Antagonistic Interplay between MicroRNA-155 and IL-10 during Lyme Carditis and Arthritis

MicroRNA-155 has been shown to play a role in immune activation and inflammation, and is suppressed by IL-10, an important anti-inflammatory cytokine. The established involvement of IL-10 in the murine model of Borrelia burgdorferi-induced Lyme arthritis and carditis allowed us to assess the interplay between IL-10 and miR-155 in vivo. As reported previously, Mir155 was highly upregulated in joints from infected severely arthritic B6 Il10^-/- mice, but not in mildly arthritic B6 mice. In infected hearts, Mir155 was upregulated in both strains, suggesting a role of miR-155 in Lyme carditis. Using B. burgdorferi-infected B6, Mir155^-/-, Il10^-/-, and Mir155^-/- Il10^-/- double-knockout (DKO) mice, we found that anti-inflammatory IL-10 and pro-inflammatory miR-155 have opposite and somewhat compensatory effects on myeloid cell activity, cytokine production, and antibody response. Both IL-10 and miR-155 were required for suppression of Lyme carditis. Infected Mir155^-/- mice developed moderate/severe carditis, had higher B. burgdorferi numbers, and had reduced Th1 cytokine expression in hearts. In contrast, while Il10^-/- and DKO mice also developed severe carditis, hearts had reduced bacterial numbers and elevated Th1 and innate cytokine expression. Surprisingly, miR-155 had little effect on Lyme arthritis. These results show that antagonistic interplay between IL-10 and miR-155 is required to balance host defense and immune activation in vivo, and this balance is particularly important for suppression of Lyme carditis. These results also highlight tissue-specific differences in Lyme arthritis and carditis pathogenesis, and reveal the importance of IL-10-mediated regulation of miR-155 in maintaining healthy immunity.     

On the hyperthermic response to d-amphetamine in the decapitated rat

Injection of d-amphetamine (10 mg/kg i.p.) in the decapitated rat causes an increase in body temperature, oxygen consumption and heart rate and a decrease in skin temperature. These effects could be reduced or blocked by administration of α- and β-blocking agents. In the decapitated rats in which the content of endogenous catecholamine was reduced by 6-hydroxydopamine treatment and/or adrenalectomy the effects of d-amphetamine were also reduced. Thus it can be concluded that d-amphetamine has a peripheral calorigenic effect due to release of catecholamines. This calorigenic effect of d-amphetamine is resistent to treatment with pimozide in contrast to the pimozide sensitive, central thermogenic effect described by other authors.     

Oligomerization of the cytoplasmic fragment from the aspartate receptor of Escherichia coli.

We have observed that a 31-kDa cloned fragment from the Escherichia coli aspartate receptor exhibits a reversible monomer-oligomer reaction. The fragment, derived from the cytoplasmic region of the receptor (c-fragment), contains the signaling functions of the receptor. The wild-type and nine missense mutant fragments were analyzed. The latter were selected by the effect of the mutations on the signaling properties of the intact receptor, which induced either persistent smooth swimming or tumbling in bacteria [Mutoh, N., Oosawa, K., & Simon, M. I. (1986) J. Bacteriol. 167, 992-998]. In pH 7.0 buffer, the mutations caused five out of the six smooth mutant c-fragments to form oligomers, while neither the three tumble mutant nor wild-type fragments exhibited significant oligomer formation. At a lower pH (5.5), all of the fragments displayed some tendency to form oligomers. The equilibria between the monomer and the oligomers were monitored by gel permeation chromatography (GPC) which resolved two to three forms with apparent molecular weights between 110,000 and 270,000. The proportions of the different forms depended on concentration, indicating an association-dissociation reaction. Static light scattering (SLS) was used to demonstrate that the solution molecular mass of the wild-type c-fragment was 31 kDa and not 110 kDa as indicated by chromatography. One oligomer-forming c-fragment (S461L) eluted as the monomer and one other form, which was determined to be a dimer by SLS. The weight-average molecular weights, calculated from GPC data as a function of protein concentration, agreed well with the weight-average molecular weights obtained by SLS for this mutant.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of N5-methyl-N5-formyl-2,5,6-triamino-4-hydroxypyrimidine as a major adduct in rat liver DNA after treatment with the carcinogens, N,N-dimethylnitrosamine or 1,2-dimethylhydrazine

Human Tamm-Horsfall urinary glycoprotein from an individual of the blood group Sd(a+) phenotype was tritium-labelled by treatment with galactose oxidase and sodium boro[3H]hydride and was then digested with endo-beta-galactosidase. A series of dialysable, labelled fragments was released from which a pentasaccharide was isolated that strongly inhibited the agglutination of Sd(a+) red cells by human anti-Sda serum and hence contained the Sda determinant structure. Reduction, methylation analysis and sequential exo-glycosidase digestion established the structure of the pentasaccharide as: GalNAc beta(1 leads to 4)[NeuAc(2 leads to 3)]Gal beta(1 leads to 4)GlcNAc beta(1 leads to 3)Gal     

Specific silver staining of experimentally undercondensed chromosome regions

Treatment of human and mouse cell cultures with DNA binding AT-specific compounds and with some base analogues induced distinct undercondensations in several heterochromatic chromosome regions. All those heterochromatic regions undercondensed by AT-specific DNA ligands (distamycin A, DAPI, Hoechst 33258) could be heavily labeled with the silver(Ag)-staining technique; but the heterochromatic regions undercondensed with the cytidine analogue 5-azacytidine were Ag-negative. In metaphase chromosomes from BrdU-treated human cell cultures, the bifilarly substituted chromatids, which show a slight undercondensation, were also Ag-negative. Cytochemical analyses of the Ag-stained undercondensed heterochromatic regions showed that the Ag-stainable material consisted of nonhistone proteins. The mechanism of Ag staining in the undercondensed heterochromatic regions was compared with Ag staining of the nucleolus organizer regions.     

The role of methylation in chemotaxis. An explanation of outstanding anomalies

The role of methylation in chemotaxis is understood generally, but several anomalies exist which bring into question the timing of methylation relative to sensing. A double mutant bacterium, deficient in both methyltransferase and methylesterase (Tr-Es-) is capable of chemotaxis even though the respective single mutants (Tr- and Es-) are not. This Tr-Es- mutant will accumulate in capillaries containing aspartic acid but not in capillaries containing serine despite the fact that both the aspartate and serine receptors are part of the methylation-dependent pathway. To understand these anomalies, a combination of theoretical analyses and experimental studies was performed. A mathematical analysis of the gradients of aspartate and serine in the capillary assay shows that outside the capillary the gradients are shallow, but just inside the mouth of the capillary they are very steep. Also, when the number of bacteria accumulated in the capillary is at a maximum, the range of attractant concentrations in the steep gradient just inside the mouth of the capillary is optimal for response and partial adaptation by the Tr-Es- mutant. We postulate that random motion brings the Tr-Es- mutant into the capillary, where it is able to move up the steep gradient. The difference in timing of the responses to serine and aspartate explains why the Tr-Es- mutant accumulates in aspartate- but not in serine-containing capillaries. A simple diffusion-capture model incorporating these concepts can account for experimental values of the number of Tr-Es- bacteria accumulating in the capillary. These studies provide a rational explanation for all of the apparent anomalies and lead to the conclusion that methylation/demethylation plays a crucial role in sensing as well as setting the zero point of the receptor.     

Symbiosis research, technology, and education: Proceedings of the 6th International Symbiosis Society Congress held in Madison Wisconsin, USA, August 2009

Symbiosis, the intimate association between two or more organisms, is a fundamental component of biological systems. Our ability to understand the processes involved in the establishment and function of Symbiosis has critical consequences for the health of humans and the world we live in. For example, a deeper understanding of how legumes and insects have harnessed the nitrogen-fixing capacity of microbes can pave the way toward novel strategies to decrease fertilizer use. Also, using insect models to elucidate links between diet, gut microbiota, and toxin sensitivity not only has implications for biological control strategies, but also will lend insights into similar links in the human gut ecosystem. These types of ideas were presented and discussed at the 6th International Symbiosis Society Congress held in Madison, Wisconsin August, 2009. Over 300 participants from 20 countries attended the 7-day event, which featured cutting-edge symbiosis research from many different perspectives and disciplines. The conference was organized thematically, with oral sessions focused on Evolution, Ecology, Metabolism, the Host-Microbe Interface, Threats to Earth Systems, Symbiosis Models and the Human Microbiome, Viruses and Organelles, and Symbiosis Education. World-renowned scientists, post-doctoral fellows, and students were given the opportunity to describe their most recent discoveries. Session chairs provided overviews of their programs which highlight how the comparative analysis of different systems reveal common trends underlying symbiotic associations, what tools and theory are being developed that may be applied more broadly in symbiosis research, how symbiosis research contributing solutions to global issues such as emerging antibiotic resistance, a need for alternative energy sources, the pursuit of sustainable agriculture and natural resources, and how symbiotic systems are ideal for educating people about the fascinating natural world around us. The following paragraphs provide an overview of the research and discussions that took place during the congress.     

Determination of serum oxalate using peroxyoxalate chemiluminescence of free oxalic acid

We describe a new sensitive and specific method for determination of oxalate in human serum. By using the chemiluminescence decay of monoperoxyoxalic acid very low concentrations of oxalate (200 nmol/L) can be determined. The mean serum oxalate level in apparently healthy controls was 14.5 ± 8.5 m?mol/L. Supplementation of ascorbic acid leads to an increase in serum oxalate level. While serum oxalate concentrations of calcium oxalate stone formers (x = 16.4 ± 9.8 m?mol/L) are not significantly different from the control group, an extreme increase of serum oxalate is evident in haemodialysis patients. The serum oxalate concentration decreased during dialysis treatment from 141.4 ± 32.1 m?mol/L to 36.4 ± 12.7 m?mol/L.     

Transcriptional Profiling of Whole Blood Identifies a Unique 5-Gene Signature for Myelofibrosis and Imminent Myelofibrosis Transformation

Identifying a distinct gene signature for myelofibrosis may yield novel information of the genes, which are responsible for progression of essential thrombocythemia and polycythemia vera towards myelofibrosis. We aimed at identifying a simple gene signature – composed of a few genes - which were selectively and highly deregulated in myelofibrosis patients. Gene expression microarray studies have been performed on whole blood from 69 patients with myeloproliferative neoplasms. Amongst the top-20 of the most upregulated genes in PMF compared to controls, we identified 5 genes (DEFA4, ELA2, OLFM4, CTSG, and AZU1), which were highly significantly deregulated in PMF only. None of these genes were significantly regulated in ET and PV patients. However, hierarchical cluster analysis showed that these genes were also highly expressed in a subset of patients with ET (n = 1) and PV (n = 4) transforming towards myelofibrosis and/or being featured by an aggressive phenotype. We have identified a simple 5-gene signature, which is uniquely and highly significantly deregulated in patients in transitional stages of ET and PV towards myelofibrosis and in patients with PMF only. Some of these genes are considered to be responsible for the derangement of bone marrow stroma in myelofibrosis. Accordingly, this gene-signature may reflect key processes in the pathogenesis and pathophysiology of myelofibrosis development.