Immunomodulatory function and in vivo properties of pediococcus pentosaceus OZF, a promising probiotic strain

Some of the important properties of probiotics are the ability to survive during gastrointestinal transit and to modulate the immune functions. The objectives of the reported study were to assess in vivo gastrointestinal survival of orally administered Pediococcus pentosaceus OZF using an animal model BALB/c mice, and to examine its effects on the immune response. Following oral administration to mice, the ability of Pediococcus pentosaceus OZF to pass and survive through the mouse gastrointestinal system was investigated by analyzing the recovery of the strain in fecal samples. Microbiological and polymerase chain reaction (PCR) methods proved that the strain OZF could overcome specific conditions in the gastrointestinal tract of mice and reach the intestine alive after ingestion. To observe the effect of oral administration on immune response, IL-6, IL-12 and IFN-γ were measured by ELISA, and the strain OZF was found to cause increases in IL-6 synthesis in regularly fed mice. However, stimulation was carried out with various concentrations of bacterial ssDNA and heat killed cells of Pediococcus pentosaceus OZF. The heat killed cells of the strain OZF were shown to produce IFN- γ independently from IL-12. On the other hand, a significant difference between control and experimental group was noticed when lipopolysaccharide, a TLR4 (toll like receptor) ligand, was used. Overall, Pediococcus pentosaceus OZF may be a valuable probiotic strain for therapeutic uses. Nevertheless, further studies on the mechanisms of immunomodulatory effect will allow for better clarification of the immune functions of this strain. © Springer-Verlag Berlin Heidelberg and the University of Milan 2012

The HELLP syndrome: Clinical issues and management. A Review

<p>Abstract</p> <p>Background</p> <p>The HELLP syndrome is a serious complication in pregnancy characterized by haemolysis, elevated liver enzymes and low platelet count occurring in 0.5 to 0.9% of all pregnancies and in 10–20% of cases with severe preeclampsia. The present review highlights occurrence, diagnosis, complications, surveillance, corticosteroid treatment, mode of delivery and risk of recurrence.</p> <p>Methods</p> <p>Clinical reports and reviews published between 2000 and 2008 were screened using Pub Med and Cochrane databases.</p> <p>Results and conclusion</p> <p>About 70% of the cases develop before delivery, the majority between the 27th and 37th gestational weeks; the remainder within 48 hours after delivery. The HELLP syndrome may be complete or incomplete. In the Tennessee Classification System diagnostic criteria for HELLP are haemolysis with increased LDH (> 600 U/L), AST (≥ 70 U/L), and platelets < 100·10⁹/L. The Mississippi Triple-class HELLP System further classifies the disorder by the nadir platelet counts. The syndrome is a progressive condition and serious complications are frequent. Conservative treatment (≥ 48 hours) is controversial but may be considered in selected cases < 34 weeks' gestation. Delivery is indicated if the HELLP syndrome occurs after the 34th gestational week or the foetal and/or maternal conditions deteriorate. Vaginal delivery is preferable. If the cervix is unfavourable, it is reasonable to induce cervical ripening and then labour. In gestational ages between 24 and 34 weeks most authors prefer a single course of corticosteroid therapy for foetal lung maturation, either 2 doses of 12 mg betamethasone 24 hours apart or 6 mg or dexamethasone 12 hours apart before delivery. Standard corticosteroid treatment is, however, of uncertain clinical value in the maternal HELLP syndrome. High-dose treatment and repeated doses should be avoided for fear of long-term adverse effects on the foetal brain. Before 34 weeks' gestation, delivery should be performed if the maternal condition worsens or signs of intrauterine foetal distress occur. Blood pressure should be kept below 155/105 mmHg. Close surveillance of the mother should be continued for at least 48 hours after delivery.</p

Cloning and expression of a plasmid-linked pediocin determinant trait of Pediococcus acidilactici F

Plasmid DNA from Pediococcus acidilactici F was prepared by lysozyme-mutanolysin method and purified by cesium chloride-ethidium bromide (CsCl-EtBr) density gradient ultracentrifugation. Agarose gel electrophoresis of plasmid DNA and plasmid-curing experiments suggested that bacteriocin activity was harboured on a small plasmid of approximately 9.1 kb (kilobasepair) in Pediococcus acidilactici F. Plasmid encoding bacteriocin production in P. acidilactici F was examined for restriction enzyme cleavage patterns and its map has been constructed. An Escherichia coli strain transformed with the recombinant plasmid, pQE322, produced and, most probably, secreted pediocin F

Comparison of three methods for determination of protein concentration in lactic acid bacteria for proteomics studies

Finding the best method of cell lysis and extraction of protein from the lysed cells is the key step in detection and identification of extra- and intra-cellular proteins in all applications of proteomics. To develop an optimized protein extraction protocol, Enterococcus faecalis V583, Lactococcus lactis NIZO 0900 and Pediococcus pentosaceus OZF strains, respectively, belonging to each genus of Enterococcus, Lactococcus and Pediococcus were used as a representative cells in a study of lactic acid bacteria (LAB). This report covers the use and comparison of three different protein extraction methods including sonication, centrifugation and rupture by glass beads (FastPrep) to get a better understanding about which methods give better extract quality and higher amount of proteins when applied to one dimensional (1D) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) and for subsequent analysis by two dimensional (2D)-PAGE. The results clearly showed that, all methods can be used to lyse LAB strains. However, a six fold greater amount of protein was obtained when FastPrep was applied to lyse LAB cells. Our results also indicate that, this fast and easy extraction method allows more spot-abundant polyacrylamide gels. More clear and consistent strips were detected by SDS-PAGE when proteins were extracted by FastPrep. These results testify to the suitability of FastPrep protein extraction protocols for 2D proteomic studies of representative strains of LAB.Key words: FastPrep, sonication, centrifugation, lactic acid bacteria (LAB)

Analysis of the genetic determinant for production of the pediocin P of Pediococcus pentosaecus Pep 1

Pediococcus pentosaceus Pep1 is a vacuum-packaged Turkish sausage isolate which produces a potentially novel bacteriocin of the pediocin (anti-Listeria) family of peptides designated as pediocin P. Curing experiments and plasmid profile analysis indicated that both bacteriocin immunity and production determinants were linked and encoded by 9.0 MDa plasmid, pHD1.0. Attempts to transform purified plasmid pHD1.0 into recipient Escherichia coli JM109 cells by electroporation were successful but none of the E. coli JM109 cells were able to express and/or release pediocin P. However. P. pentosaceus PC, a plasmid-cured variant of P. pentosaceus Pep1 was successfully transformed with pHD1.0 by electroporation and Bac(-)Bac(s) P. pentosaceus PC cells restarted to express and/or released pediocin P again as indicted by the presence of zone of growth inhibition of L. plantarum NCDO 955 around colonies