No description available.

No description available.

No description available.

The ability of sperm cells to induce specific auto- and iso-immunity was reported as early as the turn of the century. However the mechanism controlling autoreactivity to sperm is not well known. As lymphocytes constitute the major cellular components of the immune system, determination of their anatomical location within the tissues of the male genital tract may be of considerable importance in understanding immunological infertility and other urogenital disorders. A series of monoclonal antibodies that react with human lymphoreticular cells was therefore used in an indirect immunoperoxidase technique to study their distribution throughout the male genital tract. The normal human tissues investigated were: testis, epididymis, vas deferens, prostate and seminal vesicles obtained from multiorgan transplant donors. The clinical specimens examined included surgical biopsies of testis, epididymis and prostate obtained during surgical procedures directed at the investigation and treatment of subfertile males and other patients. All normal tissues, apart from the testis, were found to contain appreciable numbers of T-lymphocytes (leu 4<SUP>+</SUP>). T-cells of the suppressor/cytotoxic phenotype (leu 2a<SUP>+</SUP>) were more abundant in the intraepithelial compartment while T-cells of the helper/inducer phenotye (leu 3a<SUP>+</SUP>) were more common in the interstitial areas. With the exception of the prostate, very few B-cells were observed. Macrophages (leu M3<SUP>+</SUP>) were identified within normal testicular tissues as well as the rest of the male genital tract. HLA-DR<SUP>+</SUP> cells were also identified and the HLA-DR antigens were normally expressed on the lining epithelium of the rete testis, epididymis and vas deferens. Derangement of this pattern was observed in clinical specimens. Testicular biopsies from patients with testicular obstruction showed marked infiltration with lymphocytes mainly of the T-cell type. Biopsies from patients with benign prostatic hyperplasia showed increased infiltration with the helper/inducer T-cells and other cell types such as natural killer cells (leu llb<SUP>+</SUP>) and activated T-cells (IL<SUB>2</SUB>-r<SUP>+</SUP>). These patterns of lymphoid cells distribution could provide an insight into both normal immunohomeostatic mechanisms and pathological events within the male genital tract. The presence and distribution of lymphocyte subpopulations and macrophages within human urothelium in health and disease was also examined. T-lymphocyte subsets and macrophages were identified in normal urothelium and shown to have a similar pattern of distribution to that seen in normal epithelium of the genital tract. The existence of these cell populations may contribute to the health and protection of urothelium, particularly in resistance to infection and tumour surviellance. The presence of leucocytes and their subpopulations was also studied in the ejaculate from 69 men with an infertile marriage and 12 fertile men. Leucocytes were found in large numbers in the fertile men compared with the patients. Lymphocytes were found in 20% of the patients. There was no correlation between leucocyte counts and growth of micro-organisms. These results cast doubt on the conventional criteria of subclinical genital tract infection, namely positive culture and excess leucocyte counts.

The objective of our study was to investigate the hypothesis that carbonated soft drink components modulate detrusor muscle contraction. Strips of rat detrusor muscle were placed in an organ bath and stimulated with electrical field stimulation (EFS) in the absence and presence of atropine, and with alpha, beta methylene ATP, potassium, calcium and carbachol. The responses were repeated in the presence of carbonated soft drink components and the whole carbonated soft drink. The artificial sweeteners, ascorbic acid and critic acid (10-7 M to 10-2 M) enhanced the contractile response to 10 Hz EPS compared to control (p<0.01). The sweeteners (acesulfame K 10-6 M, aspartame 10-7M, sodium saccharin 10-7 M) increased the atropine resistant response to EFS marginally. Acesulfame K 10-6 M and sodium saccharin 10-7 M enhanced the maximum contractile response to alpha, beta methylene ATP, to KC1 and calcium significantly compared to control. Ascorbic acid significantly increased the atropine resistant response to EFS and inhibited contraction in response to carbachol. Both ascorbic acid and citric acid enhanced the contractile responses to alpha, beta methylene ATP, KC1 and calcium significantly compared to control. Whole carbonated soft drink (1:200) also enhanced the contractile responses to alpha,beta methylene ATP, KC1 and calcium. These results suggested that low concentrations of artificial sweeteners, ascorbic acid and critic acid, enhanced detrusor muscle contraction by augmenting Ca2+ influx by a mechanism yet to be defined. Western blot analysis suggested the presence of protein molecules in the rat bladder of similar size to the sweet taste receptor present in tongue. These data suggest that carbonated soft drink components may be acting via specific receptors to modulate the effects observed.

Angiotensin II (ANGII) has profound effects on renal and systemic haemodynamics and renal tubular sodium handling. A pathophysiological role has been proposed for ANGII in causing alterations in renal function in the early stages of human Type 1 (insulin-dependent) diabetes mellitus (IDDM). The studies in this thesis have examined the effect on renal function of acute changes in renin-angiotensin system (RAAS) activity induced by low dose ANGII infusion in normal man and patients with IDDM. In preliminary studies the effects of low dose ANGII infusion on whole kidney renal fucntion were defined in normal man and in IDDM. The renal haemodynamic response to ANGII was normal in IDDM patients, but whole kidney tubular sodium retention occurred in IDDM in parallel with a reduced suppression of plasma renin activity after dietary sodium loading compared to control subjects. The utility of lithium clearance as an indirect marker of tubular sodium handling was then assessed. Several problems in interpreting renal haemodynamic data after lithium pretreatment were identified in normal man and in IDDM, but supplementary studies indicated that lithium clearance remains of value as a marker of tubular sodium handling in IDDM. The data indicate that enhanced proximal tubular reabsorption of sodium is associated with a blunted proximal antinatriuretic response to ANGII infusion in IDDM, the severity of this abnormality correlating with the level of chronic glycaemic control. The urinary concentrating response to ANGII infusion is also abnormal inn IDDM. In separate studies insulin did not affect lithium clearance or interact intrarenally with angiotensin II; insulin treatment could not therefore itself account for the abnormal proximal tubular function found in the diabetic subjects. These results support the hypothesis that increased renal proximal tubular retention of sodium in stable uncomplicated Type 1 diabetes is an acquired functional defect, related to the severity of the diabetic metabolic abnormality.

Patients with IDDM, both with normal albumin excretion and microalbuminuria, have no evidence of a defect of urinary dopamine excretion. This genetically determined association with essential hypertension does not, therefore, explain the link between early diabetic nephropathy and hypertension. In hypertensive patients with IDDM, captopril and nifedipine retard have a similar effect in reducing blood pressure and albumin excretion after 8 weeks treatment. Captopril, however, retains an acute effect on renal haemodynamics, resulting in a decreased filtration fraction, and this may be of specific importance in the management of patients with diabetic nephropathy. In newly-presenting patients with NIDDM, 26% had abnormal urinary albumin excretion, this correlating with age, glycaemic control, systolic blood pressure and generalised vascular disease. Over the year following diagnosis, 16% of patients had persistent microalbuminuria. Although tubular dysfunction is commonly found in patients with microalbuminuria, the relatively poor correlation of currently available markers would seem to limit their value in the detection of early nephropathy. In patients with IDDM and normal albumin excretion, acute hypoglycaemia causes a number of changes in renal function, notably a significant fall in both effective renal plasma flow and glomerular filtration rate (GFR). The fall in GFR is more marked than in non-diabetic subjects. It is possible that hypoglycaemia may have a role in the aetiology or progression of diabetic nephropathy.

No description available.

The polypeptide nerve growth factor (NGF) has been explored extensively over the span of six decades since its detection with amazing discoveries from its neurotrophic action to tissue healing properties. In lower urinary tract there is substantial evidence linking NGF and lower urinary tract dysfunction (LUTD). Over recent years the role of urinary NGF [UrNGF] in diagnosing LUTD as well as monitoring treatment response has been investigated extensively. However, the available studies report conflicting data regarding an association between lower urinary tract symptoms [LUTS] and UrNGF, and there is limited evidence for the validity and reliability of urinary NGF assays. In a quest to explore the role of UrNGF as a LUTS biomarker, levels were measured in patients with LUTD, prolapse and asymptomatic controls. This thesis hypotheses that measurement of NGF is of no value in LUTD and prolapse. Therefore the aims were to evaluate the diagnostic and discriminant ability of UrNGF measurement in LUTD and to test the reliability of NGF assays. The other objective was to study the association between UrNGF levels and cystoscopic and histology findings of bladder inflammation in women with overactive bladder [OAB] to explore the link between NGF and inflammation. Change in UrNGF levels after cystocele repair was studied, since bladder wall stretching has been postulated as one of the causes for increased NGF levels. Finally UrNGF levels before and after antibiotic treatment for refractory OAB were measured with the aim to evaluate its role to assess treatment response. On test retest reliability analysis of 13 samples there was almost perfect reliability with an Intraclass correlation coefficient of 0.889; 95%[C.I=0.676-0.965; p<0.001]. Urinary NGF was significantly but non-specifically increased in symptomatic patients [n=205] when compared to controls [n=31](13.33 vs. 2.05 ng NGF/ g Cr, Mann Whitney test; p <0.001) However ROC analysis, demonstrated poor discriminant ability between either different symptomatic groups or urodynamic groups. Using a cut off of 13.0 ng NGF/ g creatinine the test provides a sensitivity of 81%, but a specificity of only 39 % for overactive bladder. UrNGF levels were not associated with cystoscopic or histology findings of inflammation and did not improve after anterior repair in women who had an improvement in OAB symptoms. However in the study done to explore the role of urinary NGF as a biomarker to assess treatment response, NGF levels were found to decrease significantly after six weeks of antibiotic therapy in women with refractory OAB symptoms [n=35 patients] (Wilcoxon Signed rank test; p=0.015). This was associated with improvement in OAB symptoms. UrNGF does not appear to be a good diagnostic biomarker but may have a role as a marker of treatment response, hence may have limited role in assessment of women with LUTD.

Retinoids, including the prototypic vitamin A and its main bioactive form, alltrans retinoic acid (tRA), have both anti- and pro-fibrotic effects in renal disease models. To understand and prevent the pro-fibrotic effects of retinoids it is important to establish in vitro models. This work aimed to explore the mechanisms behind the fibrogenic effects of tRA in renal fibroblasts. A picro-Sirius red-based in vitro assay was used to determine the effects of retinoids on total collagen accumulation with and without transforming growth factor (TGF)-β1 in NRK-49F normal rat kidney fibroblasts. Individual fibrotic markers and nuclear receptors were investigated using molecular biology approaches, activity assays and chemical agonists and inhibitors. tRA dose-dependently increased total collagen deposition with and without TGF-β1 in NRK-49F cells. At the level of gene expression tRA showed dual potential, down-regulating mRNAs encoding a range of extracellular matrix proteins and matrix metalloproteinases (MMPs), while up-regulating others including plasminogen activator inhibitor (PAI)-1 and transglutaminase 2 (TG2). tRA alone and additively, with TGF-β1, reduced MMP activity and increased PAI-1 protein; the PAI-1 inhibitor tiplaxtinin reduced the increase in total collagen caused by tRA and TGF-β1 treatment. TG2 protein was not modulated by tRA and a TG2 inhibitor did not reduce tRA’s pro-fibrotic effect. NRK-49F cells expressed retinoid nuclear receptors and PPARβ/δ and nuclear receptor mRNAs were differentially regulated by tRA and TGF-β1. RAR and RXR antagonists reduced tRA’s pro-fibrotic effect while a pan-RXR agonist more than a pan-RAR or PPARβ/δ agonist increased total collagen deposition. RAR isotype-selective agonists had less, if any, effect on fibrosis. In summary, an in vitro model for the pro-fibrotic effects of retinoids has been established, which is associated with modulation of MMPs, PAI-1 and RAR/ RXR. Further studies of RAR isotype-selective agonists might be of merit as they had reduced pro-fibrotic activities compared to less selective retinoids.