ABSTRACT
The effect of paraprobiotic Lacticaseibacillus casei 01 inactivated by ohmic heating (8 V/cm, 95 °C/7 min, 60 Hz) whey-grape juice drink at the postprandial glycemia was evaluated. In vitro hypoglycemic activity was assessed by the α-glucosidase and α-amylase inhibition, while in vivo activity was determined using 15 healthy subjects, which consumed bread + probiotic whey drink, bread + paraprobiotic whey drink, and bread alone as a control. The probiotic and paraprobiotic grape-flavored whey drinks showed similar α-glucosidase and α-amylase inhibition (51.2 vs 51.8% and 43.2 vs 44.2%, respectively). The consumption of both paraprobiotic and probiotic whey drinks increased the incremental glucose rate when compared to the control due to the presence of sugar in its composition, without changes in the other parameters evaluated (maximum glucose value, glucose incremental percentage, and peak blood glucose time), showing a reduced glycemic response. In addition, the consumption of the paraprobiotic drink maintained the maximum glucose increase similar to the control, while an increase in this parameter was observed after the consumption of the probiotic drink. Therefore, the paraprobiotic grape-flavored whey drink may be an effective alternative to replace the probiotic product in reducing the postprandial glycemia in healthy individuals.
Subject(s)
Vitis , Adult , Blood Glucose , Heating , Humans , Postprandial Period , WheyABSTRACT
In this work, we demonstrate that Trypanosoma cruzi Y strain epimastigotes exhibit Mg2+-dependent ecto-ATPase activity that is stimulated by heat shock. When the epimastigotes were incubated at 37°C for 2h, the ecto-ATPase activity of the cells was 43.95±0.97 nmol Pi/h×10(7) cells, whereas the ecto-ATPase activity of cells that were not exposed to heat shock stress was 16.97±0.30 nmol Pi/h×10(7) cells. The ecto-ATPase activities of cells, that were exposed or not exposed to heat shock stress had approximately the same Km values (2.25±0.26 mM ATP and 1.55±0.23 mM ATP, respectively) and different Vmax values. The heat-shocked cells had higher Vmax values (54.38±3.07 nmol Pi/h×10(7) cells) than the cells that were not exposed to heat shock (19.38±1.76 nmol Pi/h×10(7) cells). We also observed that the ecto-phosphatase and ecto-5'nucleotidase activities of cells that had been incubated at 28°C or 37°C were the same. Interestingly, cycloheximide, an inhibitor of protein synthesis, suppressed the heat shock effect of ecto-ATPase activity on T. cruzi. The Mg2+-dependent ecto-ATPase activity from the Y strain (high virulence) was approximately 2-fold higher than that of Dm28c (a clone with low virulence). In addition, these two strains presented different responses to heat shock with regard to their ecto-ATPase activities; Y strain epimastigotes had a stimulation of 2.52-fold while the Dm28c strain had a 1.71-fold stimulation. In this context, the virulent trypomastigote form of T. cruzi, Dm28c, had an ecto-ATPase activity that was more than 7-fold higher (66.67±5.98 nmol Pi/h×10(7) cells) than that of the insect epimastigote forms (8.91±0.76 nmol Pi/h×10(7) cells). This difference increased to approximately 10-fold when both forms were subjected to heat shock stress (181.14±16.48 nmol Pi/h×10(7) cells for trypomastigotes and 16.71±1.17 nmol Pi/h×10(7) cells for epimastigotes at 37°C). The ecto-ATPase activity of a plasma membrane-enriched fraction obtained from T. cruzi epimastigotes was not increased by heat treatment, which suggested that cytoplasmic components had an influence on enzyme activation by heat shock stress.
Subject(s)
Adenosine Triphosphatases/metabolism , Ca(2+) Mg(2+)-ATPase/metabolism , Heat-Shock Response/physiology , Hot Temperature/adverse effects , Trypanosoma cruzi/enzymology , 5'-Nucleotidase/genetics , 5'-Nucleotidase/metabolism , Adenosine Triphosphatases/genetics , Adenosine Triphosphate/metabolism , Blotting, Western , Ca(2+) Mg(2+)-ATPase/genetics , Cell Membrane/enzymology , Cycloheximide/pharmacology , Gene Expression Regulation, Enzymologic , Heat-Shock Response/drug effects , Humans , Hydrolysis , Protein Synthesis Inhibitors/pharmacology , Time Factors , Trypanosoma cruzi/drug effects , Trypanosoma cruzi/geneticsABSTRACT
There is increasing evidence in the literature showing that fungal pathogens express biologically active ectoenzymes. The expression of surface phosphatases at the cell surface of Cryptococcus neoformans, the etiologic agent of cryptococcosis, was evaluated in the present study. Different isolates of C. neoformans express ectophosphatase activity, which is not influenced by capsule size or serotype. The cryptococcal enzyme is an acid phosphatase, inhibited by classic inhibitors of ectophosphatases, including ammonium molybdate and sodium salts of fluoride and orthovanadate. Only the inhibition of enzyme activity caused by sodium orthovanadate has been shown to be irreversible. The cryptococcal ectoenzyme is also inhibited by Zn2+ and inorganic phosphate, the final product of reactions catalyzed by phosphatases. The ectophosphatase from C. neoformans efficiently releases phosphate groups from different phosphorylated amino acids, giving a higher rate of phosphate removal when phosphothreonine is used as a substrate. Yeast cells with irreversibly inhibited ectophosphatases are less capable of adhering to animal epithelial cells than fungi fully expressing enzyme activity, suggesting that ectoenzyme expression can contribute to the pathogenesis of C. neoformans.
