Differential effects of nutritional folic acid deficiency and moderate hyperhomocysteinemia on aortic plaque formation and genome-wide DNA methylation in vascular tissue from ApoE-/- mice

Low folate intake is associated with vascular disease. Causality has been attributed to hyperhomocysteinemia. However, human intervention trials have failed to show the benefit of homocysteine-lowering therapies. Alternatively, low folate may promote vascular disease by deregulating DNA methylation. We investigated whether folate could alter DNA methylation and atherosclerosis in ApoE null mice. Mice were fed one of six diets (n = 20 per group) for 16 weeks. Basal diets were either control (C; 4% lard) or high fat (HF; 21% lard and cholesterol, 0.15%) with different B-vitamin compositions: (1) folic acid and B-vitamin replete, (2) folic acid deficient (−F), (3) folic acid, B6 and B12 deficient (−F−B). −F diets decreased plasma (up to 85%; P < 0.05), whole blood (up to 70%; P < 0.05), and liver folate (up to 65%; P < 0.05) and hepatic SAM/SAH (up to 80%; P < 0.05). −F−B diets reduced plasma (up to 76%; P < 0.05), whole blood (up to 72%; P < 0.05), and liver B12 (up to 39%; P < 0.05) and hepatic SAM/SAH (up to 90%; P < 0.05). −F increased homocysteine 2-fold, while −F−B increased homocysteine 3.6- and 6.8-fold in the C and HF groups (P < 0.05). Plaque formation was increased 2-fold (P < 0.0001) in mice fed a HF diet. Feeding a HF–F diet increased lesion formation by 17% (P < 0.05). There was no change in 5-methyldeoxycytidine in liver or vascular tissue (aorta, periadventitial tissue and heart). These data suggest that atherogenesis is not associated with genome-wide epigenetic changes in this animal model

Sensitivity of markers of DNA stability and DNA repair activity to folate supplementation in healthy volunteers

We have previously reported that supplementation with folic acid (1.2 mg day−1 for 12 week) elicited a significant improvement in the folate status of 61 healthy volunteers. We have examined effects of this supplement on markers of genomic stability. Little is known about the effect of folate supplementation on DNA stability in a cohort, which is not folate deficient. Preintervention, there was a significant inverse association between uracil misincorporation in lymphocyte DNA and red cell folate (P<0.05). In contrast, there were no associations between folate status and DNA strand breakage, global DNA methylation or DNA base excision repair (measured as the capacity of the lymphocyte extract to repair 8-oxoGua ex vivo). Folate supplementation elicited a significant reduction in uracil misincorporation (P<0.05), while DNA strand breakage and global DNA methylation remained unchanged. Increasing folate status significantly decreased the base excision repair capacity in those volunteers with the lowest preintervention folate status (P<0.05). Uracil misincorporation was more sensitive to changes in folate status than other measures of DNA stability and therefore could be considered a specific and functional marker of folate status, which may also be relevant to cancer risk in healthy people

Signal peptide mutations in RANK prevent downstream activation of NF-κB

Familial expansile osteolysis and related disorders are caused by heterozygous tandem duplication mutations in the signal peptide region of the gene encoding receptor activator of NF-κB (RANK), a receptor critical for osteoclast formation and function. Previous studies have shown that overexpression of these mutant proteins causes constitutive activation of NF-κB signaling in vitro, and it has been assumed that this accounts for the focal osteolytic lesions that are seen in vivo. We show here that constitutive activation of NF-κB occurred in HEK293 cells overexpressing wild-type or mutant RANK but not in stably transfected cell lines expressing low levels of each RANK gene. Importantly, only cells expressing wild-type RANK demonstrated ligand-dependent activation of NF-κB. When overexpressed, mutant RANK did not localize to the plasma membrane but localized to extensive areas of organized smooth endoplasmic reticulum, whereas, as expected, wild-type RANK was detected at the plasma membrane and in the Golgi apparatus. This intracellular accumulation of the mutant proteins is probably the result of lack of signal peptide cleavage because, using two in vitro translation systems, we demonstrate that the mutations in RANK prevent cleavage of the signal peptide. In conclusion, signal peptide mutations lead to accumulation of RANK in the endoplasmic reticulum and prevent direct activation by RANK ligand. These results strongly suggest that the increased osteoclast formation/activity caused by these mutations cannot be explained by studying the homozygous phenotype alone but requires further detailed investigation of the heterozygous expression of the mutant RANK proteins. © 2011 American Society for Bone and Mineral Researc

Nutritional B vitamin deficiency alters the expression of key proteins associated with vascular smooth muscle cell proliferation and migration in the aorta of atherosclerotic apolipoprotein E null mice.

Low B vitamin status is linked with human vascular disease. We employed a proteomic and biochemical approach to determine whether nutritional folate deficiency and/or hyperhomocysteinemia altered metabolic processes linked with atherosclerosis in ApoE null mice. Animals were fed either a control fat (C; 4 % w/w lard) or a high-fat [HF; 21 % w/w lard and cholesterol (0/15 % w/w)] diet with different B vitamin compositions for 16 weeks. Aorta tissue was prepared and global protein expression, B vitamin, homocysteine and lipoprotein status measured. Changes in the expression of aorta proteins were detected in response to multiple B vitamin deficiency combined with a high-fat diet (P < 0.05) and were strongly linked with lipoprotein concentrations measured directly in the aorta adventitia (P < 0.001). Pathway analysis revealed treatment effects in the aorta-related primarily to cytoskeletal organisation, smooth muscle cell adhesion and invasiveness (e.g., fibrinogen, moesin, transgelin, vimentin). Combined B vitamin deficiency induced striking quantitative changes in the expression of aorta proteins in atherosclerotic ApoE null mice. Deregulated expression of these proteins is associated with human atherosclerosis. Cellular pathways altered by B vitamin status included cytoskeletal organisation, cell differentiation and migration, oxidative stress and chronic inflammation. These findings provide new insight into the molecular mechanisms through which B vitamin deficiency may accelerate atherosclerosis

Consumption of a recommended serving of wheat bran cereals significantly increases human faecal butyrate levels in healthy volunteers and reduces markers of inflammation ex vivo.

Wheat bran cereals are an important source of dietary fibre. The aim of the study was to investigate if a high intake (120 g) of fibre rich breakfast cereal (which delivers the UK Government guidelines for fibre intake in one serving but is three-fold higher than the manufacturers recommended serving) has additional potential health benefits compared to the recommended serving (40 g, containing 11 g of dietary fibre). To assess this, the study determined the short chain fatty acid (SCFA) profiles in human faecal, urine and plasma samples after consumption of two different servings of fibre-rich cereal. Inhibition of prostanoid production was measured (ex vivo) in human colonic fibroblast cells after cytokine (IL-1β) inflammation stimulation. Eight healthy volunteers, 18-55 years old; BMI (18-30 kg/m2) consumed the wheat bran-rich "ready to eat cereal", at both the high (120 g) serving and recommended (40 g) serving. Faecal, urine and plasma samples were collected at baseline, throughout the five-hour intervention period and approximately 24 hours following consumption. Faecal butyrate showed the largest increase (p < 0.05) of more than a two-fold change following the consumption of the recommended serving of wheat bran cereal (from 13.95 ± 9.17 to 31.63 ± 20.53 mM) and no significant change following the higher serving (from 21.96 ± 11.03 to 22.9 ± 12.69 mM). ANOVA analysis also found a weak serving effect (p = 0.046) of the portion size (high vs. recommended) only for butyrate in urine 24 hours after consumption of the bran cereal. The physiological nutritionally relevant concentrations of faecal SCFAs, as determined in the volunteers' faecal samples showed significant anti-inflammatory activity or the individual faecal SCFAs; acetate (p < 0.001), propionate (p < 0.001) and butyrate (p < 0.01), as well as in combination. Plasma folate was also increased after consumption of both wheat bran servings and was significant (p = 0.037) at the three-hour time point following consumption of the high wheat bran serving. The consumption of the recommended serving (40 g) of wheat bran cereal increased the total microbial SCFAs levels (from 96.88 to 136.96 mM) compared to the higher serving (120 g) (from 110.5 to 117.64 mM) suggesting that the intake of the higher portion size is likely to promote a faecal bulking effect and thereby decrease colonic SCFA levels. These data indicate that consumption of the recommended serving of wheat bran cereal serving would therefore be sufficient to promote microbial butyrate formation, reduce colonic inflammation and increase plasma folate levels in humans

A case study of electron precipitation fluxes due to plasmaspheric hiss

We find that during a large geomagnetic storm in October 2011 the trapped fluxes of >30, >100, and >300 keV outer radiation belt electrons were enhanced at L=3-4 during the storm main phase. A gradual decay of the trapped fluxes was observed over the following 5–7 days, even though no significant precipitation fluxes could be observed in the Polar Orbiting Environmental Satellite (POES) electron precipitation detectors. We use the Antarctic-Arctic Radiation-belt (Dynamic) Deposition - VLF Atmospheric Research Konsortium (AARDDVARK) receiver network to investigate the characteristics of the electron precipitation throughout the storm period. Weak electron precipitation was observed on the dayside for 5–7 days, consistent with being driven by plasmaspheric hiss. Using a previously published plasmaspheric hiss-induced electron energy e-folding spectrum of E0=365 keV, the observed radiowave perturbation levels at L=3-4 were found to be caused by >30 keV electron precipitation with flux ~100 el. cm−2 s−1 sr−1. The low levels of precipitation explain the lack of response of the POES telescopes to the flux, because of the effect of the POES lower sensitivity limit and ability to measure weak diffusion-driven precipitation. The detection of dayside, inner plasmasphere electron precipitation during the recovery phase of the storm is consistent with plasmaspheric hiss wave-particle interactions, and shows that the waves can be a significant influence on the evolution of the outer radiation belt trapped flux that resides inside the plasmapause

Capacity building for conservation: problems and potential solutions for sub-Saharan Africa

To successfully achieve their stated conservation goals individuals, communities and organisations need to acquire a diversity of skills, knowledge and information (capacity). Despite current efforts to build and maintain appropriate levels of conservation capacity, it has been recognised that there will need to be a significant scaling-up of these activities in sub-Saharan Africa. This is because of the rapidly growing number and extent of environmental problems in the region. This paper presents a range of socio-economic contexts relevant to four key areas of African conservation capacity building: protected area management, community engagement, effective leadership, and professional e-Learning. Under these core themes, 39 specific recommendations are presented. These were derived from multi-stakeholder workshop discussions at an international conference held in Nairobi (Kenya) in 2015. At the meeting, 185 delegates (practitioners, scientists, community groups and government agencies) represented 105 organisations from 24 African nations and 8 non-African nations. The 39 recommendations constitute five broad types of suggested action: those that recommend (i) the development of new methods, (ii) the provision of capacity building resources e.g. information or data, (iii) the communication of ideas or examples of successful initiatives, (iv) the implementation of new research or gap analyses, (v) the establishment of new structures within and between organisations, and (vi) the development of new partnerships. A number of cross-cutting issues also emerged from the discussions. For example, all four workshops highlighted the need for a greater sense of urgency in developing capacity building activities in response to ongoing and rapid socio-environmental change in the region. Delegates also felt that conservation organisations, responsible agencies and donors need to recognise capacity building as one of the most urgent conservation issues we face. The need to develop novel and cost-efficient capacity building methodologies (and associated evaluation metrics), was also identified as a key issue. However, it was stressed that future of capacity building efforts will be best served by integrating new methods with more established activities. Importantly, given the broad suite of social, cultural and economic contexts found across sub-Saharan Africa, the need to move away from ‘one-size-fits-all’ approaches was strongly recommended in all thematic areas. Lastly, it was recognised that closing the gap between capacity need and capacity provision in the region will only be achieved through multi-partner capacity initiatives and networks.Additional co-authors: Vivian Kosgei, Anthony Kuria, Chris Magero, Maaike Manten, Paul Mugo, Eduard Müller, Julie Mulonga, Leo Niskanen, Josephine Nzilani, Mary Otieno, Nisha Owen, Juliet Owuor, Stuart Paterson, Sébastien Regnaut, Richard Rono, Joseph Ruhiu, Jesse Theuri Njoka, Lucy Waruingi, Brian Waswala Olewe and Emily Wilso

Ampullary cancers harbor ELF3 tumor suppressor gene mutations and exhibit frequent WNT dysregulation

The ampulla of Vater is a complex cellular environment from which adenocarcinomas arise to form a group of histopathologically heterogenous tumors. To evaluate the molecular features of these tumors, 98 ampullary adenocarcinomas were evaluated and compared to 44 distal bile duct and 18 duodenal adenocarcinomas. Genomic analyses revealed mutations in the WNT signaling pathway among half of the patients and in all three adenocarcinomas irrespective of their origin and histological morphology. These tumors were characterized by a high frequency of inactivating mutations of ELF3, a high rate of microsatellite instability, and common focal deletions and amplifications, suggesting common attributes in the molecular pathogenesis are at play in these tumors. The high frequency of WNT pathway activating mutation, coupled with small-molecule inhibitors of β-catenin in clinical trials, suggests future treatment decisions for these patients may be guided by genomic analysis