The In Vitro Adsorption Ability of Lactobacillus acidophilus NCFM to Benzo(a)pyrene in PM2.5

The objective of this work was to explore the ability of lactic acid bacteria strains to bind benzo(a)pyrene (B(a)P) existing in PM2.5. In this study, we examined the ability of Lactobacillus acidophilus NCFM to bind B(a)P in the simulated PM2.5 environment. Among the tested 5 strains, Lactobacillus acidophilus NCFM exhibited the best capacity to bind B(a)P, and its B(a)P binding percentage was 60.00%. Simulations of organic and inorganic systems which represent PM2.5 indicated that B(a)P could be absorbed by strain L. acidophilus NCFM. For the inorganic system of pH 5, L. acidophilus NCFM bound 92.74% B(a)P with a cell concentration of 1 × 1010 cfu/mL at 37°C for 8 hr. Regarding the organic system with pH 6, 73.00% B(a)P was bound by strain L. acidophilus NCFM after this bacterium was incubated at 37°C for 10 min. A quick B(a)P binding by this probiotic bacterium took place in the organic system. The removal of B(a)P from PM2.5 was significantly related to incubation time, cultivation temperature, pH, and cell concentration. Thus, our finding shows that long-term consumption of L. acidophilus NCFM is beneficial for the reduction of B(a)P towards the population who are exposed to PM2.5, although the ability of this bacterium to adsorb B(a)P is partly affected by the differences in the origin of PM2.5.

Aflatoxin B1 and Sterigmatocystin Binding Potential of Non-Lactobacillus LAB Strains

Research on the ability of lactic acid bacteria (LAB) to bind aflatoxin B1 (AFB1) has mostly been focusing on lactobacilli and bifidobacteria. In this study, the AFB1 binding capacities of 20 Enterococcus strains belonging to E. casseliflavus, E. faecalis, E. faecium, E. hirae, E. lactis, and E. mundtii, 24 Pediococcus strains belonging to species P. acidilactici, P. lolii, P. pentosaceus, and P. stilesii, one strain of Lactococcus formosensis and L.garviae, and 3 strains of Weissella soli were investigated in MRS broth at 37 °C at 0.2 µg/mL mycotoxin concentration. According to our results, among non-lactobacilli LAB, the genera with the best AFB1 binding abilities were genus Pediococcus, with a maximum binding percentage of 7.6% by P. acidilactici OR83, followed by genus Lactococcus. For AFB1 bio-detoxification purposes, beside lactobacilli, pediococci can also be chosen, but it is important to select a strain with better binding properties than the average value of its genus. Five Pediococcus strains have been selected to compare their sterigmatocystin (ST) binding abilities to AFB1 binding, and a 2–3-fold difference was obtained similar to previous findings for lactobacilli. The best strain was P. acidilactici OR83 with 18% ST binding capacity. This is the first report on ST binding capabilities of non-Lactobacillus LAB strains.

A chromatographic approach to development of 5-aminosalicylate/folic acid fixed-dose combinations for treatment of Crohn’s disease and ulcerative colitis

Medication adherence is an important factor in inflammatory bowel disease therapy, which includes regular supplementation of malabsorbed vitamins. Absorption of folic acid is limited due to the damaging of the gastrointestinal tract, which can increase the chances to develop megaloblastic anaemia and colorectal cancer. In this work, 5-aminosalicylates (mesalazine, balsalazide, sulfasalazine and olsalazine) and folic acid were characterized regarding their pharmacokinetic related properties (hydrophobicity, phospholipid and plasma protein binding) using the biomimetic chromatographic approach. Despite the high binding percentage of 5-aminosalicylates for human serum albumin (> 61.44%), results have shown that folic acid binding to human serum albumin protein is far greater (69.40%) compared to α1-acid-glycoprotein (3.45%). Frontal analysis and zonal elution studies were conducted to provide an insight into the binding of folic acid to human serum albumin and potential competition with 5-aminosalicylates. The analytical method for the simultaneous determination of assay in proposed fixed-dose combinations was developed and validated according to ICH Q2 (R1) and FDA method validation guidelines. Separation of all compounds was achieved within 16 min with satisfactory resolution (Rs > 3.67) using the XBridge Phenyl column (150 × 4.6 mm, 3.5 µm). High linearity (r > 0.9997) and precision (RSD < 2.29%) was obtained, whilst all recoveries were within the regulatory defined range by British (100.0 ± 5.0%) and United States Pharmacopeia (100.0 ± 10.0%).

Antioxidant and Glucose Lowering Effects of Hydroethanolic Extract of Baillonella toxisperma Pulp

Hyperglycemia and oxidative stress play an important role in the pathogenesis of diabetes. Their management is a key point in the prevention and treatment of this disease which is a potential cause of mortality in the world. We evaluated the antioxidant and glucose lowering effects of hydroethanolic extract of Baillonella toxisperma pulp. The total polyphenol and flavonoid contents were determined and the antioxidant activity was evaluated by 3 mechanisms: scavenging by DPPH•, ABTS• and NO• radicals; reducing property by MoO42+ and Fe3+ reduction; metal chelation by Cu2+ and Fe2+. Glucose adsorption capacity was evaluated followed by the capacity to promote insulino-sensitivity through glucose uptake by yeast and muscle cell assays. The hydroethanolic extract of B. toxisperma pulp possessed high total polyphenols and flavonoids; 459.55 µg Equivalent Gallic Acid/mg and 252.15 µg of Equivalent Catechin /mg respectively. This showed their ability to scavenge DPPH•, ABTS• and NO• radicals with SC50 of 3.49, 3.24 and 4.28 mg/ml respectively. The extract also reduced MoO42+ and Fe3+ and chelated Cu2+ through inhibiting their capacity to induce hemolysis with the IC50 of 3.49 mg/ml. The extract showed a high glucose binding capacity with a glucose binding percentage rise of 60 %. It increased yeast cell absorption of glucose with the increasing percentage varying from 42.97 to 56.62 %. In the muscle cells, after 30 min of administration of the extract, we also noted an increased glucose absorption with the percentage glucose reducing to 22 %. We demonstrated that hydroethanolic extract of B. toxisperma pulp possess antiradical, reducing, metal chelating, glucose binding and insulino-sensitivity promoting properties. These mechanisms imply that B. toxisperma pulp is both a good antioxidant and an antihyperglycemiant, thus a potential agent in the management of diabetes and its complications.

Synthesis of Novel Nicotinic Ligands with Multimodal Action: Targeting Acetylcholine α4β2, Dopamine and Serotonin Transporters

Nicotinic acetylcholine receptors (nAChRs), serotonin transporters (SERT) and dopamine transporters (DAT) represent targets for the development of novel nicotinic derivatives acting as multiligands associated with different health conditions, such as depressive, anxiety and addiction disorders. In the present work, a series of functionalized esters structurally related to acetylcholine and nicotine were synthesized and pharmacologically assayed with respect to these targets. The synthesized compounds were studied in radioligand binding assays at α4β2 nAChR, h-SERT and h-DAT. SERT experiments showed not radioligand [3H]-paroxetine displacement, but rather an increase in the radioligand binding percentage at the central binding site was observed. Compound 20 showed Ki values of 1.008 ± 0.230 μM for h-DAT and 0.031 ± 0.006 μM for α4β2 nAChR, and [3H]-paroxetine binding of 191.50% in h-SERT displacement studies, being the only compound displaying triple affinity. Compound 21 displayed Ki values of 0.113 ± 0.037 μM for α4β2 nAChR and 0.075 ± 0.009 μM for h-DAT acting as a dual ligand. Molecular docking studies on homology models of α4β2 nAChR, h-DAT and h-SERT suggested potential interactions among the compounds and agonist binding site at the α4/β2 subunit interfaces of α4β2 nAChR, central binding site of h-DAT and allosteric modulator effect in h-SERT.

Unraveling Hepcidin Plasma Protein Binding: Evidence from Peritoneal Equilibration Testing

Peptide hormone hepcidin regulates systemic iron metabolism and has been described to be partially bound to α2-macroglobulin and albumin in blood. However, the reported degree of hepcidin protein binding varies between <3% and ≈89%. Since protein-binding may influence hormone function and quantification, better insight into the degree of hepcidin protein binding is essential to fully understand the biological behavior of hepcidin and interpretation of its measurement in patients. Here, we used peritoneal dialysis to assess human hepcidin protein binding in a functional human setting for the first time. We measured freely circulating solutes in blood and peritoneal fluid of 14 patients with end-stage renal disease undergoing a peritoneal equilibration test to establish a curve describing the relation between molecular weight and peritoneal clearance. Calculated binding percentages of total cortisol and testosterone confirmed our model. The protein-bound fraction of hepcidin was calculated to be 40% (±23%). We, therefore, conclude that a substantial proportion of hepcidin is freely circulating. Although a large inter-individual variation in hepcidin clearance, besides patient-specific peritoneal transport characteristics, may have affected the accuracy of the determined binding percentage, we describe an important step towards unraveling human hepcidin plasma protein binding in vivo including the caveats that need further research.

Investigation of specific interactions between T7 promoter and T7 RNA polymerase by force spectroscopy using atomic force microscope

The specific recognition and binding of promoter and RNA polymerase is the first step of transcription initiation in bacteria and largely determines transcription activity. Therefore, direct analysis of the interaction between promoter and RNA polymerase in vitro may be a new strategy for promoter characterization, to avoid interference due to the cell's biophysical condition and other regulatory elements. In the present study, the specific interaction between T7 promoter and T7 RNA polymerase was studied as a model system using force spectroscopy based on atomic force microscope (AFM). The specific interaction between T7 promoter and T7 RNA polymerase was verified by control experiments, and the rupture force in this system was measured as 307.2 ± 6.7 pN. The binding between T7 promoter mutants with various promoter activities and T7 RNA polymerase was analyzed. Interaction information including rupture force, rupture distance and binding percentage were obtained in vitro, and reporter gene expression regulated by these promoters was also measured according to a traditional promoter activity characterization method in vivo. Using correlation analysis, it was found that the promoter strength characterized by reporter gene expression was closely correlated with rupture force and the binding percentage by force spectroscopy. These results indicated that the analysis of the interaction between promoter and RNA polymerase using AFM-based force spectroscopy was an effective and valid approach for the quantitative characterization of promoters.

PHYTOESTROGEN IN SEVERAL FRUITS AND LEAVES

Phytoestrogen is molecularly almost similar with and acts the same as estrogen and is found a lot in several fruits and leaves grown either in tropical or subtropical countries. However, the quantity of molecular contents are not yet known exactly. In menopause and andropause, people need substitution of estrogens as a replacement therapy of sex hormone, due to the significant hormone decline and impacted disturbance of several organ functions and thus progressively causing severe organ dysfunctions. The objective of this study was to know the estrogen content by analyzing extractions of pegaga, green clover leaves, tomato and papaya fruit of which certain communites have overviewed contents of the leaves and fruit The samples which were collected for this purpose used 10 times replication in four different groups: the pegaga, green clover leaves, tomato and papaya fruit. All these groups were divided into two (2) subgroups based on the process or subspecies. All samples were made as an infusion 1:4) w/v), and then extracted after centrifugation 1000xg for 15mnts, with 1:5 petroleum ether) v/v). After it was evaporated, each extraction then was kept dry-frozen at –20° C until the analysiswas performed. Solid phase Radioimmunoassay technique was used to identify the estrogen contents, up to a total of 80 samples The binding percentage of each sample was then interpolated on a logit-log paper to find out the real concentrations.14 The lowest estrogen level was found in fresh pegaga leaves extract (Mean+SD) was 47.9+5.5 pg/g, but in dry leaves extract the level was increased twice, about 96.1+11.2 pg/g. Meanwhile the estrogen level in fresh green clover leaves extract was 538.0+30.5 pg/g, more than ten times higher compared to fresh pegaga level, but twice lower than the estrogen level compared to dry green clover leaves extract, which was 1068.0+97.2 pg/g. In the fruit group, the fibrin part of tomatoes had more or less the same estrogen content compared with Thai papaya subspecies, 1037.0+37.7 pg/g and 1175.0+67.7 pg/g, respectively. On the other hand, it was noted that the inner part/fibrin part of tomato had a higher estrogen level of four (4) times compared to the outer part which was 315.0+18.4 pg/g. While it was noted that local Java papaya besides being not so sweet, the estrogen level was also not as high or the same as that found in fresh green clover which is 530.1+50.7 pg/g and 538.0+30.5 pg/g. Based on this study so far, it can be concluded that semanggi/green clover, tomato and papaya could be suggested as a replacement therapy for certain people who are considered to have reduced estrogen content, except that pegaga leaves are not recommended. The last mentioned plant besides that it is difficult to obtain, its estrogen content is also very low.