Anti-Helicobacter pylori Activity of a Lactobacillus sp. PW-7 Exopolysaccharide

Helicobacter pylori is a cause of gastric cancer. We extracted the exopolysaccharide (EPS) of Lactobacillus plajomi PW-7 for antibacterial activity versus H. pylori, elucidating its biological activity and structural characteristics. The minimum inhibitory concentration (MIC) of EPS against H. pylori was 50 mg/mL. Disruption of the cell membranes of pathogenic bacteria by EPS was indicated via the antibacterial mechanism test and confirmed through electron microscopy. EPS also has antioxidant capacity. The IC50 of EPS for 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide anions, and hydroxyl radicals were 300 μg/mL, 180 μg/mL, and 10 mg/mL, respectively. The reducing power of EPS was 2 mg/mL, equivalent to 20 μg/mL of ascorbic acid. EPS is a heteropolysaccharide comprising six monosaccharides, with an approximate molecular weight of 2.33 × 104 Da. Xylose had a significant effect on H. pylori. EPS from L. plajomi PW-7 showed potential as an antibacterial compound and antioxidant, laying a foundation for the development of EPS-based foods.

The Regulation of Bone Metabolism and Disorders by Wnt Signaling

Wnt, a secreted glycoprotein, has an approximate molecular weight of 40 kDa, and it is a cytokine involved in various biological phenomena including ontogeny, morphogenesis, carcinogenesis, and maintenance of stem cells. The Wnt signaling pathway can be classified into two main pathways: canonical and non-canonical. Of these, the canonical Wnt signaling pathway promotes osteogenesis. Sclerostin produced by osteocytes is an inhibitor of this pathway, thereby inhibiting osteogenesis. Recently, osteoporosis treatment using an anti-sclerostin therapy has been introduced. In this review, the basics of Wnt signaling, its role in bone metabolism and its involvement in skeletal disorders have been covered. Furthermore, the clinical significance and future scopes of Wnt signaling in osteoporosis, osteoarthritis, rheumatoid arthritis and neoplasia are discussed.

The use of nanofiltration membranes for the fractionation of polyphenols from grape pomace extracts

Filtration experiments in batch concentration mode (with recycling of the retentate stream) of grape pomace extract were performed in laboratory filtration membrane equipment by using nine commercial nanofiltration (NF) membranes with an approximate molecular weight cut-off (MWCO) of 1000‒150 Da. The filtration experiments of the selected pomace extract were performed by modifying the most important operating variables: transmembrane pressure, tangential velocity, temperature, and the nature and MWCO of the membranes. The evolution of the cumulative permeate volumes and permeate fluxes with processing time was analyzed till a volume reduction factor (VRF) of 10 was reached. The effect of the mentioned operating conditions was discussed. The effectiveness of the filtration treatments was determined by the evaluation of the rejection coefficients for several families of polyphenols. Membranes possessing MWCO between 1000 and 500 Da were able to quantitatively recover polymeric proanthocyanidins in the concentrate stream and separate them from phenols that passed through the membrane into the permeate stream. On the other hand, the 600 to 300 Da membranes could also be used for the fractionation of monomeric phenolic families. The membranes were able to partially remove the anthocyanin fragments of phenolic acid derivatives and flavonols in the concentrate stream and at the same time.

Isolation and Purification of a Cyclooxygenase-2 from the Blood of a Patient Suffering from Rheumatoid Arthritis and Studying the Effect of Natural Products of the Soapwort on the Activity of Purified Enzyme

In this paper to isolate and study the properties of the cyclooxygenase-2 (EC: 1.14.99.1) enzyme in the blood of a patient suffering from rheumatoid arthritis and study the effect of natural products of the Soapwort on the activity of purified enzyme. The study involves taking 30 ml of blood from an adult woman 40 years old, who suffers from rheumatoid arthritis disease for 13 years. Serum is separated and subjected to a series of purification processes including: precipitation by ammonium sulfate, filtration by centrifugation radiator, dialysis in presence of ammonium bicarbonate, separation using the technology of ion exchange, lipholization and then estimating approximate molecular weight of the enzyme using gel filtration technique and sodium dodecyl sulfate (SDS)-page polyacrylamide gel electrophoresis. The study also includes isolating the natural products of Soapwort plant and study the effect of isolated natural products on the activity of the purified enzyme. The result of the study indicates that cycloxygenase-2 has an approximate molecular weight of 71.5 kDa and that the extracted oil of the Soapwort has a negative impact on the activity of the enzyme (r= -0.824; P=0.006), while flavonoids and Saponin have no such impact (r= -0.565; P=0.113; r= -0.634; P=0.067 respectively).

Jungle Honey Enhances Immune Function and Antitumor Activity

Jungle honey (JH) is collected from timber and blossom by wild honey bees that live in the tropical forest of Nigeria. JH is used as a traditional medicine for colds, skin inflammation and burn wounds as well as general health care. However, the effects of JH on immune functions are not clearly known. Therefore, we investigated the effects of JH on immune functions and antitumor activity in mice. Female C57BL/6 mice were injected with JH (1 mg/mouse/day, seven times intra-peritoneal). After seven injections, peritoneal cells (PC) were obtained. Antitumor activity was assessed by growth of Lewis Lung Carcinoma/2 (LL/2) cells. PC numbers were increased in JH-injected mice compared to control mice. In Dot Plot analysis by FACS, a new cell population appeared in JH-injected mice. The percent of Gr-1 surface antigen and the intensity of Gr-1 antigen expression of PC were increased in JH-injected mice. The new cell population was neutrophils. JH possessed chemotactic activity for neutrophils. Tumor incidence and weight were decreased in JH-injected mice. The ratio of reactive oxygen species (ROS) producing cells was increased in JH-injected mice. The effective component in JH was fractionized by gel filtration using HPLC and had an approximate molecular weight (MW) of 261. These results suggest that neutrophils induced by JH possess potent antitumor activity mediated by ROS and the effective immune component of JH is substrate of MW 261.

Extraction and Antioxidant Activity of Collagen from the Chinese Soft-Shelled Turtle (Pelodiscus sinensis)

Collagen is regarded as one of the most useful biomaterials. Collagen was extracted from wild Chinese soft-shelled turtles (Pelodiscus sinensis) and its molucular patterns and antioxidant activities were investigated. The SDS-PAGE analysis revealed that the collagen had three α bands with an approximate molecular weight of 400-410 kDa, which was similar to type V collagen with a molecular pattern of α1(V)α2(V)α3(V) or α1(V)[α2(V)]2. The antioxidant activity of collagen were examined by the radical-scavenging activity (RSA) assay and reducing power (RP) methods respectively. In the RSA assay, the DPPH free radical scavenging capability of the sample collagen was positively correlated with its concentration with a IC50 of 1.95mg/ml. The same trend was found in the RP assay. The reducing power of the sample collagen was also dependent with its dose employed.

Characterization of a Fourth Tungsten-Containing Enzyme from the Hyperthermophilic Archaeon Pyrococcus furiosus

ABSTRACT Pyrococcus furiosus grows optimally near 100°C using peptides and carbohydrates as carbon sources, and it reduces elemental sulfur (S0), if present, to H2S. Tungsten (W), an element rarely used in biology, is required for optimal growth, and three different tungsten-containing enzymes have been previously purified from this organism. They all oxidize aldehydes of various types and are thought to play primary roles in the catabolism of sugars or amino acids. Here, the purification of a fourth tungsten-containing enzyme, termed WOR 4, from cell extracts of P. furiosus grown with S0 is described. This was achieved by monitoring through multiple chromatography steps the W that is not associated with the three characterized tungstoenzymes. The N-terminal sequence of WOR 4 and the approximate molecular weight of its subunit determined electrophoretically (69,000) correspond to the product of an ORF (PF1961, wor4) present in the complete genome sequence of P. furiosus. WOR 4 is a homodimer and contains approximately one W, three Fe, three or four acid-labile sulfide, and one Ca atom per subunit. The visible and electron paramagnetic resonance spectra of the oxidized and reduced enzyme indicate the presence of an unusual iron-sulfur chromophore. WOR 4 does not oxidize aliphatic or aromatic aldehydes or hydroxy acids, nor does it reduce keto acids. Consistent with prior microarray data, the protein could not be purified from P. furiosus cells grown in the absence of S0, suggesting that it may have a role in S0 metabolism.

Effect of ABA and GA3 on protein mobilization in embryos and cotyledons of angico [Anadenanthera peregrina (L.) speg] seeds during germination

In this work, a woody species [A. peregrina (L.) Speg.] was studied in order to observe the effect of ABA and GA3 at the biochemical level during the process of seed germination. Embryos incubated in sucrose solution containing ABA and/or GA3 were analyzed through SDS-PAGE to observe the mobilization pattern of storage proteins during the beginning of germination. Cotyledons isolated from seeds incubated in aqueous solutions containing ABA and/or GA3, were also analyzed through SDS-PAGE and by PAGE/Activity Gels (polyacrylamide gels copolymerized with substrate for enzymes) to observe the mobilization pattern of storage proteins and protease activity after the beginning of the germination. Results of these experiments show that ABA blocks protein mobilization by inhibiting protease activity in cotyledons. This inhibition is not sufficient to prevent germination showing that the effect of ABA on germination is not dependent on protease activity. The blockage of storage protein mobilization was also observed in embryos, but no protease activity inhibition was clearly detected. ABA was able to induce the synthesis of proteins in cotyledons but not in embryos. A polypeptide with an approximate molecular weight of 17 kD, was degraded within 6 hours in control embryos, but this degradation was blocked by ABA and GA3. Using the same concentrations of ABA and GA3 on embryos and cotyledons, the effect of ABA was counteracted by GA3 in embryos, but not in cotyledons. Although the effects of ABA and GA3 were not so different from those shown in the literature, the behavior of 17 kD-polypeptide contradicts these reports suggesting that specific studies should be performed.

Anti-oxidant enzymes in Cryptosporidium parvum oocysts

Oocysts of Cryptosporidium parvum showed relatively low levels of SOD activity. The SOD which had a pI of 4.8 and an approximate molecular weight of 35 kDa appeared to be iron dependent. Catalase, glutathione transferase, glutathione reductase and glutathione peroxidase activity could not be detected, nor could trypanothione reductase. No NADH or NADPH oxidase activity could be detected, nor could peroxidase activity be demonstrated using o-dianisidine, guaiacol, NADPH or NADH as co-substrates. However, an NADPH-dependent H2O2 scavenging system was detected in the insoluble fraction.

Isolation and Purification of Enterocin EL1, a Bacteriocin Produced by a Strain of Enterococcus faecium†

A meat isolate, identified as Enterococcus faecium L1, was found to produce a bacteriocin designated enterocin EL1 Enterocin EL1 was active against a narrow spectrum of microorganisms, inhibiting all tested strains of Listeria. Identification of the producer strain was determined phenotypically by biochemical and morphological tests. Enterocin EL1 was heat stable, sensitive to several proteolytic enzymes, and stable from pH 2 to 11. Adsorption of the bacteriocin to producer cells was dependent on ionic interaction of the bacteriocin and the cell surface at various pHs. By changing the pH of the extraction buffer, enterocin EL1 was extracted from E. faecium L1 cells in a concentrated form. Enterocin EL1 isolated by cell extraction was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a protein with an approximate molecular weight of 2,300. Partially purified enterocin EL1 added to sensitive cells of Listeria ivanovii was bactericidal; however, the bacteriocin did not inhibit the producer strain L1.