Macrophages overexpressing tartrate-resistant acid phosphatase show altered profile of free radical production and enhanced capacity of bacterial killing

Activated macrophages and osteoclasts express high amounts of tartrate-resistant acid phosphatase (TRACP, acp5). TRACP has a binuclear iron center with a redox-active iron that has been shown to catalyze the formation of reactive oxygen species (ROS) by Fenton’s reaction. Previous studies suggest that ROS generated by TRACP may participate in degradation of endocytosed bone matrix products in resorbing osteoclasts and degradation of foreign compounds during antigen presentation in activated macrophages. Here we have compared free radical production in macrophages of TRACP overexpressing (TRACP+) and wild-type (WT) mice. TRACP overexpression increased both ROS levels and superoxide production. Nitric oxide production was increased in activated macrophages of WT mice, but not in TRACP+ mice. Macrophages from TRACP+ mice showed increased capacity of bacterial killing. Recombinant TRACP enzyme was capable of bacterial killing in the presence of hydrogen peroxide. These results suggest that TRACP has an important biological function in immune defense system.     

TLR2 mediates neuroinflammation and neuronal damage

Innate immunity relies on pattern recognition receptors to detect the presence of infectious pathogens. In the case of Gram-positive bacteria, binding of bacterial lipopeptides to TLR2 is currently regarded as an important mechanism. In the present study, we used the synthetic bacterial lipopeptide Pam3CysSK4, a selective TLR2 agonist, to induce meningeal inflammation in rodents. In a 6-h rat model, intrathecal application of Pam3CysSK4 caused influx of leukocytes into the cerebrospinal fluid (CSF) and induced a marked increase of regional cerebral blood flow and intracranial pressure. In wild-type mice, we observed CSF pleocytosis and an increased number of apoptotic neurons in the dentate gyrus 24 h after intrathecal challenge. Inflammation and associated neuronal loss were absent in TLR2 knockout mice. In purified neurons, cytotoxicity of Pam3CysSK4 itself was not observed. Exposure of microglia to Pam3CysSK4 induced neurotoxic properties in the supernatant of wild-type, but not TLR2-deficient microglia. We conclude that TLR2-mediated signaling is sufficient to induce the host-dependent key features of acute bacterial meningitis. Therefore, synthetic lipopeptides are a highly specific tool to study mechanisms of TLR2-driven neurodegeneration in vivo.     

Coordination sphere of the third metal site is essential to the activity and metal selectivity of alkaline phosphatases

Alkaline phosphatases (APs) are commercially applied enzymes that catalyze the hydrolysis of phosphate monoesters by a reaction involving three active site metal ions. We have previously identified H135 as the key residue for controlling activity of the psychrophilic TAB5 AP (TAP). In this article, we describe three X‐ray crystallographic structures on TAP variants H135E and H135D in complex with a variety of metal ions. The structural analysis is supported by thermodynamic and kinetic data. The AP catalysis essentially requires octahedral coordination in the M3 site, but stability is adjusted with the conformational freedom of the metal ion. Comparison with the mesophilic Escherichia coli, AP shows differences in the charge transfer network in providing the chemically optimal metal combination for catalysis. Our results provide explanation why the TAB5 and E. coli APs respond in an opposite way to mutagenesis in their active sites. They provide a lesson on chemical fine tuning and the importance of the second coordination sphere in defining metal specificity in enzymes. Understanding the framework of AP catalysis is essential in the efforts to design even more powerful tools for modern biotechnology.     

Unattended monitoring of potentially toxic phytoplankton species in the Baltic Sea in 1993

Variabilities in chlorophyll a fluorescence, temperature andsalinity in the surface were recorded unattended on board twomerchant ships in the Baltic Sea. When these recordings werecomplemented by automated water sampling, it was possible toanalyze the phytoplankton species composition in 426 samples.In total, 22 potentially toxic phytoplankton species or generawere detected. Nodularia spumigena was the only species thatformed extensive blooms The system has proved to be an effectiveearly warning method for exceptional and eventually harmfulalgal blooms. The possibilities for using this method as analternative, or a complement, to conventional methods in marinephytoplankton monitoring are discussed.     

Records of chrysophytes with siliceous scales (Mallomonadaceae and Paraphysomonadaceae) from Finnish inland waters

Seventy-three taxa of silica-scaled chrysophytes were found with LM and TEM in 264 samples from 141 Finnish water bodies, including the genera Mallomonas (40 taxa), Synura (9), Chrysosphaerella (3), Spiniferomonas (8) and Paraphysomonas (13). More than half of the taxa are recorded for the first time from Finland and five (Mallomonas actinoloma v. actinoloma, M. favosa, M. mangofera f. gracilis, M. scrobiculata and Spiniferomonas serrata) are new to Europe. Most taxa are illustrated with TEM micrographs and the taxonomy of some interesting species is discussed in detail. The seasonal occurrence of the commonest species is described.     

Effects of environmental factors on the phytoplankton community in the Gulf of Finland – unattended high frequency measurements and multivariate analyses

We studied life history and secondary production of Cheumatopsyche spp. (Trichoptera: Hydropsychidae) in StroublesCreek, a 3rd order stream near Blacksburg, Virginia to compare lifehistories and production dynamics in two reaches with differentland-uses. The upstream reach was influenced by agriculturalactivities and more annual degree-days accumulated in this sectionof the stream. The downstream reach was surrounded by forest,received less thermal input than the upstream reach, and showedlittle evidence of human disturbance. Differences between the twosites included a shift in dominant taxa and changes in voltinismand production of hydropsychids. Cheumatopsyche pettiti(Banks) was dominant at the upstream site, while C. pettitiand Cheumatopsyche oxa Ross were both abundant downstream.Cheumatopsyche larvae and other aquatic insects had higherdensities at the forested downstream site. C. pettiti wasbivoltine at the upstream site, but C. pettiti and C. oxa were partially bivoltine downstream, with a small summergeneration overlapping with continuous emergence of overwinteringlarvae. Cheumatopsyche production at the two sites inStroubles Creek was higher than in forested, low-order streams ofNorth America, but similar to some values obtained at streams inagricultural areas and large rivers. Production was higher at thedownstream site, but the superior bivoltinism at the upstream sitecounterbalanced, to some extent, the increase in larval densitythat caused higher production downstream. The adjustment in lifehistory and production dynamics, in the two reaches, showed theecological plasticity of Cheumatopsyche and the importance ofhydropsychids in the energy flow of moderately disturbedstreams.