Bacterial growth in space flight: logistic growth curve parameters for Escherichia coli and Bacillus subtilis

Previous investigations have reported that bacterial suspension cultures grow to higher stationary concentrations in space flight than on Earth; however, none of these investigations included extensive ground controls under varied inertial conditions. This study includes extensive controls and cell-growth data taken at several times during lag phase, log phase, and stationary phase of Escherichia coli and Bacillus subtilis. The Marquardt-Levenberg, least-squares fitting algorithm was used to calculate kinetic growth parameters from the logistic bacterial growth equations for space-flight and control growth curves. Space-flight cultures grew to higher stationary-phase concentrations and had shorter lag-phase durations. Also, evidence was found for increased exponential growth rate in space. Received: 27 February 1998 / Received revision: 21 August 1998 / Accepted: 3 September 1998     

Toll-like receptor 9 mediates CpG oligonucleotide-induced cellular invasion

Toll-like receptor 9 (TLR9) belongs to the innate immune system and recognizes microbial and vertebrate DNA. We showed previously that treatment with the TLR9-agonistic ODN M362 (a CpG sequence containing oligonucleotide) induces matrix metalloproteinase-13-mediated invasion in TLR9-expressing human cancer cell lines. Here, we further characterized the role of the TLR9 pathway in this process. We show that CpG oligonucleotides induce invasion in macrophages from wild-type C57/B6 and MyD88 knockout mice and in human MDA-MB-231 breast cancer cells lacking MyD88 expression. This effect was significantly inhibited in macrophages from TLR9 knockout mice and in human MDA-MB-231 breast cancer cells stably expressing TLR9 small interfering RNA or dominant-negative tumor necrosis factor receptor-associated factor 6 (TRAF6). Sequence modifications to the CpG oligonucleotides that targeted the stem loop and other secondary structures were shown to influence the invasion-inducing effect in MDA-MB-231 cells. In contrast, methylation of the cytosine residues of the parent CpG oligonucleotide did not affect the TLR9-mediated invasion compared with the unmethylated parent CpG oligonucleotide. Finally, expression of TLR9 was studied in clinical breast cancer samples and normal breast epithelium with immunohistochemistry. TLR9 staining localized in epithelial cells in both cancer and normal samples. The mean TLR9 staining intensity was significantly increased in the breast cancer cells compared with normal breast epithelial cells. In conclusion, our results suggest that TLR9 expression is increased in breast cancer and CpG oligonucleotide-induced cellular invasion is mediated via TLR9 and TRAF6, independent of MyD88. Further, our findings suggest that the structure and/or stability of DNA may influence the induction of TLR9-mediated invasion in breast cancer.     

A state and transition model for the eastern Monte Phytogeographycal Province in Rio Negro

We developed a functional model for a shrub steppe vegetation of the eastern Monte Phytogeographical Province in Río Negro (Argentina) with six stable states and 12 transitions, based on the woody encroachment degree, characteristics of the herbaceous layer and the soil surface, and biological crust cover. Information was obtained from letters of naturalists and travelers along the region since the late eighteenth century, reports of old settlers, and our own research work. On the Monte, different functional groups can be distinguished. Our work focused on the three most conspicuous to analyze the dynamics of the system: shrubs, grasses and biological crust. The shrub layer is only affected by large-scale disturbances (fire, mechanical clearing) which maintain the balance between grasses and shrubs in the system. Mechanical clearing with soil removal also causes a decrease in grass cover and vegetation diversity. This layer, however, is most frequently affected by intensive and continuous grazing, which reduces the occurrence of fires and shifts the equilibrium toward the shrubs. All disturbances in general deteriorate the biological crust that covers the soil, which offers situations of facilitation for the regeneration of the herbaceous layer. Grazing and mechanical clearing reduce soil cover and promote the development of soil compaction, which reduce water infiltration and germination and/or seedling establishment, and this creates deteriorated states difficult to reverse. Those states can also be produced by tillage for agriculture and stubble grazing. Some of the transitions that we describe can be favored through grassland management strategies, in order to reach higher states of pastoral value and reverse deteriorating situations.     

Onion yield as affected by plant density,nitrogen level and loss of leaf area

Onion (Allium cepa) is an important horticulture crop because of its value as a food with a long shelf life being a relatively non-perishable product. It is very helpful to understand the growth response of the seeded onion crop to conduct appropriate field practices in attaining the highest or optimum yields. A three year field experiment was conducted using a variety of onion Valcatorce INTA, in a randomized block design with five replicates. Treatments were two plant densities and three rates of N application. The bulb growth followed a classical sigmoid curve. During the rapid growing period, the crop had the greatest leaf area (LA) with at least six leaves per plant. Increasing plant density increased yield in kg/ha, but decreased bulb size. Defoliating 40 to 60% of the LA had a significant impact on bulb production only at early growth stages. Late in the growing period, the remaining LA was apparently large enough for producing sufficient amounts of metabolites to feed new leaves, increasing their photosynthesis efficiency for the benefit of bulb production.     

IGF-1 modulates gene expression of proteins involved in inflammation,cytoskeleton, and liver architecture

Even though the liver synthesizes most of circulating IGF-1, it lacks its receptor under physiological conditions. However, according to previous studies, a damaged liver expresses the receptor. For this reason, herein, we examine hepatic histology and expression of genes encoding proteins of the cytoskeleton, extracellular matrix, and cell-cell molecules and inflammation-related proteins. A partial IGF-1 deficiency murine model was used to investigate IGF-1’s effects on liver by comparing wild-type controls, heterozygous igf1^+/?, and heterozygous mice treated with IGF-1 for 10 days. Histology, microarray for mRNA gene expression, RT-qPCR, and lipid peroxidation were assessed. Microarray analyses revealed significant underexpression of igf1 in heterozygous mice compared to control mice, restoring normal liver expression after treatment, which then normalized its circulating levels. IGF-1 receptor mRNA was overexpressed in Hz mice liver, while treated mice displayed a similar expression to that of the controls. Heterozygous mice showed overexpression of several genes encoding proteins related to inflammatory and acute-phase proteins and underexpression or overexpression of genes which coded for extracellular matrix, cytoskeleton, and cell junction components. Histology revealed an altered hepatic architecture. In addition, liver oxidative damage was found increased in the heterozygous group. The mere IGF-1 partial deficiency is associated with relevant alterations of the hepatic architecture and expression of genes involved in cytoskeleton, hepatocyte polarity, cell junctions, and extracellular matrix proteins. Moreover, it induces hepatic expression of the IGF-1 receptor and elevated acute-phase and inflammation mediators, which all resulted in liver oxidative damage.     

Fungi associated with rice at entre rios province, Argentina. toxigenic capacity offusarium graminearum andmicrodochium nivale isolates

A mycological survey was carried out on rice samples harvested in 1997 and 1998 from Entre Ríos province, belonging to the main production area of Argentina. The relative density and isolation frequency of the prevalent fungi were statistically compared between locations and harvest seasons. The genusAlternaria was the most prevalent component of the internal seedborne mycoflora in the two harvest seasons. Fungi belonging to the generaPhoma, Fusarium, Microdochium, Penicillium andAspergillus were also isolated. The predominant species of these genera wereAlternaria alternata, Phoma glomerata, Fusarium graminearum, Microdochium nivale, Penicillium citrinum and Aspergillus flavus.     

Molecular dynamics of synthetic leucine-serine ion channels in a phospholipid membrane

Molecular dynamics calculations were carried out on models of two synthetic leucine-serine ion channels: a tetrameric bundle with sequence (LSLLLSL)(3)NH(2) and a hexameric bundle with sequence (LSSLLSL)(3)NH(2). Each protein bundle is inserted in a palmitoyloleoylphosphatidylcholine bilayer membrane and solvated by simple point charge water molecules inside the pore and at both mouths. Both systems appear to be stable in the absence of an electric field during the 4 ns of molecular dynamics simulation. The water motion in the narrow pore of the four-helix bundle is highly restricted and may provide suitable conditions for proton transfer via a water wire mechanism. In the wider hexameric pore, the water diffuses much more slowly than in bulk but is still mobile. This, along with the dimensions of the pore, supports the observation that this peptide is selective for monovalent cations. Reasonable agreement of predicted conductances with experimentally determined values lends support to the validity of the simulations.     

Genetic engineering of Bacillus subtilis for the commercial production of riboflavin

Recombinant DNA engineering was combined with mutant selection and fermentation improvement to develop a strain of Bacillus subtilis that produces commercially attractive levels of riboflavin. The B. subtilis riboflavin production strain contains multiple copies of a modified B. subtilis riboflavin biosynthetic operon (rib operon) integrated at two different sites in the B. subtilis chromosome. The modified rib operons are expressed constitutively from strong phage promoters located at the 5′ end and in an internal region of the operon. The engineered strain also contains purine analog-resistant mutations designed to deregulate the purine pathway (GTP is the precursor for riboflavin), and a riboflavin analog-resistant mutation in ribC that deregulates the riboflavin biosynthetic pathway. Received 22 June 1998/ Accepted in revised form 6 November 1998