Isolation, Nucleotide Sequence, and Physiological Relevance of the Gene Encoding Triose Phosphate Isomerase from Kluyveromyces lactis

Lack of triose phosphate isomerase activity (TIM) is of special interest because this enzyme works at an important branch point of glycolytic flux. In this paper, we report the cloning and sequencing of the Kluyveromyces lactis gene encoding TIM. Unlike Saccharomyces cerevisiae ΔTPI1 mutants, the K. lactis mutant strain was found to be able to grow on glucose. Preliminary bioconversion experiments indicated that, like the S. cerevisiae TIM-deficient strain, the K. lactis TIM-deficient strain is able to produce glycerol with high yield.     

Time-dependent attachment mechanism of bacterial pathogen during ice-ice infection in Kappaphycus alvarezii (Gigartinales, Rhodophyta)

The mechanism of infection by Vibrio sp. P11 promoting the ice-ice disease in Kappaphycus alvarezii was investigated in vitro. Its intensity of infection differs from that of another ice-ice promoter (Cytophaga sp. P25) by promoting the disease much faster. However, when secondary infection by other bacteria starts, its ability to compete with these bacteria gradually diminishes, whereas, infection by P25, although not displaying such drastic effects as P11, shows consistent competitive ability against other bacteria. Time-series infection experiments with application of polyclonal antibodies to specifically detect Vibrio sp. P11 revealed that this bacterium has a high affinity for the seaweed especially when the latter is stressed. It promotes the disease after a rapid increase in cell density of up to 10⁷ g⁻¹ (wet wt.) in the first 24 h. This bacterial cell build-up may take only 1–2 h on stressed thalli, but takes about 24 h on non-stressed thalli. Build-up is not sustainable in non-stressed thalli as high density is usually followed by a sudden decline in cell number believed to result from an algal defence against potential pathogens. Inoculation of the bacterium on thalli incubated in continuous culture system extends the time of bacterial attachment due to laminar flow and, possibly, competition by existing bacteria on the seaweed surface and in ambient seawater medium. Motility-driven cell attachment by this bacterium is suggested as an important factor for infection. This revised version was published online in June 2006 with corrections to the Cover Date.     

Glyphosate affects chlorophyll,photosynthesis and water use of four Intacta RR2 soybean cultivars

The planting of RR2 Intacta soybeans by farmers has been expanding strongly. However, some visual injuries have been noted after glyphosate application. The aim of this study was to evaluate the influence of glyphosate application on chlorophyll, photosynthesis and water use of four Intacta RR2 soybean cultivars. The experiment was conducted in a greenhouse, in a randomized block design with a 3 × 4 factorial scheme, consisting of three glyphosate rates and four soybean cultivars. The glyphosate formula used was isopropylamine salt + potassium salt. The parameters analyzed were phytotoxicity at 7, 14, 21 and 28 days after application, and total chlorophyll index at 0, 3, 7, 14, 21, 28, 35, 42 and 49 days after application. Furthermore, 40 days after application, the net CO2 assimilation rate (A), transpiration rate (E), stomatal conductance (G), and internal CO₂ concentration (Ci) were evaluated as well. The water-use efficiency (WUE) and carboxylation efficiency were calculated. The data were submitted to analysis of variance and compared by the Tukey’s test (p ≤ 0.05), followed by regression analysis. The phytotoxicity influence could be seen until 21 days after application, in which Monsoy 6210 IPRO cultivar was the most injured. The increasing doses promoted a reduction of the chlorophyll level up to 35 days after application in Monsoy 6410 IPRO. The cultivars tested here showed similar chlorophyll index values. On the 3rd, 7th and 14th DAA (Fig. 5a–c), there was a significant linear decline in the chlorophyll index with rising glyphosate dose for all four cultivars. The chlorophyll index cultivars were not influenced by the doses on the 42nd and 49th DAA. There was no difference in water use and carboxylation efficiency. The parameters A, E and A/Ci showed a positive correlation as the doses increased, while Ci declined, in both cultivars. The application of glyphosate on these soybean cultivars causes different injuries according to the sensitivity. In general, RR2 soybeans have the ability to recover from visual intoxication injuries and reestablish the normal chlorophyll production and photosynthetic parameters after glyphosate application.     

Growth and development of conifer pollen tubes

Conifer pollen tubes are an important but underused experimental system in plant biology. They represent a major evolutionary step in male gametophyte development as an intermediate form between the haustorial pollen tubes of cycads and Ginkgo and the structurally reduced and faster growing pollen tubes of flowering plants. Conifer pollen grains are available in large quantities, most can be stored for several years, and they grow very well in culture. The study of pollen tube growth and development furthers our understanding of conifer reproduction and contributes towards our ability to improve on their productivity. This review covers taxonomy and morphology to cell, developmental, and molecular biology. It explores recent advances in research on conifer pollen and pollen tubes in vivo, focusing on pollen wall structure, male gametophyte development within the pollen wall, pollination mechanisms, pollen tube growth and development, and programmed cell death. It also explores recent research in vitro, including the cellular mechanisms underlying pollen tube elongation, in vitro fertilization, genetic transformation and gene expression, and pine pollen tube proteomics. With the ongoing sequencing of the Pinus taeda genome in several labs, we expect the use of conifer pollen tubes as an experimental system to increase in the next decade.     

Synthesis and biological affinity of new imidazo- and indol-arylpiperazine derivatives: further validation of a pharmacophore model for alpha(1)-adrenoceptor antagonists

In the continuing search for selective alpha(1)-adrenoceptor (AR) antagonists, new alkoxyarylpiperazinylalkylpyridazinone derivatives were designed and synthesized. The new compounds were tested for their affinity toward alpha(1)-AR, alpha(2)-AR and 5-HT(1A) receptors. The ability of these compounds to inhibit the serotonin transporters (SERT) was also determined. The pharmacological data confirm that increasing the size of the ortho alkoxy substituent on the phenyl ring of the arylpiperazine moiety afforded compounds with enhanced affinity toward the alpha(1)-AR. The isopropoxy group, the largest group evaluated, led the best alpha(1)-AR affinity profile. In contrast, the compounds which have an amide group within of the o-alkoxy-phenylpiperazine fragment showed low affinity toward the receptors studied. Similar results were obtained when the amide group was present in the linker of the junction between the two major constituents of the molecule.     

Soil–vegetation relationships in cerrados under different fire frequencies

Fire is an important ecological factor that structures savannas, such as the cerrado, by selecting plant species and altering soil nutrient content. In Emas National Park, central Brazil, we compared soils under three different fire regimes and their relationship to the cerrado species they support. We collected 25 soil and vegetation samples at each site. We found differences in soil characteristics (p?<?0.05), with fertility and fire frequency positively related: in the annually burned site we found higher values of organic matter, nitrogen, and clay, whereas in the protected site we detected lower values of pH and higher values of aluminum. We also observed differences in plant community structure, with distinct floristic compositions in each site. Floristic composition was more related to sand proportion (intra-set correlation?=?0.834). Different fire frequencies increase environmental heterogeneity and beta diversity in the Brazilian cerrado.     

Insulin induces the low density lipoprotein receptor‐related protein 1 (LRP1) degradation by the proteasomal system in J774 macrophage‐derived cells

Low‐density lipoprotein receptor‐related protein 1 (LRP1) is an endocytic receptor, which binds and internalizes diverse ligands such as activated α₂‐macroglobulin (α₂M*). LRP1 promotes intracellular signaling, which downstream mediates cellular proliferation and migration of different types of cells, including macrophages. Unlike the LDL receptor, LRP1 expression is not sensitive to cellular cholesterol levels but appears to be responsive to insulin. It has been previously demonstrated that insulin increases the cell surface presentation of LRP1 in adipocytes and hepatocytes, which is mediated by the intracellular PI₃K/Akt signaling activation. The LRP1 protein distribution is similar to other insulin‐regulated cell surface proteins, including transferring receptor (Tfr). However, in macrophages, the insulin effect on the LRP1 distribution and expression is not well characterized. Considering that macrophages play a central role in the pathogenesis of atherosclerosis, herein we evaluate the effect of insulin on the cellular expression of LRP1 in J774 macrophages‐derived cells using Western blot and immunofluorescence microscopy. Our data demonstrate that insulin induces a significant decrease in the LRP1 protein content, without changing the specific mRNA level of this receptor. Moreover, insulin specifically affected the protein expression of LRP1 but not Tfr. The insulin‐induced protein degradation of LRP1 in J774 cells was mediated by the activation of the PI₃K/Akt pathway and proteasomal system by an enhanced ubiquitin–receptor conjugation. The decreased content of LRP1 induced by insulin affected the cellular internalization of α₂M*. Thus, we propose that the protein degradation of LRP‐1 induced by insulin in macrophages could have important effects on the pathogenesis of atherosclerosis. J. Cell. Biochem. 106: 372–380, 2009. © 2008 Wiley‐Liss, Inc.     

Effects of pasture implantation on the termite (Isoptera) fauna in the Central Brazilian Savanna (Cerrado)

The advance of agricultural frontier may cause the Cerrado (Brazilian savanna) to disappear before 2030. This work focuses on measuring the impact of pasture implantation on a cerrado’s termite fauna. Termites were sampled in a cerrado sensu stricto and a pasture, originally cerrado. All species were classified as their feeder group, accumulation curves were made and Shannon-Wiener indexes and β diversity were calculated for both areas. Cerrado was richer than pasture and species composition differed considerably, leading β diversity to a high value. The humivorous was the most representative species, followed by grass/litter feeders, xylophagous and, less representative, the intermediates. There were more xylophagous and intermediates species on cerrado than in pasture; the grass/litter feeders were more abundant in pasture, but didn’t differed in number or species; and humivorous didn’t differed neither in richness nor in abundance. This work shows that the simplification of the habitat is indeed causing the extinction of populations that depend on some specifics resource.