Steroidogenic activity of liposomal methylated resveratrol analog 3,4,5,4'-tetramethoxystilbene (DMU-212) in human luteinized granulosa cells in a primary three-dimensional in vitro model.

PURPOSE: Steroid hormone secretion is one of the key functions of granulosa cells (GCs). Resveratrol is a natural polyphenol, known for its beneficial health effects, such as improving reproductive health. However, its application is limited due to poor bioavailability. The methoxy derivative of resveratrol (DMU-212) was demonstrated to be more lipophilic, and therefore of greater bioavailability. However, since the addition of methoxy groups to the stilbene scaffold was found to make the molecule insoluble in water, DMU-212 was loaded into liposomes. This study aimed to evaluate how the liposomal formulation of DMU-212 (lipDMU-212) alters estradiol and progesterone secretion of human ovarian GCs in a primary three-dimensional cell culture model. METHODS: DMU-212-loaded liposomes were prepared by thin film hydration followed by extrusion. Cell viability was measured after exposure of GCs spheroids to the liposomal formulation of DMU-212 using CellTiter-Glo® 3D Cell Viability Assay. The secretion of estradiol and progesterone was determined using commercial ELISA kits. RT-qPCR was conducted to analyze the expression of steroidogenesis-related genes. Finally, the western blot technique was used to analyze the effect of lipDMU-212 and FSH treatments on CYP11A1 and HSD3B1 protein levels. RESULTS: lipDMU-212 was found to significantly increase estradiol and progesterone secretion in a dose-dependent manner by enhancing the expression of CYP11A1, HSD3B1, StAR, CYP17A1, CYP19A1, and HSD17B1 genes. We have also shown that lipDMU-212, used alone and in combination with FSH, significantly increased the expression of the HSD3B1 and CYP11A1 proteins in GCs. Furthermore, our study suggests that lipDMU-212 increases FSH activity. CONCLUSIONS: This is the first study to describe the steroidogenic activity of liposomal formulation of DMU-212, possibly through increasing the StAR and CYP19A1 expression. These findings suggest that lipDMU-212 might have a beneficial effect in the treatment of disorders related to estrogen deficiency and hyperandrogenism, such as PCOS.

Effects of phenolic compounds on 3ß-hydroxysteroid dehydrogenase activity in human and rat placenta: Screening, mode of action, and docking analysis.

Human 3ß-hydroxysteroid dehydrogenase type I (HSD3B1) and rat type IV (HSD3B4) in placentas catalyze the conversion of pregnenolone to progesterone, which plays a key role in maintaining pregnancy. Many phenolic compounds potentially inhibit HSD3B in placentas as endocrine disruptors. In this study, the effects of 16 phenolic compounds on the activity of human HSD3B1 and rat HSD3B4 were determined and the structure-activity relationship was compared. HSD3B1 in human placental microsomes and HSD3B4 in rat placental microsomes were used to measure their activities and pregnenolone and NAD+ were used as substrates. Of the 16 phenolic compounds, 4-nonylphenol, pentabromophenol, and 2-bromophenol resulted in residual human HSD3B1 activity lower than 50 % and 4-nonylphenol and pentabromophenol resulted in residual rat HSD3B4 activity lower than 50 %. 4-Nonylphenol, pentabromophenol, and 2-bromophenol were mixed inhibitors of human HSD3B1, with Ki values of 2.31, 3.58 and 4.86 µM, respectively, while 4-nonylphenol and pentabromophenol were noncompetitive inhibitors of rat HSD3B4 with Ki values of 20.86 and 141.8 µM. Molecular docking showed that 4-nonylphenol, pentabromophenol, and 2-bromophenol docked to the active sites of human HSD3B1 and rat HSD3B4, and the shift of residue S125 in human HSD3B1 to T125 in rat HSD3B4 could explain the species-dependent difference in their inhibitory potency and mode of action. This study demonstrates that 4-nonylphenol, pentabromophenol, and 2-bromophenol are mixed inhibitors of human placental HSD3B1, while 4-nonylphenol and pentabromophenol are noncompetitive inhibitors of rat HSD3B4, possibly blocking the placental steroidogenesis.

Tetramethyl bisphenol A stimulates proliferation but inhibits fetal Leydig cell function in male rats by targeting estrogen receptor α after in utero exposure.

Tetramethyl bisphenol A (TMBPA) is a widely used flame retardant. TMBPA has been a toxic to Leydig cells in puberty, but it remains unclear whether TMBPA has a similar inhibitor effect on fetal Leydig cells (FLCs). This study reported morphological and functional alterations of FLCs in the testes of male offspring at birth after in utero exposure to TMBPA. Pregnant Sprague Dawley rats were dosed via continuous gavage of TMBPA (0, 10, 50, and 200 mg/kg/day) from gestational day 14 to 21. TMBPA markedly raised serum total testosterone level, testicular volume, and FLC number of male offspring at 200 mg/kg dose. The up-regulation of Insl3, Star, and Cyp11a1 mRNAs was observed after 200 mg/kg TMBPA exposure. After normalization to the number of FLCs, TMBPA significantly reduced Lhcgr and Hsd3b1 expressions at 10 mg/kg, and Cyp17a1 at 200 mg/kg paralleling with their protein levels. TMBPA compromised the expression of Esr1, while increased the expression of Cdk2 and Cdk4 as well as their protein levels. TMBPA particularly increased the phosphorylation of AKT1 and AKT2 at 200 mg/kg. In conclusion, the present study suggests that TMBPA may promote FLC proliferation via ESR1-CDK2/4-AKT pathway, while inhibits the function of FLCs by reducing steroidogenic enzyme activity.

Hydrolytic Enzymes as Potentiators of Antimicrobials against an Inter-Kingdom Biofilm Model.

Biofilms are recalcitrant to antimicrobials, partly due to the barrier effect of their matrix. The use of hydrolytic enzymes capable to degrade matrix constituents has been proposed as an alternative strategy against biofilm-related infections. This study aimed to determine whether hydrolytic enzymes could potentiate the activity of antimicrobials against hard-to-treat interkingdom biofilms comprising two bacteria and one fungus. We studied the activity of a series of enzymes alone or in combination, followed or not by antimicrobial treatment, against single-, dual- or three-species biofilms of Staphylococcus aureus, Escherichia coli, and Candida albicans, by measuring their residual biomass or culturable cells. Two hydrolytic enzymes, subtilisin A and lyticase, were identified as the most effective to reduce the biomass of C. albicans biofilm. When targeting interkingdom biofilms, subtilisin A alone was the most effective enzyme to reduce biomass of all biofilms, followed by lyticase combined with an enzymatic cocktail composed of cellulase, denarase, and dispersin B that proved previously active against bacterial biofilms. The subsequent incubation with antimicrobials further reduced the biomass. Enzymes alone did not reduce culturable cells in most cases and did not interfere with the cidal effects of antimicrobials. Therefore, this work highlights the potential interest of pre-exposing interkingdom biofilms to hydrolytic enzymes to reduce their biomass besides the number of culturable cells, which was not achieved when using antimicrobials alone. IMPORTANCE Biofilms are recalcitrant to antimicrobial treatments. This problem is even more critical when dealing with polymicrobial, interkingdom biofilms, including both bacteria and fungi, as these microorganisms cooperate to strengthen the biofilm and produce a complex matrix. Here, we demonstrate that the protease subtilisin A used alone, or a cocktail containing lyticase, cellulase, denarase, and dispersin B markedly reduce the biomass of interkingdom biofilms and cooperate with antimicrobials to act upon these recalcitrant forms of infection. This work may open perspectives for the development of novel adjuvant therapies against biofilm-related infections.

Magnetic combi CLEA for inhibition of bacterial biofilm: A green approach.

In the present study different enzymes (α- amylase, trypsin, cellulase, horse-radish peroxidase and pectinex ultra clear) were studied for bacterial biofilm inhibition and Pectinex ultra clear showed best inhibition. So, m-combi-CLEA of Pectinex ultra clear was developed by cross linked enzyme aggregate (CLEA) formation on APTES (3-aminopropyltriethoxysilane) modified iron oxide nanoparticles. Different parameters were optimized and it was observed that 0.4 mg/ml of protein (containing 25 U/mg cellulase activity), 0.5 mg/ml BSA and 10 mM glutaraldehyde when incubated for 3 h gives 100% enzyme activity using ethanol as the precipitant. The CLEA formed were thermally more stable as compared to free enzyme. m-combi-CLEA of Pectinex ultra clear shows 75-78% biofilm inhibition of E. coli and S. aureus. Furthermore, m-combi-CLEA can be reused till 4 cycles with same efficiency. The carbohydrate contents of E. coli biofilm decreased from 64.629 µg to 6.23 µg and for S. aureus biofilm, it decreased from 58.46 µg to 5.52 µg when treated with m-combi CLEA in comparison to untreated biofilms. FTIR, darkfield illumination Fluorescence Microscopy, and Scanning Electron Microscopy was further used for characterization.

Comparative analysis of physical and functional properties of cellulose nanofibers isolated from alkaline pre-treated wheat straw in optimized hydrochloric acid and enzymatic processes.

Two methods, HCl and enzymatic treatments, were evaluated for diversification of morphological and functional properties of cellulose nanofibers (CNF) from two- stage-alkaline pre-treated wheat straw (WS). The extraction conditions were optimized by a central composite designed experimental approach varying time (4-8 h) and temperature (80-120 °C) for the HCl-based treatment and time (4-8 h), and FiberCare dosage (50-100 endo-1,4-ß-glucanase unit/g) and Viscozyme (10-20 fungal ß-glucanase units/g) for the enzyme-based treatment. The CNF yields, morphological (polydispersity index (PdI), length and diameter), and functional (crystallinity and thermal degradation) properties were compared. The CNF produced by the HCl (HCN) and enzymatically (ECN) attained diameters ~17 nm had PdI, length, and crystallinity of 0.53, 514 nm & 70%, and 0.92, 1.0 µm & 48%, respectively. Thus, the HCN morphology suits homogenous nano-applications, whereas that of the ECN, would suit heterogenous nano-applications. The HCN and ECN yields were similar (~20%) with optimal production time of 7.41 and 4.64 h, respectively. Both the HCN & ECN can be classified as thermally stable nanocolloids with maximum thermal degradation temperatures of ~380 °C and Zeta potential ~-16 mV. The two CNF production methods have potential synergetic effects on CNF production, morphological, and functional properties.

Influence of multi-enzyme preparation supplemented with sodium butyrate on growth performance blood profiles and economic benefit of growing rabbits.

The present study was carried out to explore the impacts of dietary supplementation of enzyme mixture with sodium butyrate on the growth performance, carcass traits, blood profile and economic benefit in two breeds of weanling rabbits adapted to survive in Egypt (New Zealand White and Rex). One-hundred and twenty weaned male rabbits (New Zealand White and Rex) of 6 weeks of age and 770.5 ± 20 g body weight were allotted randomly into four groups in a factorial arrangement. The obtained results indicated that there were non-significant differences in all growth performance traits, blood profile and economic parameters due to the breed effect. However, there were significant differences in most of carcass traits due to the breed effect except total giblets and New Zealand White breed showed the highest value of these parameters including dressing % (p < .01), forequarter and loin % (p < .001) and hindquarter % (p < .003) compared with Rex breed counterparts. The effect of the treatment and its interaction with the breed significantly (p < .05) improved body weight gain, feed consumption and carcass traits (percentage of dressing, forequarter, hind quarter and lion). However, final body weight and feed conversion ratio were not significantly influenced. Supplementing a diet with treatment significantly decreased blood triglycerides, cholesterol and the ratio between albumin and globulin (A/G ratio), while increased blood total protein and globulin. Although higher feed cost and total costs in treated groups than control ones in each breed, they showed higher total return and net return. Rex non-treated rabbit breed showed the lowest profitability measures compared with other groups. In conclusion, dietary supplementation of multi-enzyme with sodium butyrate is highly recommended in growing rabbits due to their beneficial effects on the growth performance and profitability.

The efficacy of lyticase and ß-glucosidase enzymes on biofilm degradation of Pseudomonas aeruginosa strains with different gene profiles.

BACKGROUND: Pseudomonas aeruginosa is a nosocomial pathogen that causes severe infections in immunocompromised patients. Biofilm plays a significant role in the resistance of this bacterium and complicates the treatment of its infections. In this study, the effect of lyticase and ß-glucosidase enzymes on the degradation of biofilms of P. aeruginosa strains isolated from cystic fibrosis and burn wound infections were assessed. Moreover, the decrease of ceftazidime minimum biofilm eliminating concentrations (MBEC) after enzymatic treatment was evaluated. RESULTS: This study demonstrated the effectiveness of both enzymes in degrading the biofilms of P. aeruginosa. In contrast to the lyticase enzyme, ß-glucosidase reduced the ceftazidime MBECs significantly (P < 0.05). Both enzymes had no cytotoxic effect on the A-549 human lung carcinoma epithelial cell lines and A-431 human epidermoid carcinoma cell lines. CONCLUSION: Considering the characteristics of the ß-glucosidase enzyme, which includes the notable degradation of P. aeruginosa biofilms and a significant decrease in the ceftazidime MBECs and non-toxicity for eukaryotic cells, this enzyme can be a promising therapeutic candidate for degradation of biofilms in burn wound patients, but further studies are needed.

Safety and Efficacy of N-SORB®, a Proprietary KD120 MEC Metabolically Activated Enzyme Formulation: A Randomized, Double-Blind, Placebo-Controlled Study.

Background: Enzymes are crucial for all aspects of metabolic function. Digestive enzymes from natural sources have been credited with beneficial effects in the digestion and absorption of food. N-SORB is a novel KD120 multienzyme complex (MEC) of metabolically activated enzymes composed of proteases, amylases, lipases, alpha-galactosidase, and glucoamylase from natural sources. These enzymes are encapsulated in a SK713 SLP (non-GMO soy lecithin phospholipid) absorption technology (Prodosome®). Objective: This randomized, double-blind placebo-controlled investigation assessed the safety and efficacy of N-SORB KD120 MEC in healthy male and female volunteers on various parameters of the blood, immunity, body composition, physical health, and quality of life following a 90-day intervention. Methods: Forty-six male and female (mean age: 25.8 ± 12.1 years) healthy volunteers were randomly assigned to receive either N-SORB (1 mL, twice daily) or placebo for 90 consecutive days. Complete blood count, as well as blood glucose, liver enzymes, and lipid profile were assessed pre- and post-intervention. Serum cytokine levels were determined by using a Bio-Plex Pro Human Cytokine 8-plex assay and enzyme linked immunosorbent assay (ELISA). Whole body composition analysis was performed by dual-energy x-ray absorptiometry (DEXA) to determine body fat mass, lean mass, and android and gynoid fat. Body weight, blood pressure, and physical health were assessed. Changes in quality of life were examined using the World Health Organization Quality of Life-abbreviated version and sleep quality was assessed using the 24-item Pittsburgh Sleep Quality Index (PSQI) questionnaire. Adverse events were monitored before, during, and after completion of the study. Results: Of the 46 subjects enrolled, a total of 40 subjects successfully completed the study. Compared to placebo, changes in blood cell counts including hematocrit, hemoglobin, mean corpuscular volume, platelets, and lymphocytes provide evidence of some improvement. Quality of life (QOL) parameters showed a small but significant improvement in the N-SORB group. A significant increase was observed in aspartate aminotransferase level in the placebo group at the end of 90 days of treatment; however, no increase was observed in the N-SORB group. No significant changes in blood urea nitrogen, serum creatinine, alkaline phosphatase, alanine aminotransferase, and lipid profile were observed between the placebo and treatment groups before and following intervention. No adverse effects were reported. Conclusions: This randomized, double blind, placebo-controlled clinical study demonstrates that short-term intervention with N-SORB improves the QOL and PSQI in healthy volunteers and did not significantly alter cardiometabolic parameters, lipid profile, or body composition.

Molecular characterization of bmyC gene of the mosquito pupicidal bacteria, Bacillus amyloliquefaciens (VCRC B483) and in silico analysis of bacillomycin D synthetase C protein.

A strain of Bacillus amyloliquefaciens (VCRC B483) exhibiting mosquito pupicidal, keratinase and antimicrobial activities was isolated from mangrove forest ecosystem of Andaman and Nicobar Islands. Molecular characterization of the strain showed the presence of lipopeptide encoding bmyC gene. Phylogenetic tree based on protein sequence of this gene exhibited homology with mycosubtilin synthetase of Bacilus atropheus and Iturin synthetase of Bacillus subtilis and B. amyloliquefaciens. This is the first report on the evolutionary conservation of amino acids concerned with the function and structure of bmyC protein of B. amyloliquefaciens. The presence of valine at the 1197th position in our strain was found to be unique and different from the existing strains of B. subtilis and B. amyloliquefaciens. Molecular modelling studies revealed significant changes in the structure of epimerization domain of the bmyC protein with A1197V variation. Crude metabolite of this strain exhibited antifungal activity against Fusarium sp. and Carvularia sp.

Assessment of methods of detection of water estrogenicity for their use as monitoring tools in a process of estrogenicity removal.

Methods of monitoring of estrogenicity in water were gathered, compared, and tested within the context of their practical use as measurement and design tools, in the development of a process of degradation of estrogenic endocrine disruptors. In this work, the focus was put on in vitro assays, with the use of analytical techniques as additional analysis when possible. Practically, from a literature review, four methods that seemed most suitable to practical use required in a process development were tested: the Yeast Estrogen Screen assay, the Lyticase-assisted Yeast Estrogen Screen assay (LYES), the MMV-LUC assay and the HPLC-UV analytical method. Dose-response curves in response to estrogenic standard 17ß-estradiol were compared. Bisphenol A estrogenicity was measured by the methods as well. The model for the calculation of estradiol equivalents as measurements units was adapted. The methods were assessed in terms of ranges of detection, time of experiment, cost, ease of the experiment, reproducibility, etc. Based on that assessment, the LYES assay was selected and successfully applied to the monitoring of estrogenicity removal from 17ß-estradiol and bisphenol A. More precisely, the bioassay allowed the acquisition of kinetic curves for a laboratory-scaled process of estrogenicity removal by immobilized enzymes in a continuous packed-bed reactor. The LYES assay was found to have a real methodological potential for scale-up and design of a treatment process. The HPLC-UV method showed good complementarity with the LYES assay for the monitoring of bisphenol A concentrations in parallel with estrogenicity, reporting no significant estrogenicity from degradation byproducts, among others.

Design, synthesis and antimalarial screening of some hybrid 4-aminoquinoline-triazine derivatives against pf-DHFR-TS.

Existing antifolate antimalarial drugs have shown resistance due to the mutations at some amino acid positions of Plasmodium falciparum DHFR-TS. In the present study, to overcome this resistance, a new series of hybrid 4-aminoquinoline-triazine derivatives were designed and docked into the active site of Pf-DHFR-TS (PDB i.d. 1J3K) using validated CDOCKER protocol. Binding energy was calculated by applying CHARMm forcefield. Binding energy and the pattern of interaction of the docked compounds were analysed. Fifteen compounds were selected for synthesis based on their binding energy values and docking poses. Synthesized compounds were characterised by FTIR, (1)H NMR, (13)C NMR, mass spectroscopy and were screened for antimalarial activity against 3D7 strain of Plasmodium falciparum.

Using dielectrophoresis to study the dynamic response of single budding yeast cells to Lyticase.

Budding yeast cells are quick and easy to grow and represent a versatile model of eukaryotic cells for a variety of cellular studies, largely because their genome has been widely studied and links can be drawn with higher eukaryotes. Therefore, the efficient separation, immobilization, and conversion of budding yeasts into spheroplast or protoplast can provide valuable insight for many fundamentals investigations in cell biology at a single cell level. Dielectrophoresis, the induced motion of particles in non-uniform electric fields, possesses a great versatility for manipulation of cells in microfluidic platforms. Despite this, dielectrophoresis has been largely utilized for studying of non-budding yeast cells and has rarely been used for manipulation of budding cells. Here, we utilize dielectrophoresis for studying the dynamic response of budding cells to different concentrations of Lyticase. This involves separation of the budding yeasts from a background of non-budding cells and their subsequent immobilization onto the microelectrodes at desired densities down to single cell level. The immobilized yeasts are then stimulated with Lyticase to remove the cell wall and convert them into spheroplasts, in a highly dynamic process that depends on the concentration of Lyticase. We also introduce a novel method for immobilization of the cell organelles released from the lysed cells by patterning multi-walled carbon nanotubes (MWCNTs) between the microelectrodes.

Effects of dietary supplementation of multi-enzyme on growth performance, nutrient digestibility, small intestinal digestive enzyme activities, and large intestinal selected microbiota in weanling pigs.

Two experiments were conducted to assess the effects of dietary supplementation of an exogenous multi-enzyme (EME) preparation to 35- to 65-d-old piglets on apparent total tract digestibility (ATTD), growth performance, digestive enzyme activities, and selected microbial populations in feces. In Exp.1, twenty eight 35-d-old piglets were randomly assigned to 7 dietary treatments (corn-soybean based diet supplemented with 0, 100, 150, 200, 250, 300, or 350 mg EME/kg) in a 14-d digestibility study. Piglets fed the diets supplemented with EME had greater ATTD of DM, CP, and GE (P = 0.001, 0.005, and 0.009, respectively) than those fed the diet without EME supplementation, and those ATTD values increased linearly and quadratically (P < 0.001) as the levels of supplemented EME increased. In Exp. 2, two hundred 35-d-old weanling piglets were randomly allocated to 20 pens. The pens were then randomly assigned to 5 dietary treatments (corn-soybean based diet supplemented with 0, 100, 150, 250, or 350 mg EME/kg) with 4 pens per treatment in a 30-d feeding experiment. Piglets has ad libitum access to diets and water, and they were weighed at the beginning (35-d-old), middle (50-d-old), and end (65-d-old) of the experiment. Fecal samples were grabbed directly from the rectum and digesta samples from duodenum, jejunum, and ileum were taken at the end of the experiment for the analysis of selected bacteria populations and digestive-enzyme activities. The ADG and ADFI tended to be greater with the increasing levels of supplemented EME in both periods, whereas G:F was improved (P = 0.012 and 0.017) by EME in the period of 35 to 50 d of age and during the overall experimental period. Furthermore, inclusion of EME in diet increased the counts of Lactobacilli spp. and Bacillus subtilis spp., but reduced the populations of Salmonella spp. and Escherichia coli spp. in the feces. The EME supplementation also enhanced (P < 0.05) the activities of amylase, lipase, and protease in the small intestine. The growth performance-enhancing effects of EME appeared to be mediated by the age of the piglet and the dose of EME used. Supplementation of corn-soybean meal diets for 35- to 65-d-old piglets with EME has a potential to enhance gut health condition, increase nutrient digestion, and increase growth performance.

Efficient inhibition of human leukocytic elastase by means of α1-antitrypsin/peptide complexes.

α1 -Antitrypsin (AT), a serine protease inhibitor that specifically targets hydrolytic enzymes, plays a significant role in the termination of tissue inflammation and can therefore represent a key factor in chronic incidences as chronic obstructive pulmonary disease (COPD) or chronic hepatitis. A local and low-dose therapy for the treatment of acquired chronic inflammatory processes which are characterized by insufficient AT amounts but also of genetically conditioned AT deficiencies is supposed to be more effective and less cost-intensive compared to current therapies. In this study, a noncovalent complex formation between the cell-penetrating peptide carrier hCT(18-32)-k7 and AT was performed. The complex was applied to HEK293T/17 cells, as proof-of-principle, and polymorphonuclear leukocytes (PMN), which are responsible for tissue destruction and the perpetuation of inflammation in chronic processes. Both cell species show a successful uptake and subsequently both, an intracellular dot-shaped and homogeneous distribution of the complex demonstrating phagolysosomal as well as cytoplasmic availability. Furthermore, a decreased human leukocytic elastase (HLE) activity was observed after the direct complex administration to PMN. Since the application did not cause an enhanced vitality loss, the complex could facilitate an improvement in direct, local and low-dose treatment of chronically proceeding processes in order to attenuate protease-mediated tissue destruction.

Organ weight, intestinal morphology, and fasting whole-body oxygen consumption in growing pigs fed diets containing distillers dried grains with solubles alone or in combination with a multienzyme supplement.

The effects of adding a multienzyme complex to a diet containing distillers dried grains with solubles (DDGS) produced from a 1:1 mixture of corn and wheat on visceral organ weight, intestinal morphology, and fasting whole-body oxygen consumption (FWBOC) were investigated in growing pigs in a 28-d trial. Twenty-four pigs (BW = 19.9 ± 0.5 kg) were individually housed in floor pens and randomly assigned to 3 experimental diets (8 pigs per diet). The diets contained corn and soybean meal with 0% (control) or 30% DDGS (DDGS diet); the third diet was supplemented with a multienzyme complex in addition to the 30% DDGS (DDGS + enzyme diet). All diets had similar nutrient concentrations and met the 1998 NRC nutrient requirements for growing pigs. Pigs were fed at 4% of their BW once daily. On d 15, 4 pigs from each dietary treatment were randomly selected for measurement of FWBOC during the 24- to 30-h postprandial period using an open-circuit indirect calorimeter. At the end of the study, pigs were killed to determine visceral organ weights, ileal and cecal digesta viscosity, and intestinal morphology. There was no effect (P > 0.05) of dietary treatment on final BW, WBFOC, or digesta viscosity. Empty BW of pigs fed the control diet was heavier (P = 0.02) than that of pigs fed the DDGS diet, but the empty BW of pigs fed the DDGS + enzyme diet was not different (P > 0.05) from that of pigs fed the control or DDGS diet. There were no differences (P > 0.05) in empty BW of liver, spleen, pancreas, heart, stomach, small intestine, and cecum among dietary treatments on a per kilogram basis. However, pigs fed the DDGS diet had heavier (P < 0.05) colon plus rectum and portal-drained viscera (PDV) than pigs fed the control diet, but weights of colon plus rectum and PDV in pigs fed the DDGS + enzyme diet were not different (P > 0.05) from those of pigs fed the control diet. Although morphological data showed no differences (P > 0.05) in the duodenum, jejunum, and colon segments among dietary treatments, the DDGS diet tended to decrease (P < 0.10) villous height and villous height to crypt depth in the ileum. The results of this experiment indicated that pigs fed a diet containing 30% DDGS have reduced dressing percentage and increased visceral organ mass compared with pigs fed a corn-soybean meal diet. However, the addition of a multienzyme complex to the DDGS diet resulted in pigs having a dressing percentage and visceral organ mass that are not different from those of pigs fed a corn-soybean meal diet.

[Progress in the ligands and their complex structures of farnesoid X receptor].

Farnesoid X receptor (FXR) belongs to the nuclear receptor superfamily. It is highly related to the formation of metabolic syndrome and the glucose homeostasis, and therefore represents an important drug target against metabolic diseases and diabetes. In recent years, great progress has been made in the agonists, antagonists, and crystal structures of FXR. The diverse FXR ligands and their structure-activity relationship are reviewed in this article. The advances in the crystal structures of FXR in complex with different ligands are also introduced.

Effects of exogenous enzyme supplementation to corn- and soybean meal-based or complex diets on growth performance, nutrient digestibility, and blood metabolites in growing pigs.

Two experiments were conducted to determine the effects of dietary supplementation of exogenous enzymes on growth performance, apparent total tract digestibility (ATTD) of energy and nutrients, blood metabolites, fecal VFA, and fecal ammonia-N in growing pigs (Sus scrofa) fed a corn (Zea mays L.)- and soybean [Glycine max (L.) Merr.] meal (SBM)-based diet. In Exp. 1, 240 growing barrows (initial BW: 55.6 ± 0.9 kg) were randomly allotted to 5 treatments on the basis of BW. There were 4 replicates in each treatment with 12 pigs per replicate. The 5 treatments consisted of a corn-SBM-based control diet and 4 additional diets were similar to the control diet, with the exception that 0.05% ß-mannanase (M), α-amylase + ß-mannanase (AM), ß-mannanase + protease (MPr), or α-amylase + ß-mannanase + protease (AMP) was added to the diets, which were fed for 28 d. Pigs fed the AM, MPr, or AMP diet had greater (P < 0.05) ADG than pigs fed the control diet. Pigs fed the AMP diet also had greater (P < 0.05) ADG than pigs fed the M, AM, or MPr diet. Pigs fed the AMP diet had greater (P < 0.05) G:F than pigs fed the control diet. The G:F of the pigs fed the M, AM, or MPr diet were not different (P > 0.05) from the G:F in pigs fed the AMP or control diet. The ADFI, ATTD of nutrients, blood metabolites, and fecal VFA and ammonia-N concentrations were not different among treatments. In Exp. 2, 192 growing barrows (initial BW: 56.9 ± 1.0 kg) were allotted to 4 treatments. There were 4 replicates in each treatment with 12 pigs per replicate. Pigs were fed a corn-SBM-based diet (CSD) or a complex diet (CD) that contained corn, SBM, 3% rapeseed (Brassica napus L.) meal, 3% copra (Cocos nucifera L.) meal, and 3% palm (Elaeis guineensis Jacq.) kernel meal. Each diet was prepared without exogenous enzymes or with 0.05% AMP and all diets were fed for 28 d. The ADG and G:F of pigs fed the CSD were greater (P < 0.05) than pigs fed the CD. However, the type of diet had no effect on the ATTD of nutrients, blood metabolites, or fecal VFA and ammonia-N, and there was no diet × enzyme interaction for any of the measured variables. Supplementation of diets with exogenous enzymes resulted in greater (P < 0.05) ADG, G:F, ATTD of DM, GE, and CP, and blood urea nitrogen (BUN) concentration. These results indicate that supplementation of 0.05% of AMP enzymes to a corn-SBM diet or a complex diet may improve the performance of growing pigs.

In vitro antimalarial activity and molecular docking analysis of 4-aminoquinoline-clubbed 1,3,5-triazine derivatives.

AIMS: Present report describes the in vitro antimalarial activity and docking analysis of seven 4-aminoquinoline-clubbed 1,3,5-triazine derivatives on pf-DHFR-TS. METHODS AND RESULTS: The antimalarial activity was evaluated in vitro against chloroquine-sensitive 3D7 strain of Plasmodium falciparum. Compounds were docked onto the active site of pf-DHFR-TS using docking server to explicate necessary structural requirements for antimalarial activity. CONCLUSION: Title molecules demonstrated considerable bioactivity against the malaria parasite. Docking analysis revealed deep engulfment of the molecules into the inner groove of pf-DHFR-TS active site by making stable ligand-receptor posses. Hydrophobic interaction was identified as the only major interacting force playing a role between ligand-receptor interaction and minor with hydrogen bonds. SIGNIfiCANCE AND IMPACT OF THE STUDY: The study provided the novel insight into the necessary structural requirement for rationale-based antimalarial drug discovery.

Digestible energy values of feed ingredients with or without addition of enzymes complex in growing pigs.

The DE values and digestible nutrients content of 6 diets were measured in 60-kg male growing pigs fed restricted amount of feed. Diets were prepared from 5 ingredients [wheat (Triticum aestivum), corn (Zea mays), barley (Hordeum vulgare), wheat bran, and soybean (Glycine max) meal; inclusion levels of ingredients were not correlated] with or without carbohydrose enzyme (Rovabio Excel AP; 3300 endo-ß-1,4-xylanase visco units and 300 endo-1,3(4)-ß-glucanase units/kg of feed; 150 g/t of feed) according to a 6 × 2 factorial arrangement; dietary NDF ranged from 10.6 to 20.1% of DM. Pigs (5 per treatment) were placed in metabolism cages that allowed total collections of feces and urine for 10 d after a 11-d adaptation. Samples of feed, urine, and feces were analyzed for GE, ash, and N. Digestibility of GE, N, and OM were calculated. The effects of diet and enzyme (Enz) were evaluated by ANOVA. In addition, the DE and digestible nutrient contents of ingredients were calculated by regression of nutritive values of diets on level of ingredient inclusions. Apparent total tract digestibility of OM, N, and GE of diets were associated with dietary NDF content (r = -0.97; P < 0.001) and were increased (P < 0.05) by Enz addition by 0.4, 1.6, and 0.5%-units (a difference between two percentage values) for OM, N, and GE digestibility, respectively. Improvement in DE value due to Enz averaged 0.09 MJ/kg DM (15.11 vs. 15.02 MJ/kg DM; P < 0.05). The ADG (891 vs. 850 g/d; P < 0.05) was also increased by Enz addition. The calculated DE content without Enz addition averaged 16.3, 16.4, 14.9, 10.5, and 17.2 MJ/kg DM for wheat, corn, barley, wheat bran, and soybean meal, respectively. The Enz addition increased the DE value of ingredients similarly, but the best response was observed for wheat (0.33 MJ/kg DM).