Biology subjects

Dhillon, B.

Publications and source records attributed to Dhillon, B..

6 recordsLinked to original sources

bioRxiv2026

Burkholderia cenocepacia physiology and molecular adaptations to the acidic pH of the CF nutritional environment

Burkholderia cenocepacia is an opportunistic pathogen associated with increased disease severity and mortality in cystic fibrosis (CF) patients. We have previously shown that elevated iron and acidic pH in the CF nutritional environment increases B. cenocepacia growth rate and decreases its susceptibility to some of the antimicrobials used clinically to treat CF infections. Here, we aimed to characterize B. cenocepacia physiology and its molecular response under acidic pH and increased zinc and iron concentrations using a modified synthetic CF sputum media (SCFM-FeZn). By investigating B. cenocepacia internal pH homeostasis, we found that it maintains a neutral internal pH when exposed to mildly acidic media at pH 5.5. We also assessed the effect of B. cenocepacia growth on the pH of SCFM-FeZn. When cultured at pH 6.8, B. cenocepacia maintained a media pH of [~]6.5. In contrast, when the culture pH value was initially 5.5, it increased to 6.5 during growth. Using comparative transcriptomics and metabolomics analysis, we identified 990 differentially expressed genes, and 23 differentially abundant metabolites in supernatants at acidic compared to neutral pH. Some of these genes and metabolites were involved in aromatic amino acid metabolism including the upregulated trpE gene, encoding a tryptophan biosynthetic enzyme. A tryptophan auxotrophic trpE deletion strain grew slower in SCFM-FeZn. Overall, this work identifies mechanisms involved in B. cenocepacia adaptation to acidic pH under conditions to model the CF nutritional environment. Some of these mechanisms are also associated with pathogenicity and virulence. ImportancePathogenic bacteria can be exposed to acidic pH inside and outside the host. Their ability to adapt to pH fluctuations contributes to success in host colonization. B. cenocepacia can grow at acidic pH ([~]3.5) and has been recovered from intracellular acidic compartments of amoebas and macrophages. Adaptation to acidic pH depends on molecular mechanisms that maintain a near optimal pH inside the cell for the function of vital processes. A few mechanisms that contribute to its adaptation to acidic pH have been described, but not in conditions reflecting the CF nutritional environment. Here, we identified multiple differentially-regulated systems that are associated with bacterial susceptibility to antimicrobials and pathogenesis. This research provides a better understanding of the role of acidic pH on B. cenocepacia physiology in the CF nutritional context and highlights possible systems that should be further characterized.

microbiology↗

bioRxiv2023

THE POSSIBILITY OF AN EXTENSION OF CATARACT TREATMENT BEYOND EYE SURGERY: A NOVEL NON-INVASIVE PHOTOBLEACHING OF THE HUMAN LENS BY LED

As discussed below cataract is the world-leading cause of blindness and impaired vision with only eye surgery as a treatment. There is major requirement for an alternative which this research has provided via Genetically Modified pigs with natural cataract making live animal experiments of LED photobleaching successful monitored by fluorescence spectra. The latter were shown by EBS as spectrally identical in pigs and humans. EBS developed a miniature fluorescence spectrometer via tunable interference filters with fluorescence excited by an LED. A second LED in the blue/violet region provided a 20mW treatment beam which was shown to reverse the fluorescence signal and dramatically improve the transmission of pigs eyes. These results led to permission for early Human Trials. The results were judged by independent optometrists by LOCS and fluorescence in pigs and in Human Trials by LOCS and Visual Acuity. Adequate positive results have led to full length Human Trials being conducted in four European countries. Prospects of worldwide application have been therefore indicated.

biophysics↗

bioRxiv2021

Quantitative Action Spectroscopy Reveals ARPE-19 Sensitivity to Long-Wave Ultraviolet Radiation at 350 nm and 380 nm

The role of ultraviolet radiation (UVR) exposure in the pathology of age-related macular degeneration (AMD) has been debated for decades with epidemiological evidence failing to find a clear consensus for or against it playing a role. A key reason for this is a lack of foundational research into the response of living retinal tissue to UVR in regard to AMD-specific parameters of tissue function. We therefore explored the response of cultured retinal pigmented epithelium (RPE), the loss of which heralds advanced AMD, to specific wavelengths of UVR across the UV-B and UV-A bands found in natural sunlight. Using a bespoke in vitro UVR exposure apparatus coupled with bandpass filters we exposed the immortalised RPE cell line, ARPE-19, to 10nm bands of UVR between 290 and 405nm. Physical cell dynamics were assessed during exposure in cells cultured upon specialist electrode culture plates which allow for continuous, non-invasive electrostatic interrogation of key cell parameters during exposure such as monolayer coverage and tight-junction integrity. UVR exposures were also utilised to quantify wavelength-specific effects using a rapid cell viability assay and a phenotypic profiling assay which was leveraged to simultaneously quantify intracellular reactive oxygen species (ROS), nuclear morphology, mitochondrial stress, epithelial integrity and cell viability as part of a phenotypic profiling approach to quantifying the effects of UVR. Electrical impedance assessment revealed unforeseen detrimental effects of UV-A, beginning at 350nm, alongside previously demonstrated UV-B impacts. Cell viability analysis also highlighted increased effects at 350nm as well as 380nm. Effects at 350nm were further substantiated by high content image analysis which highlighted increased mitochondrial dysfunction and oxidative stress. We conclude that ARPE-19 cells exhibit a previously uncharacterised sensitivity to UV-A radiation, specifically at 350nm and somewhat less at 380nm. If upheld in vivo, such sensitivity will have impacts upon geoepidemiological risk scoring of AMD.

pharmacology and toxicology↗

bioRxiv2019

Variable Genome Evolution in Fungi After Transposon-Mediated Amplification of a Housekeeping Gene

BackgroundTransposable elements (TEs) can be key drivers of evolution, but the mechanisms and scope of how they impact gene and genome function are largely unknown. Previous analyses revealed that TE-mediated gene amplifications can have variable effects on fungal genomes, from inactivation of function to production of multiple active copies. For example, a DNA methyltransferase gene in the wheat pathogen Zymoseptoria tritici (synonym Mycosphaerella graminicola) was amplified to tens of copies, all of which were inactivated by Repeat-Induced Point mutation (RIP) including the original, resulting in loss of cytosine methylation. In another wheat pathogen, Pyrenophora tritici-repentis, a histone H3 gene was amplified to tens of copies with little evidence of RIP, leading to many potentially active copies. To further test the effects of transposon-aided gene amplifications on genome evolution and architecture, the repetitive fraction of the significantly expanded Pseudocercospora fijiensis genome was analyzed in greater detail. ResultsThese analyses identified a housekeeping gene, histone H3, which was captured and amplified to hundreds of copies by a hAT DNA transposon, all of which were inactivated by RIP, except for the original. In P. fijiensis the original H3 gene probably was not protected from RIP, but most likely was maintained intact due to strong purifying selection. Comparative analyses revealed that a similar event occurred in five additional genomes representing the fungal genera Cercospora, Pseudocercospora and Sphaerulina. ConclusionsThese results indicate that the interplay of TEs and RIP can result in different and unpredictable fates of amplified genes, with variable effects on gene and genome evolution.

genomics↗

bioRxiv2018

Retinal microvasculature and cerebral small vessel disease in the Lothian Birth Cohort 1936 and Mild Stroke Study

Research has suggested that the retinal vasculature may act as a surrogate marker for diseased cerebral vessels. Retinal vascular parameters were measured using Vessel Assessment and Measurement Platform for Images of the Retina (VAMPIRE) software in two cohorts: (i) community-dwelling older subjects of the Lothian Birth Cohort 1936; and (ii) patients with recent minor ischaemic stroke of the Mild Stroke Study. Imaging markers of small vessel disease (SVD) (white matter hyperintensities [WMH] on structural MRI, visual scores and volume; perivascular spaces; lacunes and microbleeds), and vascular risk measures were assessed in both cohorts. We assessed associations between retinal and brain measurements using structural equation modelling and regression analysis. In the Lothian Birth Cohort 1936 arteriolar fractal dimension accounted for 4% of the variance in WMH load. In the Mild Stroke Study lower arteriolar fractal dimension was associated with deep WMH scores (odds ratio [OR] 0.53; 95% CI, 0.32-0.87). No other retinal measure was associated with SVD. Reduced fractal dimension, a measure of vascular complexity, is related to SVD imaging features in older people. The results provide some support for the use of the retinal vasculature in the study of brain microvascular disease.

neuroscience↗

bioRxiv2017

Novel Locus Influencing Retinal Venular Tortuosity Is Also Associated With Risk Of Coronary Artery Disease

Structural variation in retinal blood vessels is associated with global vascular health in humans and may provide a readily accessible indicator of several diseases of vascular origin. Increasing evidence suggests variation in retinal vasculature is highly heritable. This study aimed to identify genetic determinants of retinal vascular traits. We reported a meta-analysis of genome-wide association studies (GWAS) for quantitative retinal vascular traits derived using semi-automatic image analysis of digital retinal photographs from the Genetics of Diabetes Audit and Research in Tayside (GoDARTS) (n=1736) and the Orkney Complex Disease Study (ORCADES) (n=1358) cohorts. We identified a novel genome-wide significant locus at 19q13 (ACTN4/CAPN12) for retinal venular tortuosity (TortV), and one at 13q34 (COL4A2) for retinal arteriolar tortuosity (TortA); these two loci were subsequently confirmed in three independent cohorts (n=1413). In the combined analysis in ACTN4/CAPN12 the lead single nucleotide polymorphism (SNP) was rs1808382 (n=4507; Beta=-0.109; standard error (SE) =0.015; P=2.39x10-13) and in COL4A2 it was rs7991229 (n=4507; Beta=0.103; SE=0.015; P=4.66x10-12). Notably, the ACTN4/CAPN12 locus associated with retinal TortV is also associated with coronary artery disease and heart rate. Our findings demonstrate the contribution of genetics in retinal tortuosity traits, and provide new insights into cardiovascular diseases.\n\nAuthor SummaryRetinal vascular features are associated with wide range of diseases related to vascular health and provide an opportunity to understand early structural changes in vasculature which may help to predict disease risk. Emerging evidence indicates that retinal tortuosity traits are both associated with vascular health and highly heritable. However, the genetic architecture of retinal vascular tortuosity has not been investigated. We therefore performed a genome-wide association study on retinal arteriolar tortuosity (TortA) and retinal venular tortuosity trait (TortV) using data from two independent discovery cohorts of 3094 individuals of European-heritage. We found a novel associations at 19q13 (ACTN4/CAPN12) for TortV, and one at 13q34 (COL4A2) for TortA at discovery stage and validated in three independent cohorts. A significant association was subsequently found between lead SNPs at 19q13 and coronary artery disease, cardiovascular vascular risk factors and heart rate. We also performed genome-wide association studies for retinal vascular calibres and optic disc radius (ODradius) and replicated previously reported locus at 10q21.3 for ODradius. Our findings highlight genetic impacts on retinal venular tortuosity and it is association with cardiovascular disease. This may provide a molecular pathophysiological foundation for use of retinal vascular traits as biomarkers for cardiovascular diseases.

genomics↗