl-5-pyrazolone) pre-column derivatization and high performance liquid chromatography (HPLC) testing, the results showed that RPS consist of 8 known monosaccharides, including D-mannose (Man), Rhamnose (Rha), D-glucuronic acid (GlcUA), D-galacturonic acid (GalUA), D-glucose (Glc), D-galactose (Gal), Arabinose (Ara), and Fucose (Fuc), with the total content of 171.70 mg g−1 and Glc has the highest content of 45.40 mg g−1. The phagocytic ability of mouse peritoneal macrophages was investigated after RPS stimulating alone and combined with lipopolysaccharide (LPS). RPS played an important role in the engulfment of mouse peritoneal macrophages and can significantly enhance the phagocytic ability of macrophages. However, no synergistic effects were observed when RPS combined with LPS.

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