scholarly journals Highly Efficient Biosynthesis of Glycyrrhetinic Acid Glucosides by Coupling of Microbial Glycosyltransferase to Plant Sucrose Synthase

2021 ◽  
Vol 9 ◽  
Author(s):  
Mohamed Yassin Ali ◽  
Qing Chang ◽  
Quande Yan ◽  
Zheng Qian ◽  
Xiang Guo ◽  
...  
Keyword(s):  
Sucrose Synthase ◽  
Large Scale ◽  
Water Solubility ◽  
Human Colon ◽  
Glycyrrhetinic Acid ◽  
Uridine Diphosphate ◽  
Pentacyclic Triterpenoid ◽  
Solubility Improvement ◽  
Therapeutic Compounds ◽  
Dose Dependent

Glycyrrhetinic acid (GA) is a principal bioactive pentacyclic triterpenoid from Glycyrrhiza uralensis. Uridine diphosphate-dependent glycosyltransferases (UGTs) have been widely used to catalyze glycosylation of diverse nature products for the development of potential therapeutic compounds. In this study, we have characterized a UGT109A3 from Bacillus subtilis, which can glycosylate both the free C3 hydroxyl and C30 carboxyl groups of GA to yield a unique 3, 30-O-β-D-diglucoside-GA. By coupling the microbial UGT109A3 to plant sucrose synthase (SUS), GA-diglucoside could be biosynthesized in an efficient and economical way. With a fed-batch glycosylation, a large scale of GA-diglucoside (6.26 mM, 4.98 g/L in 8 h) could be enzymatically transformed from GA. The obtained GA-diglucoside showed a significant water solubility improvement of around 3.4 × 103 fold compared with that of the parent GA (29 μM). Moreover, it also exhibited dose-dependent cytotoxicity toward human colon carcinoma Caco-2 cell line according to MTT assay, having an IC50 at 160 μM. This study not only establishes efficient platform for producing GA-glucosides, but is also valuable for developing further the biosynthesis of other complex glycosylated natural products.

scholarly journals An engineered IL-2 reprogrammed for anti-tumor therapy using a semi-synthetic organism

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jerod L. Ptacin ◽  
Carolina E. Caffaro ◽  
Lina Ma ◽  
Kristine M. San Jose Gall ◽  
Hans R. Aerni ◽  
...  
Keyword(s):  
Nk Cells ◽  
Large Scale ◽  
Tumor Therapy ◽  
Covalent Modification ◽  
Synthetic Dna ◽  
Expansion Characteristic ◽  
Synthetic Organisms ◽  
Dose Dependent ◽  
Receptor Beta

AbstractThe implementation of applied engineering principles to create synthetic biological systems promises to revolutionize medicine, but application of fundamentally redesigned organisms has thus far not impacted practical drug development. Here we utilize an engineered microbial organism with a six-letter semi-synthetic DNA code to generate a library of site-specific, click chemistry compatible amino acid substitutions in the human cytokine IL-2. Targeted covalent modification of IL-2 variants with PEG polymers and screening identifies compounds with distinct IL-2 receptor specificities and improved pharmacological properties. One variant, termed THOR-707, selectively engages the IL-2 receptor beta/gamma complex without engagement of the IL-2 receptor alpha. In mice, administration of THOR-707 results in large-scale activation and amplification of CD8+ T cells and NK cells, without Treg expansion characteristic of IL-2. In syngeneic B16-F10 tumor-bearing mice, THOR-707 enhances drug accumulation in the tumor tissue, stimulates tumor-infiltrating CD8+ T and NK cells, and leads to a dose-dependent reduction of tumor growth. These results support further characterization of the immune modulatory, anti-tumor properties of THOR-707 and represent a fundamental advance in the application of synthetic biology to medicine, leveraging engineered semi-synthetic organisms as cellular factories to facilitate discovery and production of differentiated classes of chemically modified biologics.

scholarly journals Effects of D-allulose on glucose tolerance and insulin response to a standard oral sucrose load: results of a prospective, randomized, crossover study

2021 ◽  
Vol 9 (1) ◽  
pp. e001939
Author(s):  
Francesco Franchi ◽  
Dmitry M Yaranov ◽  
Fabiana Rollini ◽  
Andrea Rivas ◽  
Jose Rivas Rios ◽  
...  
Keyword(s):  
Crossover Study ◽  
Plasma Glucose ◽  
Large Scale ◽  
Insulin Response ◽  
Mean Difference ◽  
Dietary Guidelines ◽  
Sugar Intake ◽  
Time Points ◽  
Insulin Levels ◽  
Dose Dependent

IntroductionCurrent dietary guidelines recommend limiting sugar intake for the prevention of diabetes mellitus (DM). Reduction in sugar intake may require sugar substitutes. Among these, D-allulose is a non-calorie rare monosaccharide with 70% sweetness of sucrose, which has shown anti-DM effects in Asian populations. However, there is limited data on the effects of D-allulose in other populations, including Westerners.Research design and methodsThis was a prospective, randomized, double-blind, placebo-controlled, crossover study conducted in 30 subjects without DM. Study participants were given a standard oral (50 g) sucrose load and randomized to placebo or escalating doses of D-allulose (2.5, 5.0, 7.5, 10.0 g). Subjects crossed-over to the alternate study treatment after 7–14 days of wash out. Plasma glucose and insulin levels were measured at five time points: before and at 30, 60, 90 and 120 min after ingestion.ResultsD-allulose was associated with a dose-dependent reduction of plasma glucose at 30 min compared with placebo. In particular, glucose was significantly lower with the 7.5 g (mean difference: 11; 95% CI 3 to 19; p=0.005) and 10 g (mean difference: 12; 95% CI 4 to 20; p=0.002) doses. Although glucose was not reduced at the other time points, there was a dose-dependent reduction in glucose excursion compared with placebo, which was significant with the 10 g dose (p=0.023). Accordingly, at 30 min D-allulose was associated with a trend towards lower insulin levels compared with placebo, which was significant with the 10 g dose (mean difference: 14; 95% CI 4 to 25; p=0.006). D-allulose did not reduce insulin at any other time point, but there was a significant dose-dependent reduction in insulin excursion compared with placebo (p=0.028), which was significant with the 10 g dose (p=0.002).ConclusionsThis is the largest study assessing the effects of D-allulose in Westerners demonstrating an early dose-dependent reduction in plasma glucose and insulin levels as well as decreased postprandial glucose and insulin excursion in subjects without DM. These pilot observations set the basis for large-scale investigations to support the anti-DM effects of D-allulose.Trial registration numberNCT02714413.

scholarly journals New inhibitors of glucosylceramide synthase and their effect on cell fate

2014 ◽  
Vol 7 (2) ◽  
pp. 99-104 ◽  
Author(s):  
Katarína Turáková ◽  
Boris Lakatoš ◽  
Andrej Ďuriš ◽  
Daniela Moravčíková ◽  
Dušan Berkeš
Keyword(s):  
Cell Viability ◽  
Cell Fate ◽  
Calcium Transport ◽  
Large Scale ◽  
Process Development ◽  
Pharmaceutical Research ◽  
Pathological Process ◽  
Uridine Diphosphate ◽  
Induction Of Apoptosis

Abstract Glucosylceramide (GlcCer) is an essential glycosylated lipid found in organisms ranging from fungi to mammals. It is composed of a hydrophilic β-linked glucose and a hydrophobic ceramide, with a predominant content of sphingosine in mammals (d18:1). GlcCer is the precursor of a large scale of different glycosphingolipids. This cerebrozide is synthesized from uridine diphosphate-glucose and ceramide by a GlcCer synthase (UDP-glucose:ceramide glucosyltransferase; UGCG, EC 2.4.1.80). GlcCer-based sphingolipids have been identified as important mediators of a variety of cellular functions and their disequilibrium leads to pathological process development and may induce several diseases progression. Therefore, design of UGCG inhibitor represents an important topic for pharmaceutical research. In this paper, we aimed to study effects of newly synthesized derivatives of (±)-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol (PPMP, known UGCG inhibitor) on: i) activity of UGCG in vitro; ii) thymocytes viability; iii) calcium transport through plasma membrane of thymocytes; iv) induction of apoptosis and autophagy in thymocytes. Thymocytes were isolated from thymus of three to seven weeks old mice (ICR strain). The key factors influencing the effect of PPMP analogues were their concentration, chemical structure and incubation time. Derivatives were able to change Ca2+ transport already after 15 min of cultivation, but their effects on cell viability were manifested at least after 12 h of cultivation. Four from fifteen studied compounds affected UGCG activity after four hour lasting cultivation, - but without correlation with data relating to effects on calcium transport and/or cell viability. Most potent UGCG inhibitor was chosen and applied for induction of apoptosis and autophagy in thymocytes. This inhibitor induced typical DNA fragmentation and upregulation of LC3B protein as autophagy marker, after 2 h and 4 h cultivation, respectively.

scholarly journals Association of Mesothelioma Deaths With Cumulated Neighborhood Exposures Due to a Large-Scale Asbestos Cement Plant in Amagasaki City, Japan: A Nested Case-control Study

2021 ◽  
Author(s):  
Yuri Kitamura ◽  
Ling Zha ◽  
Rong Liu ◽  
Masayuki Shima ◽  
Tomoki Nakaya ◽  
...  
Keyword(s):  
Occupational Exposure ◽  
Large Scale ◽  
Case Control Study ◽  
Conditional Logistic Regression ◽  
Asbestos Exposure ◽  
Case Control ◽  
Asbestos Cement ◽  
Nested Case Control ◽  
Dose Dependent ◽  
Control Study

Abstract BackgroundAlthough a causal relationship between mesothelioma and asbestos exposure is well known, few studies have shown a relationship to non-occupational exposure, including neighborhood exposure, most likely because of the large effect size of occupational exposure. The aim of this study was to quantify the risk of malignant mesothelioma death associated with neighborhood asbestos exposure due to a large-scale asbestos-cement (AC) plant in Amagasaki, Japan, by properly adjusting for occupational exposure. MethodsThis was a nested case-control study in which a fixed population of 143,929 residents who had been living in Amagasaki City between 1975 and 2002 were followed from 2002 to 2015. All 133 cases and 403 matched controls were interviewed about their occupational, domestic, household, and neighborhood asbestos exposures. Odds ratios (ORs) for mesothelioma death associated with neighborhood exposure were estimated by a conditional logistic-regression model that adjusted for other asbestos exposures. We adopted cumulative indices that considered residence-specific asbestos (crocidolite) concentrations and durations during the potential exposure period of 1957-1975 to evaluate individual neighborhood exposures.ResultsThere was an increasing, dose-dependent risk of mesothelioma death associated with neighborhood exposure, demonstrated by ORs in the highest quintile category that were 21.4 (95%CI: 5.8 to 79.2) for all, 23.7 (95% CI: 3.8-147.2) for males and 26.0 (95% CI: 2.8-237.5) for females, compared to the lowest quintile, respectively. These results clearly demonstrated no substantial differences between males and females in relation to the magnitude of risk from neighborhood exposure.Our findings suggest that the risk of mesothelioma death associated with neighborhood exposure persists and will not be diminished for many years, even though it has been decades since the AC plant closed. ConclusionsBy adjusting for occupational and other asbestos exposures, a dose-dependent relationship was demonstrated between mesothelioma death and neighborhood asbestos exposure from a large-scale AC plant.

scholarly journals Considerable Improvement of Ursolic Acid Water Solubility by Its Encapsulation in Dendrimer Nanoparticles: Design, Synthesis and Physicochemical Characterization

Nanomaterials ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2196 ◽  
Author(s):  
Silvana Alfei ◽  
Anna Maria Schito ◽  
Guendalina Zuccari
Keyword(s):  
Ursolic Acid ◽  
Water Solubility ◽  
Drug Loading ◽  
Physicochemical Characterization ◽  
Systemic Toxicity ◽  
Water Soluble ◽  
Design Synthesis ◽  
Pentacyclic Triterpenoid

Ursolic acid (UA) is a pentacyclic triterpenoid found in many medicinal plants and aromas endowed with numerous in vitro pharmacological activities, including antibacterial effects. Unfortunately, UA is poorly administered in vivo, due to its water insolubility, low bioavailability, and residual systemic toxicity, thus making urgent the development of water-soluble UA formulations. Dendrimers are nonpareil macromolecules possessing highly controlled size, shape, and architecture. In dendrimers with cationic surface, the contemporary presence of inner cavities and of hydrophilic peripheral functions, allows to encapsulate hydrophobic non-water-soluble drugs as UA, to enhance their water-solubility and stability, and to promote their protracted release, thus decreasing their systemic toxicity. In this paper, aiming at developing a new UA-based antibacterial agent administrable in vivo, we reported the physical entrapment of UA in a biodegradable not cytotoxic cationic dendrimer (G4K). UA-loaded dendrimer nanoparticles (UA-G4K) were obtained, which showed a drug loading (DL%) much higher than those previously reported, a protracted release profile governed by diffusion mechanisms, and no cytotoxicity. Also, UA-G4K was characterized by principal components analysis (PCA)-processed FTIR spectroscopy, by NMR and elemental analyses, and by dynamic light scattering experiments (DLS). The water solubility of UA-G4K was found to be 1868-fold times higher than that of pristine UA, thus making its clinical application feasible.

Loading AKBA on surface of silver nanoparticles to improve their sedative-hypnotic and anti-inflammatory efficacies

Nanomedicine ◽  
2019 ◽  
Vol 14 (21) ◽  
pp. 2783-2798
Author(s):  
Ajmal Khan ◽  
Ahmed Al-Harrasi ◽  
Najeeb Ur Rehman ◽  
Rizwana Sarwar ◽  
Touqeer Ahmad ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Silver Nanoparticles ◽  
Water Solubility ◽  
Boswellic Acid ◽  
Transmission Electron ◽  
Anti Inflammatory ◽  
Compound Materials ◽  
Dose Dependent ◽  
Scanning Electron

Aim: Acetyl-11-keto- β-boswellic acid (AKBA) is a potent anti-inflammatory compound limited by its low water solubility and bioavailability. To load AKBA on silver nanoparticles (AgNPs) to improve bioavailability and water solubility of the compound. Materials & methods: AKBA-AgNPs were chemically synthesized and characterized by UV–Vis spectrophotometry, Fourier transform infrared spectroscopy, scanning electron microscopy and transmission electron microscopy. AKBA and AKBA-Ag were studied for their sedative-hypnotic and anti-inflammatory efficacies. Results: Pretreatment with AKBA or AKBA-Ag caused significant dose-dependent sedative-hypnotic effects at 5 and 10 mg/kg intraperitoneal. The effects of AKBA-loaded AgNPs caused pronounced changes in mice compared with those of AKBA, and the AKBA-AgNPs demonstrated anti-inflammatory effects that were superior to those of AKBA. Conclusion: The loading of AKBA on nanoparticles improved its pharmacokinetic effects, and capacity for drug delivery.

Rapid isolation, reliable characterization, and water solubility improvement of polymethoxyflavones from cold-pressed mandarin essential oil

2016 ◽  
Vol 39 (11) ◽  
pp. 2018-2027 ◽  
Author(s):  
Marina Russo ◽  
Francesca Rigano ◽  
Adriana Arigò ◽  
Danilo Sciarrone ◽  
Maria Luisa Calabrò ◽  
...  
Keyword(s):  
Essential Oil ◽  
Water Solubility ◽  
Rapid Isolation ◽  
Solubility Improvement ◽  
Cold Pressed
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