Heme Amplifies the Innate Immune Response to Microbial Molecules through Spleen Tyrosine Kinase (Syk)-dependent Reactive Oxygen Species Generation

Infectious diseases that cause hemolysis are among the most threatening human diseases, because of severity and/or global distribution. In these conditions, hemeproteins and heme are released, but whether heme affects the inflammatory response to microorganism molecules remains to be characterized. Here, we show that heme increased the lethality and cytokine secretion induced by LPS in vivo and enhanced the secretion of cytokines by macrophages stimulated with various agonists of innate immune receptors. Activation of nuclear factor κB (NF-κB) and MAPKs and the generation of reactive oxygen species were essential to the increase in cytokine production induced by heme plus LPS. This synergistic effect of heme and LPS was blocked by a selective inhibitor of spleen tyrosine kinase (Syk) and was abrogated in dendritic cells deficient in Syk. Moreover, inhibition of Syk and the downstream molecules PKC and PI3K reduced the reactive oxygen species generation by heme. Our results highlight a mechanism by which heme amplifies the secretion of cytokines triggered by microbial molecule activation and indicates possible pathways for therapeutic intervention during hemolytic infectious diseases.     

Adjuvant effect of the Propionibacterium acnes and its purified soluble polysaccharide on the immunization with plasmidial DNA containing a Trypanosoma cruzi gene

In the present work we investigated the role of killed Propionibacterium acnes or a soluble polysaccharide extracted from bacterium cell wall in modulated experimental immunization with plasmidial DNA. We used a plasmid, p154/13, containing a gene-encoding catalytic domain of Trypanosoma cruzi (T. cruzi) trans-sialidase. As previously described, immunization of BALB/c mice with p154/13 elicited humoral, cell-mediated and protective immune responses against T. cruzi infection. In this study we describe that both P. acnes and its soluble polysaccharide fraction have the ability to modulate the immune response elicited by p154/13. Treatment with these adjuvants enhanced specific trans-sialidase Th1 immune response, as revealed by a lower IgG1/IgG2a ratio and stronger in vitro IFN-gamma synthesis by CD4+ T cells. The most important fact was that treatment with P. acnes or its soluble polysaccharide fraction in the presence of p154/13 significantly reduced the peak of parasitemia observed 7 to 8 days after T. cruzi challenge. These data suggest that P. acnes or its soluble polysaccharide fraction may improve the protective potential of a DNA vaccine against experimental T. cruzi infection.     

The rising incidence of type 1 diabetes in south-eastern Poland. A study of the 0-29 year-old age group, 1980-1999

INTRODUCTION: Studies carried out over the last few years have provided information about the increase in the incidence of type 1 diabetes in different parts of the world including the European countries bordering Poland. THE AIM OF STUDY: The aim of this study was to determine the long-term trends in the incidence of type 1 diabetes over the 20 years between 1980 and 1999 and to compare the incidence during the decades preceding and following the 1989 economic and political transformation in Poland. MATERIAL AND METHODS: The registration of type 1 diabetes among people aged 0-29 was drawn up according to the DERI recommendations using three data sources. We calculated the age-standardised incidence rates for five-year age groups and determined the long-term trend in the incidence of type 1 diabetes in south-eastern Poland. RESULTS: A significant growth in the incidence of type 1 diabetes was observed among people aged 0-29 in the Rzeszów Province in the period between 1980 and 1999. The mean age-standardised incidence rate was 6.1/100,000, and a statistically significant difference was noted between the 1980s and the 1990s (5.3 [95%CI 4.5-6.0] and 6.8 [95%CI 5.9-7.6]). The male incidence of 6.7 significantly exceeded that for females--5.5/100,000. There was also a higher incidence in the group aged 0-14 in comparison with the group aged 15-29 (6.4 and 5.8/100,000 respectively). The highest incidence was found in boys aged 10-14 (11.5/100,000) and a significantly rising trend was observed in children of 0-4 years old. CONCLUSIONS: The mean incidence of type 1 diabetes among the study population was low. Nevertheless, we demonstrated a significantly increasing trend in the incidence during the 20-year observation period. The incidence in the 1990s, both in general and for males, was significantly higher when compared to the 1980s.     

Rehardening of caries-like lesions in root surfaces by saliva substitutes

Objective: This study sought to evaluate whether root dentine caries‐like lesions could be remineralised by saliva substitutes. Methods: Root dentine slabs (3 × 3 × 2 mm) were cut from bovine incisors, ground flat, polished and pre‐tested for Knoop microhardness (KHN) at five locations spaced 500 μm apart and 500 μm from the left edge of each sectioned piece. After 60 out of the 100 slabs had been selected based upon their KHN values, specimens were coated with wax except for their outer surface. Specimens were then cycled through a highly cariogenic challenge model to induce caries‐like lesions, whose formation was confirmed by KHN measurements located 500 μm from the right edge of the specimen. According to a randomised complete block design, the experimental units (n = 15) were exposed to 1.5 ml of saliva substitutes, based on either mucin (MC) or carboxymethylcellulose (CM), to natural human saliva (HS) or to 100% relative humidity (RH) over 20 days. Remineralisation was verified by KHN measurements located 1000 μm apart from the right edge of the specimen. Results: Analysis of variance indicated a significant (p < 0.0001) difference among the KHN values attained by the carious root dentine after exposure to the remineralising agents. Tukey's test ascertained that remineralisation was greatest with MC, intermediate with CM and least with HS, but rehardening did not reach the pre‐caries lesion formation values. Conclusion: Saliva substitutes may provide partial remineralisation to preformed caries‐like lesions in root dentine.     

Trypanosoma cruzi modulates the expression of Rabs and alters the endocytosis in mouse cardiomyocytes in vitro.

Chagas disease is an incurable illness caused by the protozoan Trypanosoma cruzi. Cardiomyocytes represent important targets for the parasite infection and alterations in their physiology were reported. Because endocytosis is involved in different cellular events and guanosine triphosphatase (GTPase) Rab proteins play important roles in various aspects of the membrane traffic, our aim was to characterize the expression of Rab proteins in T. cruzi-infected cardiomyocytes, which displayed a downregulation of Rab7 and Rab11, whereas the expression of Rab5a was maintained in the infected cultures even after longer periods of parasite internalization, but early endosome antigen 1 was partially downregulated. The parasite infection also decreased the uptake of fluid phase ligands by the cardiac cultures. The regulation of GTPase proteins and effector molecules can contribute to the altered physiology of the host cells by modifying the normal incoming of nutrients as well as interfering with other important events related to the endocytic pathway.     

The heterogeneity of ion channels in chromaffin granule membranes

Chromaffin granules are involved in catecholamine synthesis and traffic in the adrenal glands. The transporting membrane proteins of chromaffin granules play an important role in the ion homeostasis of these organelles. In this study, we characterized components of the electrogenic ⁸⁶Rb⁺ flux observed in isolated chromaffin granules. In order to study single channel activity, chromaffin granules from the bovine adrenal medulla were incorporated into planar lipid bilayers. Four types of cationic channel were found, each with a different conductance. The unitary conductances of the potassium channels are 360 ± 10 pS, 220 ± 8 pS, 152 ± 8 pS and 13 ± 3 pS in a gradient of 450/150 mM KCl, pH 7.0. A multiconductance potassium channel with a conductivity of 110 ± 8 pS and 31 ± 4 pS was also found. With the exception of the 13 pS conductance channel, all are activated by depolarizing voltages. One type of chloride channel was also found. It has a unitary conductance of about 250 pS in a gradient of 500/150 mM KCl, pH 7.0.     

Describing the organization of dominance relationships by dominance-directed tree method

Methods to describe dominance hierarchies are a key tool in primatology studies. Most current methods are appropriate for analyzing linear and near-linear hierarchies; however, more complex structures are common in primate groups. We propose a method termed "dominance-directed tree." This method is based on graph theory and set theory to analyze dominance relationships in social groups. The method constructs a transitive matrix by imposing transitivity to the dominance matrix and produces a graphical representation of the dominance relationships, which allows an easy visualization of the hierarchical position of the individuals, or subsets of individuals. The method is also able to detect partial and complete hierarchies, and to describe situations in which hierarchical and nonhierarchical principles operate. To illustrate the method, we apply a dominance tree analysis to artificial data and empirical data from a group of Cebus apella.     

Clinical presentation of a patient with congenital polycythemia

The authors describe a 19 year old male with an isolated but absolute erythrocytosis with iron deficiency without evidence for polycythemia vera as well as another causes of erythrocytosis. The polycythemia was due to a recently described von Hippel-Lindau (VHL) mutation. By stopping iron therapy there was no more requirements for phlebotomy in this patient.     

Variations in mugwort (Artemisia spp.) airborne pollen concentrations at three sites in central Croatia, in period from 2002 to 2003

In spite of the low atmospheric pollen levels, Artemisia sensitisation and allergy has been reported widely. The aim of the study was to determine the length of pollen season, intradiurnal, daily and monthly pollen variation, and the effect of some meteorological parameters on atmospheric pollen concentrations in Central Croatia. Seven-day Hirst volumetric pollen and spore traps were used for pollen sampling. The Artemisia pollen season lasted from the end of July until the end of September with the highest concentrations in August. The percentage of the total pollen count ranged from 0.52% to 0.92%. The intradiurnal peak occurred between 10 a.m. and 12 a.m. Statistical analysis showed a significant correlations between higher air temperature and high pollen concentration as well as high precipitation and low pollen concentration. Results of this study are expected to help in preventing the symptoms of allergic reaction in individuals with Artemisia pollen hypersensitivity.