Immunomodulatory effects of Lactobacillous plantarum and Helicobacter pylori CagA⁺ on the expression of selected superficial molecules on monocyte and lymphocyte and the synthesis of cytokines in whole blood culture.

Helicobacter pylori (H. pylori) infections are usually superficial and clinically asymptomatic, but in approximately 10-20% cases it can be more aggressive and associated with other pathologies. The reason for weak or strong pro-inflammatory responses in gastric mucosa that occur during H. pylori infection is not understood. Combined treatment, including antibiotic therapy with administration of probiotic bacteria along, considerably improves the effectiveness of H. pylori eradication and reduces the relapse rate. Thus, the aim of this study was to analyze the effect of Lactobacillus plantarum (L. plantarum) and/or H. pylori CagA(+) on leucocytes in whole blood cultures. This study revealed how selected strains of H. pylori and L. plantarum modulate expression of chosen membrane markers of monocytes and lymphocytes, and the cytokine synthesis of in vitro cultures. The level of IFN-γ was higher in cultures stimulated with L. plantarum than in combination of this two examinated strains. We also observe the tendency to increase the level of IFN-γ by L. planatrum in relation to cells stimulated by H. pylori. In contrast, both H. pylori alone and in combination with L. plantarum had a strong modulatory effect on the synthesis of interleukin-10. Moreover lymphocytes with higher expression of CD25 and CD58 receptors was observed only in those cultures that were stimulated with L. plantarum strain alone or in combination with H. pylori. Effects exerted on the immune system, both in terms of natural and adaptive response, constitute the only functional criterion of probiotic bacteria. The immunostimulant effects documented in this study suggest that Lactobacillus spp. can restore immune function of mucosal membrane during symptomatic infection with H. pylori.

Hyperthermia-induced reorganization of microtubules and microfilaments and cell killing in CHO AA8 cell line.

The aim of this study was to elucidate the effects of hyperthermic treatment on cell morphology and the cytoskeleton in CHO AA8 cell line. The effects of exposure to elevated temperature were analyzed in CHO AA8 cell line by fluorescence microscopy and flow cytometry. The 30 min, at 44.5 degrees C heat shock treatment resulted in the collapse of microtubules (MTs) and microfilaments (MFs) around the nucleus followed by their recovery 24 h after heating. The initial collapse of these cytoskeletal systems, observed 15 min after treatment, was accompanied by the appearance of cells with reduction of volume, shrunken cytoplasm and condensed chromatin. 24 h afterwards, there was the increase in the number of cells with restored and extended MT and MF cytoskeletons. Most of them were larger in size compared to the control cells and had multiple nuclei. 48 h after heat shock the highest number of the giant cells with alternation in nuclear morphology was seen. Flow cytometry analysis revealed the increase in the number of cells with externalized phosphatidylserine 24 h and 48 h after hyperthermic treatment. These results suggest that following heat shock, CHO AA8 cells undergo mitotic catastrophe that presumably represents one of the events resulting in apoptosis.

Combined effect of different lactic acid bacteria strains on the mode of cytokines pattern expression in human peripheral blood mononuclear cells.

The balance between immunogenic and tolerogenic activities in human immune system strongly depends on microflora-induced pro-and anti-inflammatory activities. Lactic acid bacteria (LAB) are important components of microflora. The interactions of the different strains of LAB and the cells of immune system are largely unknown. To assess if LAB strains composition would have an effect on the cellular responses profile (proliferation, cytokines synthesis) peripheral blood mononuclear cells (PBMC) model system was used. PBMC were induced by three different strains of LAB: Lactobacillus acidophilus, Lactobacillus delbrueckii spp. bulgaricus, Bifidobacterium bifidum. Tested strains were mixed together, in combinations with each other (pairs) or alone. Both, the LAB mixture as well as the pairs and the single LAB strains induced low lymphocyte proliferation (about 10% of ConA-induced response). However, the single LAB strains and their combinations were quite different cytokines inducers. First, L. acidophilus was much stronger IFN-gamma inducer than the LAB mixture, being a few times higher IL-12 stimulator than L. bulgaricus and B. bifidum. Second, L. bulgaricus and B. bifidum suppressed L.acidophilus-induced IFN-gamma synthesis to the level equal to that induced by the LAB mixture, limiting IL-12 production by about 30% and 70%, respectively. Third, the LAB strains were good IL-10 and TNF-alpha inducers, irrespectively of their combinations used. We conclude that LAB strains' pro or anti-inflammatory potentials are at least in part dependent on their composition. Low LAB mixture-induced IL-12 and IFN-gamma production and relatively high IL-10 and TNF-alpha expression may represent cellular activities normally induced in vivo by a combined action of bacterial antigens. Their presence is important to limit pro-inflammatory reactions (via IL-10) and to provide protection against infections (via TNF-alpha).

Cardiovascular disease and diabetes: modifying risk factors other than glucose control.

Patients with type 2 diabetes have a significantly increased risk of developing cardiovascular disease. Atherosclerosis kills more diabetic patients than all other causes combined. Multiple risk factors tend to cluster in some patients in a syndrome termed insulin resistance syndrome or "Syndrome X." Increasing evidence has changed the recommended management of diabetes from simple glucose control to aggressive lipid management and control of the other components of the metabolic syndrome to prevent development of cardiovascular disease. One mechanism linking hyperglycemia and atherogenesis is nonenzymatic glycation of proteins. Hyperglycemia increases the linkage of glucose to proteins producing insoluble complexes, termed advanced glycation end products, that cause endothelial cell changes. Glycation of lipoproteins increases their atherogenic potential. It is not clear whether intensive glucose control in diabetic patients significantly lowers the rate of long-term macrovascular complications, and glucose control by itself may not be sufficient to prevent cardiovascular disease. Elevated triglyceride levels in diabetic patients are risk factors for cardiovascular disease. Though LDL-cholesterol levels are not necessarily elevated in type 2 diabetes, higher levels (or LDL phenotype B) are shown to be more atherogenic. The association between obesity and hypertension is well documented, and obesity can worsen other risk factors. Glycemic control may not always normalize lipid and lipoprotein levels, particularly in type 2 diabetes. Trials of intensive glycemic control have not shown a significant reduction in coronary events despite significant decreases in microvascular complications. Medical nutrition therapy and exercise remain the cornerstone for nonpharmacologic treatment with a goal of improved insulin sensitivity.

Intracellular mechanisms underlying prostaglandin F2alpha-stimulated phasic myometrial contractions.

These studies sought to test the hypothesis that prostaglandin F2alpha (PGF2alpha)-stimulated phasic myometrial contractions are characterized by the activation of the phosphatidylinositol-signaling pathway resulting in the generation of cytosolic calcium oscillations. For the experiments described in this report rat myometrial tissue was used, after the tissue was loaded with fura 2, to perform cytosolic calcium imaging studies and to perform computer-digitalized in vitro isometric contraction studies. Consistent with the above hypothesis, the cytosolic calcium-imaging studies demonstrated PGF2alpha-stimulated cytosolic calcium oscillations occurring simultaneously with phasic contractions. The in vitro isometric contraction studies confirmed that previously reported inhibitors of the phosphatidylinositol-signaling pathway and cytosolic calcium oscillation mechanisms resulted in significant inhibition of PGF2alpha-stimulated phasic myometrial contractions. In summary, these studies have provided substantial support for the hypothesis that PGF2alpha-stimulated phasic myometrial contractions are generated by intracellular signaling mechanisms involving activation of the phosphatidylinositol-signaling pathway and the production of cytosolic calcium oscillation-like phenomena.

Effects of sodium and calcium channel blockade on cytosolic calcium oscillations and phasic contractions of myometrial tissue.

OBJECTIVE: These studies sought to test the hypothesis that agonist-stimulated cytosolic calcium oscillations and phasic myometrial contractions are dependent on calcium influx through dihydropyridine-sensitive calcium channels, but not sodium influx through tetrodotoxin-sensitive sodium channels. METHODS: Cytosolic calcium imaging studies and in vitro isometric contraction studies were performed using uterine tissue from proestrus/estrus Sprague-Dawley rats. The calcium imaging studies were performed after loading partial thickness strips of myometrium with Fura-2. For the in vitro isometric contraction studies, the contraction data were computer digitalized, analyzed for contraction area, and normalized for cross-section area. The effects of nifedipine (1.0-5 mumol/L), a calcium channel blocker, were compared to tetrodotoxin (0.01-1 mumol/L), a sodium channel blocker. RESULTS: Oxytocin-stimulated simultaneous cytosolic calcium oscillations and phasic contractions were completely inhibited by 1 mumol/L nifedipine; in contrast, 1 mumol/L tetrodotoxin had no effect on the oxytocin-stimulated calcium oscillations and contractions. Oxytocin, aluminum fluoride, potassium chloride, and ionomycin stimulated in vitro phasic myometrial contractions. Tetrodotoxin had no effect on these agonist-stimulated phasic contractions, whereas nifedipine produced a significant, dose-related inhibition of the phasic contractile activity. CONCLUSIONS: The studies described in this report support the hypothesis that the influx of extracellular calcium is an important component of the cellular mechanisms responsible for the cytosolic calcium oscillations occurring during phasic myometrial contractions. In contrast, sodium influx through tetrodotoxin-sensitive sodium channels does not appear to play a comparably important role.

(+)cis-dioxolane stimulation of cytosolic calcium oscillations and phasic contractions of myometrial smooth muscle.

These studies sought to determine the intracellular mechanisms underlying muscarinic receptor stimulated phasic contractions of myometrial smooth muscle. Utilizing cytosolic calcium imaging studies with Fura-2 loaded muscle strips, along with in vitro isometric contraction studies, we have demonstrated that phasic myometrial contractions stimulated in response to (+)cis-dioxolane, a muscarinic selective acetylcholine agonist, are mediated by activation of the M3 receptor subtype resulting in stimulation of the phosphatidylinositol signaling pathway and the generation of cytosolic calcium oscillations.

The effects of ruthenium red, an inhibitor of calcium-induced calcium release, on phasic myometrial contractions.

Ruthenium red inhibits calcium-induced calcium release from the ryanodine-sensitive intracellular calcium stores; this study sought to evaluate the effects of ruthenium red on agonist-stimulated phasic myometrial contractions. Ruthenium red was found to significantly inhibit in vitro isometric contractions stimulated in response to oxytocin, prostaglandin F2 alpha, aluminum fluoride, potassium chloride, ionomycin, and Bay K 8644. These observations provide support for the hypothesis that calcium-induced calcium release from ryanodine-sensitive calcium stores is an important event during agonist-stimulated phasic myometrial contractions.

Cancer epidemiology in the Philippines.

Based on 16,492 cancer cases recorded at the Central Tumor Registry of the Philippines from July 1968 to June 1973, an epidemiologic analysis was conducted. Age-adjusted incidence rates for cancer of all sites in the Philippines, the United States, and Japan were similar. Cancers of the lung and breast were the leading sites in males and females, respectively. Age-specific incidence rates by each site were compared for the Philippines, the United States, and Japan. Cancers of the oral cavity, nasopharynx, liver, lung, breast, cervix, ovary, and thyroid and malignant lymphoma occurred with higher frequency in the Philippines. The more education people had, the more likely they were to develop cancers of the lung, pancreas, bladder, prostate, breast, and ovary, whereas cancers of the stomach, skin, esophagus, oropharynx, tongue, and mouth were more common in individuals who had not completed high school. Among smokers, neoplasms of the lung, larynx, tongue, mouth, liver, esophagus, and oropharynx occurred with significantly higher frequency. Epidemiologic implications and significance of these results for cancer control were discussed.