Equol: a comparison of the effects of the racemic compound with that of the purified S-enantiomer on the growth, invasion, and DNA integrity of breast and prostate cells in vitro

It has been postulated that the R- and S-equol enantiomers have different biological properties given their different binding affinities for the estrogen receptor. S-(-)equol is produced via the bacterial conversion of the soy isoflavone daidzein in the gut. We have compared the biological effects of purified S-equol to that of racemic (R and S) equol on breast and prostate cancer cells of varying receptor status in vitro. Both racemic and S-equol inhibited the growth of the breast cancer cell line MDA-MB-231 (> or = 10 microM) and the prostate cancer cell lines LNCaP (> or = 5 microM) and LAPC-4 (> or = 2.5 microM). The compounds also showed equipotent effects in inhibiting the invasion of MDA-MB-231 and PC-3 cancer cells through matrigel. S-equol (1, 10, 30 microM) was unable to prevent DNA damage in MCF-7 or MCF-10A breast cells following exposure to 2-hydroxy-4-nonenal, menadione, or benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide. In contrast, racemic equol (10, 30 microM) prevented DNA damage in MCF-10A cells following exposure to 2-hydroxy-4-nonenal or menadione. These findings suggest that racemic equol has strong antigenotoxic activity in contrast to the purified S-equol enantiomer implicating the R-, rather than the S-enantiomer as being responsible for the antioxidant effects of equol, a finding that may have implications for the in vivo chemoprotective properties of equol.

Fimbriae- and flagella-mediated association with and invasion of cultured epithelial cells by Salmonella enteritidis

Salmonella enteritidis expresses flagella and several finely regulated fimbriae, including SEF14, SEF17 and SEF21 (type 1). A panel of mutants was prepared in three strains of S. enteritidis to elucidate the role of these surface appendages in the association with and invasion of cultured epithelial cells. In all assays, the naturally occurring regulatory-defective strain 27655R associated with tissue culture cells significantly more than wild-type progenitor strains LA5 and S1400/94. Compared with wild-type strains, SEF14 mutants had no effect on association and invasion, whereas SEF17, SEF21 and aflagellate mutants showed significant reductions in both processes. Histological examination suggested a role for SEF17 in localized, aggregative adherence, which could be specifically blocked by anti-SEF17 sera and purified SEF17 fimbriae. SEF21-mediated association was neutralized by mannose and a specific monoclonal antibody, although to observe enhanced association it was necessary for the bacteria to be in fimbriate phase prior to infection. Additionally, aflagellate mutants associated and invaded less than motile bacteria. This study demonstrated the potential for multifactorial association and invasion of epithelial cells which involved SEF17 and SEF21 fimbriae, and flagella-mediated motility.

Contribution of fimbriae and flagella of Salmonella enteritidis to colonization and invasion of chicks

Isogenic mutants of Salmonella enteritidis defective for the elaboration of fimbrial types SEF14, SEF17, SEF21 and flagella were used to study the contribution these organelles made to colonization, invasion and lateral transfer in young chicks. The caecum, liver and spleen were colonized within 24 h following oral inoculation of 1-day-old chicks with 10(5) wild-type S. enteritidis strain LA5. However, for some mutants, the numbers of organisms recovered from internal organs was reduced significantly, particularly at 24 h post-inoculum, which supported the hypothesis that the organelles contribute to invasion and dissemination to internal organs. Specifically, mutations affecting SEF17, SEF21 and flagella contributed to a delay in colonization of the spleen, and those affecting SEF21 and flagella delayed colonization of the liver. Lower numbers of bacteria were recovered from the caecum with mutants deficient in elaboration of SEF21. Sentinel birds were colonized by LA5 or EAV40 (14(-), 17(-), 21(-), fla(-)) directly from the environment within 2 days, although a consistent slight delay was observed with the multiple mutant. Overall, our data suggest a collective role for SEF17, SEF21 and flagella, but not SEF14, in the early stages of colonization and invasion of young chicks by S. enteritidis, but these surface appendages appear unnecessary for colonization of birds from their immediate environment.

The role of fimbriae and flagella in the colonization, invasion and persistence of Escherichia coli O78 : K80 in the day-old-chick model

To understand the role of flagella and fimbriae of Escherichia coli O78:K80 in avian colibacillosis, day-old chicks were dosed orally with defined afimbriate and or aflagellate mutants and colonization, invasion and persistence compared with that of the wild-type. In an invasion model, chicks were dosed with 1 x 10(5) c.f.u. of a single strain and mutants defective for type 1 fimbriae, curli fimbriae or flagella colonized livers by 24 h although the numbers of bacteria present were significantly less than the wild-type, Mutants colonized between 50 and 75 % of spleens whereas the wild-type colonized 100 % of spleens. Additionally, the numbers of mutant bacteria in colonized spleens were significantly less than the wild-type. Surprisingly, mutants defective for the elaboration of more than one appendage were no more attenuated than single mutants. In a persistence model, chicks were dosed with 1 x 10(2) c.f.u. of a single strain and mutants defective for type 1 or curli or flagella or any combination thereof persisted as assessed by cloacal swabbing for 5 weeks of the experiment less well than the wild-type. In an additional persistence model, chicks were dosed with 5 x 10(2) c.f.u. of each of wild-type and one mutant together. All mutants were significantly less persistent than the wild-type (P < 0.001) and one mutant which lacked type 1, curli and flagella, was eliminated within 2 weeks. Analysis of the trends of elimination indicated that flagella contributed to persistence more than curli, which contributed more than type 1 fimbriae. Here was evidence for a major role in colonization, invasion and persistence played by type 1, curli and flagella.

Membrane ruffling and invasion of human and avian cell lines is reduced for aflagellate mutants of Salmonella enterica serotype Enteritidis

Independent studies have demonstrated that flagella are associated with the invasive process of Salmonella enterica serotypes, and aflagellate derivatives of Salmonella enterica serotype Enteritidis are attenuated in murine and avian models of infection. One widely held view is that the motility afforded by flagella, probably aided by chemotactic responses, mediates the initial interaction between bacterium and host cell. The adherence and invasion properties of two S. Enteritidis wild-type strains and isogenic aflagellate mutants were assessed on HEp-2 and Div-1 cells that are of human and avian epithelial origin, respectively. Both aflagellate derivatives showed a significant reduction of invasion compared with wild type over the three hours of the assays. Complementation of the defective fliC allele recovered partially the wild-type phenotype. Examination of the bacterium-host cell interaction by electron and confocal microscopy approaches showed that wild-type bacteria induced ruffle formation and significant cytoskeletal rearrangements on HEp-2 cells within 5 minutes of contact. The aflagellate derivatives induced fewer ruffles than wild type. Ruffle formation on the Div-1 cell line was less pronounced than for HEp-2 cells for wild-type S. Enteritidis. Collectively, these data support the hypothesis that flagella play an active role in the early events of the invasive process.

The boreal forest and the polar front

The analysis presented in this paper suggests that the larger heating over the boreal forest in the spring and summer, as contrasted with weaker heating over the adjacent tundra, results in a preferred position of the polar front along the northern edge of the boreal forest. This positioning is well documented in the literature (see, for example, Bryson, 1966; Barry and Hare, 1974; Kreps and Barry, 1970). This heating results from the lower albedo of the boreal forest which is not compensated by an increase in transpiration, even with the larger leaf area index of the forest. The warmer temperatures are mixed upward by the deep boundary layer over the forest and mesoscale circulations which result from the patchiness of heating associated with the heterogeneous landscapes of the forest. Thus in contrast to previous assumptions in which the arctic front position in the summer determines the northern limit of the boreal tree line, our study suggests the boreal forest itself significantly influences the preferred position of the front. This conclusion reinforces the findings of Bonan et al. (1992) and Foley et al. (1994) on the important role of boreal forest-tundra interactions with climate.

Daidzein, R-(+)equol and S-(-)equol inhibit the invasion of MDA-MB-231 breast cancer cells potentially via the down-regulation of matrix metalloproteinase-2

PURPOSE: Soy isoflavones may inhibit tumor cell invasion and metastasis via their effects on matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). The current study investigates the effects of daidzein, R- and S-equol on the invasion of MDA-MB-231 human breast cancer cells and the effects of these compounds on MMP/TIMP expression at the mRNA level. METHODS: The anti-invasive effects of daidzein, R- and S-equol (0, 2.5, 10, 50 μM) on MDA-MB-231 cells were determined using the Matrigel invasion assay following 48-h exposure. Effects on MMP-2, MMP-9, TIMP-1 and TIMP-2 expression were assessed using real-time PCR. Chiral HPLC analysis was used to determine intracellular concentrations of R- and S-equol. RESULTS: The invasive capacity of MDA-MB-231 cells was significantly reduced (by approximately 50-60 %) following treatment with 50 μM daidzein, R- or S-equol. Anti-invasive effects were also observed with R-equol at 2.5 and 10 μM though overall equipotent effects were induced by all compounds. Inhibition of invasion induced by all three compounds at 50 μM was associated with the down-regulation of MMP-2, while none of the compounds tested significantly affected the expression levels of MMP-9, TIMP-1 or TIMP-2 at this concentration. Following exposure to media containing 50 μM R- or S-equol for 48-h intracellular concentrations of R- and S-equol were 4.38 ± 1.17 and 3.22 ± 0.47 nM, respectively. CONCLUSION: Daidzein, R- and S-equol inhibit the invasion of MDA-MB-231 human breast cancer cells in part via the down-regulation of MMP-2 expression, with equipotent effects observed for the parent isoflavone daidzein and the equol enantiomers.

SPI-23 of S.Derby: role in adherence and invasion of porcine tissues

Salmonella enterica serovars Derby and Mbandaka are isolated from different groups of livestock species in the UK. S. Derby is predominantly isolated from pigs and turkeys and S. Mbandaka is predominantly isolated from cattle and chickens. Alignment of the genome sequences of two isolates of each serovar led to the discovery of a new putative Salmonella pathogenicity island, SPI-23, in the chromosome sequence of S. Derby isolates. SPI-23 is 37 kb in length and contains 42 ORFs, ten of which are putative type III effector proteins. In this study we use porcine jejunum derived cell line IPEC-J2 and in vitro organ culture of porcine jejunum and colon, to characterise the association and invasion rates of S. Derby and S. Mbandaka, and tissue tropism of S. Derby respectively. We show that S. Derby invades and associates to an IPEC-J2 monolayer in significantly greater numbers than S. Mbandaka, and that S. Derby preferentially attaches to porcine jejunum over colon explants. We also show that nine genes across SPI-23 are up-regulated to a greater degree in the jejunum compared to the colon explants. Furthermore, we constructed a mutant of the highly up-regulated, pilV-like gene, potR, and find that it produces an excess of surface pili compared to the parent strain which form a strong agglutinating phenotype interfering with association and invasion of IPEC-J2 monolayers. We suggest that potR may play a role in tissue tropism.

Virgin olive oil phenolics extract inhibit invasion of HT115 human colon cancer cells in vitro and in vivo

The decreased cancer risk associated with consumption of olive oil may be due to the presence of phenolics which can modulate pathways including apoptosis and invasion that are relevant to carcinogenesis. We have previously shown that a virgin olive oil phenolics extract (OVP) inhibited invasion of HT115 colon cancer cells in vitro. In the current study we assessed the in vitro effects of OVP (25 μg mL(-1)) on HT115 cell migration, spreading and integrin expression. Furthermore, the anti-metastatic activity of OVP - at a dose equivalent to 25 mg per kg per day for 2, 8 or 10 weeks - was assessed in a Severe Combined ImmunoDeficiency (SCID) Balb-c mouse model. After 24 h OVP did not inhibit cell migration but significantly reduced cell spreading on fibronectin (65% of control; p < 0.05) and expression of a range of α and β integrins was modulated. In vivo, OVP by gavage significantly (p < 0.05) decreased not only tumour volume but also the number of metastases in SCID Balb-c mice. Collectively, the data suggest that - possibly through modulation of integrin expression - OVP decreases invasion in vitro and also inhibits metastasis in vivo.

Gallic war songs (II): Marcus Cicero, Quintus Cicero, and Caesar’s invasion of Britain

This article assembles and examines the evidence for the poetic outputs of Marcus and Quintus Cicero related to Caesar’s invasion of Britain. Following the establishment of a relative chronology of the evidence for their work, it is argued that Quintus Cicero most likely produced a fabula praetexta (not an epic poem, as commonly assumed). His brother, in turn, wrote an epic, based on Quintus’ eye-witness reports. Careful analysis of the ancient discourse about this piece reveals insights in Cicero’s poetic workshop and the creation of ‘archival truth’ through narrativising historical events in epic poetry. Finally, a case is made for greater attention to financial affairs between Caesar and the Ciceros that happen to coincide with the drafting process of their respective literary works.

Secondary cyclogenesis along an occluded front leading to damaging wind gusts: windstorm Kyrill, January 2007

Windstorm Kyrill affected large parts of Europe in January 2007 and caused widespread havoc and loss of life. In this study the formation of a secondary cyclone, Kyill II, along the occluded front of the mature cyclone Kyrill and the occurrence of severe wind gusts as Kyrill II passed over Germany are investigated with the help of high-resolution regional climate model simulations. Kyrill underwent an explosive cyclogenesis south of Greenland as the storm crossed polewards of an intense upper-level jet stream. Later in its life cycle secondary cyclogenesis occurred just west of the British Isles. The formation of Kyrill II along the occluded front was associated (a) with frontolytic strain and (b) with strong diabatic heating in combination with a developing upper-level shortwave trough. Sensitivity studies with reduced latent heat release feature a similar development but a weaker secondary cyclone, revealing the importance of diabatic processes during the formation of Kyrill II. Kyrill II moved further towards Europe and its development was favored by a split jet structure aloft, which maintained the cyclone’s exceptionally deep core pressure (below 965 hPa) for at least 36 hours. The occurrence of hurricane force winds related to the strong cold front over North and Central Germany is analyzed using convection-permitting simulations. The lower troposphere exhibits conditional instability, a turbulent flow and evaporative cooling. Simulation at high spatio-temporal resolution suggests that the downward mixing of high momentum (the wind speed at 875 hPa widely exceeded 45 m s-1) accounts for widespread severe surface wind gusts, which is in agreement with observed widespread losses.

At the Front: common traitors in West German war films of the 1950s

The article looks at the figure of the traitor in 1950s’ West German films about World War II. It focuses on the representation of Wehrmacht soldiers who entertain relations with the Soviet enemy and are therefore seen to betray their nation. The discussion of three well-known films – 08/15, Der Arzt von Stalingrad, and Unruhige Nacht – shows these ‘traitors’ to have a highly ambivalent function: their narrative punishment is part of German post-war exculpation, yet they are also reminders of German guilt and ethical responsibility towards the ‘other’.

Synergy of extreme drought and shrub invasion reduce ecosystem functioning and resilience in water-limited climates

Extreme drought events and plant invasions are major drivers of global change that can critically affect ecosystem functioning and alter ecosystem-atmosphere exchange. Invaders are expanding worldwide and extreme drought events are projected to increase in frequency and intensity. However, very little is known on how these drivers may interact to affect the functioning and resilience of ecosystems to extreme events. Using a manipulative shrub removal experiment and the co-occurrence of an extreme drought event (2011/2012) in a Mediterranean woodland, we show that native shrub invasion and extreme drought synergistically reduced ecosystem transpiration and the resilience of key-stone oak tree species. Ecosystem transpiration was dominated by the water use of the invasive shrub Cistus ladanifer, which further increased after the extreme drought event. Meanwhile, the transpiration of key-stone tree species decreased, indicating a competitive advantage in favour of the invader. Our results suggest that in Mediterranean-type climates the invasion of water spending species and projected recurrent extreme drought events may synergistically cause critical drought tolerance thresholds of key-stone tree species to be surpassed, corroborating observed higher tree mortality in the invaded ecosystems. Ultimately, this may shift seasonally water limited ecosystems into less desirable alternative states dominated by water spending invasive shrubs.

Differential epigenetic reprogramming in response to specific endocrine therapies promotes cholesterol biosynthesis and cellular invasion

Endocrine therapies target the activation of the oestrogen receptor alpha (ERα) via distinct mechanisms, but it is not clear whether breast cancer cells can adapt to treatment using drug-specific mechanisms. Here we demonstrate that resistance emerges via drug-specific epigenetic reprogramming. Resistant cells display a spectrum of phenotypical changes with invasive phenotypes evolving in lines resistant to the aromatase inhibitor (AI). Orthogonal genomics analysis of reprogrammed regulatory regions identifies individual drug-induced epigenetic states involving large topologically associating domains (TADs) and the activation of super-enhancers. AI-resistant cells activate endogenous cholesterol biosynthesis (CB) through stable epigenetic activation in vitro and in vivo. Mechanistically, CB sparks the constitutive activation of oestrogen receptors alpha (ERα) in AI-resistant cells, partly via the biosynthesis of 27-hydroxycholesterol. By targeting CB using statins, ERα binding is reduced and cell invasion is prevented. Epigenomic-led stratification can predict resistance to AI in a subset of ERα-positive patients

Re-examining the U.K.'s greatest tornado outbreak: forecasting the limited extent of tornadoes along a cold front

On 23 November 1981, a strong cold front swept across the U.K., producing tornadoes from the west to the east coasts. An extensive campaign to collect tornado reports by the Tornado and Storm Research Organisation (TORRO) resulted in 104 reports, the largest U.K. outbreak. The front was simulated with a convection-permitting numerical model down to 200-m horizontal grid spacing to better understand its evolution and meteorological environment. The event was typical of tornadoes in the U.K., with convective available potential energy (CAPE) less than 150 J kg-1, 0-1-km wind shear of 10-20 m s-1, and a narrow cold-frontal rainband forming precipitation cores and gaps. A line of cyclonic absolute vorticity existed along the front, with maxima as large as 0.04 s-1. Some hook-shaped misovortices bore kinematic similarity to supercells. The narrow swath along which the line was tornadic was bounded on the equatorward side by weak vorticity along the line and on the poleward side by zero CAPE, enclosing a region where the environment was otherwise favorable for tornadogenesis. To determine if the 104 tornado reports were plausible, first possible duplicate reports were eliminated, resulting in as few as 58 tornadoes to as many as 90. Second, the number of possible parent misovortices that may have spawned tornadoes is estimated from model output. The number of plausible tornado reports in the 200-m grid-spacing domain was 22 and as many as 44, whereas the model simulation was used to estimate 30 possible parent misovortices within this domain. These results suggest that 90 reports was plausible.