The biological effects of slow release implantable tablets for the delivery of anti-scarring agents following glaucoma filtration surgery

The scarring response that follows glaucoma filtration surgery (GFS), an incisional surgery aimed at reducing glaucoma-related pressure increases within the eye, often prevents a successful outcome leading to further disease progression. Whilst a number of drugs have been shown to regulate scarring, a number of side effects are also induced largely related to the delivery method. Following on from previous work initiated in the lab whereby drugs (including the cytotoxic drugs, 5-fluorouracil (5-FU) and mitomycin-C (MMC), the anti-vascular endothelial growth factor (VEGF) monoclonal antibody, bevacizumab, and the matrix metalloproteinase inhibitor, ilomastat) had been fabricated into solid, slow release, implantable tablets, a number of questions remained pertaining to their effectiveness within a biological context. Using 5-FU, it was shown that concentrations at which the drug is released from the tablet can inhibit fibroblast activity (such as proliferation) as effectively as when using conventional concentrations for up to a 30 day period. Furthermore, it has been shown, for the first time, how gene expression within healing tissue (in-vivo) is altered upon the application of 5-FU at currently used clinical concentrations. Results from this study indicated that genes involved in apoptosis such as TP53, RelA, Bax, MYC and TXN were downregulated indicative of a response by the cells to limit the effects of apoptosis on tissue exposed to 5-FU. Secondly this thesis focuses on angiogenesis where it was shown that solid-dosage bevacizumab tablets functioned as effectively as bevacizumab solution in-vitro, with excipients of this drug such as trehalose shown to have implications in modulating wound healing. Furthermore, VEGF was shown for the first time to be a positiveregulator of fibroblast activity, such as in its ability to promote matrix metalloproteinase (MMP) production, presenting mechanisms by which anti-VEGF drugs may function to inhibit the wound healing process. Finally this thesis focuses on the effects of exposing cells and tissue to ilomastat, whereby microarray screening was utilised to uncover novel changes in gene expression that both regulate the direct activity of ilomastat, as well as highlight secondary effects of the drug. For example, a dampening of the immune response (such as in the downregulation of IL-1a, IL-6, IL-8, Bip and XBP1) was observed following ilomastat injection. Importantly however, these experiments also highlighted a previously uncharacterised foreign body response that was observed upon implantation of the ilomastat tablet in-vivo, with the widespread upregulation of genes involved in this process observed including TNF, FN1, IL-1β and IL-6. As such, whilst these experiments exposed novel mechanisms through which these drugs function, care will be required in moving forward with the use of such agents and delivery methods to ensure thatmaximum biocompatibility is achieved

Groundwater vulnerability assessment: A review including new statistical and hybrid methods

The concept of groundwater vulnerability was first introduced in the 1970s in France to recognize sensitive areas in which surface pollution could affect groundwater, and to enable others to develop management methods for groundwater protection against surface pollutants. Since this time, numerous methods have been developed for groundwater vulnerability assessment (GVA). These can be categorized into four groups: (i) overlay and index-based methods, (ii) process-based simulation models, (iii) statistical methods, and (iv) hybrid methods. This work provides a comprehensive review of modern GVA methods, which in contrast to previous reviews, examines the last two categories in detail. First, the concept of groundwater vulnerability is defined, then the major GVA methods are introduced and classified. This includes detailed accounts of statistical methods, which can be subdivided into orthodox statistical, data-driven and Bayesian methods, and their advantages and disadvantages, as well as modern hybrid methods. It is concluded that Bayesian inference offers many advantages compared with other GVA methods. It combines theory and data to give the posterior probabilities of different models, which can be continually updated with new data. Furthermore, using the Bayesian approach, it is possible to calculate the probability of a proposition, which is exactly what is needed to make decisions. However, despite the advantages of Bayesian inference, its applications to date have been very limited

Comparison of DRASTIC and DRASTICL groundwater vulnerability assessments of the Burdekin Basin, Queensland, Australia

In the Burdekin Basin, Queensland, Australia, groundwater contamination due to agricultural activities has led to concerns over its impacts on globally significant ecosystems such as the Great Barrier Reef. An appropriate method for groundwater vulnerability assessment is essential for the sustainable use of this groundwater resource and its longer-term environmental management. The aim of this study is to apply and assess the suitability of the standard DRASTIC index-based method for groundwater vulnerability assessment of the Burdekin Basin. The intrinsic groundwater vulnerability is calculated in ArcGIS, using data for the period 2010 to 2021. The results are compared to available water quality data. The calculated DRASTIC scores are found to be only weakly correlated with water quality parameters, including the nitrate concentration (R = 0.07), which should behave as a proxy measure of groundwater vulnerability. To address this, a modified DRASTICL method containing a land use parameter is also implemented, to assess the specific groundwater vulnerability. The correlation between DRASTICL scores and nitrate levels (R = 0.2) is more significant but is still relatively weak. From this study, it is recommended that alternative methods be developed to assess groundwater vulnerability in the Burdekin Basin, and other comparable aquifer systems

Interactive effects of inbreeding and endocrine disruption on reproduction in a model laboratory fish

This is the final version of the article. Available from Wiley via the DOI in this record.Inbreeding depression is expected to be more severe in stressful environments. However, the extent to which inbreeding affects the vulnerability of populations to environmental stressors, such as chemical exposure, remains unresolved. Here we report on the combined impacts of inbreeding and exposure to an endocrine disrupting chemical (the fungicide clotrimazole) on zebrafish (Danio rerio). We show that whilst inbreeding can negatively affect reproductive traits, not all traits are affected equally. Inbreeding depression frequently only became apparent when fish were additionally stressed by chemical exposure. Embryo viability was significantly reduced in inbred exposed fish and there was a tendency for inbred males to sire fewer offspring when in direct competition with outbred individuals. Levels of plasma 11-ketotestosterone, a key male sex hormone, showed substantial inbreeding depression that was unaffected by addition of the fungicide. In contrast, there was no effect of inbreeding or clotrimazole exposure on egg production. Overall, our data provide evidence that stress may amplify the effects of inbreeding on key reproductive traits, particularly those associated with male fitness. This may have important implications when considering the consequences of exposure to chemical pollutants on the fitness of wild populations.Thanks to NERC's Post Genomics & Proteomics Programme NE/F0077871/1 and AstraZeneca's Safety, Health and Environment Research Programme for funding this work. We thank Alexander Scott (11-ketotestosterone radioimmunoassay) at the Centre for Environment, Fisheries and Aquaculture Science, Jan Shears and Luanne Wilkes at University of Exeter, Gareth Readman, Vicki Cammack, Kate Hurd and Yohanna Glennon at Brixham Environmental Laboratory for their assistance

Indigenous food sources as vectors of Escherichia coli and antibiotic resistance

The contamination of surface waters by fecal bacteria, measured by the number of Escherichia coli, is a significant public health issue. When these bacteria are also resistant to antimicrobials, infections are more complicated to treat. While water is regularly tested at recreational sites, wild-harvested foods, known as mahinga kai by the indigenous Māori people of Aotearoa New Zealand, are commonly overlooked as a source of exposure to potential pathogens and antimicrobial resistance (AMR). We investigate two likely sources of risk from harvesting aquatic wild foods. The first is water contact, and the second is contact with/ingestion of the harvest. We used E. coli as a proxy for microbial water quality at harvesting sites. Two popular mahinga kai species were also harvested and assessed. We found antibiotic-resistant bacteria on watercress (Nasturtium officinale) and cockles (Austrovenus stutchburyi). One-third of E. coli isolates were conjugative donors of at least one resistance phenotype. Tank experiments were used to track the internalization of E. coli by Greenshell/lip mussels (Perna canaliculus). Greenshell mussels kept at environmentally relevant concentrations of E. coli were colonized to levels considered unsafe for human consumption in 24 h. Finally, we measured horizontal gene transfer between bacteria within the shellfish, what we termed ‘intra-shellular’ conjugation. The transmission frequency of plasmid RP4 was significantly higher in mussels than in water alone. Our results indicate that shellfish could promote the dissemination of antibiotic resistance. They highlight the need to limit or reduce human pathogenic bacteria where food is gathered

Antiepileptic drugs’ tolerability and safety – a systematic review and meta-analysis of adverse effects in dogs

<p>Various anti-epileptic drugs (AEDs) are used for the management of idiopathic epilepsy (IE) in dogs. Their safety profile is an important consideration for regulatory bodies, owners and prescribing clinicians. However, information on their adverse effects still remains limited with most of it derived from non-blinded non-randomized uncontrolled trials and case reports.</p><p><span>This poster won third place, which was presented at the Veterinary Evidence Today conference, Edinburgh November 1-3, 2016. </span></p><br /> <img src="https://www.veterinaryevidence.org/rcvskmod/icons/oa-icon.jpg" alt="Open Access" /

Multibeam bathymetric surveys of submarine volcanoes and mega-pockmarks on the Chatham Rise, New Zealand

Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of Taylor & Francis for personal use, not for redistribution. The definitive version was published in New Zealand Journal of Geology and Geophysics 54 (2011): 329-339, doi:10.1080/00288306.2011.589860.Multibeam bathymetric surveys east of the South Island of New Zealand present images of submarine volcanoes and pockmarks west of Urry Knolls on the Chatham Rise, and evidence of submarine erosion on the southern margin of the Chatham Rise. Among numerous volcanic cones, diameters of the largest reach ~2000 m, and some stand as high as 400 m above the surrounding seafloor. The tops of most of the volcanic cones are flat, with hints of craters, and some with asymmetric shapes may show flank collapses. There are hints of both northeast-southwest and northwest-southeast alignments of volcanoes, but no associated faulting is apparent. Near and to the west of these volcanoes, huge pockmarks, some more than ~1 km in diameter, disrupt bottom topography. Pockmarks in this region seem to be confined to sea floor shallower than ~1200 m, but we see evidence of deeper pockmarks at water depths of up to 2100 m on profiles crossing the Bounty Trough. The pockmark field on the Chatham Rise seems to be bounded on the south by a trough near 1200 m depth; like others, we presume that contour currents have eroded the margin and created the trough.This research was supported by the National Science Foundation under grants EAR-0409564, EAR-0409609, and EAR-0409835.2012-08-3

Mre11-Rad50 Promotes Rapid Repair of DNA Damage in the Polyploid Archaeon Haloferax volcanii by Restraining Homologous Recombination

Polyploidy is frequent in nature and is a hallmark of cancer cells, but little is known about the strategy of DNA repair in polyploid organisms. We have studied DNA repair in the polyploid archaeon Haloferax volcanii, which contains up to 20 genome copies. We have focused on the role of Mre11 and Rad50 proteins, which are found in all domains of life and which form a complex that binds to and coordinates the repair of DNA double-strand breaks (DSBs). Surprisingly, mre11 rad50 mutants are more resistant to DNA damage than the wild-type. However, wild-type cells recover faster from DNA damage, and pulsed-field gel electrophoresis shows that DNA double-strand breaks are repaired more slowly in mre11 rad50 mutants. Using a plasmid repair assay, we show that wild-type and mre11 rad50 cells use different strategies of DSB repair. In the wild-type, Mre11-Rad50 appears to prevent the repair of DSBs by homologous recombination (HR), allowing microhomology-mediated end-joining to act as the primary repair pathway. However, genetic analysis of recombination-defective radA mutants suggests that DNA repair in wild-type cells ultimately requires HR, therefore Mre11-Rad50 merely delays this mode of repair. In polyploid organisms, DSB repair by HR is potentially hazardous, since each DNA end will have multiple partners. We show that in the polyploid archaeon H. volcanii the repair of DSBs by HR is restrained by Mre11-Rad50. The unrestrained use of HR in mre11 rad50 mutants enhances cell survival but leads to slow recovery from DNA damage, presumably due to difficulties in the resolution of DNA repair intermediates. Our results suggest that recombination might be similarly repressed in other polyploid organisms and at repetitive sequences in haploid and diploid species

AKT overactivation can suppress DNA repair via p70S6 kinase-dependent downregulation of MRE11

Deregulated AKT kinase activity due to PTEN deficiency in cancer cells contributes to oncogenesis by incompletely understood mechanisms. Here, we show that PTEN deletion in HCT116 and DLD1 colon carcinoma cells leads to suppression of CHK1 and CHK2 activation in response to irradiation, impaired G2 checkpoint proficiency and radiosensitization. These defects are associated with reduced expression of MRE11, RAD50 and NBS1, components of the apical MRE11/RAD50/NBS1 (MRN) DNA damage response complex. Consistent with reduced MRN complex function, PTEN-deficient cells fail to resect DNA double-strand breaks efficiently after irradiation and show greatly diminished proficiency for DNA repair via the error-free homologous recombination (HR) repair pathway. MRE11 is highly unstable in PTEN-deficient cells but stability can be significantly restored by inhibiting mTORC1 or p70S6 kinase (p70S6K), downstream kinases whose activities are stimulated by AKT, or by mutating a residue in MRE11 that we show is phosphorylated by p70S6K in vitro. In primary human fibroblasts, activated AKT suppresses MRN complex expression to escalate RAS-induced DNA damage and thereby reinforce oncogene-induced senescence. Taken together, our data demonstrate that deregulation of the PI3K-AKT/ mTORC1/ p70S6K pathways, an event frequently observed in cancer, exert profound effects on genome stability via MRE11 with potential implications for tumour initiation and therapy