Effects of amoebae on the growth of microbes isolated from moisture-damaged buildings

Dampness, moisture, and mold in buildings are associated with adverse health outcomes. In addition to fungi and bacteria, amoebae have been found in moisture-damaged building materials. Amoebae and a growing list of bacteria have been shown to have mutual effects on each other's growth, but the interactions between amoebae and microbes common in moisture-damaged buildings have not been reported. We co-cultivated the amoeba Acanthamoeba polyphaga with bacteria and fungi isolated from moisture-damaged buildings in laboratory conditions for up to 28 days. The microbes selected were the bacteria Streptomyces californicus, Bacillus cereus, and Pseudomonas fluorescens, and the fungi Stachybotrys chartarum, Aspergillus versicolor, and Penicillium spinulosum. Fungi and bacteria generally benefited from the presence of the amoebae, whereas the growth of amoebae was hindered by Streptomyces californicus, Stachybotrys chartarum, and Bacillus cereus. Pseudomonas fluorescens slightly enhanced amoebae viability. Amoebae were indifferent to the presence of Aspergillus versicolor and Penicillium spinulosum. Thus, our results show that amoebae can alter the survival and growth of some microbes in moisture-damaged buildings.     

Chaotic neovascularization induced by aggressive fibrosarcoma cells overexpressing S-adenosylmethionine decarboxylase

S-adenosylmethionine decarboxylase is a key enzyme in the biosynthesis of polyamines essential for cell proliferation. Overexpression of S-adenosylmethionine decarboxylase in rodent fibroblasts led to aggressive transformants (Amdc-s cells) that had unforeseen high invasive capacity in nude mice, invading rapidly from the subcutaneous injection site into the peritoneal cavity and its organs. In vitro, these cells were much more invasive than Ras-oncogene-transformed fibroblasts, or human HT-1080 fibrosarcoma and MDA-MB-231 breast cancer cells. In immunohistological characterization, Amdc-s-induced tumors showed chaotic neovascularization, with abundant pleomorphic vessel-like structures that had noncontiguous or totally missing laminin (basement membrane) and CD31 (endothelial cell) immunoreactivity. Gene expression and protein analyses of Amdc-s cells showed them to overexpress several pro-angiogenic molecules, including vascular endothelial growth factor (VEGF-A), and to exhibit profound down-regulation of the anti-angiogenic thrombospondin-1 (TSP-1). By reintroduction of TSP-1 into Amdc-s cells, the high invasiveness was efficiently inhibited in vitro. Interestingly, Amdc-s cells showed up-regulation of hepatocyte growth factor (HGF) and also expressed the MET receptor, creating thus an autocrine loop able to regulate VEGF-A and TSP-1 levels. Further, we found Amdc-s cells to express increased amounts of matrix metalloproteinase-2 (MMP-2) and the large isoform of tenascin-C (TN-C), which may also contribute to the angiogenic switch and invasiveness. Consequently, Amdc-s cells offer an excellent model to sort out the key molecules of aggressive tumor growth, and thereby help in designing rational, novel anti-vascular and other cancer therapies.     

The unknown northern green: evaluation of a national forest biodiversity research program

The taxonomic knowledge gap and lack of knowledge on species-level diversity has been a global concern within conservation biology through the recent two decades. As a national response, Finland has funded a Research Program of Poorly Known and Threatened Forest Species from the year 2003. The program is an essential part of the knowledge base applied in Finnish forest policy. The present paper evaluates the outcomes of the program, covering scientific publication, training, taxonomic coverage, increased knowledge on boreal forest biodiversity and awareness rising. The ongoing program has been funded by 6.5 million Euros to date including 59 projects. The program has produced 19 master’s theses and six doctoral theses. Due to their expertise the graduated biologists have been employed in environmental administration or become researchers. The program has produced 163 refereed scientific articles, 104 of which are recognized by the Web of Science; with an average impact factor of 1.81. The results of the program include 15 genera and 348 species new to science, and 60 genera and 1,664 species to Finland’s list of known species. Due to new knowledge gathered in the program 3,000–4,000 species could be included in the most recent National Red List assessment. The publishing of identification books on local language has proved as the most effective way of advancing general species knowledge among the public and end-users. The high number of new species to science from a biodiversity-poor boreal country describes the scale of the huge work still be done in describing the global species-level diversity. The program has achieved its goals in many ways, but the program leaves out a large part of the species that do not occur in forest biotopes. Moreover, the funding has decreased through the years despite the international goal of halting the world’s biodiversity loss has not been met.     

The daily rhythms of melatonin and free fatty acids in goats under varying photoperiods and constant darkness

The purpose of the study was to explore parallel and divergent features of the daily rhythms of melatonin and plasma free fatty acids (FFA) in goats exposed to different lighting conditions. From these features, we attempted to analyze whether the endogenous melatonin rhythm plays any role in the maintenance of the FFA rhythm. Seven Finnish landrace goats were kept under artificial lighting that simulated the annual changes of photoperiod at 60°N (longest photoperiod, 18 h; shortest, 6 h). The ambient temperature and feeding regimen were kept constant. Blood samples were collected 6 times a year at 2 h intervals for 2 d, first in the prevailing light‐dark (LD) conditions and then after 3 d in constant darkness (DD). In LD conditions, the melatonin levels always increased immediately after lights‐off and declined around lights‐on, except in winter (18 h darkness), when the low daytime levels were restored clearly before lights‐on. The FFA levels also displayed a consistent rhythmicity, with low levels at night and a transient peak around lights‐on. In DD conditions, the melatonin profiles were very similar to those found in the habitual LD conditions, but the rhythm tended to advance. The FFA rhythm persisted also in DD, and the morning peak tended to advance. There was an overall parallelism between the two rhythms, with one significant exception. In winter in LD conditions, the morning rise in FFA levels coincided with lights‐on and not with the declining phase of melatonin, whereas in DD conditions, the FFA peak advanced several hours and coincided with the declining phase of melatonin. From this finding and comparisons of the calculated rhythm characteristics, i.e., phase‐shifts, phase differences, and correlations, we conclude that the daily rhythm of FFA levels is most probably generated by an endogenous oscillator, primarily adjusted by dawn, whereas the melatonin rhythm in this species is regulated by an oscillator primarily adjusted by dusk. The results did not exclude a modulatory effect of melatonin on the daily FFA profiles, but melatonin secretion, alone, does not explain the patterns sufficiently.     

Locus for quantitative HDL-cholesterol on chromosome 10q in Finnish families with dyslipidemia

Decreased HDL-cholesterol (HDL-C) and familial combined hyperlipidemia (FCHL) are the two most common familial dyslipidemias predisposing to premature coronary heart disease (CHD). These dyslipidemias share many phenotypic features, suggesting a partially overlapping molecular pathogenesis. This was supported by our previous pooled data analysis of the genome scans for low HDL-C and FCHL, which identified three shared chromosomal regions for a qualitative HDL-C trait on 8q23.1, 16q23.3, and 20q13.32. This study further investigates these regions as well as two other loci we identified earlier for premature CHD on 2q31 and Xq24 and a locus for high serum triglycerides (TGs) on 10q11. We analyzed 67 microsatellite markers in an extended study sample of 1,109 individuals from 92 low HDL-C or FCHL families using both qualitative and quantitative lipid phenotypes. These analyses provided evidence for linkage (a logarithm of odds score of 3.2) on 10q11 using a quantitative HDL-C trait. Importantly, this region, previously linked to TGs, body mass index, and obesity, provided evidence for association for quantitative TGs (P = 0.0006) and for a combined trait of HDL-C and TGs (P = 0.008) with marker D10S546. Suggestive evidence for linkage also emerged for HDL-C on 2q31 and for TGs on 20q13.32. Finnish families ascertained for dyslipidemias thus suggest that 10q11, 2q31, and 20q13.32 harbor loci for HDL-C and TGs.     

Fluorescence Resonance Energy Transfer (FRET) and Proximity Ligation Assays Reveal Functionally Relevant Homo- and Heteromeric Complexes among Hyaluronan Synthases HAS1, HAS2, and HAS3

In vertebrates, hyaluronan is produced in the plasma membrane from cytosolic UDP-sugar substrates by hyaluronan synthase 1–3 (HAS1–3) isoenzymes that transfer N-acetylglucosamine (GlcNAc) and glucuronic acid (GlcUA) in alternative positions in the growing polysaccharide chain during its simultaneous extrusion into the extracellular space. It has been shown that HAS2 immunoprecipitates contain functional HAS2 homomers and also heteromers with HAS3 (Karousou, E., Kamiryo, M., Skandalis, S. S., Ruusala, A., Asteriou, T., Passi, A., Yamashita, H., Hellman, U., Heldin, C. H., and Heldin, P. (2010) The activity of hyaluronan synthase 2 is regulated by dimerization and ubiquitination. J. Biol. Chem. 285, 23647–23654). Here we have systematically screened in live cells, potential interactions among the HAS isoenzymes using fluorescence resonance energy transfer (FRET) and flow cytometric quantification. We show that all HAS isoenzymes form homomeric and also heteromeric complexes with each other. The same complexes were detected both in Golgi apparatus and plasma membrane by using FRET microscopy and the acceptor photobleaching method. Proximity ligation assays with HAS antibodies confirmed the presence of HAS1-HAS2, HAS2-HAS2, and HAS2-HAS3 complexes between endogenously expressed HASs. C-terminal deletions revealed that the enzymes interact mainly via uncharacterized N-terminal 86-amino acid domain(s), but additional binding site(s) probably exist in their C-terminal parts. Of all the homomeric complexes HAS1 had the lowest and HAS3 the highest synthetic activity. Interestingly, HAS1 transfection reduced the synthesis of hyaluronan obtained by HAS2 and HAS3, suggesting functional cooperation between the isoenzymes. These data indicate a general tendency of HAS isoenzymes to form both homomeric and heteromeric complexes with potentially important functional consequences on hyaluronan synthesis.     

Trebouxia aggregata undGloeocapsa sanguinea,Phycobionten inEuopsis granatina (Lichinaceae)

The green algaTrebouxia aggregata (Archibald)Gärtner (Chlorellales) and the procaryotic blue-green alga (cyanobacterium)Gloeocapsa sanguinea (C. Agardh)Kütz. emend.Jaag (Chroococcales) are reported to be the phycobionts inEuopsis granatina (Sommerf.)Nyl. (Lichinaceae). The ultrastructure of the two organisms, studied in the lichen thallus, is described.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

The Effects of Inorganic Elements on the Reduction of Phytophthora Stem Rot Disease of Soybean, the Growth Rate and Zoospore Release of Phytophthora sojae

The effects of several inorganic elements contained in B5 medium on Phytophthora stem rot disease reduction of Glycine max (L.) Merr. cv. Chusei‐Hikarikuro, fungal growth of Phytophthora sojae isolate and zoospore release were investigated. Application of B5 solution and macro inorganic nutrients in the B5 medium prior to inoculation significantly inhibited infection, compared with controls. Various concentrations of KNO₃, (NH₄)₂SO₄, MgSO₄, CaCl₂ and NaH₂PO₄ in the presence of macro inorganic nutrients were investigated in an effort to determine the elements most effective in suppressing the incidence of disease. A concentration of 2.47–24.7 mm KNO₃ and 0.1–10.2 mm CaCl₂ greatly inhibited infection. Although mycelium growth of the isolate was affected by the potassium and calcium concentration, no significant relationship was observed between inhibition of the growth rate and disease reduction at 2.47 mm KNO₃ and 0.1–5.1 mm CaCl₂ application. Disease suppression recorded in laboratory experiments using pathogen mycelium was due to the response of plant tissues rather than a direct inhibition of pathogen fungal growth by the application of potassium or calcium. The extent of disease reduction was related to an increased potassium and calcium uptake by plants, suggesting that the effective elements in reducing Phytophthora stem rot were potassium and calcium. The presence of 2.47–247 mm KNO₃ and 5.1–10.2 mm CaCl₂ decreased the release of zoospores, although 0.1–2.5 mm CaCl₂ significantly induced zoospore release. These results suggest that applying a solution containing more than 2.47 mm of potassium and 5.1 mm of calcium can decrease the incidence of disease in agricultural fields by the inhibition of zoospore release.