Identification of VEGF Isoforms in Mouse, Rat, and Zebrafish Using RT-qPCR.

Reverse transcription quantitative PCR (RT-qPCR) to quantify gene expression is a key molecular technique for the detection of mRNAs due to amplification of transcripts that may be present in low abundance. The technique also permits detection of mRNAs for different isoforms of proteins due to the exquisite specificity of carefully designed primers. Here I describe the detection of mRNAs for VEGF isoforms in mouse, rat and zebrafish.

Novel urinary biomarkers for the detection of bladder cancer: A systematic review.

Background: Urinary biomarkers for the diagnosis of bladder cancer represents an area of considerable research which has been tested in both patients presenting with haematuria and non-muscle invasive bladder cancer patients requiring surveillance cystoscopy. In this systematic review, we identify and appraise the diagnostic sensitive and specificity of reported novel biomarkers of different 'omic' class and highlight promising biomarkers investigated to date.

Methods: A MEDLINE/Pubmed systematic search was performed between January 2013 and July 2017 using the following keywords: (bladder cancer OR transitional cell carcinoma OR urothelial cell carcinoma) AND (detection OR diagnosis) AND urine AND (biomarker OR assay).

The role of circulating tumour cells and nucleic acids in blood for the detection of bladder cancer: A systematic review.

Background: Blood-based biomarkers are a neglected resource in bladder cancer, where the mainstay of focus has been on urinary biomarkers. However, blood-based biomarkers are gaining popularity in other solid cancers, particularly circulating tumour cells (CTCs) and circulating nucleic acids. In this systematic review, we identify and discuss the diagnostic value of CTC, cell-free DNA and RNA based biomarkers in bladder cancer.

DETECT I & DETECT II: a study protocol for a prospective multicentre observational study to validate the UroMark assay for the detection of bladder cancer from urinary cells.

Background: Haematuria is a common finding in general practice which requires visual inspection of the bladder by cystoscopy as well as upper tract imaging. In addition, patients with non-muscle invasive bladder cancer (NMIBC) often require surveillance cystoscopy as often as three monthly depending on disease risk. However, cystoscopy is an invasive procedure which is uncomfortable, requires hospital attendance and is associated with a risk of urinary tract infection.

UroMark-a urinary biomarker assay for the detection of bladder cancer.

Background: Bladder cancer (BC) is one of the most common cancers in the western world and ranks as the most expensive to manage, due to the need for cystoscopic examination. BC shows frequent changes in DNA methylation, and several studies have shown the potential utility of urinary biomarkers by detecting epigenetic alterations in voided urine. The aim of this study is to develop a targeted bisulfite next-generation sequencing assay to diagnose BC from urine with high sensitivity and specificity.

CSN1 Somatic Mutations in Penile Squamous Cell Carcinoma.

Other than an association with HPV infection, little is known about the genetic alterations determining the development of penile cancer. Although penile cancer is rare in the developed world, it presents a significant burden in developing countries. Here, we report the findings of whole-exome sequencing (WES) to determine the somatic mutational landscape of penile cancer.

Epigenetics markers of metastasis and HPV-induced tumorigenesis in penile cancer.

Purpose: Penile cancer is a rare malignancy in the developed world with just more than 1,600 new cases diagnosed in the United States per year; however, the incidence is much higher in developing countries. Although HPV is known to contribute to tumorigenesis, little is known about the genetic or epigenetic alterations defining penile cancer.

Experimental Design: Using high-density genome-wide methylation arrays, we have identified epigenetic alterations associated with penile cancer.

An ongoing role of α-calcitonin gene-related peptide as part of a protective network against hypertension, vascular hypertrophy, and oxidative stress.

α-Calcitonin gene-related peptide (αCGRP) is a vasodilator, but there is limited knowledge of its long-term cardiovascular protective influence. We hypothesized that αCGRP protects against the onset and development of angiotensin II-induced hypertension and have identified protective mechanisms at the vascular level. Wild-type and αCGRP knockout mice that have similar baseline blood pressure were investigated in the angiotensin II hypertension model for 14 and 28 days.

Epithelial cells promote fibroblast activation via IL-1alpha in systemic sclerosis.

Systemic sclerosis (SSc) is a disorder of systemic and dermal fibrosis of uncertain etiology. Recently, we found that SSc epidermis is abnormal, taking on an activated phenotype observed during wound healing and tissue repair. As epithelial-fibroblast interactions are important during wound repair and in fibrosis in general, we investigated further the phenotype of the SSc epidermis, and tested whether the SSc epidermis provides a pro-fibrotic stimulus to fibroblasts.

Decreased platelet nitric oxide contributes to increased circulating monocyte-platelet aggregates in hypertension.

Aims: The aim of this study was to determine the effect of blood pressure (BP) on platelet nitric oxide (NO) signalling and on formation of circulating monocyte-platelet aggregates (MPA), as well as the role of platelet NO in modulating MPA in hypertension.

Methods And Results: We first examined platelet NO signalling in 23 untreated hypertensive (UH) and 23 normotensive (NT) subjects. Platelets from hypertensives exhibited reduced NO synthase activation by albuterol or collagen, as well as suppressed basal and stimulated NO-attributable cyclic guanosine-3',5'-monophosphate, compared with NT.

Connective tissue growth factor: structure-function relationships of a mosaic, multifunctional protein.

Connective tissue growth factor (CTGF) is a member of the CCN family of six small secreted, cysteine-rich growth factors. The unique modular structure encompasses distinct functional domains which enable CTGF to interact with growth factors, surface receptors and matrix components. Widely expressed, CTGF has critical roles in embryonic development and the maintenance of normal cell and connective tissue function.

The transient receptor potential vanilloid 1 (TRPV1) receptor protects against the onset of sepsis after endotoxin.

Transient potential vanilloid 1 (TRPV1) receptor is an ion channel receptor primarily localized on sensory nerves and activated by specific stimuli to initiate and amplify pain and inflammation, as typified by murine models of scald and arthritis. Little is known of the role of TRPV1 in sepsis, an infective disease associated with inflammation. Through use of a sublethal murine model of lipopolysaccharide-induced peritoneal sepsis, we provide novel evidence that genetic deletion of TRPV1 leads to an enhanced onset of various pathological components of systemic endotoxemia.

The effects of intracellular signalling pathway inhibitors on phagocytosis by haemocytes of Manduca sexta.

The intracellular signalling pathways activated during phagocytosis by larval haemocytes of a lepidopteran, Manduca sexta, were investigated. Using fluorescein-labelled Escherichia coli as bioparticles, a fluorescence-based assay was used to quantify phagocytosis by haemocytes in monolayers in vitro, and the intracellular signalling pathways involved in phagocytosis were examined using inhibitors. Pathways known to be involved in phagocytosis by mammalian cells were selected for the study in haemocytes, and the amino acid sequences of human isoforms of the selected protein targets were used to conduct searches of two completed databases of insect proteins, those of Drosophila melanogaster and Anopheles gambiae and EST databases of moths Bombyx mori and M.

Activation of endothelial nitric oxide synthase by dietary isoflavones: role of NO in Nrf2-mediated antioxidant gene expression.

The endothelium plays a key role in the maintenance of vascular homeostasis, and increased oxidative stress in vascular disease leads to reduced nitric oxide bioavailability and impaired endothelium-dependent relaxation of resistance vessels. Although epidemiological evidence suggests that diets containing high amounts of natural antioxidants afford protection against coronary heart disease (CHD), antioxidant supplementation trials have largely reported only marginal health benefits. There is controversy concerning the cardiovascular benefits of prolonged estrogen/progestin or soy isoflavone therapy for postmenopausal women and patients with an increased risk of CHD.

Nrf2/ARE regulated antioxidant gene expression in endothelial and smooth muscle cells in oxidative stress: implications for atherosclerosis and preeclampsia.

Increased generation of reactive oxygen species (ROS) in vascular diseases such as atherosclerosis, diabetes, chronic renal failure and preeclampsia readily leads to impaired endothelium-dependent relaxation and vascular injury. To counteract ROS- and electrophile-mediated injury, cells can induce a number of genes encoding phase II detoxifying enzymes and antioxidant proteins. A cis-acting transcriptional regulatory element, designated as antioxidant response element (ARE) or electrophile response element (EpRE), mediates the transcriptional activation of genes such as heme oxygenase-1, gamma-glutamylcysteine synthethase, thioredoxin reductase, glutathione-S-transferase and NAD(P)H:quinone oxidoreductase.

Cardiovascular targets for estrogens and phytoestrogens: transcriptional regulation of nitric oxide synthase and antioxidant defense genes.

The benefits of dietary antioxidants against cardiovascular disease have been examined in numerous clinical trials and animal models, yet there is limited evidence linking consumption of soy isoflavones with enhanced expression of antioxidant defense genes in the vasculature. Epidemiological evidence that populations consuming soy products, rich in isoflavones genistein and daidzein, have a lower incidence of cardiovascular disease has led to the suggestion that isoflavones may be beneficial for cardiovascular health. However, population-based studies cannot prove causality and may be confounded by other dietary influences.

Patients' preferences for the management of non-metastatic prostate cancer: discrete choice experiment.

Objective: To establish which attributes of conservative treatments for prostate cancer are most important to men.

Design: Discrete choice experiment.

Setting: Two London hospitals.