Cecil S. Thompson

Purinergic receptor‐mediated effects of ATP in high‐grade bladder cancer

To assess whether the antineoplastic action of extracellular ATP seen in hormone‐refractory prostate cancer extends to other aggressive urological malignancies by investigating its effect in high‐grade bladder cancer cells in vitro and in vivo.

Effect of extracellular ATP on the growth of hormone-refractory prostate cancer in vivo.

To investigate whether the antineoplastic action of ATP on hormone‐refractory prostate carcinoma (HRPC) cells in vitro also occurs in vivo, by examining the effect of ATP in vivo on tumours resulting from implanted HRPC cells in mice.

Characterization of calcium-independent purinergic receptor-mediated apoptosis in hormone-refractory prostate cancer.

To investigate the nature of purinergic signalling in hormone‐refractory prostate cancer (HRPC) cells in vitro, as extracellular ATP inhibits the growth of HRPC in vitro via the activation of P2 purinergic receptors, and to characterize which P2 receptors subtypes and secondary mechanisms are involved.

The Management of Phosphodiesterase-5 (PDE5) Inhibitor Failure

The oral phosphodiesterase type 5 (PDE5) inhibitors have made a valuable contribution to the treatment of erectile dysfunction (ED). PDE5 inhibitors enhance cavernosal smooth muscle relaxation, vasodilatation and penile erection. However, PDE5 inhibitors are not always effective. Decreased efficacy, cost, incorrect administration, lack of sexual stimulation, vascular risk factors associated with ED and vascular or neurogenic diseases are causes of PDE5 inhibitor failure. Tachyphylaxis may also occur. This is defined as reduced tissue responsiveness to a drug in the presence of a constant concentration of this drug. Treatment failure may cause considerable distress. If dose titration, more attempts and continuous dosing of PDE5 inhibitors (taken on a daily basis) fail to resolve the initial PDE5 inhibitor failure, clinicians need to consider alternative treatments. These include sublingual apomorphine, intracavernosal/intraurethral pharmacotherapy, vacuum devices, the insertion of a prosthesis and penile vascular surgery. Combination therapy like prostaglandin E(1) (PGE(1)) with doxazosin (dox; an alpha-1-blocker) or ketanserin (ketan; a 5-HT(2) antagonist) as well as other pro-erection agents, like Endothelin-1 antagonists, angiotensin II antagonists (valsartan/losartan), adrenomedullin, Rho kinase inhibitors and nitric oxide (NO) donors may be beneficial in the treatment of ED. However, these combination therapies need to be validated. Adding an androgen to a PDE5 inhibitor may help when circulatory testosterone levels are low. The early use of PDE5 inhibitors in patients with hypertension, hyperlipidaemia or diabetes with concomitant ED and treating these risk factors may improve corporeal blood flow and lead to long-term preservation of cavernosal function. Therefore, the efficacy of PDE5 inhibitors may be maintained. Targeting the risk factors of ED (similar to those for arteriosclerosis) in the early stages of the disease may prevent the development or decrease the severity of ED.

Smooth Muscle and Purinergic Contraction of the Human, Rabbit, Rat, and Mouse Testicular Capsule

Abstract The smooth-muscle cells of the testicular capsule (tunica albuginea) of man, rat, and mouse were examined by electron microscopy. They were characteristically flattened, elongated, branching cells and diffusely incorporated into the collagenous matrix and did not form a compact muscle layer. Contractile and synthetic smooth-muscle cell phenotypes were identified. Nerve varicosities in close apposition to smooth muscle were seen in human tissue. Contractions induced by adenosine 5′-triphosphate (ATP), alpha, beta-methylene ATP, noradrenaline (NA), acetylcholine (ACh), and electrical field stimulation (EFS) of autonomic nerves were investigated. Nerve-mediated responses of the rabbit and human tunica albuginea were recorded. The EFS-induced human responses were completely abolished by prazosin. In the rabbit, EFS-induced contractile responses were reduced by pyridoxalphosphate-6-azophenyl-2′,4′-disulphonic acid by 36% and by prazosin by 77%. Both antagonists together almost completely abolished all EFS-induced contractions. The human tunica albuginea was contracted by NA, ATP, and alpha, beta-methylene ATP, but not by ACh. The rabbit and rat tunica albuginea were contracted by NA, ATP, alpha, beta-methylene ATP, and ACh. The mouse tunica albuginea was contracted by ACh, ATP, and alpha, beta-methylene ATP, but relaxed to NA. Immunohistochemical studies showed that P2X1 (also known as P2RX1) and P2X2 (also known as P2RX2) receptors were expressed on the smooth muscle of the rodent testicular capsule, expression being less pronounced in man. The testicular capsule of the rat, mouse, rabbit, and man all contain contractile smooth muscle. ATP, released as a cotransmitter from sympathetic nerves, can stimulate the contraction of rabbit smooth muscle. Human, rat, and mouse testicular smooth muscle demonstrated purinergic responsiveness, probably mediated through the P2X1 and/or P2X2 receptors.

Endothelin-1 and Nitric Oxide in the Pathogenesis of Urinary Tract Disorders Secondary to Bladder Outlet Obstruction

Bladder outlet obstruction (BOO) is a common disorder that is associated with urinary tract symptoms. Nitric oxide (NO), synthesized by NO synthase (NOS) is a potent vasodilator that is present throughout the urinary tract and the corpus cavernosum. Endothelin-1 (ET-1) conversely is a potent vasoconstrictor peptide that is similarly distributed throughout the urinary tract. ET-1 and NO as well as possessing opposing actions regulate each others synthesis. The disruption of the balance between ET-1 and NO is associated with various vascular pathologies. However, their potential roles in the pathogenesis of urinary tract disorders, secondary to BOO, is not well established. New Zealand White rabbits with BOO are considered to be a suitable model of the human condition. Hence, using this model, we systematically investigated the potential roles of ET-1 and NO in the pathogenesis of the various urological disorders associated with BOO. In this review we discuss the results of our studies, which support the concept that an imbalance between ET-1 and NO may be associated with the pathogenesis of urinary tract disorders secondary to BOO. We also discuss the potential clinical implications of this association. This review is based on the Bard Silver Medal Lecture given (by MAK) at the 2002 British Association of Urological Surgeons (BAUS) annual meeting.

Animal models of diabetes mellitus: relevance to vascular complications.

The prevalence of diabetes mellitus is increasing worldwide at an alarming rate due to population growth, obesity, sedentary lifestyle and aging. Consequently, diabetic microvascular complications (retinopathy and nephropathy) and macrovascular complications (coronary heart disease, peripheral arterial disease and cerebrovascular disease) are also rising. Traditional oral hypoglycaemic agents only partially prevent the development of these complications. This suggests that selective treatment options that target specific biological pathways (i.e. metabolic factors, intracellular signaling proteins and growth factors) may be a more effective strategy. Type 1 and Type 2 diabetic animal models have been produced spontaneously by selective inbreeding or by genetic modification, as well as, pharmacological induction. These models have become a safe and reliable option to test the therapeutic potential of novel drugs. They also help to understand the pathophysiology of diabetes mellitus. This review highlights the most commonly used animal models for the treatment of diabetic micro and macrovascular complications.

Direct measurement of hepatic tissue hypoxia by using a novel tcpO2/pCO2 monitoring system in comparison with near-infrared spectroscopy.

Abstract: Background/Aims: Hepatic hypoxia occurs during liver surgery and transplantation and it may also appear within liver tumours, correlating with prognosis and efficacy of the treatment. The present study measured liver tissue hypoxia by directly using near‐infrared spectroscopy (NIRS) and a novel tcpO2/pCO2 monitoring system. Methods: Graded hypoxia was achieved in a rabbit model by a stepwise reduction of the fraction of inspired oxygen (FiO2) from 0.3 to 0.0. Animals were allowed to recover from hypoxia at FiO2 of 3.0 indicated by normalised arterial blood gas values. Hepatic tissue oxyhaemoglobin (HbO2), deoxyhaemoglobin (Hb), cytochrome oxidase (Cyt Ox), oxygen partial pressure (pO2) and carbon dioxide partial pressure (pCO2) were measured continuously with the help of NIRS and a Clark‐type surface tcpO2/pCO2 monitoring system, throughout the period of hypoxaemia.

A functional study of purinergic signalling in the normal and pathological rabbit corpus cavernosum

To examine rabbit cavernosal smooth muscle (CSM) relaxation to ATP, ADP and UTP in normal rabbits and in models of conditions that predispose to erectile dysfunction (ED), diabetes mellitus (DM; induced for 6u2003months) and bladder outlet obstruction (BOO, 6u2003weeks after surgery).

Possible role of endothelin-1 in the rabbit urinary bladder hyperplasia secondary to partial bladder outlet obstruction.

Objectives: Urinary bladder hypertrophy and hyperplasia are common features of bladder outlet obstruction (BOO). The urinary bladder is known to synthesize endothelin-1 (ET-1), which is a potent vasoconstrictor peptide with mitogenic properties. Using an animal model of partial BOO, we investigated the potential role of ET-1 and its receptor subtypes (ETA and ETB) in bladder smooth muscle cell (SMC) proliferation. Materials and methods: Partial BOO was produced in adult male New Zealand White rabbits. After 3 weeks, the bladder was removed and SMCs from the dome and bladder neck were grown using standard explant methodology. At passage 2, the cells were made quiescent and then further incubated in foetal calf serum (FCS), control age-matched rabbit serum (CRS) or partial BOO serum (BRS) in the presence or absence of ETA-antagonist (BQ123) or ETB-antagonist (BQ788). SMC proliferation was then measured 24 h later with 5-bromo-2deoxy-uracil and by cell counting using a haemocytometer at 48 h. Immunostaining for alpha-actin was performed on detrusor and bladder neck cells to confirm the presence of smooth muscle cells. Results: BQ123 and BQ788 did not influence detrusor or bladder neck SMC proliferation in FCS or CRS. However, in the presence of BRS, BQ123 and BQ788 (100 nmol/L) significantly (p = 0.008) inhibited detrusor and bladder neck SMC proliferation. Cell counts were significantly reduced from the detrusor (p = 0.03, p = 0.01 with BQ123 and BQ788, respectively) and bladder neck (p = 0.01 for both BQ123 and BQ78). Conclusions: These results suggest that ET antagonists may have a role in preventing SMC hyperplasia associated with partial BOO.