DMSO Represses Inflammatory Cytokine Production from Human Blood Cells and Reduces Autoimmune Arthritis

Dimethyl sulfoxide (DMSO) is currently used as an alternative treatment for various inflammatory conditions as well as for cancer. Despite its widespread use, there is a paucity of data regarding its safety and efficacy as well as its mechanism of action in human cells. Herein, we demonstrate that DMSO has ex-vivo anti-inflammatory activity using Escherichia coli- (E. coli) and herpes simplex virus-1 (HSV-1)-stimulated whole human blood. Specifically, we found that between 0.5%– 2%, DMSO significantly suppressed the expression of many pro-inflammatory cytokines/chemokines and prostaglandin E₂ (PGE₂). However, a significant reduction in monocyte viability was also observed at 2% DMSO, suggesting a narrow window of efficacy. Anti-inflammatory concentrations of DMSO suppressed E. coli-induced ERK1/2, p38, JNK and Akt phosphorylation, suggesting DMSO acts on these signaling pathways to suppress inflammatory cytokine/chemokine production. Although DMSO induces the differentiation of B16/F10 melanoma cells in vitro, topical administration of DMSO to mice subcutaneously implanted with B16 melanoma cells was ineffective at reducing tumor growth, DMSO was also found to block mouse macrophages from polarizing to either an M1- or an M2-phenotype, which may contribute to its inability to slow tumor growth. Topical administration of DMSO, however, significantly mitigated K/BxN serum-induced arthritis in mice, and this was associated with reduced levels of pro-inflammatory cytokines in the joints and white blood cell levels in the blood. Thus, while we cannot confirm the efficacy of DMSO as an anti-cancer agent, the use of DMSO in arthritis warrants further investigation to ascertain its therapeutic potential.     

Tetrad analysis shows that gene conversion is the major mechanism involved in mutation at the human minisatellite MS1 integrated in Saccharomyces cerevisiae

Minisatellites are arrays of tandemly repeated DNA sequences which occur at thousands of locations in the human genome. They are frequently hypervariable with respect to allele length as a result of high rates of complex and incompletely understood recombination-based germline mutation events that alter the repeat copy number. MS1 is one of the most variable minisatellites so far isolated from the human genome. We have integrated MS1, flanked by synthetic markers, in the vicinity of a hot spot for meiotic double-strand breaks upstream of the LEU2 locus in chromosome III of Saccharomyces cerevisiae. Here we present the first tetrad analysis of mutations at a human minisatellite locus. The data showed that mutant alleles occur as single mutants in one of the spores in a tetrad, also when the mutant structure was the result of a combination of intra- and inter-allelic rearrangements. The conversional transfer of repeat units from one allele to the other was associated with flanking marker conversion which always involved the same flank of the minisatellite. The results demonstrate that conversion is the predominant mechanism by which minisatellite alleles mutate to new lengths, and also support the assumption that cis-acting elements are involved in the regulation of the mutational process in humans.     

Reddish spring colouring of deciduous leaves: a sign of ecotype?

Several deciduous woody plant species produce anthocyanins during leaf development in spring and again during leaf senescence in autumn. The leaves of Betula pendula Roth (silver birch) commonly exhibit transient reddening in juvenile leaves under northern growing conditions, with the intensity of the red colour varying among individual trees. The objective of our study was to test the hypothesis that the accumulation of foliar anthocyanins during spring in leaves of B. pendula is an ecotypic response. Chlorophyll fluorescence ratio (Fv/Fm), leaf reflectance and anthocyanin concentrations were measured, in relation to phenology in spring, summer and autumn from birches used for landscaping with either red or green-emergent leaves. The results suggest that (1) the trees with green- or red-emergent juvenile leaves represent different populations, and (2) that the red-emergent leaves senesced earlier, indicating that (3) trees with red-emerging leaves belong to a more northern ecotype than the trees with green-emerging leaves. The role of anthocyanin synthesis in a northern radiation environment is discussed.     

Simple and inexpensive immunoassay-based diagnostic tests

Simple and inexpensive yet sensitive and robust diagnostic tests are critically needed for resource-poor settings to enable timely diagnosis and effective use of limited health care resources. Current tests are often too expensive, too slow, or have compromised clinical performance, and they often require health care professional to perform the test. In addition, most assays are not intended to be used in extreme environmental conditions, but their performance may be affected by high temperatures and humidity often encountered in resource-poor settings. This review provides an overview of current immunoassay technologies and their advantages and limitations with respect to their feasibility to resource-poor settings. Future trends of immunoassay development for decentralized testing are also discussed. Homogeneous assays as such are out of the scope of this article because they are generally not yet sensitive enough or otherwise less feasible for inexpensive rapid diagnostic tests.     

Variation in carbon footprint of milk due to management differences between Swedish dairy farms

To identify mitigation options to reduce greenhouse gas (GHG) emissions from milk production (i.e. the carbon footprint (CF) of milk), this study examined the variation in GHG emissions among dairy farms using data from previous CF studies on Swedish milk. Variations between farms in these production data, which were found to have a strong influence on milk CF, were obtained from existing databases of 1051 dairy farms in Sweden in 2005. Monte Carlo (MC) analysis was used to analyse the impact of variations in seven important parameters on milk CF concerning milk yield (energy-corrected milk (ECM) produced and delivered), feed dry matter intake (DMI), enteric CH4 emissions, N content in feed DMI, N-fertiliser rate and diesel used on farm. The largest between-farm variations among the analysed production data were N-fertiliser rate (kg/ha) and diesel used (l/ha) on farm (CV = 31% to 38%). For the parameters concerning milk yield and feed DMI, the CV was approximately 11% and 8%, respectively. The smallest variation in production data was found for N content in feed DMI. According to the MC analysis, these variations in production data led to a variation in milk CF of between 0.94 and 1.33 kg CO2 equivalents (CO2e)/kg ECM, with an average value of 1.13 kg CO2e/kg ECM. We consider that this variation of ±17%, which was found to be based on the used farm data, would be even greater if all Swedish dairy farms were included, as the sample of farms in this study was not totally unbiased. The variation identified in milk CF indicates that a potential exists to reduce GHG emissions from milk production on both the national and farm levels through changes in management. As milk yield and feed DMI are two of the most influential parameters for milk CF, feed conversion efficiency (i.e. units ECM produced/unit DMI) can be used as a rough key performance indicator for predicting CF reductions. However, it must be borne in mind that feeds have different CF due to where and how they are produced.     

Not4 enhances JAK/STAT pathway-dependent gene expression in Drosophila and in human cells

The JAK/STAT pathway is essential for organogenesis, innate immunity, and stress responses in Drosophila melanogaster. The JAK/STAT pathway and its associated regulators have been highly conserved in evolution from flies to humans. We have used a genome-wide RNAi screen in Drosophila S2 cells to identify regulators of the JAK/STAT pathway, and here we report the characterization of Not4 as a positive regulator of the JAK/STAT pathway. Overexpression of Not4 enhanced Stat92E-mediated gene responses in vitro and in vivo in Drosophila. Specifically, Not4 increased Stat92E-mediated reporter gene activation in S2 cells; and in flies, Not4 overexpression resulted in an 8-fold increase in Turandot M (TotM) and in a 4-fold increase in Turandot A (TotA) stress gene activation when compared to wild-type flies. Drosophila Not4 is structurally related to human CNOT4, which was found to regulate interferon-γ- and interleukin-4-induced STAT-mediated gene responses in human HeLa cells. Not4 was found to coimmunoprecipitate with Stat92E but not to affect tyrosine phosphorylation of Stat92E in Drosophila cells. However, Not4 is required for binding of Stat92E to its DNA recognition sequence in the TotM gene promoter. In summary, Not4/CNOT4 is a novel positive regulator of the JAK/STAT pathway in Drosophila and in humans.