1,721,066 research outputs found
The Epithelial Barrier Model Shows That the Properties of VSL#3 Depend from Where It Is Manufactured
No full textBackground: VSL#3 has been extensively investigated and is currently recommended for the prevention and treatment of chronic pouchitis and ulcerative colitis. Nonetheless, in vitro and in vivo studies have recently shown variability in the VSL#3 efficacy often attributed to the manufacturing process. Objective: The aim was to comparatively study the in vitro effects of two VSL#3 preparations produced in different sites (named US- and Italy-made VSL#3) on CaCo-2 epithelial barrier model in terms of trans-epithelial electrical resistance (TEER), dextran flux and expression of TJ proteins i.e. zonulin-1 (ZO-1) and occludin, in the absence or presence of a heat stress-related damage of monolayer. Methods: TEER was evaluated on CaCo-2 differentiated monolayers. Epithelial permeability of polarized monolayers was assessed by measuring the FITC-labeled dextran flux from the apical to basolateral chambers. ZO-1/occludin levels were analyzed by western blotting analysis. A set of experiments was performed to compare the effects of both VSL#3 on TEER values, dextran flux and ZO- 1/occludin expression in CaCo-2 monolayers after heat stress exposure. Results: US- and Italy-made VSL#3 have opposing effects on TEER values, dextran flux, and ZO- 1/occludin expression, being all these parameters negatively influenced just by Italy-made product. US-made probiotic did not affect baseline TEER, dextran flux and ZO-1 expression and strongly increased occludin levels. Of note, pre-treatment of monolayer with US-made VLS#3, but not Italy-made product, totally prevented the heat-induced epithelial barrier integrity loss. Conclusion: Our data trigger the need for reassessing efficacy or safety of the Italy-made VSL#3 considering intestinal epithelial barrier plays an important role in maintaining host health
Effect of vaginal probiotic lactobacilli on in vitro-induced sperm lipid peroxidation and its impact on sperm motility and viability
No full text"A combination of three selected strains of lactobacilli (Lactobacillus brevis [CD2], L. salivarius [FV2], and L. plantarum [FV9]), whose effectiveness in treating bacterial vaginosis in the form of vaginal tablets has been reported recently, prevented sperm lipid peroxidation that was induced in vitro by a ferrous ion promoter, thus preserving sperm motility and viability. This finding suggests the potential of vaginal probiotic lactobacilli for protecting human spermatozoa from radical oxygen species in the presence of vaginal disorders, thereby improving the fertilization potential of the female host..
Induction of apoptosis by quercetin: different response of human chronic myeloid (K562) and acute lymphoblastic (HSB-2) leukemia cells
No full textThis work shows that 25 lM quercetin caused a marked inhibition of K562 cells growth together with a mild cytotoxicity,
while HSB-2 cells were practically unaffected. Moreover, quercetin induced caspase-3 and cytochrome c-dependent apoptosis
almost exclusively in the former cell line. Exposure of K562 cells to quercetin caused also a significant increase of cells in G2/
M phase that reached the maximum peak at 24 h (4-fold with respect to the basal value). The major sensitivity exhibited by
K562 cells was only in part imputable to their higher glutathione content, as compared to HSB-2 cells, thus confirming
previous reports describing the formation of intracellular quercetin–thiol toxic adducts in cells exposed to the flavonoid. In fact,
after induction of intracellular glutathione increase we detected in both cell lines a significant rise of apoptotic cells, again more
marked in K562 cells. By contrast, glutathione-depleted cells, failed to show a decrease of apoptosis in both cell lines, thus
contradicting our previous findings and literature data. Since the yet unresolved question about the anti-oxidant or the prooxidant
capacity of quercetin, we investigated which of these two properties worked in our experimental model. Interestingly,
not only quercetin did not produce reactive oxygen species but also prevented their formation, as observed in cells exposed to
the oxidizing agent ter-butylhydroperoxide, acting as an efficient oxygen radicals scavenger. This result indicates that quercetin
exhibited, in these cell lines, anti-oxidant more than pro-oxidant ability
Biological control of vaginosis to improve reproductive health
No full textAbstract
The human vaginal microbiota plays an important role in the maintenance of a woman's health, as well as of her partner's and newborns'. When this predominantly Lactobacillus community is disrupted, decreased in abundance and replaced by different anaerobes, bacterial vaginosis (BV) may occur. BV is associated with ascending infections and obstetrical complications, such as chorioamnionitis and preterm delivery, as well as with urinary tract infections and sexually transmitted infections. In BV the overgrowth of anaerobes produces noxious substances like polyamines and other compounds that trigger the release of pro-inflammatory cytokines interleukin (IL)-1 β and IL-8. BV can profoundly affect, with different mechanisms, all the phases of a woman's life in relation to reproduction, before pregnancy, during fertilization, through and at the end of pregnancy. BV can directly affect fertility, since an ascending dissemination of the involved species may lead to tubal factor infertility. Moreover, the increased risk of acquiring sexually transmitted diseases contributes to damage to reproductive health. Exogenous strains of lactobacilli have been suggested as a means of re-establishing a normal healthy vaginal flora. Carefully selected probiotic strains can eliminate BV and also exert an antiviral effect, thus reducing viral load and preventing foetal and neonatal infection. The administration of beneficial microorganisms (probiotics) can aid recovery from infection and restore and maintain a healthy vaginal ecosystem, thus improving female health also in relation to reproductive health
Erk-dependent cytosolic phospholipase A2 activity is induced by CD95 ligand cross-linking in the mouse derived Sertoli cell line TM4 and is required to trigger apoptosis in CD95 bearing cells
No full textIn the present study we demonstrated that CD95L cross-linking generated reverse signalling in the mouse derived Sertoli cell line TM4. Treatment of TM4 cells with mAb anti-CD95L induced activation of the cytosolic phospholipase A2 (cPLA2). Cytosolic PLA2 activation was controlled by the MAPK pathway as indicated by the ability of the specific MEK inhibitor, PD098059, to abolish cPLA2 activation. In addition, Western blot experiments showed a rapid increase in phosphorylated Erk1/2 following CD95L cross-linking, while no effect on the phosphorylation of other MAPK, p38 or JNK, was observed. CD95L cross-linking by mAb increased the levels of soluble CD95L and apoptotic activity of TM4 cell supernatants, which was blocked by co-incubation with the PLA2 inhibitor, AACOCF3 or PD098059. Finally, pre-treatment of TM4 cells with AACOCF3 or PD098059 completely abolished TM4-induced apoptosis of Jurkat T cells, thus indicating that the Erk/cPLA2 pathway is required for CD95L-induced apoptosis
Increase of skin-ceramide levels in aged subjects following a short-term topical application of bacterial sphingomyelinase from Streptococcus thermophilus
No full textSeveral studies have demonstrated that ceramides play an essential role in both the barrier and water-holding functions of healthy stratum corneum, suggesting that the dysfunction of the stratum corneum associated with ageing as well that observed in patients with several skin diseases could result from a ceramide deficiency. In a previous study our group reported a significant increase in skin ceramide levels in healthy subjects after treatment in vivo with a cream containing a preparation of Streptococcus thermophilus. The presence of high levels of neutral sphingomyelinase activity in this organism was responsible for the observed increase of stratum corneum ceramide levels, thus leading to an improvement in barrier function and maintenance of stratum corneum flexibility. The aim of the present work is to investigate the effects of the topical treatment of a Streptococcus thermophilus-containing cream on ceramide levels of stratum corneum of healthy elderly women. The ceramide levels, transepidermal water loss and capacitance were evaluated on stratum corneum sheets from the forearms of 20 healthy female subjects treated with a base cream or the same cream containing a sonicated preparation of the lactic acid bacterium Streptococcus thermophilus. A 2-week topical application of a sonicated Streptococcus thermophilus preparation led to significant and relevant increase of stratum corneum ceramide levels. Moreover, the hydration values of the treated forearm of each subject was significantly higher than control sites. These results suggest that the experimental cream was able to improve the lipid barrier and to increase a resistance against ageing-associated xerosis
Sphingolipids and the immune system
No full textThe importance of sphingolipids, not only as components of plasma membranes but also as key players in different physiological and pathophysiological cellular events, is now emerging. This review gathers together what the authors feel are the most relevant data, present in the literature, regarding the roles and the effects of sphingolipids, such as ceramide, ceramide-1-phosphate (C1P), sphingosine (SP) and sphingosine-1-phosphate (S1P), on the development, activation and regulation of the immune system
Immunophenotypic characterization of seminal macrophages in ejaculates from subfertile men and relationship with sperm quality and DNA fragmentation
No full textInvolvement of mitochondrial activity in mediating ELF-EMF stimulatory effect on human sperm motility
No full textIt has recently been reported that the exposure of human spermatozoa to an extremely low frequency (ELF) electromagnetic field (EMF) with a square waveform of 5 mT amplitude and frequency of 50 Hz improves sperm motility. The functional relationship between the energy metabolism and the enhancement of human sperm motility induced by ELF-EMF was investigated. Sperm exposure to ELF-EMF resulted in a progressive and significant increase of mitochondrial membrane potential and levels of ATP, ADP and NAD(+) that was associated with a progressive and significant increase in the sperm kinematic parameters. No significant effects were detected on other parameters such as ATP/ADP ratio and energy charge. When carbamoyl cyanide m-chlorophenylhydrazone (CICCP) was applied to inhibit the oxidative phosphorylation in the mitochondria, the values of energy parameters and motility in the sperm incubated in the presence of glucose and exposed to ELF-EMF did not change, thus indicating that the glycolysis was not involved in mediating ELF-EMF stimulatory effect on motility. By contrast, when pyruvate and lactate were provided instead of glucose, the energy status and motility increased significantly in ELF-EMF-treated sperm. Under these culture conditions, the inhibition of glycolitic metabolism by 2-deoxy-D-glucose (DOG) again resulted in increased values of energy and kinematic parameters, indicating that gluconeogenesis was not involved in producing glucose for use in glycolysis. We concluded that the key role in mediating the stimulatory effects exerted by ELF-EMF on human sperm motility is played by mitochondrial oxidative phosphorylation rather than glycolysis
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