Tissue invasion and metastasis: Molecular, biological and clinical perspectives.

Cancer is a key health issue across the world, causing substantial patient morbidity and mortality. Patient prognosis is tightly linked with metastatic dissemination of the disease to distant sites, with metastatic diseases accounting for a vast percentage of cancer patient mortality. While advances in this area have been made, the process of cancer metastasis and the factors governing cancer spread and establishment at secondary locations is still poorly understood. The current article summarizes recent progress in this area of research, both in the understanding of the underlying biological processes and in the therapeutic strategies for the management of metastasis. This review lists the disruption of E-cadherin and tight junctions, key signaling pathways, including urokinase type plasminogen activator (uPA), phosphatidylinositol 3-kinase/v-akt murine thymoma viral oncogene (PI3K/AKT), focal adhesion kinase (FAK), ß-catenin/zinc finger E-box binding homeobox 1 (ZEB-1) and transforming growth factor beta (TGF-ß), together with inactivation of activator protein-1 (AP-1) and suppression of matrix metalloproteinase-9 (MMP-9) activity as key targets and the use of phytochemicals, or natural products, such as those from Agaricus blazei, Albatrellus confluens, Cordyceps militaris, Ganoderma lucidum, Poria cocos and Silybum marianum, together with diet derived fatty acids gamma linolenic acid (GLA) and eicosapentanoic acid (EPA) and inhibitory compounds as useful approaches to target tissue invasion and metastasis as well as other hallmark areas of cancer. Together, these strategies could represent new, inexpensive, low toxicity strategies to aid in the management of cancer metastasis as well as having holistic effects against other cancer hallmarks.

Effects of the Ras homolog Rhes on Akt/protein kinase B and glycogen synthase kinase 3 phosphorylation in striatum.

G protein-coupled receptors (GPCR) signal not only through heterotrimeric G proteins, but also through alternate pathways. Thus, dopamine D2 receptors in the striatum signal through Gαi/o and also by promoting formation of a multi-protein complex containing ß-arrestin2, protein phosphatase 2A (PP2A), and Akt in order to dephosphorylate Akt. Lithium, on the other hand, disrupts this complex to increase Akt phosphorylation. Rhes is a striatally enriched GTP-binding protein that has been shown to inhibit dopamine receptor-mediated behavior and signaling through heterotrimeric G proteins. Therefore, our objective was to test whether Rhes similarly affects signaling through the Akt/GSK3 pathway in the striatum. Rhes(-/-) mice showed basally increased Akt and GSK3ß phosphorylation relative to rhes(+/+) mice that was not further enhanced by lithium treatment. Furthermore, they responded to the D1/D2 agonist apomorphine with increased Akt and GSK3 phosphorylation. Co-immunoprecipitation experiments revealed that apomorphine treatment recruits PP 2A-C to Akt in both rhes(+/+) and rhes(-/-) mice. Lithium did not disrupt their interaction in rhes(-/-) mice as there was little basal interaction. Rhes co-immunoprecipitated with ß-arrestins, suggesting that it is integral to the multi-protein complex. Thus, Rhes is necessary for Akt dephosphorylation by the striatal multi-protein complex, and in its absence, a lithium-treated phenotype results.

Purification, primary structure, and properties of Euphorbia characias latex purple acid phosphatase.

A purple acid phosphatase was purified to homogeneity from Euphorbia characias latex. The native protein has a molecular mass of 130 ± 10 kDa and is formed by two apparently identical subunits, each containing one Fe(III) and one Zn(II) ion. The two subunits are connected by a disulfide bridge. The enzyme has an absorbance maximum at 540 nm, conferring a characteristic purple color due to a charge-transfer transition caused by a tyrosine residue (Tyr172) coordinated to the ferric ion. The cDNA nucleotide sequence contains an open reading frame of 1392 bp, and the deduced sequence of 463 amino acids shares a very high degree of identity (92-99%) to other purple acid phosphatases isolated from several higher plants. The enzyme hydrolyzes well p-nitrophenyl phosphate, a typical artificial substrate, and a broad range of natural phosphorylated substrates, such as ATP, ADP, glucose-6-phosphate, and phosphoenolpyruvate. The enzyme displays a pH optimum of 5.75 and is inhibited by molybdate, vanadate, and Zn2+, which are typical acid phosphatase inhibitors.

Activity and structural changes of Euphorbia characias peroxidase in the presence of trifluoroethanol.

Activity assays, conformational changes and transitional switches between secondary structures of a peroxidase from Euphorbia characias were studied in the presence of trifluoroethanol and in the presence or absence of calcium ions. The addition of trifluoroethanol up to 10-20% first induced a drastic decrease of alpha-helix content followed by an increase of tryptophan fluorescence emission intensity, a progressive re-induction of the formation of alpha-helical elements concomitant with loss of enzyme activity. In the presence of calcium ions, the fluorescence of the enzyme almost remained unchanged in the trifluoroethanol concentration range 5-20%. Further increase in trifluoroethanol concentration led to a protein structure characterized by a progressive re-induction of alpha-helical elements, a remarkable increase of the tryptophan fluorescence and a loss of enzyme activity. These results indicate that calcium ions in Euphorbia peroxidase play an essential role in maintaining the hydrophobic interactions on the protein structure preserving enzymatic activity.

Estrogen-induced proliferation in cultured hepatocytes involves cyclin D1, p21(Cip1) and p27(Kip1).

The purpose of this study was to establish if estrogen-induced hepatocyte proliferation in vitro involves the cell cycle regulators cyclin D1, p21(Cip1), and p27(Kip1). Male rat hepatocytes were cultured in presence of 17-beta-estradiol (E2) +/- ICI-182780, a pure estrogen antagonist, and [3H]-thymidine, as required. DNA synthesis as well as p21(Cip1), p27(Kip1), and cyclin D1mRNA and protein levels were evaluated at different times (12, 24, 36, and 48 hours) of incubation. E2-increased DNA synthesis was correlated with cyclin D1 and p21(Cip1) (mRNA and protein) variations that were reversed by the addition of ICI-182780. p27(Kip1) protein levels progressively increased regardless of the presence of E2 or ICI-182780. Our data confirm that estrogens' stimulatory effect is related to their ability to increase cyclin D1 levels. The increase of p21(Cip1) is probably related to the reentry of hepatocytes in the quiescent state. p27(Kip1) protein is not able to arrest hepatocyte proliferation.

An improved model for determining degree-day values from daily temperature data.

Although using hourly weather data offers the greatest accuracy for estimating growing degree-day values, daily maximum and minimum temperature data are often used to estimate these values by approximating the diurnal temperature trends. This paper presents a new empirical model for estimating the hourly mean temperature. The model describes the diurnal variation using a sine function from the minimum temperature at sunrise until the maximum temperature is reached, another sine function from the maximum temperature until sunset, and a square-root function from then until sunrise the next morning. The model was developed and calibrated using several years of hourly data obtained from five automated weather stations located in California and representing a wide range of climate conditions. The model was tested against an additional data-set at each location. The temperature model gave good results, the rootmean-square error being less than 2.0 degrees C for most years and locations. The comparison with published models from the literature showed that the model was superior to the other methods. Hourly temperatures from the model were used to calculate degree-day values. A comparison between degree-day estimates determined from the model and those obtained other selected methods is presented. The results showed that the model had the best accuracy in general regardless of the season.

Temperature data for phenological models.

In an arid environment, the effect of evaporation on energy balance can affect air temperature recordings and greatly impact on degree-day calculations. This is an important consideration when choosing a site or climate data for phenological models. To our knowledge, there is no literature showing the effect of the underlying surface and its fetch around a weather station on degree-day accumulations. In this paper, we present data to show that this is a serious consideration, and it can lead to dubious models. Microscale measurements of temperature and energy balance are presented to explain why the differences occur. For example, the effect of fetch of irrigated grass and wetting of bare soil around a weather station on diurnal temperature are reported. A 43-day experiment showed that temperature measured on the upwind edge of an irrigated grass area averaged 4% higher than temperatures recorded 200 m inside the grass field. When the single-triangle method was used with a 10 degrees C threshold and starting on May 19, the station on the upwind edge recorded 900 degree-days on June 28, whereas the interior station recorded 900 degree-days on July 1. Clearly, a difference in fetch can lead to big errors for large degree-day accumulations. Immediately after wetting, the temperature over a wet soil surface was similar to that measured over grass. However, the temperature over the soil increased more than that over the grass as the soil surface dried. Therefore, the observed difference between temperatures measured over bare soil and those over grass increases with longer periods between wettings. In most arid locations, measuring temperature over irrigated grass gives a lower mean annual temperature, resulting in lower annual cumulative degree-day values. This was verified by comparing measurements over grass with those over bare soil at several weather stations in a range of climates. To eliminate the effect of rainfall frequency, using temperature data collected only over irrigated grass, is recommended for long-term assessment of climate change effects on degree-day accumulation. In high evaporative conditions, a fetch of at least 100 m of grass is recommended. Our results clearly indicate that weather stations sited over bare soil have consistently higher degree-day accumulations. Therefore, especially in arid environments, phenology models based on temperature collected over bare soil are not transferable to those based on temperature recorded over irrigated grass. At a minimum, all degree-day-based phenology models reported in the literature should clearly describe the weather station site.