Temperature dependence of the effects of some monohydric alcohols on the oxygen affinity of hemoglobin: Determination and analysis of thermodynamic parameters

We studied the effects of methanol, ethanol, iso-propanol, and n-propanol on the reaction of hemoglobin with oxygen at various temperatures. The analysis of the results in terms of the Monod-Wyman-Changeux model allowed determination of the overall contribution of the alcohols to the standard enthalpy and entropy differences between the T and R states of hemoglobin. A phenomenological approach allowed us to obtain separately the contributions related to the variations of the bulk dielectric constant of the solvent (bulk electrostatic contributions) and the contributions related to other effects (non-bulk-electrostatic contributions). The values of non-bulk-electrostatic contributions to ΔΔH and ΔΔS supported the suggestion that these contributions are mainly related to protein-solvent hydrophobic interactions.     

Identification of a novel transporter for dicarboxylates and tricarboxylates in plant mitochondria. Bacterial expression, reconstitution, functional characterization, and tissue distribution

A cDNA from Arabidopsis thaliana and four related cDNAs from Nicotiana tabacum that we have isolated encode hitherto unidentified members of the mitochondrial carrier family. These proteins have been overexpressed in bacteria and reconstituted into phospholipid vesicles. Their transport properties demonstrate that they are orthologs/isoforms of a novel mitochondrial carrier capable of transporting both dicarboxylates (such as malate, oxaloacetate, oxoglutarate, and maleate) and tricarboxylates (such as citrate, isocitrate, cis-aconitate, and trans-aconitate). The newly identified dicarboxylate-tricarboxylate carrier accepts only the single protonated form of citrate (H-citrate2-) and the unprotonated form of malate (malate2-) and catalyzes obligatory, electroneutral exchanges. Oxoglutarate, citrate, and malate are mutually competitive inhibitors, showing K(i) close to the respective K(m). The carrier is expressed in all plant tissues examined and is largely spread in the plant kingdom. Furthermore, nitrate supply to nitrogen-starved tobacco plants leads to an increase in its mRNA in roots and leaves. The dicarboxylate-tricarboxylate carrier may play a role in important plant metabolic functions requiring organic acid flux to or from the mitochondria, such as nitrogen assimilation, export of reducing equivalents from the mitochondria, and fatty acid elongation.     

Human chromosomes 9, 12, and 15 contain the nucleation sites of stress-induced nuclear bodies

We previously reported the identification of a novel nuclear compartment detectable in heat-shocked HeLa cells that we termed stress-induced Src-activated during mitosis nuclear body (SNB). This structure is the recruitment center for heat shock factor 1 and for a number of RNA processing factors, among a subset of Serine-Arginine splicing factors. In this article, we show that stress-induced SNBs are detectable in human but not in hamster cells. By means of hamster>human cell hybrids, we have identified three human chromosomes (9, 12, and 15) that are individually able to direct the formation of stress bodies in hamster cells. Similarly to stress-induced SNB, these bodies are sites of accumulation of hnRNP A1-interacting protein and heat shock factor 1, are usually associated to nucleoli, and consist of clusters of perichromatin granules. We show that the p13-q13 region of human chromosome 9 is sufficient to direct the formation of stress bodies in hamster>human cell hybrids. Fluorescence in situ hybridization experiments demonstrate that the pericentromeric heterochromatic q12 band of chromosome 9 and the centromeric regions of chromosomes 12 and 15 colocalize with stress-induced SNBs in human cells. Our data indicate that human chromosomes 9, 12, and 15 contain the nucleation sites of stress bodies in heat-shocked HeLa cells.     

In vitro T cell-mediated killing of Pseudomonas aeruginosa. I. Evidence that a lymphokine mediates killing

Previous studies have demonstrated in vivo that T cells can provide protective immunity, in the absence of antibody, against infection with the extracellular Gram-negative bacterium Immunotype 1 (IT-1) Pseudomonas aeruginosa. We established an in vitro system in which immune T cells, after reexposure to bacterial antigens and to macrophages, secrete a product that kills the bacteria. Although macrophages are required for in vitro killing, they function neither as antigen-presenting nor as phagocytic cells in this system. T cells from animals immunized against a different P. aeruginosa immunotype will not kill IT-1 organisms; but the supernatants produced by IT-1 immune T cells after exposure to macrophages and IT-1 P. aeruginosa organisms are nonspecifically effective in killing unrelated bacteria. Because the supernatants from immune T cells lose their bactericidal properties upon minimal dilution, we conclude that if this mechanism is active in vivo, it must play a role in local immunity.     

Characterization of Nine Novel Mutations in the CD40 Ligand Gene in Patients with X-Linked Hyper IgM Syndrome of Various Ancestry

X-linked immunodeficiency with hyper-IgM (HIGMX-1) is a rare disorder caused by defective expression of the CD40 ligand (CD40L) by activated T lymphocytes, resulting in inefficient T-B cell cooperation and failure of B cells to undergo immunoglobulin isotype switch. In the present work, we describe nine patients of various ancestry who bear different mutations in the X chromosome–specific CD40L gene. Two of the mutations were nonsense mutations, one each resulting in premature stop codons at amino acid residues 39 and 140. Three patients had single point missense mutations, one each at codons 126, 140, and 144. Another patient had a 4-bp genomic deletion in exon 2, resulting in a frameshift and premature termination. Three patients showed insertions, one each of 1, 2, and 4 nt, probably because of polymerase slippage, resulting in frameshift mutation and premature termination. Overall, these observations confirm the heterogeneity of mutations in HIGMX-1. However, the identification of two patients whose mutation involves codon 140 (previously shown to be altered in two other unrelated subjects) suggests that this may be a hotspot of mutation in HIGMX-1. In two additional patients with clinical and immunological features indistinguishable from canonical HIGMX-1, no mutation was detected in the coding sequence, in the 5' flanking region, or in the 3' UTR.     

Factors affecting the distribution of<Emphasis Type="Italic">Sorex samniticus</Emphasis>, an endemic Italian shrew,in an heterogeneous landscape

We studied the factors affecting the distribution of an endemic shrewSorex samniticus Altobello, 1926 in the Province of Siena, central Italy. Distribution data were obtained by examining pellets from 31 barn owlTyto alba sites (roosts) over a period spanning from 1974 to 2005. We constructed a model using logistic regression analysis on presence/absence data. Results show that an increase in forest dominated byQuercus cerris andCastanea sativa led to the local extinction of this species. Results were confirmed through the examination of one box, with 3044 prey items deposited in layers, that documents the expansion of the species in that area following increased logging ofQuercus cerris andCastanea sativa forests. Cessation of logging has again led to the absence of the species from the area. We discuss these results from the perspective of ecological network planning, showing that utilisation of non-detailed maps, such as Corine Land Cover, that do not distinguish between the various kind of broadleaved forests is inadequate to describe the finer grade of habitat selection of this small mammal.     

Playing both sides: nucleophosmin between tumor suppression and oncogenesis

A question preoccupying many researchers is how signal transduction pathways control metabolic processes and energy production. A study by Jang et al. (Jang, C., G. Lee, and J. Chung. 2008. J. Cell Biol. 183:11–17) provides evidence that in Drosophila melanogaster a signaling network controlled by the LKB1 tumor suppressor regulates trafficking of an Sln/dMCT1 monocarboxylate transporter to the plasma membrane. This enables cells to import additional energy sources such as lactate and butyrate, enhancing the repertoire of fuels they can use to power vital activities.     

Soil Carbon, Nitrogen and Phosphorus Dynamics as Affected by Solarization Alone or Combined with Organic Amendment

Soil solarization, alone or combined with organic amendment, is an increasingly attractive approach for managing soil-borne plant pathogens in agricultural soils. Even though it consists in a relatively mild heating treatment, the increased soil temperature may strongly affect soil microbial processes and nutrients dynamics. This study aimed to investigate the impact of solarization, either with or without addition of farmyard manure, in soil dynamics of various C, N and P pools. Changes in total C, N and P contents and in some functionally-related labile pools (soil microbial biomass C and N, K₂SO₄-extractable C and N, basal respiration, KCl-exchangeable ammonium and nitrate, and water-soluble P) were followed across a 72-day field soil solarization experiment carried out during a summer period on a clay loam soil in Southern Italy. Soil physico-chemical properties (temperature, moisture content and pH) were also monitored. The average soil temperature at 8-cm depth in solarized soils approached 55 °C as compared to 35 °C found in nonsolarized soil. Two-way ANOVA (solarization×organic amendment) showed that both factors significantly affected most of the above variables, being the highest influence exerted by the organic amendment. With no manure addition, solarization did not significantly affect soil total C, N and P pools. Whereas soil pH, microbial biomass and, at a greater extent, K₂SO₄-extractable N and KCl-exchangeable ammonium were greatly affected. An increased release of water-soluble P was also found in solarized soils. Yet, solarization altered the quality of soluble organic residues released in soil as it lowered the C-to-N ratio of both soil microbial biomass and K₂SO₄-extractable organic substrates. Additionally, in solarized soils the metabolic quotient (qCO₂) significantly increased while the microbial biomass C-to-total organic C ratio (microbial quotient) decreased over the whole time course. We argued that soil solarization promoted the mineralization of readily decomposable pools of the native soil organic matter (e.g. the microbial biomass) thus rendering larger, at least over a short-term, the available fraction of some soil mineral nutrients, namely N and P forms. However, over a longer prospective solarization may lead to an over-exploitation of labile organic resources in agricultural soils. Manure addition greatly increased the levels of both total and labile C, N and P pools. Thus, addition of organic amendments could represent an important strategy to protect agricultural lands from excessive soil resources exploitation and to maintain soil fertility while enhancing pest control.