Inhibition of cytokines expression in human microglia infected by virulent and non-virulent mycobacteria

The pathogenesis of tuberculosis (TBC) meningitis is still unknown. As shown by previous studies, human microglia can be the target of mycobacteria, but no data are available about their cellular response to infection. Consequently, we studied the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) and IL-10 in human microglia pure cultures infected with the two variants of Mycobacterium avium (domed-opaque (SmD) and transparent (SmT)) and with Mycobacterium tuberculosis. Results showed that microglia was productively infected by mycobacteria which could grow inside the cells. Mycobacteria internalization was more rapid for M. avium, but M. tuberculosis infection turned out to be more efficient due to the incorporation of densely packed bacteria. TNF-alpha expression was not affected by M. avium, whereas an increase followed by a decrease was observed in M. tuberculosis. Both IL-1 and IL-10 cytokine expression was rapidly inhibited by infection with the more virulent bacteria, whereas the non-pathogenic one had almost no effect. Also, the expression of the co-stimulatory molecule CD137, a member of tumor necrosis factor receptor family, was affected by infection with virulent mycobacteria. Our results show that microglia response to mycobacterial infection is modulated in correlation with virulence, mainly toward inhibition of inflammatory response. This observation might be one of the mechanisms by which non-pathogenic mycobacteria are quickly eliminated, explaining one of the bases of virulence.     

Novel angular furo and thieno-quinolinones: synthesis and preliminary photobiological studies

A number of new furo and thienoquinolinones carrying an electron-withdrawing function or unsubstituted at the position 3 were synthesized in order to obtain new potential photochemotherapeutic agents with increased antiproliferative activity and decreased toxic side effects. Our interest in studying the SAR of these derivatives also prompted us to investigate the influence of N-methylation on biological activity, by preparing N-methyl derivatives. The antiproliferative activity of all the newly synthesized compounds was evaluated and compared to 8-methoxypsoralen (8-MOP), the drug widely used in PUVA-therapy. The 3-unsubstituted thienoquinolinones were generally the most potent derivatives, followed by the furo-analogues. In particular, the unsubstituted thieno[2,3-h]quinoline-2(1H)one showed the highest activity in T2 bacteriophage, HeLa cells and Ehrlich cells tests. All the compounds, assayed on Escherichia coli WP2 TM9, showed a similar mutagenic activity, very close to that of 8-MOP. Except for 2-oxo-1,2-dihydrothieno[2,3-h]quinoline-3-carboxylic acid, which appeared to be very effective, all compounds generated singlet oxygen to slightly larger amounts when compared to 8-MOP. The N-methyl analogues only induced moderate skin erythemas on albino guinea pigs, while all other derivatives appeared to be entirely inactive. On the basis of these results, the unsubstituted thieno[2,3h]quinoline 2(1H)one seems to be the most interesting potential drug for PUVA photochemotherapy and photopheresis.     

Calcium regulates scallop muscle by changing myosin flexibility

Muscle myosins are molecular motors that convert the chemical free energy available from ATP hydrolysis into mechanical displacement of actin filaments, bringing about muscle contraction. Myosin cross-bridges exert force on actin filaments during a cycle of attached and detached states that are coupled to each round of ATP hydrolysis. Contraction and ATPase activity of the striated adductor muscle of scallop is controlled by calcium ion binding to myosin. This mechanism of the so-called “thick filament regulation” is quite different to vertebrate striated muscle which is switched on and off via “thin filament regulation” whereby calcium ions bind to regulatory proteins associated with the actin filaments. We have used an optically based single molecule technique to measure the angular disposition adopted by the two myosin heads whilst bound to actin in the presence and absence of calcium ions. This has allowed us to directly observe the movement of individual myosin heads in aqueous solution at room temperature in real time. We address the issue of how scallop striated muscle myosin might be regulated by calcium and have interpreted our results in terms of the structures of smooth muscle myosin that also exhibit thick filament regulation. This paper is not being submitted elsewhere and the authors have no competing financial interests     

Biodiversity and resilience of arthropod communities after fire disturbance in temperate forests

Changes in ecosystem functions following disturbances are of central concern in ecology and a challenge for ecologists is to understand the factors that affect the resilience of community structures and ecosystem functions. In many forest ecosystems, one such important natural disturbance is fire. The aim of this study was to understand the variation of resilience in six functional groups of invertebrates in response to different fire frequencies in southern Switzerland. We measured resilience by analysing arthropod species composition, abundance and diversity in plots where the elapsed time after single or repeated fires, as determined by dendrochronology, varied. We compared data from these plots with data from plots that had not burned recently and defined high resilience as the rapid recovery of the species composition to that prior to fire. Pooling all functional groups showed that they were more resilient to single fires than to repeated events, recovering 6–14 years after a single fire, but only 17–24 years after the last of several fires. Flying zoophagous and phytophagous arthropods were the most resilient groups. Pollinophagous and epigaeic zoophagous species showed intermediate resilience, while ground-litter saprophagous and saproxylophagous arthropods clearly displayed the lowest resilience to fire. Their species composition 17–24 years post-burn still differed markedly from that of the unburned control plots. Depending on the fire history of a forest plot, we found significant differences in the dominance hierarchy among invertebrate species. Any attempt to imitate natural disturbances, such as fire, through forest management must take into account the recovery times of biodiversity, including functional group composition, to ensure the conservation of multiple taxa and ecosystem functions in a sustainable manner.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

An EPR Study of NO Bonding to Erythrocruorin from the Earthworm Octolasium Complanatum

The EPR properties of the nitric oxide derivative of Octolasium complanatum erythrocruorin have been investigated as a function of the concentration of protons and cations which are known to affect the oxygen-linked allosteric equilibrium. The EPR spectrum has a rhombic shape with g_x = 2.08, g_z = 2.005, and g_y = 1.99, and remains unchanged under all the experimental conditions used. A supernyperfine pattern consisting of nine equally spaced lines is present in the g_z region indicating an interaction with two nonequivalent nitrogen atoms, one contributed by the nitric oxide and the other by the proximal histidine. The constancy of the EPR spectrum suggests that changes in the allosteric equilibrium do not involve differences in the strain of the Fe(II)-histidine bond as in tetrameric hemoglobins.     

Decrease in N-Acetyl-l-Aspartic Acid in Brain of Myodystrophic Mice

Brain tissue from myodystrophic mice (male 129/ReJ-dy and female 129 B6F1/J-dy) was examined to determine whether CNS abnormalities accompany the known muscular defects. Brain N-acetyl-L-aspartate, aspartate, and glutamate were significantly lower than in brain from control mice. These amino acids are only slightly reduced in brains of female dystrophic mice.     

Fine needle aspiration cytopathology of phyllodes tumor. Differential diagnosis with fibroadenoma

The differential diagnosis of benign or borderline phyllodes tumors of the breast in fine needle aspiration biopsy smears was studied. Smears from five histologically proven cases and 20 proven cases of fibroadenoma were evaluated with regard to their cytologic features. The findings indicate that the presence of a high cellularity of stromal fragments (including bipolar naked nuclei), clusters of hyperplastic ductal cells and giant cells and the absence of apocrine metaplasia may suggest a diagnosis of phyllodes tumor, as opposed to fibroadenoma. The borderline phyllodes tumor showed a sarcomatous atypia and/or small intranuclear cytoplasmic invaginations in some cells. The differential diagnosis of this tumor with malignant phyllodes tumor and plasma-cell mastitis is also discussed.     

N-andO-glycosylation of glycoprotein C synthesized by Herpes simplex virus type 1-infected ricin resistant cells

The progeny of Herpes simplex virus type 1 (HSV-1) grown in ricin-resistant 14 cells (Ric^R14) lackingN-acetylglucosaminyltransferase I was released in the extracellular medium at a very low rate. By using a monoclonal antibody immobilized on Sepharose we purified from HSV-1-infected Ric^R14 cells a viral glycoprotein (gC), which carries bothN-andO-linked oligosaccharides. Glycopeptides obtained from [³H]mannoselabeled gC by Pronase digestion were entirely susceptible to endo--N-acetylglucosaminidase H, and the major oligosaccharide released was Man₄GlcNAc. The accumulation of this high-mannose species was related to the enzymic defect of the host cells and to the long retention of the viral glycoprotein within the cells. The extent ofO-glycosylation evaluated in [¹⁴C]glucosamine-labeled gC from Ric^R14 cells as compared to that of gC from wild type cells did not appear to be significantly modified.Abbreviations Con A concanavalin A - BHK cells baby hamster kidney cells - HSV Herpes simplex virus