Metastasis-inducing DNA Regulates the Expression of the Osteopontin Gene by Binding the Transcription Factor Tcf-4

Small 1,000-bp fragments of genomic DNA obtained from human malignant breast cancer cell lines when transfected into a benign rat mammary cell line enhance transcription of the osteopontin gene and thereby cause the cells to metastasize in syngeneic rats. To identify the molecular events underlying this process, transient cotransfections of an osteopontin promoter-reporter construct and fragments of one metastasis-inducing DNA (Met-DNA) have identified the active components in the Met-DNA as the binding sites for the T-cell factor (Tcf) family of transcription factors. Incubation of cell extracts with active DNA fragments containing the sequence CAAAG caused retardation of their mobilities on polyacrylamide gels, and Western blotting identified Tcf-4, beta-catenin, and E-cadherin in the relevant DNA complexes in vitro. Transfection of an expression vector for Tcf-4 inhibited the stimulated activity of the osteopontin promoter-reporter construct caused by transiently transfected active fragments of Met-DNA or permanently transfected Met-DNA. This stimulated activity of the osteopontin promoter-reporter construct is accompanied by an increase in endogenous osteopontin mRNA but not in fos or actin mRNAs in the transfected cells. Permanent transfection of the benign rat mammary cell line with a 20-bp fragment from the Met-DNA containing the Tcf recognition sequence CAAAG caused an enhanced permanent production of endogenous osteopontin protein in vitro and induced the cells to metastasize in syngeneic rats in vivo. The corresponding fragment without the CAAAG sequence was without either effect. Therefore, the regulatory effect of the C9-Met-DNA is exerted, at least in part, by a CAAAG sequence that can sequester the endogenous inhibitory Tcf-4 and thereby promote transcription of osteopontin, the direct effector of metastasis in this system.

Partial structure of the phylloxin gene from the giant monkey frog, Phyllomedusa bicolor: parallel cloning of precursor cDNA and genomic DNA from lyophilized skin secretion

Phylloxin is a novel prototype antimicrobial peptide from the skin of Phyllomedusa bicolor. Here, we describe parallel identification and sequencing of phylloxin precursor transcript (mRNA) and partial gene structure (genomic DNA) from the same sample of lyophilized skin secretion using our recently-described cloning technique. The open-reading frame of the phylloxin precursor was identical in nucleotide sequence to that previously reported and alignment with the nucleotide sequence derived from genomic DNA indicated the presence of a 175 bp intron located in a near identical position to that found in the dermaseptins. The highly-conserved structural organization of skin secretion peptide genes in P. bicolor can thus be extended to include that encoding phylloxin (plx). These data further reinforce our assertion that application of the described methodology can provide robust genomic/transcriptomic/peptidomic data without the need for specimen sacrifice.

Prograded Holocene beach-ridges with superimposed dunes in north-east Ireland: mechanisms and timescales of fine and coarse beach sediment decoupling and deposition

Two depositional models to account for Holocene gravel-dominated beach ridges covered by dunes, occurring on the northern coast of Ireland, are considered in the light of infrared-stimulated luminescence ages of sand units within beach ridges, and 14C ages from organic horizons in dunes. A new chronostratigraphy obtained from prograded beach ridges with covering dunes at Murlough, north-east Ireland, supports a model of mesoscale alternating sediment decoupling (ASD) on the upper beach, rather than macroscale sequential sediment sourcing to account for prograded beach ridges and covering dunes. The ASD model specifies storm or fair-weather sand beach ridges forming at high-tide positions (on an annual basis at minimum), which acted as deflationary sources for landward foredune development. Only a limited number of such late-Holocene beach ridges survive in the observed prograded series. Beach ridges only survive when capped by storm-generated gravel beaches that are deposited on a mesoscale time spacing of 50–130 years. The morphodynamic shift from a dissipative beach face for dune formation to a reflective beach face for gravel capping appears to be controlled by the beach sand volume falling to a level where reflective conditions can prevail. Sediment volume entering the beach is thought to have fluctuated as a function of a forced regression associated with the falling sea level from the mid-Holocene highstand (ca. 6000 cal. yr BP) identified in north-east Ireland. The prograded beach ridges dated at ca. 3000 to 2000 cal. yr BP indicate that the Holocene highstand’s regressive phase may have lasted longer than previously specified.

Genetically increased risk of sleep disruption in alzheimer's disease

STUDY OBJECTIVES: To investigate the role of a monoamine A oxidase promoter polymorphism in sleep disruption in Alzheimer's disease (AD). DESIGN: A case-control association analysis. SETTING: Sleep disturbance in AD is common, is extremely stressful for caregivers, and increases the risk of institutionalisation. It remains unclear why only some patients develop sleep disturbance; neuropathologic changes of AD are not typically seen in the areas of the brain responsible for sleep. We hypothesized that the risk of sleep disturbance is, at least in part, influenced by the availability of serotonin used for melatonin synthesis secondary to polymorphic variation at the enzyme monoamine oxidase A (MAO-A). PATIENTS: Patients with AD diagnosed according to standard criteria. INTERVENTIONS: Data were collected using the Sleep domain of the Neuropsychiatric Inventory with Caregiver Distress. Patients' cognition and function were assessed using the Mini-Mental State Examination and the Functional Assessment Staging. Genotyping of apolipoprotein E (APOE) and of the 30 bp variable number tandem repeat of the MAO-A promoter was by standard methods. MEASUREMENTS AND RESULTS: Of 426 patients surveyed, 54% experienced sleep disturbance. We found that the high-activity 4-repeat allele of the MAO-A VNTR promoter polymorphism confers increased susceptibility to sleep disturbance (p = .008). A quantitative sleep disturbance score was significantly higher in the patients possessing MAO-A 4-repeat allele genotypes. APOE had no influence on the development of an altered sleep phenotype. CONCLUSIONS: We conclude that sleep disturbance in AD is common and distressing and is associated with genetic variation at MAO-A.

Differential expression of components of the cardiomyocyte adrenomedullin/intermedin receptor system following blood pressure reduction in nitric oxide-deficient hypertension.

Adrenomedullin (AM) and intermedin (IMD; adrenomedulln-2) are vasodilator peptides related to calcitonin gene-related peptide (CGRP). The actions of these peptides are mediated by the calcitonin receptor-like receptor (CLR) in association with one of three receptor activity-modifying proteins. CGRP is selective for CLR/receptor activity modifying protein (RAMP)1, AM for CLR/RAMP2 and -3, and IMD acts at both CGRP and AM receptors. In a model of pressure overload induced by inhibition of nitric-oxide synthase, up-regulation of AM was observed previously in cardiomyocytes demonstrating a hypertrophic phenotype. The current objective was to examine the effects of blood pressure reduction on cardiomyocyte expression of AM and IMD and their receptor components. Nomega-nitro-L-arginine methyl ester (L-NAME) (35 mg/kg/day) was administered to rats for 8 weeks, with or without concurrent administration of hydralazine (50 mg/kg/day) and hydrochlorothiazide (7.5 mg/kg/day). In left ventricular cardiomyocytes from L-NAME-treated rats, increases (-fold) in mRNA expression were 1.6 (preproAM), 8.4 (preproIMD), 3.4 (CLR), 4.1 (RAMP1), 2.8 (RAMP2), and 4.4 (RAMP3). Hydralazine/hydrochlorothiazide normalized systolic blood pressure (BP) and abolished mRNA up-regulation of hypertrophic markers sk-alpha-actin and BNP and of preproAM, CLR, RAMP2, and RAMP3 but did not normalize cardiomyocyte width nor preproIMD or RAMP1 mRNA expression. The robust increase in IMD expression indicates an important role for this peptide in the cardiac pathology of this model but, unlike AM, IMD is not associated with pressure overload upon the myocardium. The concordance of IMD and RAMP1 up-regulation indicates a CGRP-type receptor action; considering also a lack of response to BP reduction, IMD may, like CGRP, have an anti-ischemic function.

Social identification and prevalence of PTSD in Northern Ireland

Background Understanding of the psychological impact of politically motivated violence is poor. Aims To examine the prevalence of post-traumatic symptoms subsequentto the ‘troubles’ in Northern Ireland. Method A telephone survey of 3000 adults, representative of the population in Northern Ireland and the border counties of the Irish Republic, examined exposure to political violence, post-traumatic stress disorder (PTSD) and national identity. Results Ten per cent of respondents had symptoms suggestive of clinical PTSD. These people were most likely to come from low-income groups, rate national identity as relatively unimportant and have higher overall experience of the ‘troubles’than other respondents. Conclusions Direct experience of violence and poverty increase the risk of PTSD, whereas strong national identification appears to reduce this risk.

A Holocene record of human induced and natural environmental change from Lake Forsyth (Te Wairewa), New Zealand

A 1.2 m sediment core from Lake Forsyth, Canterbury, New Zealand, records the development of the catchment/lake system over the last 7000 years, and its response to anthropogenic disturbance following European settlement c. 1840 AD. Pollen was used to reconstruct catchment vegetation history, while foraminifera, chironomids, Trichoptera, and the abundance of Pediastrum simplex colonies were used to infer past environmental conditions within the lake. The basal 30 cm of core records the transition of the Lake Forsyth Basin from a tidal embayment to a brackish coastal lake. Timing of closure of the lake mouth could not be accurately determined, but it appears that Lake Forsyth had stabilised as a slightly brackish, oligo mesotrophic shallow lake by about 500 years BP. Major deforestation occurred on Banks Peninsula between 1860 AD and 1890 AD. This deforestation is marked by the rapid decline in the main canopy trees (Prumnopitys taxifolia (matai) and Podocarpus totara/hallii (totara/mountain totara), an increase in charcoal, and the appearance of grasses. At around 1895 AD, pine appears in the record while a willow (Salix spp.) appears somewhat later. Redundancy analysis (RDA) of the pollen and aquatic species data revealed a significant relationship between regional vegetation and the abundance of aquatic taxa, with the percentage if disturbance pollen explaining most (14.8%) of the constrained variation in the aquatic species data. Principle components analysis (PCA) of aquatic species data revealed that the most significant period of rapid biological change in the lakes history corresponded to the main period of human disturbance in the catchment. Deforestation led to increased sediment and nutrient input into the lake which was accompanied by a major reduction in salinity. These changes are inferred from the appearance and proliferation of freshwater algae (Pediastrum simplex), an increase in abundance and diversity of chironomids, and the abundance of cases and remains from the larvae of the caddisfly, Oecetis unicolor. Eutrophication accompanied by increasing salinity of the lake is inferred from a significant peak and then decline of P. simplex, and a reduction in the abundance and diversity of aquatic invertebrates. The artificial opening of the lake to the Pacific Ocean, which began in the late 1800s, is the likely cause of the recent increase in salinity. An increase in salinity may have also encouraged blooms of the halotolerant and hepatotoxic cyanobacteria Nodularia spumigena.

Chironomid-based reconstructions of summer air temperature from lake deposits in Lyndon Stream, New Zealand spanning the MIS 3/2 transition

We present chironomid-based temperature reconstructions from lake sediments deposited between ca 26,600 cal yr BP and 24,500 cal yr BP from Lyndon Stream, South Island, New Zealand. Summer (February mean) temperatures averaged 1 1C cooler, with a maximum inferred cooling of 3.7 1C. These estimates corroborate macrofossil and beetle-based temperature inferences from the same site and suggest climate amelioration (an interstadial) at this time. Other records from the New Zealand region also show a large degree of variability during the late Otiran glacial sequence (34,000–18,000 cal yr BP) including a phase of warming at the MIS 2/3 transition and a maximum cooling that did not occur until the global LGM (ca 20,000 cal yr BP). The very moderate cooling identified here at the MIS 2/3 transition confirms and enhances the long-standing discrepancy in New Zealand records between pollen and other proxies. Low abundances (o20%) of canopy tree pollen in records from late MIS 3 to the end of MIS 2 cannot be explained by the minor (o5 1C) cooling inferred from this and other studies unless other environmental parameters are considered. Further work is required to address this critical issue.

Cox-2 Promotes Chromogranin A Expression and Bioactivity: Evidence for a PGE2-Dependent Mechanism and the Involvement of a Proximal cAMP-Responsive Element.

Abstract The prostanoid biosynthetic enzyme cyclooxygenase-2 (Cox-2) is upregulated in several neuroendocrine tumors. The aim of the current study was to employ a neuroendocrine cell (PC12) model of Cox-2 over-expression to identify gene products that might be implicated in the oncogenic and/or inflammatory actions of this enzyme in the setting of neuroendocrine neoplasia. Expression array and real-time PCR analysis demonstrated that levels of the neuroendocrine marker chromogranin A (CGA) were 2-fold and 3.2-fold higher, respectively, in Cox-2 over-expressing cells (PCXII) vs their control (PCMT) counterparts. Immunocytochemical and immunoblotting analyses confirmed that both intracellular and secreted levels of CGA were elevated in response to Cox-2 induction. Moreover, exogenous addition of prostaglandin E2 (1u�­M), mimicked this effect in PCMT cells, while treatment of PCXII cells with the Cox-2 selective inhibitor NS-398 (100 nM) reduced CGA expression levels, thereby confirming the biospecificity of this finding. Levels of neurone specific enolase (NSE) were similar in the two cell lines, suggesting that the effect of Cox-2 on CGA expression was specific and not due to a global enhancement of neuroendocrine marker expression/differentiation. Cox-2-dependent CGA upregulation was associated with significantly increased chromaffin granule number and intracellular and secreted levels of dopamine. CGA promoter-driven reporter gene expression studies provided evidence that prostaglandin E2-dependent upregulation required a proximal cAMP-responsive element (CRE; -71 - -64 bp). This study is the first to demonstrate that Cox-2 upregulates both CGA expression and bioactivity in a neuroendocrine cell line and has major implications for the role of this polypeptide in the pathogenesis of neuroendocrine cancers in which Cox-2 is upregulated.

IntCal04 terrestrial radiocarbon age calibration, 0-26 cal kyr BP

A new calibration curve for the conversion of radiocarbon ages to calibrated (cal) ages has been constructed and internationally ratified to replace IntCal98, which extended from 0-24 cal kyr BP (Before Present, 0 cal BP = AD 1950). The new calibration data set for terrestrial samples extends from 0-26 cal kyr BP, but with much higher resolution beyond 11.4 cal kyr BP than IntCal98. Dendrochronologically-dated tree-ring samples cover the period from 0-12.4 cal kyr BP. Beyond the end of the tree rings, data from marine records (corals and foraminifera) are converted to the atmospheric equivalent with a site-specific marine reservoir correction to provide terrestrial calibration from 12.4-26.0 cal kyr BP. A substantial enhancement relative to IntCal98 is the introduction of a coherent statistical approach based on a random walk model, which takes into account the uncertainty in both the calendar age and the (super 14) C age to calculate the underlying calibration curve (Buck and Blackwell, this issue). The tree-ring data sets, sources of uncertainty, and regional offsets are discussed here. The marine data sets and calibration curve for marine samples from the surface mixed layer (Marine04) are discussed in brief, but details are presented in Hughen et al. (this issue a). We do not make a recommendation for calibration beyond 26 cal kyr BP at this time; however, potential calibration data sets are compared in another paper (van der Plicht et al., this issue).

The Cost of Sustainability Capital and the Creation of Sustainable Value by Companies

We develop and apply a valuation methodology to calculate the cost of sustainability capital, and, eventually, sustainable value creation of companies. Sustainable development posits that decisions must take into account all forms of capital rather than just economic capital. We develop a methodology that allows calculation of the costs that are associated with the use of different forms of capital. Our methodology borrows the idea from financial economics that the return on capital has to cover the cost of capital. Capital costs are determined as opportunity costs, that is, the forgone returns that would have been created by alternative investments. We apply and extend the logic of opportunity costs to the valuation not only of economic capital but also of other forms of capital. This allows (a) integrated analysis of use of different forms of capital based on a value-based aggregation of different forms of capital, (b) determination of the opportunity cost of a bundle of different forms of capital used in a company, called cost of sustainability capital, (c) calculation of sustainability efficiency of companies, and (d) calculation of sustainable value creation, that is, the value above the cost of sustainability capital. By expanding the well-established logic of the valuation of economic capital in financial markets to cover other forms of capital, we provide a methodology that allows determination of the most efficient allocation of sustainability capital for sustainable value creation in companies. We demonstrate the practicability of the methodology by the valuation of the sustainability performance of British Petroleum (BP).

Area factors and suicide: 5-year follow-up of the Northern Ireland population

Background: Suicide rates vary markedly between areas but it is unclear whether this is due to differences in population composition or to contextual factors operating at an area level.
Aims: To determine if area factors are independently related to suicide risk after adjustment for individual and family characteristics.
Method: A 5-year record linkage study was conducted of 1 116 748 non-institutionalised individuals aged 16-74 years, enumerated at the 2001 Northern Ireland census.
Results: The cohort experienced 566 suicides during follow-up. Suicide risks were lowest for women and for those who were married or cohabiting. Indicators of individual and household disadvantage and economic and health status at the time of the census were also strongly related to risk of suicide. The higher rates of suicide in the more deprived and socially fragmented areas disappeared after adjustment for individual and household factors. There was no significant relationship between population density and risk of suicide.
Conclusions: Differences in rates of suicide between areas are predominantly due to population characteristics rather than to area-level factors, which suggests that policies targeted at area-level factors are unlikely to significantly influence suicides rates.

Postglacial formation and dynamics of North Patagonian Rainforest in the Chonos Archipelago, Southern Chile

Pollen analysis of continuous sediment cores from two lakes in the northern Chonos Archipelago (44S) in southern Chile shows a complete postglacial record of vegetation change. The fossil records indicate that deglaciation was complete in the northern Chonos by at least 13,600 14Cyr BP. Ericaceous heath and grassland persisted for more than 600 years after deglaciation under the influence of dry/cold climates and frequent burning. Nothofagus-Pilgerodendron-Podocarpus forest, with modern analogues in the southern Chonos Archipelago, was established across the northern islands by 12,400 14Cyr BP under increasingly warm and wet climates. There is no evidence for a return to cooler climates during the Younger Dryas chronozone. The rise of Tepualia stipularis and Weinmannia trichosperma as important forest components between 10,600 and 6000 14Cyr BP may be associated withclimates that were warmer than present. The collapse of Pilgerodendron communities during this time may have been triggered by a combination of factors related to disturbance frequency including tephra deposition events, fire and climate change. After 6000 14Cyr BP Pilgerodendron recovers and Nothofagus-Pilgerodendron-Tepualia forest persists until the present. European logging and burning activity may have increased the susceptibility of North Patagonian Rainforest to invasion by introduced species and to future collapse of the long-lived Pilgerodendron communities.

Long-term ecology of native pinewood communities in East Glen Affric, Scotland

Fossil pollen, stomata and charcoal were examined from a lake sedimentary sequence in the Glen Affric National Nature Reserve, one of the largest areas of remnant native pine woodland in Scotland, in order to assess ecosystem dynamics over the last 11 000 years. Results reveal that pinewood communities have been continuously present in East Glen Affric for the last 8300 years. Pinus sylvestris fi rst arrived in the area around 9900 cal. BP, but occurred in only low abundance for the subsequent 1600 years. Pine populations expanded around 8300 cal. BP and remained in relatively constant abundance throughout the remainder of the Holocene. There is no evidence of a hypothesized regional mid-Holocene ‘ pine decline ’ at the site. Charcoal results reveal that pinewood communities in East Glen Affric do not appear to have been dependent on fire for either their establishment or their maintenance as has previously been suggested.