BACKGROUND: Obesity and related metabolic diseases are becoming a worldwide epidemic, leading to increased mortality and heavy medical costs. Our Chinese herbal formula Xiao-Gao-Jiang-Zhuo (XGJZ) has remarkable effects on curing obese patients in the clinic, but the cellular and molecular basis remains unknown. This study aimed to reveal the molecular mechanism involved in adipogenesis in vitro. METHODS: Chinese herbal formula XGJZ-containing serum was prepared from XGJZ-treated obesity model rats. The function of XGJZ-containing serum was validated in 3T3-L1 preadipocytes. Oil O staining was performed to determine intracellular lipid accumulation in differentiated 3T3-L1 cells. The expression of pro-adipogenic transcription factors was measured to further validate the adipogenesis of 3T3-L1 adipocytes. The contents of triglyceride (TG), free fatty acid (FFA), and glycerin, along with the activities of lipid metabolism-related enzymes (including FAT, FATP1, DGAT, GPAT, ATGL, and HSL) were measured to study the lipogenesis in 3T3-L1 adipocytes. RESULTS: XGJZ-containing serum inhibited 3T3-L1 differentiation, decreased intracellular lipid accumulation, and suppressed the expression of pro-adipogenic transcription factors in differentiated 3T3-L1 cells. The contents of TG, FFA, and glycerin were decreased when treated with XGJZ-containing serum, which also modulated lipid metabolism-related enzyme activities. The activities of fatty acid transporters (FAT, FATP1) and lipid mobilization enzymes (ATGL, HSL) were promoted, while activities of triglyceride biosynthesis enzymes (DGAT, GPAT) were attenuated in differentiated 3T3-L1 cells. CONCLUSION: XGJZ-containing serum has inhibitory effects on adipogenesis in 3T3-L1 preadipocytes, affirming the effect of XGJZ in treating obesity. It provides evidence for the mechanism of obesity.

• MeSH
• adipogeneze * MeSH
• buňky 3T3-L1 MeSH
• glycerol * metabolismus   farmakologie   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• obezita MeSH
• PPAR gama metabolismus   MeSH
• transkripční faktory MeSH
• triglyceridy MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Trypanosoma brucei is a causative agent of the Human and Animal African Trypanosomiases. The mammalian stage parasites infect various tissues and organs including the bloodstream, central nervous system, skin, adipose tissue and lungs. They rely on ATP produced in glycolysis, consuming large amounts of glucose, which is readily available in the mammalian host. In addition to glucose, glycerol can also be used as a source of carbon and ATP and as a substrate for gluconeogenesis. However, the physiological relevance of glycerol-fed gluconeogenesis for the mammalian-infective life cycle forms remains elusive. To demonstrate its (in)dispensability, first we must identify the enzyme(s) of the pathway. Loss of the canonical gluconeogenic enzyme, fructose-1,6-bisphosphatase, does not abolish the process hence at least one other enzyme must participate in gluconeogenesis in trypanosomes. Using a combination of CRISPR/Cas9 gene editing and RNA interference, we generated mutants for four enzymes potentially capable of contributing to gluconeogenesis: fructose-1,6-bisphoshatase, sedoheptulose-1,7-bisphosphatase, phosphofructokinase and transaldolase, alone or in various combinations. Metabolomic analyses revealed that flux through gluconeogenesis was maintained irrespective of which of these genes were lost. Our data render unlikely a previously hypothesised role of a reverse phosphofructokinase reaction in gluconeogenesis and preclude the participation of a novel biochemical pathway involving transaldolase in the process. The sustained metabolic flux in gluconeogenesis in our mutants, including a triple-null strain, indicates the presence of a unique enzyme participating in gluconeogenesis. Additionally, the data provide new insights into gluconeogenesis and the pentose phosphate pathway, and improve the current understanding of carbon metabolism of the mammalian-infective stages of T. brucei.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• fosfofruktokinasy metabolismus   MeSH
• glukoneogeneze * genetika   MeSH
• glukosa metabolismus   MeSH
• glycerol metabolismus   MeSH
• lidé MeSH
• savci MeSH
• transaldolasa metabolismus   MeSH
• Trypanosoma brucei brucei * genetika   metabolismus   MeSH
• uhlík metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Inadequate blood supply is one of the major risk factors for anastomotic leak after low anterior rectal resection. Early detection of ischemia might predict complicated healing and enable anastomotic salvage, which is associated with better outcomes. A microdialysis offers a real-time evaluation of adequate bowel perfusion through monitoring of tissue metabolism. In this experimental study, we assessed the role of microdialysis in detecting early ischemia after colorectal anastomosis. MATERIALS AND METHODS: Colorectal anastomosis was performed in six miniature pigs. A microdialysis catheter was placed on the bowel serosa 5 mm proximal to the anastomosis. Metabolic changes were monitored hourly before and after initiating ischemia, which was induced by ligation of the inferior mesenteric artery and skeletonization of the mesocolon. RESULTS: Significant increase in tissue levels of lactate was detected after 60 min of ischemia (13.6 [10.4-16.1] versus 6.75 [1.8-9.6] mmol/L at baseline; P < 0.005). The lactate/pyruvate ratio increased accordingly. The concentration of glycerol increased significantly after 2 hours-from a baseline value of 29.5 (3-84) to 125 (79-201) mmol/L (P < 0.005). The decrease in glucose levels was also significant after 60 minutes-0 (0-0.2) versus 4.7 (3.3-6.8) mmol/L at baseline. However, its values started to decline before ischemia. CONCLUSIONS: Surface microdialysis can detect ischemic changes early and may be a promising method in postoperative monitoring of colorectal anastomosis. Copyright © 2020 Elsevier Inc. All rights reserved.

• MeSH
• anastomóza chirurgická * škodlivé účinky   MeSH
• glycerol metabolismus   MeSH
• ischemie * MeSH
• krevní glukóza analýza   MeSH
• kyselina mléčná metabolismus   MeSH
• mikrodialýza * metody   MeSH
• miniaturní prasata MeSH
• pooperační komplikace * MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH

Strains of Limosilactobacillus reuteri are used as starter and bioprotective cultures and contribute to the preservation of food through the production of fermentation metabolites lactic and acetic acid, and of the antimicrobial reuterin. Reuterin consists of acrolein and 3-hydroxypropionaldehyde (3-HPA), which can be further metabolized to 1,3-propanediol and 3-hydroxypropionic acid (3-HP). While reuterin has been the focus of many investigations, the contribution of 3-HP to the antimicrobial activity of food related reuterin-producers is unknown. We show that the antibacterial activity of 3-HP was stronger at pH 4.8 compared to pH 5.5 and 6.6. Gram-positive bacteria were in general more resistant against 3-HP and propionic acid than Gram-negative indicator strains including common food pathogens, while spoilage yeast and molds were not inhibited by ≤ 640 mM 3-HP. The presence of acrolein decreased the minimal inhibitory activity of 3-HP against E. coli indicating synergistic antibacterial activity. 3-HP was formed during the growth of the reuterin-producers, and by resting cells of L. reuteri DSM 20016. Taken together, this study shows that food-related reuterin producers strains synthesize a second antibacterial compound, which might be of relevance when strains are added as starter or bioprotective cultures to food products.

• MeSH
• antiinfekční látky chemie   metabolismus   farmakologie   MeSH
• Bacteria účinky léků   růst a vývoj   MeSH
• fermentace MeSH
• glyceraldehyd analogy a deriváty   chemie   metabolismus   MeSH
• glycerol metabolismus   MeSH
• koncentrace vodíkových iontů MeSH
• kyselina mléčná analogy a deriváty   chemie   metabolismus   farmakologie   MeSH
• kyselina octová metabolismus   MeSH
• Lactobacillaceae chemie   růst a vývoj   metabolismus   MeSH
• potravinářská mikrobiologie MeSH
• propan chemie   metabolismus   MeSH
• stabilita léku MeSH
• Publikační typ
• časopisecké články MeSH

Saccharomyces kudriavzevii is a nonconventional and rather osmosensitive yeast with a high potential of use in fermentation processes. To elucidate the basis of its relative osmosensitivity, the role of the STL1 gene encoding a putative glycerol uptake system was studied. Under higher osmotic pressure, the addition of a low amount of glycerol to the growth medium improved the growth of S. kudriavzevii and the expression of the STL1 gene was highly induced. Deletion of this gene decreased the strain's ability to grow in the presence of higher concentrations of salts and other solutes. Moreover, the mutant had a disturbed homeostasis of intracellular pH. Expression of the SkSTL1 gene in Saccharomyces cerevisiae complemented the osmosensitivity of the S. cerevisiae hog1Δ stl1Δ mutant, and the gene's tagging with GFP localized its product to the plasma membrane. Altogether, a deficiency in glycerol uptake did not seem to be the reason for S. kudriavzevii's low osmotolerance; its Stl1 transporter properly contributes to the regulation of intracellular pH and is crucial to its survival of osmotic stress. SIGNIFICANCE AND IMPACT OF THE STUDY: An increasing demand for food products with benefits for human health turns the attention to less-exploited nonconventional yeasts with interesting traits not found in Saccharomyces cerevisiae. Among them, Saccharomyces kudriavzevii has good potential for aroma-compound production, fermentations and other biotechnological applications, but it is less adapted to stressful industrial conditions. This report studied S. kudriavzevii relative osmosensitivity and its capacity for active glycerol uptake. The results obtained (on the activity and physiological function of S. kudriavzevii glycerol transporter) may contribute to a further engineering of this species aiming to improve its osmotolerance.

• MeSH
• biologický transport genetika   fyziologie   MeSH
• buněčná membrána metabolismus   MeSH
• delece genu MeSH
• fermentace MeSH
• fyziologický stres fyziologie   MeSH
• glycerol metabolismus   MeSH
• membránové transportní proteiny genetika   MeSH
• mitogenem aktivované proteinkinasy genetika   MeSH
• osmotický tlak fyziologie   MeSH
• Saccharomyces cerevisiae - proteiny genetika   MeSH
• Saccharomyces cerevisiae klasifikace   genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

The aim of the study was to screen Yarrowia lipolytica strains for keto acid production and determine optimal conditions for pyruvic acid biosynthesis from glycerol by the best producer. The analyzed parameters were thiamine concentration, medium pH, stirring speed, and substrate concentration. The screening was performed in flask cultures, whereas pyruvic acid production was carried out in 5-L stirred-tank reactor with 2 L of working volume. In total, 24 Y. lipolytica strains were compared for their abilities to produce pyruvic and α-ketoglutaric acids. The total concentration of both acids ranged from 0.1 to 15.03 g/L. Ten strains were selected for keto acid biosynthesis in bioreactor. The Y. lipolytica SKO 6 strain was identified as the best producer of pyruvic acid. In the selected conditions (thiamine concentration 1.5 μg/L, pH 4.0, stirring speed 800 rpm, 150 g/L of glycerol), the strain Y. lipolytica SKO 6 produced 99.3 g/L of pyruvic acid, with process yield of 0.63 g/g and volumetric production rate of 1.18 g/L/h. Higher titer of pyruvic acid was obtained during fed-batch culture with 200 g/L of glycerol, reaching 125.8 g/L from pure glycerol (yield 0.68 g/g) and 124.4 g/L from crude glycerol (yield 0.62 g/g). Results obtained for the strain Y. lipolytica SKO 6 proved the suitability of microbial production of pyruvic acid at industrial scale.

• MeSH
• bioreaktory MeSH
• glycerol analýza   metabolismus   MeSH
• kultivační média chemie   MeSH
• kyselina pyrohroznová analýza   metabolismus   MeSH
• kyseliny ketoglutarové analýza   metabolismus   MeSH
• techniky vsádkové kultivace MeSH
• thiamin analýza   MeSH
• Yarrowia růst a vývoj   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

Dekkera bruxellensis is important for lambic beer fermentation but is considered a spoilage yeast in wine fermentation. We compared two D. bruxellensis strains isolated from wine and found that they differ in some basic properties, including osmotolerance. The genomes of both strains contain two highly similar copies of genes encoding putative glycerol-proton symporters from the STL family that are important for yeast osmotolerance. Cloning of the two DbSTL genes and their expression in suitable osmosensitive Saccharomyces cerevisiae mutants revealed that both identified genes encode functional glycerol uptake systems, but only DbStl2 has the capacity to improve the osmotolerance of S. cerevisiae cells.

• MeSH
• Dekkera genetika   izolace a purifikace   metabolismus   fyziologie   MeSH
• druhová specificita MeSH
• fungální proteiny genetika   metabolismus   MeSH
• genom bakteriální genetika   MeSH
• glycerol metabolismus   MeSH
• osmoregulace genetika   MeSH
• protony MeSH
• rekombinantní proteiny genetika   metabolismus   MeSH
• Saccharomyces cerevisiae genetika   MeSH
• symportéry genetika   metabolismus   MeSH
• testy genetické komplementace MeSH
• víno mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

S. kudriavzevii has potential for fermentations and other biotechnological applications, but is sensitive to many types of stress. We tried to increase its tolerance and performance via the expression of various transporters from different yeast species. Whereas the overexpression of Z. rouxii fructose uptake systems (ZrFfz1 and ZrFsy1) or a glycerol importer (ZrStl1) did not improve the ability of S. kudriavzevii to consume fructose and survive osmotic stress, the expression of alkali-metal-cation exporters (ScEna1, ScNha1, YlNha2) improved S. kudriavzevii salt tolerance, and that of ScNha1 also the fermentation performance. The level of improvement depended on the type and activity of the transporter suggesting that the natural sensitivity of S. kudriavzevii cells to salts is based on a non-optimal functioning of its own transporters.

• MeSH
• fermentace MeSH
• fruktosa metabolismus   MeSH
• glycerol metabolismus   MeSH
• iontový transport fyziologie   MeSH
• osmotický tlak fyziologie   MeSH
• proteiny přenášející kationty genetika   metabolismus   MeSH
• Saccharomyces metabolismus   MeSH
• sušené kvasnice MeSH
• tolerance k soli fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH

CONTEXT: Mucoadhesive oral films, with their prolonged residence time at the site of application, offer a promising approach for protection of the oral lesion surface. The addition of sodium hyaluronate of different molecular weights as a second mucoadhesive polymer into the film matrix could positively influence the physico-mechanical and mucoadhesive properties of films. OBJECTIVE: The aim of this study was to investigate the formulation of a monolayered film matrix containing varying amounts of sodium hyaluronate and to test the properties of such matrices by applying different characterization methods. MATERIALS AND METHODS: Film matrix was composed of two mucoadhesive polymers, carmellose sodium and sodium hyaluronate, plasticized with glycerol. Resulting films were characterized with regard to their viscosity and physico-mechanical properties. RESULTS AND DISCUSSION: Multivariate data analysis was employed to evaluate the influence of varying amounts of mucoadhesive polymers on the main mucoadhesive oral films' properties. The lower content of sodium hyaluronate caused improvements in mechanical properties and residence time on the artificial oral mucosa, both of which are the main characteristics that determine the quality of the final product. CONCLUSIONS: The best results were obtained by samples containing carmellose sodium with a small amount of sodium hyaluronate (about 0.5% in casting dispersion).

• MeSH
• adheziva chemie   metabolismus   MeSH
• adhezivita MeSH
• glycerol chemie   metabolismus   MeSH
• kyselina hyaluronová chemie   metabolismus   MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• multivariační analýza MeSH
• orální absorpce MeSH
• pevnost v tahu MeSH
• polymery MeSH
• sodná sůl karboxymethylcelulosy chemie   metabolismus   MeSH
• ústní sliznice metabolismus   MeSH
• uvolňování léčiv MeSH
• viskozita MeSH
• změkčovadla chemie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

As Pichia pastoris (syn. Komagataella sp.) yeast can secrete pure recombinant proteins at high rates, it is a desirable production system. The function of a novel synthetic variant of the AOX1 promoter was characterised comprehensively using a strain secreting Candida antarctica lipase B (CALB) as a model. A new time-saving approach was introduced to determine, in only one experiment, the hitherto unknown relationship between specific product formation rate (q p) and specific growth rate (μ). Tight control of recombinant protein formation was possible in the absence of methanol, while using glycerol as a sole carbon/energy source. CALB was not synthesised during batch cultivation in excess glycerol (>10 g l(-1)) and at a growth rate close to μ max (0.15 h(-1)). Between 0.017 and 0.115 h(-1) in glycerol-limited fedbatch cultures, basal levels of q p > 0.4 mg g(-1) h(-1) CALB were reached, independent of the μ at which the culture grew. At μ > 0.04 h(-1), an elevated q p occurred temporarily during the first 20 h after changing to fedbatch mode and decreased thereafter to basal. In order to accelerate the determination of the q p(μ) relationship (kinetics of product formation), the entire μ range was covered in a single fedbatch experiment. By linearly increasing and decreasing glycerol addition rates, μ values were repeatedly shifted from 0.004 to 0.074 h(-1) and vice versa. Changes in q p were related to changes in μ. A rough estimation of μ range suitable for production was possible in a single fedbatch, thus significantly reducing the experimental input over previous approaches comprising several experiments.

• MeSH
• fermentace MeSH
• fungální proteiny genetika   metabolismus   MeSH
• glycerol metabolismus   farmakologie   MeSH
• kinetika MeSH
• lipasa genetika   metabolismus   MeSH
• methanol analýza   MeSH
• Pichia genetika   růst a vývoj   metabolismus   MeSH
• promotorové oblasti (genetika) * MeSH
• rekombinantní proteiny metabolismus   MeSH
• techniky vsádkové kultivace MeSH
• Publikační typ
• časopisecké články MeSH