Β-glucuronidase Activity Research Articles

Genomic-, phenotypic-, and toxicity-based safety assessment and probiotic potency of Bacillus coagulans IDCC 1201 isolated from green malt

Probiotics are beneficial microorganisms, and the evaluation of their safety for human use in the food industry has become critical. This study examines the safety of Bacillus coagulans IDCC 1201 isolated from green malt by analyzing its genomic and phenotypic characteristics and determining its toxicity. The presence of antibiotic resistance and toxigenic genes and gene transferability were investigated using whole-genome analysis. The strain's hemolytic and enzyme activities, minimum inhibitory concentrations of antibiotics, and biogenic amine and D-lactate production were also examined. Furthermore, the principal properties of B. coagulans IDCC 1201 as probiotics, such as resistance to abiotic stress and intestinal adhesion, were studied. The whole-genome analysis demonstrated that B. coagulans IDCC 1201 had no antibiotic resistance or toxigenic genes; the strain was susceptible to the nine antibiotics proposed by the European Food Safety Authority. Moreover, this strain lacked hemolytic and β-glucuronidase activities. Additionally, it was confirmed that B. coagulans IDCC 1201 produced undesirable metabolites, including biogenic amines or D-lactate, at a safe level. Finally, the strain exhibited functional potential as a probiotic in terms of abiotic tolerance, such as bile tolerance and intestinal adhesion in in vitro experiments. In conclusion, B. coagulans IDCC 1201 can be considered as a safe probiotic with regard to human health.

Influence of Supplementation of Lactoferrin, Melittin and Cecropin A to Rat Diet on Changes in Faecal Ammonia Concentrations, Short-Chain Fatty Acid Concentrations and Activities of Bacterial Enzymes.

Simple SummaryIn the present study conducted on Wistar laboratory rats, the effects of two selected insect antimicrobial peptides (AMPs), melittin and cecropin A, were investigated and compared to those attributed to well-known antibacterial action of lactoferrin. It was hypothesised that the dietary presence of lactoferrin, melittin or cecropin A strongly affects the rat large gut microbial activity at the time of protein/peptide administration and the durability of the effects may differ after their withdrawal from a diet. The experiment was conducted on living animals (without their euthanasia) and the dynamics of changes in pH, microbial enzyme activity, ammonia and short-chain fatty acids concentrations were investigated in the faeces during and after the dietary treatments with lactoferrin, melittin or cecropin A. The results suggested that the faecal intensity of microbial fermentation processes in rats was quickly reduced upon dietary addition of two AMPs and lactoferrin after two days of treatment, on average. The strongest suppression effect was observed on the 5th day of treatment and persisted on days 5–8. The changes caused by the supplemented lactoferrin and AMPs were reversible after 15 days, i.e., 10 days after the withdrawal of lactoferrin, melittin and cecropin A from the diet.We hypothesised that the dietary addition of the bioactive antimicrobial protein lactoferrin (LF) and peptides melittin (MT) or cecropin A (CR) at a dosage of 100 mg/kg to the diet of Wistar rats would result in strong modulatory effects on faecal microbial enzymatic activity, short-chain fatty acid and ammonia concentrations. To date, the changes in bacterial extracellular and intracellular enzymatic activities upon addition of dietary AMPs have not yet been studied. This experiment lasted 15 days; during the first 5 day period, the rats were fed the control diet (S) and diets supplemented with LF, MT or CR. On days 6–15, all rats were fed the control S diet. The faecal fermentation processes were substantially stopped after two days of treatment, on average, in all rats receiving LF and two AMPs. The deepest suppression effect was observed on the last day of treatment (day 5) and persisted through days 5–8. The highest decreases in faecal bacterial β-glucosidase and β-glucuronidase activities as well as in SCFA and ammonia concentrations were observed in the rats fed the CR diet. Only in the CR animals did the mechanism of suppressed microbial fermentation involve diminished enzyme release from bacterial cells to the digesta.

Inhibition Properties of Arylsulfatase and β-Glucuronidase by Hydrogen Peroxide, Hypochlorite, and Peracetic Acid.

Arylsulfatase and β-glucuronidase are two important enzymes in humans, which play an important role in the dynamic equilibrium of steroidal estrogens. This work probably for the first time reported that hydrogen peroxide (H2O2), hypochlorite, and peracetic acid (PAA) could effectively inhibit the activities of arylsulfatase and/or β-glucuronidase. The 50% of inhibitions (IC50) of H2O2, hypochlorite, and PAA on arylsulfatase were found to be 142.90 ± 9.00, 91.83 ± 10.01, and 43.46 ± 2.92 μM, respectively. The corresponding IC50 values of hypochlorite and PAA on β-glucuronidase were 704.90 ± 41.40 and 23.26 ± 0.82 μM, whereas H2O2 showed no inhibition on β-glucuronidase. The inhibitions of arylsulfatase and/or β-glucuronidase by these three chemicals were pH-dependent. It was further revealed that the inhibitions of hypochlorite on both arylsulfatase and β-glucuronidase were irreversible. On the contrary, the inhibitions by H2O2 and PAA were reversible. In addition, the inhibition by H2O2 was competitive and that by PAA was noncompetitive. In general, H2O2 and hypochlorite can be endogenously produced in humans, which suggested that the two compounds are potential endocrine disruption compounds (EDCs) as they can cause endocrine disruption via the inhibition of arylsulfatase and β-glucuronidase. This work further indicated that any agent that can induce the production of H2O2 or hypochlorite in humans is a potential EDC, which explains why some EDCs with very weak or no estrogenic potency can cause endocrine disruption, which is confirmed in epidemiological studies.

Inducing new bioactive metabolites production from coculture of Pestalotiopsis sp. and Penicillium bialowiezense

Coculturing two or more fungi is a useful strategy to awaken the silent genes to produce structurally diverse and bioactive natural products. Through the coculture of Pestalotiopsis sp. and Penicillium bialowiezense, six new isoprenylated chromane derivatives, including two pairs of enantiomeric ones (1a/1b–2a/2b) and two optical pure ones (3–4), two new isoprenylated phenol glucoside derivatives (6–7), as well as eight known structural analogues (5 and 8–14), were obtained. The structures of these new compounds were characterized by NMR spectroscopy, single-crystal X-ray crystallography, and ECD calculation. The Δ10,11 double bond of pestaloficin D (5) was revised to E-configurated based on the extensive spectroscopic analyses. Compounds 1a/1b and 2a/2b were the first examples of enantiomeric isoprenylated chromane derivatives, which were successfully separated by chiral HPLC. Additionally, all the isolated compounds were evaluated for the in vitro β-glucuronidase (GUS) and butyrylcholinesterase (BChE) inhibitory activities. Compounds 1a and 1b showed significant β-glucuronidase inhibitory potency with IC50 values of 7.6 and 10.3 μM, respectively. Compound 14 exhibited moderate BChE inhibitory activity with an IC50 value of 21.3 μM. In addition, the structure-enzyme inhibitory activity relationship of compounds 1–14 is discussed.

Gut microbiota profiles and fecal beta-glucuronidase activity in kidney transplant recipients with and without post-transplant diarrhea.

Post-transplant diarrhea is a common complication after solid organ transplantation and is frequently attributed to the widely prescribed immunosuppressant mycophenolate mofetil (MMF). Given recent work identifying the relationship between MMF toxicity and gut bacterial β-glucuronidase activity, we evaluated the relationship between gut microbiota composition, fecal β-glucuronidase activity, and post-transplant diarrhea. We recruited 97 kidney transplant recipients and profiled the gut microbiota in 273 fecal specimens using 16S rRNA gene sequencing. We further characterized fecal β-glucuronidase activity in a subset of this cohort. Kidney transplant recipients with post-transplant diarrhea had decreased gut microbial diversity and decreased relative gut abundances of 12 genera when compared to those without post-transplant diarrhea (adjusted p value<.15, Wilcoxon rank sum test). Among the kidney transplant recipients with post-transplant diarrhea, those with higher fecal β-glucuronidase activity had a more prolonged course of diarrhea (≥7days) compared to patients with lower fecal β-glucuronidase activity (91% vs 40%, p=.02, Fisher's exact test). Our data reveal post-transplant diarrhea as a complex phenomenon with decreased gut microbial diversity and commensal gut organisms. This study further links commensal bacterial metabolism with an important clinical outcome measure, suggesting fecal β-glucuronidase activity could be a novel biomarker for gastrointestinal-related MMF toxicity.

Data for "Commensal intestinal microbiota regulates host luminal proteolytic activity and intestinal barrier integrity through β-glucuronidase activity"

Data for "Commensal intestinal microbiota regulates host luminal proteolytic activity and intestinal barrier integrity through β-glucuronidase activity"

Lactic acid bacteria in kimchi might be a cause for carcinogen production in intestine

The objective of this study was to evaluate if kimchi may be related to carcinogenesis by evaluating the activities of β-glucuronidase and β-glucosidase of lactic acid bacteria isolated from kimchi, which play a role in carcinogenesis. Thirty-six lactic acid bacteria (22 Lactobacillus spp., 11 Pediococcus pentosaceus, 2 Leuconostoc spp., and 1 Weisella cibaria) isolated from various kimchi were incubated in 10 mL of lactobacilli MRS broth at 35 °C for 20 h. Following incubation, 100 μL of each culture was suspended in 10 mL phosphate-buffered saline. The suspensions of 36 lactic acid bacteria strains were diluted with distilled water until OD500 = 1.048, and 65 μL aliquots of the suspensions were inoculated into the wells, where contained substrates to evaluate the activities of β-glucuronidase and β-glucosidase, followed by incubation at 35 °C for 4 h for the enzyme reactions. Of 36 isolates, 19 strains (52.8%) produced β-glucuronidase, 3 strains (13.9%) produced β-glucosidase, and two strains (5.6%) produced both enzymes. This result indicates that consumption of kimchi may have a potential possibility for a carcinogen production in the gut by producing the pro-carcinogenic enzymes (β-glucuronidase and β-glucosidase) of the lactic acid bacteria in kimchi.

Nutritional Value and Biological Activity of Gluten-Free Bread Enriched with Cricket Powder.

Cricket powder, described in the literature as a source of nutrients, can be a valuable ingredient to supplement deficiencies in various food products. Work continues on the implementation of cricket powder in products that are widely consumed. The aim of this study was to obtain gluten-free bread with a superior nutritional profile by means of insect powder addition. Gluten-free breads enriched with 2%, 6%, and 10% of cricket (Acheta domesticus) powder were formulated and extensively characterized. The nutritional value, as well as antioxidant and β-glucuronidase activities, were assessed after simulated in vitro digestion. Addition of cricket powder significantly increased the nutritional value, both in terms of the protein content (exceeding two-, four-, and seven-fold the reference bread (RB), respectively) and above all mineral compounds. The most significant changes were observed for Cu, P, and Zn. A significant increase in the content of polyphenolic compounds and antioxidant activity in the enriched bread was also demonstrated; moreover, both values additionally increased after the digestion process. The total polyphenolic compounds content increased about five-fold from RB to bread with 10% CP (BCP10), and respectively about three-fold after digestion. Similarly, the total antioxidant capacity before digestion increased about four-fold, and after digestion about six-fold. The use of CP also reduced the undesirable activity of β-glucuronidase by 65.9% (RB vs. BCP10) in the small intestine, down to 78.9% in the large intestine. The influence of bread on the intestinal microflora was also evaluated, and no inhibitory effect on the growth of microflora was demonstrated, both beneficial (Bifidobacterium and Lactobacillus) and pathogenic (Enterococcus and Escherichia coli). Our results underscore the benefits of using cricket powder to increase the nutritional value and biological activity of gluten-free food products.

Isolation and Functional Characterization of the Promoters of Miltiradiene Synthase Genes, TwTPS27a and TwTPS27b, and Interaction Analysis with the Transcription Factor TwTGA1 from Tripterygium wilfordii.

Miltiradiene synthase (MS) genes, TwTPS27a and TwTPS27b, are the key diterpene synthase genes in the biosynthesis of triptolide, which is an important medicinally active diterpenoid in Tripterygium wilfordii. However, the mechanism underlying the regulation of key genes TwTPS27a/b in triptolide biosynthesis remains unclear. In this study, the promoters of TwTPS27a (1496 bp) and TwTPS27b (1862 bp) were isolated and analyzed. Some hormone-/stress-responsive elements and transcription factor (TF) binding sites were predicted in both promoters, which might be responsible for the regulation mechanism of TwTPS27a/b. The β-glucuronidase (GUS) activity analysis in promoter deletion assays under normal and methyl jasmonate (MeJA) conditions showed that the sequence of −921 to −391 bp is the potential core region of the TwTPS27b promoter. And the TGACG-motif, a MeJA-responsive element found in this core region, might be responsible for MeJA-mediated stress induction of GUS activity. Moreover, the TGACG-motif is also known as the TGA TF-binding site. Yeast one-hybrid and GUS transactivation assays confirmed the interaction between the TwTPS27a/b promoters and the TwTGA1 TF (a MeJA-inducible TGA TF upregulating triptolide biosynthesis in T. wilfordii), indicating that TwTPS27a/b are two target genes regulated by TwTGA1. In conclusion, our results provide important information for elucidating the regulatory mechanism of MS genes, TwTPS27a and TwTPS27b, as two target genes of TwTGA1, in jasmonic acid (JA)-inducible triptolide biosynthesis.

Inhibitory effect of 4-ethylcatechol on β-glucuronidase activity

Inhibitory effect of 4-ethylcatechol on β-glucuronidase activity

Evaluation of designed consortium SNH-1 for efficient hydrolysis of agriculture waste to benefit bioethanol production

Abstract An environment friendly and cost-effective approach is a major bottleneck in biofuels production from agriculture waste residue. The present study unravels the enzymatic hydrolytic potential of a designed bacterial consortium (SNH-1) comprised of camel rumen derived B. safensis CRN 13, B. subtilis CRN 16, C. braakii CRN 21, B. circulans CRN 24, and P. dendritiformis CRN18. The consortium SNH-1 was evaluated for exo-glucanase (0.29 U/ml), β-glucosidase (0.69U/ml), β-glucuronidase (0.29U/ml), endoxylanase (0.79U/ml), arabinosidase (0.62U/ml), α-galactosidase (0.36 U/ml), and endoglucanase (0.51U/ml) activity. Genome analysis of consortium members identified 660 carbohydrate-active enzymes (CAZyme) responsible for breaking the structural intricacy. These include 296 glycoside hydrolase, 19 auxiliary activity, 186 glycosyl transferases, and 37 carbohydrate-binding modules genes. The designed consortium hydrolyzed sugarcane bagasse, wheat straw, and rice straw effectively. Response surface methodology (RSM) was applied to optimize the pH, temperature, and substrate concentration, which has resulted in a 23% higher saccharification of wheat straw. Further, 6.2% bioethanol production was estimated from wheat straw hydrolysate using S. cerevisiae. The wheat straw saccharification by the designed consortium is a cost-effective, chemical-free, cleaner, and promising approach for producing lignocellulosic bioethanol.

Cytochemically Demonstrable B-Glucuronidase Activity in Normal and Neoplastic Human Lymphoid Cells

Mononuclear cell suspensions were prepared from 40 normal peripheral blood and lymphoid tissue specimens and 42 neoplastic specimens obtained from patients with malignant lymphoma and lymphocytic leukemia. These suspensions were analyzed for la antigens, surface immunoglobulin (Slg), sheep erythrocyte (E) rosette formation and, in some instances, acid alpha-naphthyl acetate esterase (ANAE) activity. The results of these studies were correlated with the expression of cytochemically demonstrable BG activity. The percentage of BG+ lymphocytes was found to be comparable, within 10%, to the percentage of E+ (T) cells in the majority of normal, non-neoplastic peripheral blood, tonsil, spleen, and lymph node specimens examined. Occasionally, the percentage of E+ cells exceeded the percentage of BG+ cells by 20% or more, suggesting the presence of an E+BG- T cell subpopulation. BG+ B lymphocytes were only demonstrated in 1 of 40 non-neoplastic lymphoid specimens. The neoplastic B cells in each of 14 B cell (la+Slg+E-) lymphomas were BG-. However, a variable proportion of the neoplastic cells isolated from 6 cases of B cell chronic lymphocytic leukemia and neoplastic plasma cells isolated from 7 cases of multiple myeloma expressed BG activity. Thus, it appears that both normal and neoplastic BG- and BG+ B lymphocyte populations exist; the latter may be related to a state of activation or a stage of B cell differentiation. The neoplastic cells isolated from 4 T cell (la-Slg-E+) malignancies were BG+ while those isolated from 3 T cell malignancies were BG-. The variable expression of BG activity by T cell malignancies may be related to T cell differentiation. Investigation of BG expression by T cell derived malignancies may prove useful in sorting out T cell phenotypes.

Correlation of endogenous β-glucuronidase activity with differentiation of in vitro cultures of Scutellaria baicalensis

The study examined the relationship between the contents of principal flavones of Scutellaria baicalensis Georgi and the activity of endogenous b-glucuronidase (sGUS) in differentiated and undifferentiated in vitro cultures grown on solid and liquid medium. Using RP-HPLC, it was shown that sGUS activity did not correlate with the increase in biomass of cultures, but depended on tissue differentiation. In hairy roots cultured both on liquid and solid medium, it was ten times higher than in undifferentiated cultures. At the same time, the share of aglycone forms in the total content of flavones during the culture cycle depended on the growing conditions. It was significantly higher on agar medium than on liquid medium. A correlation between sGUS activity and total aglycone content, as well as wogonin content, was observed only in hairy roots grown in the liquid medium.

Dietary Lactobacillus plantarum LS/07 and inulin in the management of chronic disease risk factors.

The aim of this study was to investigate the possibilities of modification of chronic disease risk factors with probiotic strain Lactobacillus plantarum LS/07 and prebiotic inulin in rats with western high fat diet. The Sprague-Dawley rats were divided into four groups: control group (CG group), group with high fat diet (HFD group), group receiving high fat diet in combination with Lactobacillus plantarum LS/07 (HFD+PRO group), and group receiving high fat diet in combination with oligofructose enriched inulin (HFD+PRE group). The activity of β-glucuronidase, lipid parameters, bile acids, oxLDL, short chain fatty acids, and counts of coliforms and lactobacilli were determined. High fat diet as akey risk factor of chronic diseases had adverse effect on expression of metabolic and biochemical parameters. Dietary intake of Lactobacillus plantarum LS/07 (HFD+PRO group) and inulin (HFD+PRE group) suppressed weight gain of rats. In HFD+PRO group, the level of total cholesterol (P<0.001), LDL-CH (P<0.05), oxLDL (P<0.001), total bile acids (P<0.001) were statistically significantly decreased, while the production of short chain fatty acids was enhanced. Changes in the selected parameters exhibited a similar tendency also in the HFD+PRE group. Activity of β-glucuronidase was statistically significantly decreased (P<0.001) in the HFD+PRE group. Lactobacillus plantarum LS/07 and inulin caused astatistically significant increase in the count of lactobacilli (P<0.001) and a decrease in the number of coliforms (P<0.001). These results indicate Lactobacillus plantarum LS/07 and inulin could be used in diet for human and animals as an important nutritional supplement or in medicinal products.

Magnoflorine prevent the skeletal muscle atrophy via Akt/mTOR/FoxO signal pathway and increase slow-MyHC production in streptozotocin-induced diabetic rats

Ethnopharmacological relevanceTinospora cordifolia (TC) is being used as a blood purifier in Ayurveda since ancient time. It is a very popular immunomodulator and holds anti-inflammatory and anti-oxidative potential, hence anti-aging properties. Therefore, it is also known as ‘Amrita’ in Ayurveda and is widely used to treat diabetes mellitus type II (T2DM) and its secondary complications; however, its underlying mechanism was not expedited to date. Aim-To explore the in vivo therapeutic efficiency and mechanism of action of TC and its secondary constitute magnoflorine on the skeletal muscle atrophy in the rat model of T2DM. MethodAnimal model of T2DM was developed using streptozotocin (STZ) injection followed by intervention with TC, metformin, and magnoflorine for three weeks. Confirmation of T2DM and abrogation of atrophic markers and possible mechanisms on supplementation of TC and magnoflorine were explored using histology, bio-assays, Western blotting, and q-PCR. ResultTC and Magnoflorine supplementations significantly (p ≤ 0.05) decreased the fasting blood glucose (FBG) levels in T2DM rats. Both treatments prevented the lean body, individual skeletal muscle mass, and myotubes diameter loss (p ≤ 0.05). Magnoflorine significantly reduced the degradation of the protein indicated by biochemical markers of atrophy i.e. decreased serum creatine kinase (CK) levels and increased myosin heavy chain-β (MyHC-β) levels in muscles. Q-PCR and western blotting supported the findings that magnoflorine significantly increased the mRNA and protein abundances (~3 fold) of MyHC-β.TC and magnoflorine efficiently decreased the expression of ubiquitin-proteasomal E3-ligases (Fn-14/TWEAK, MuRF1, and Atrogin 1), autophagy (Bcl-2/LC3B), and caspase related genes along with calpains activities in T2DM rats. Both TC and magnoflorine also increased the activity of superoxide dismutase, GSH-Px, decreased the activities of β-glucuronidase, LPO, and prevented any alteration in the catalase activity. In contrast, magnoflorine increased expression of TNF-α and IL-6 whereas TC and metformin efficiently decreased the levels of these pro-inflammatory cytokines (p ≤ 0.05). However, magnoflorine was found to increase phosphorylation of Akt more efficiently than TC and metformin. ConclusionTC, and magnoflorine are found to be effective to control fasting blood glucose levels significantly in T2DM rats. It also promoted the Akt phosphorylation, suppressed autophagy and proteolysis that might be related to blood glucose-lowering efficacy of magnoflorine and TC. However, increased muscle weight, specifically of the soleus muscle, expression of IL-6, and slow MyHC indicated the increased myogenesis in response to magnoflorine and independent from its hypoglycemic activity.

Genotypic and Phenotypic Characterization of Treponema phagedenis from Bovine Digital Dermatitis.

This study aimed to isolate and characterize Treponema spp. from bovine digital dermatitis (BDD)-infected dairy cattle. Seven isolates were characterized in this study. Isolates exhibited slow growth, and colonies penetrated the agar and exhibited weak β-hemolysis. Round bodies were observed in old and antibiotic-treated cultures. Cells ranged from 9–12 µm in length, 0.2–2.5 µm in width, and were moderately spiraled. The 16S rRNA analysis revealed the isolates as Treponema phagedenis with >99% sequence homology. Isolates had alkaline phosphatase, acid phosphatase, β-galactosidase, N-acetyl-β-glucosaminidase, esterase (C4), esterase lipase (C8), naphthol-AS-BI-phosphohydrolase, and β-glucuronidase activities. Low concentrations of ampicillin, erythromycin, and tetracycline were required to inhibit the growth of isolates. Formic, acetic, and butyric acids were produced, while propionic acid was significantly utilized, indicating its essentiality for treponemal growth. The isolates shared the same characteristics and, therefore, were considered as a single strain. Isolate HNL4 was deposited as a representative isolate (Treponema phagedenis KS1). The average nucleotide identity of strain KS1 showed a small difference with the human strain (99.14%) compared with bovine strain (99.72%). This study was the first to isolate and characterize Treponema phagedenis from BDD in Korea and, hence, it delivered pathogenicity-related insights and provided valuable information that can be used for the management of BDD.

Molecular Screening of β-glucuronidase and Class 1 Integron of Escherichia coli from Ready-to-Eat Foods in Tiruchirappalli, Tamil Nadu

Ready-to-eat food products procured from different roadside shops in Tiruchirappalli, Tamil Nadu were screened for Escherichia coli. A total of 500 samples from 250 vegetable and 250 meat products were collected from different hotels, restaurants and street food vendors in Tiruchirappalli, Tamilnadu. Out of 500 ready-to-eat food samples, 162 (32.4%) E. coli strains were isolated. The ready-to-eat meat products had higher bacterial count than the vegetable food samples collected due to unhygienic handling, improper storage, inadequate temperature to maintain processed meat and improper cooking. Biochemically identified E. coli colonies were screened for housekeeping gene uidA and 139 (85.8%) E. coli isolates were confirmed to possess β-glucuronidase activity. In addition, antibiotic susceptibility assay was performed using 12 antibiotics. From 139 E. coli strains, 96 (69.1%) isolates showed multidrug resistance. Among them, 16.7% showed 100% resistance to all the antibiotics tested. Whereas, multidrug resistant E. coli isolates showed increased resistance (75.9%) to streptomycin followed by 70-50% level of resistance to ceftriaxone, ampicillin, cefixime, ciprofloxacin, tetracycline, gentamicin, doxycycline, co-trimoxazole, norfloxacin, ofloxacin and chloramphenicol. Furthermore, drug resistant E. coli isolates 56 (58.3%) were detected with the presence of intI1. The source of contamination was found to be water and human handling. Drinking water supply from corporation might have been contaminated with fecal waste source is being discharged into Cauvery river which might disseminate horizontal gene transfer.

Polyphenolic rich extract of Oroxylum indicum alleviate β-glucuronidase activity via down-regulate oxidative stress: Experimental and computational studies

Oroxilum indicum has an extensive history in the Indian traditional medicine system. The root and stem bark of this plant is well accepted for the treatment, prevention, and cure of various types of diseases. The bark of the plant has a high content of flavonoids and other phenolics, which are known to be responsible for exhibiting various therapeutic effects. The role of O. indicum in liver disease is well known, but until now, the mechanism has not been well studied. In the present study, the stem bark of O. indicum was extracted with ethanol using a hot extraction method and their phyto-chemical analysis was performed. Further, the plant extract was also used to evaluate their role in reducing oxidative stress and β-glucuronidase enzyme activity. The antioxidant study showed that extract has the capability to down-regulate oxidative stress, while β-glucuronidase assay displayed IC50 at 174.36 μg/mL. Additionally, we selected the major polyphenolic phyto-compounds derived from the bark of O. indicum for in silico ADME and docking studies. The ADME study reflected the optimal pharmacokinetic profile of the compounds. However, the docking experiments confirmed that all the selected phyto-compounds have tendency to interact with β-glucuronidase receptor.

Synbiotic VSL#3 and yacon-based product modulate the intestinal microbiota and prevent the development of pre-neoplastic lesions in a colorectal carcinogenesis model.

Colorectal cancer is a public health problem, with dysbiosis being one of the risk factors due to its role in intestinal inflammation. Probiotics and synbiotics have been used in order to restore the microbiota balance and to prevent colorectal carcinogenesis. We aimed to investigate the effects of the probiotic VSL#3® alone or in combination with a yacon-based prebiotic concentrate on the microbiota modulation and its influence on colorectal carcinogenesis in an animal model. C57BL/6J mice were divided into three groups: control (control diet), probiotic (control diet + VSL#3®), and synbiotic (yacon diet + VSL#3®). The diets were provided for 13 weeks and, from the third one, all animals were subjected to induction of colorectal cancer precursor lesions. Stool samples were collected to evaluate organic acids, feces pH, β-glucuronidase activity, and microbiota composition. The colon was used to count pre-neoplastic lesions and to determine the cytokines. The microbiota composition was influenced by the use of probiotic and synbiotic. Modifications were also observed in the abundance of bacterial genera with respect to the control group, which confirms the interference of carcinogenesis in the microbiota. Pre-neoplastic lesions were reduced by the use of the synbiotic, but not with the probiotic. The protection provided by the synbiotic can be attributed to the modulation of the intestinal inflammatory response, to the inhibition of a pro-carcinogenic enzyme, and to the production of organic acids. The modulation of the composition and activity of the microbiota contributed to beneficial changes in the intestinal microenvironment, which led to a reduction in carcinogenesis. KEY POINTS: • Synbiotic reduces the incidence of colorectal cancer precursor lesions. • Synbiotic modulates the composition and activity of intestinal microbiota. • Synbiotic increases the abundance of butyrate-producing bacteria.

DREB2 Family Transcription Factors Promote the Expression of Phytocystatin1 in Arabidopsis thaliana During Seed Germination

Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors of cysteine proteases (CPs) that regulate protein turnover and defense responses. Here, we characterized a member of the PhyCYS gene family in Arabidopsis thaliana, AtCYS1, during seed germination. AtCYS1 transcript level and AtCYS1 promoter-driven β-glucuronidase (GUS) activity gradually increased during germination, as measured by quantitative reverse transcription PCR (qRT-PCR) and GUS staining assays, respectively. The level and activity of the AtCYS1 protein were up-regulated during germination, as shown by western blot and CP inhibitory activity analyses, respectively. In addition, a canonical dehydration-responsive element (DRE) was identified in the AtCYS1 promoter. Electrophoretic mobility shift assays (EMSAs) revealed that the DRE-binding factor 2C (DREB2C) binds directly to the AtCYS1 promoter. Transient transactivation assays using Arabidopsis leaf protoplasts showed that DREB2A and DREB2C act as transcriptional activators of AtCYS1. In DREB2C overexpression lines, AtCYS1 protein levels were elevated, while the endogenous CP activity was reduced. However, AtCYS1 overexpression lines (35S:AtCYS1) and the cys1-1 null mutant, characterized by a quantitative imbalance in the AtCYS1 protein level, showed delayed germination under normal growth conditions. These results suggest that DREB2 proteins act as transcriptional activators of the AtCYS1 gene, and an imbalance in the level of AtCYS1 inhibits seed germination.