Does the addition of chlorhexidine to glass ionomer cements influence its antimicrobial effect and survival rate? A systematic review.

PURPOSE: To evaluate the influence of the addition of chlorhexidine on the antimicrobial effect and on the survival of restorations performed with glass ionomer cement. METHODS: Nine databases were used to search for randomized clinical trials that compared the survival rate and the antimicrobial effect of glass ionomer cement (GIC) restorations with and without the incorporation of chlorhexidine (CHX), without restrictions on year or language. Cochrane Collaboration's Risk of Bias 2 was used to assess the risk of bias. The GRADE approach was used to assess the certainty of evidence. RESULTS: From 593 studies found, seven met the inclusion criteria. The concentration of CHX varied between 0.5 and 2%. In general, the addition of CHX to GIC promoted reductions in Streptococcus mutans and Lactobacillus acidophilus burdens when compared to those without CHX. No study showed a difference in the survival of restorations between GIC with CHX and conventional GIC. Individual risk of bias varied from low to high and the certainty of evidence was classified as very low. CONCLUSIONS: Based on a very low level of certainty, the evidence suggests that the incorporation of CHX in GIC might improve the antimicrobial effects for a short time, in addition to having little influence on the survival of the restoration.

Increased expression of clp genes in Lactobacillus delbrueckii UFV H2b20 exposed to acid stress and bile salts.

The ability to survive in harsh environments is an important criterion to select potential probiotics strains. The objective of this study was to identify and carry out phylogenetic and expression analysis by quantitative real-time PCR of the clpP, clpE, clpL and clpX genes in the probiotic strain Lactobacillus delbrueckii UFV H2b20 exposed to the conditions prevailing in the gastrointestinal tract (GIT). Phylogenetic trees reconstructed by Bayesian inference showed that the L. delbrueckii UFV H2b20 clpP, clpL and clpE genes and the ones from L. delbrueckii ATCC 11842 were grouped. The exposure of cells to MRS broth of pH 3.5 for 30 and 60 min resulted in an increased expression of the four genes. Exposure of the L. delbrueckii UFV H2b20 cells for 30 and 60 min to MRS broth containing 0.1% bile salts increased the expression of the clpP and clpE genes, while the expression level of the clpL and clpX genes increased only after 30 min of exposure. The involvement of the studied genes in the responses to acid stress and bile salts suggests a possible central role of these genes in the survival of L. delbrueckii UFV H2b20 during the passage through the GIT, a characteristic necessary for probiotic strains.

Survival of Lactobacillus delbrueckii UFV H2b20 in fermented milk under simulated gastric and intestinal conditions.

The survival of Lactobacillus delbrueckii UFV H2b20 was assessed in fermented milk, both during the storage period and after exposure to simulated gastric and intestinal juices, as well the detection of the gene fbpA involved in adherence to human gastrointestinal tract. L. delbrueckii UFV H2b20 remained stable and viable for 28 days under refrigerated storage conditions. After one day of storage, that strain exhibited a one-log population reduction following exposure in tandem to simulated gastric and intestinal juices. After 14 days of storage, a two-log reduction was observed following 90 min of exposure to the simulated gastric conditions. However, the strain did not survive following exposure to the simulated intestinal juice. The observed tolerance to storage conditions and resistance to the simulated gastric and intestinal conditions confirm the potential use of L. delbrueckii UFV H2b20 as a probiotic, which is further reinforced by the detection of fbpA in this strain.

Genes involved in lactose catabolism and organic acid production during growth of Lactobacillus delbrueckii UFV H2b20 in skimmed milk.

There are three main reasons for using lactic acid bacteria (LAB) as starter cultures in industrial food fermentation processes: food preservation due to lactic acid production; flavour formation due to a range of organic molecules derived from sugar, lipid and protein catabolism; and probiotic properties attributed to some strains of LAB, mainly of lactobacilli. The aim of this study was to identify some genes involved in lactose metabolism of the probiotic Lactobacillus delbrueckii UFV H2b20, and analyse its organic acid production during growth in skimmed milk. The following genes were identified, encoding the respective enzymes: ldh - lactate dehydrogenase, adhE - Ldb1707 acetaldehyde dehydrogenase, and ccpA-pepR1 - catabolite control protein A. It was observed that L. delbrueckii UFV H2b20 cultivated in different media has the unexpected ability to catabolyse galactose, and to produce high amounts of succinic acid, which was absent in the beginning, raising doubts about the subspecies in question. The phylogenetic analyses showed that this strain can be compared physiologically to L. delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis, which are able to degrade lactose and can grow in milk. L. delbrueckii UFV H2b20 sequences have grouped with L. delbrueckii subsp. bulgaricus ATCC 11842 and L. delbrueckii subsp. bulgaricus ATCC BAA-365, strengthening the classification of this probiotic strain in the NCFM group proposed by a previous study. Additionally, L. delbrueckii UFV H2b20 presented an evolutionary pattern closer to that of probiotic Lactobacillus acidophilus NCFM, corroborating the suggestion that this strain might be considered as a new and unusual subspecies among L. delbrueckii subspecies, the first one identified as a probiotic. In addition, its unusual ability to metabolise galactose, which was significantly consumed in the fermentation medium, might be exploited to produce low-browning probiotic Mozzarella cheeses, a desirable property for pizza cheeses.

Genes involved in protein metabolism of the probiotic lactic acid bacterium Lactobacillus delbrueckii UFV H2b20.

A basic requirement for the prediction of the potential use of lactic acid bacteria (LAB) in the dairy industry is the identification of specific genes involved in flavour-forming pathways. The probiotic Lactobacillus delbrueckii UFV H2b20 was submitted to a genetic characterisation and phylogenetic analysis of genes involved in protein catabolism. Eight genes belonging to this system were identified, which possess a closely phylogenetic relationship to NCFM strains representative, as it was demonstrated for oppC and oppBII, encoding oligopeptide transport system components. PepC, PepN, and PepX might be essential for growth of LAB, probiotic or not, since the correspondent genes are always present, including in L. delbrueckii UFV H2b20 genome. For pepX gene, a probable link between carbohydrate catabolism and PepX expression may exists, where it is regulated by PepR1/CcpA-like, a common feature between Lactobacillus strains and also in L. delbrueckii UFV H2b20. The well conserved evolutionary history of the ilvE gene is evidence that the pathways leading to branched-chain amino acid degradation, such as isoleucine and valine, are similar among L. delbrueckii subsp. bulgaricus strains and L. delbrueckii UFV H2b20. Thus, the involvement of succinate in flavour formation can be attributed to IlvE activity. The presence of aminopeptidase G in L. delbrueckii UFV H2b20 genome, which is absent in several strains, might improve the proteolytic activity and effectiveness. The nucleotide sequence encoding PepG revealed that it is a cysteine endopeptidase, belonging to Peptidase C1 superfamily; sequence analysis showed 99% identity with L. delbrueckii subsp. bulgaricus ATCC 11842 pepG, whereas protein sequence analysis revealed 100% similarity with PepG from the same organism. The present study proposes a schematic model to explain how the proteolytic system of the probiotic L. delbrueckii UFV H2b20 works, based on the components identified so far.

Effects of maternal ageing and dietary antioxidant supplementation on ovulation, fertilisation and embryo development in vitro in the mouse.

The present study aims to ascertain whether dietary supplementation with a mixture of vitamins C and E may prevent the maternal-age-associated decrease in both the number of ovulated oocytes after exogenous ovarian stimulation and embryo development in vitro in the mouse. Experimental females were fed a standard diet supplemented with i) high doses of vitamins C and E from the first day of weaning until 12 or 40 weeks of age; or ii) moderate doses of vitamins C and E from the first day of weaning until 12 weeks of age or from 22 to 33 weeks of age. The age-related reduction in ovulation rate was partially prevented by supplementing diet with high doses of vitamins C and E from the first day of weaning. Shorter periods of treatment and lower doses of vitamins C and E were also efficient in preventing the maternal-age-associated reduction in ovulation rate after exogenous ovarian stimulation. No effect of maternal diet on fertilisation and embryo development was observed until the blastocyst stage. Although any extrapolation to human fertility should be made with caution, these findings may have direct implications for preventing or delaying maternal-age-associated infertility in humans.

Effect of intracellular Ca2+ chelation with the acetoxymethyl ester-derived form of bis(o-aminophenoxy)ethane-N,N,N,N',N'-tetraacetic acid on meiotic division and chromosomal segregation in mouse oocytes.

PURPOSE: Our purpose was to ascertain the effect of intracellular Ca2+ chelation on the chromosomal distribution and segregation of mouse oocytes during maturation in vitro. METHODS: Germinal vesicle oocytes were loaded with the acetoxymethyl ester-derived form of bis(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Chromosomal distribution and segregation of control and BAPTA-AM-treated metaphase II (MII) oocytes were evaluated at 16 hr, and intracellular ATP content at 0, 1, and 16 hr after BAPTA-AM loading. RESULTS: BAPTA-AM treatment decreased (P < or = 0.05) the potential for in vitro maturation, increased (P < or = 0.0001) the percentage of oocytes displaying an abnormal distribution of metaphase II chromosomes in the meiosis II spindle and aneuploidy, and decreased (P < or = 0.005) the ATP content at 0, 1, and 16 hr of culture compared to the control groups. CONCLUSIONS: These findings raise some concern about any other condition/drug that may directly or indirectly decrease the intracellular Ca2+ concentration in human oocytes.

Ascorbate-supplemented media in short-term cultures of human embryos.

The present study aimed to evaluate whether ascorbate, a reactive oxygen species (ROS) scavenger, can improve fertilization and development of human embryos in vitro when added to the simple salt solution human tubal fluid (HTF) or the complex tissue culture medium Ham's F-10, which contains iron and copper in its formulation. Human oocytes, spermatozoa and embryos from 83 infertile IVF couples were randomly allocated and cultured in the presence or absence of 62.5 microM ascorbate in HTF medium (39 couples) or Ham's F-10 medium (44 couples). No significant effect of ascorbate on fertilization, number of cells and embryo grade per embryo on days 2 and 3 after insemination, or percentage of embryos showing developmental block on day 3 (those embryos that were still at the 2-cell stage) was observed when data were analysed together or divided into several groups according to the cause of infertility, quality of semen sample used for insemination and women's age in either of the two media tested. Despite these results, a positive effect of ascorbate on fertilization and embryo development in vitro cannot be totally ruled out until the effects of other, non-physiological concentrations of ascorbate and longer-term embryo cultures (to the blastocyst stage) have been tested.