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Purpose: Prostate cancer (PCa) is a leading cause of cancer-related death. Upon androgen-deprivation therapy, the disease may progress further to castration-resistant PCa (CRPC) with a poor prognosis. MicroRNAs (miRNAs) are small non-coding RNAs, which play crucial roles in gene regulation. The aim of our study is to find CRPC-associated miRNAs and to evaluate their functional role. Methods: In this study, 23 benign prostatic hyperplasia (BPH), 76 primary PCa, and 35 CRPC specimens were included. Total RNA extracted from tissue sections was used for miRNA profiling on the Affymetrix GSC 3000 platform. Subsequently, stem-loop RT-qPCR analysis was performed to validate the expression levels of selected miRNAs. PCa cell lines were transfected with miRNA mimics or inhibitors to evaluate the effects on cell proliferation, cell migration and cell invasion. Results: In our profiling study, several miRNAs were found to be deregulated in CRPC compared to primary PCa tissue, of which miR-205 (− 4.5-fold; p = 0.0009), miR-92b (− 3.1 fold; p < 0.0001) were downregulated and miR-3195 (5.6-fold; p < 0.0001), miR-3687 (8.7-fold; p = 0.0006) and miR-4417 (5.0-fold; p = 0.0005) were most upregulated. While KLK3, miR-21 and miR-141 expression levels in androgen-treated VCaP and LNCaP cells were increased, the expression levels of miR-3687 and miR-4417 were reduced. None of the miRNAs were androgen-regulated in the AR-negative PC3 cell line. Overexpression of miR-3687 reduced cell migration and cell invasion, whilst miR-3195 enhanced cell migration. Conclusion: We have identified several novel deregulated miRNAs in CRPC tissue, including two microRNAs that are potentially involved in tumor invasion. Our data support the hypothesized involvement of miRNAs in PCa tumorigenesis and progression to CRPC. The applicability of these miRNAs as novel biomarkers for CRPC remains to be further investigated. [ABSTRACT FROM AUTHOR]

Aims: To determine whether glycosaminoglycan (GAG) replenishment is able to improve recovery of a deficient urothelial barrier, chondroitin sulfate (CS) instillations were tested using an in vitro model. Porcine urothelial cells (Ucells) were terminally differentiated in culture conditions to construct a urothelial layer with a functional barrier. This layer was damaged to compromise barrier function to simulate a key characteristic of bladder pain syndrome/interstitial cystitis. The functional effect of subsequent treatment with CS was evaluated. Methods: Primary porcine Ucells were isolated and cultured on inserts. Differentiation of cells was evaluated with immunohistochemical analysis for the presence of umbrella cells, tight junctions and CS. Transepithelial electrical resistance (TEER) measurements were performed to evaluate barrier function. Protamine was used to simulate mild urothelial damage. CS 0.2% (vol/vol), a GAG, was subsequently instilled in the treatment group. The recovery of barrier function was further evaluated with TEER measurements. The Student t test was used for the analysis of results. Results: After induction of differentiation, the Ucells expressed barrier markers and a functional barrier was established (measured by high TEER). TEER decreased significantly after instillation with protamine. CS instillation improved recovery of TEER significantly measured after 7 hours (84% vs 22% in controls). After 24 hours; however, the TEER was comparable in both experimental groups. Conclusion: CS instillation improves the recovery of the urothelial barrier after damage in vitro. This functional experiment shows that CS improves recovery of damaged urothelial function, which supports the hypothesis behind the mechanism of action of GAG‐replenishment therapy. [ABSTRACT FROM AUTHOR]

Aim This review critically evaluates the evidence on mechanoreceptors and pathways in the bladder urothelium that are involved in normal bladder filling signalling. Methods Evidence from in vitro and in vivo studies on (i) signalling pathways like the adenosine triphosphate pathway, cholinergic pathway and nitric oxide and adrenergic pathway, and (ii) different urothelial receptors that are involved in bladder filling signalling like purinergic receptors, sodium channels and TRP channels will be evaluated. Other potential pathways and receptors will also be discussed. Results Bladder filling results in continuous changes in bladder wall stretch and exposure to urine. Both barrier and afferent signalling functions in the urothelium are constantly adapting to cope with these dynamics. Current evidence shows that the bladder mucosa hosts essential pathways and receptors that mediate bladder filling signalling. Intracellular calcium ion increase is a dominant factor in this signalling process. However, there is still no complete understanding how interacting receptors and pathways create a bladder filling signal. Currently, there are still novel receptors investigated that could also be participating in bladder filling signalling. Conclusions Normal bladder filling sensation is dependent on multiple interacting mechanoreceptors and signalling pathways. Research efforts need to focus on how these pathways and receptors interact to fully understand normal bladder filling signalling. [ABSTRACT FROM AUTHOR]

Aim The molecular interactions between transient receptor potential vanilloid subtype 4 channels ( TRPV4) and cell junction formation were investigated in the human and mouse urogenital tract. Materials and Methods A qualitative study was performed to investigate TRPV4 channels, adherence junctions ( AJs) and tight junctions ( TJs) in kidney, ureter and bladder tissues from humans and wild-type and transgenic TRPV4 knockout (−/−) mice with immunohistochemistry, Western blotting, immunoprecipitation and reverse trasnscription- PCR. Cell junction formation in the wild-type and TRPV4 knockout (−/−) mouse was evaluated with immunohistochemistry and transmission electron microscope ( TEM) techniques. Results TRPV4 channels are predominantly located in membranes of epithelial cells of the bladder, ureter and the collecting ducts of the kidney. There is a molecular interaction between the TRPV4 channel and the AJ. TEM evaluation showed that AJ formation is disrupted in the TRPV4 −/− mouse resulting in deficient intercellular connections and integrity of the epithelium. Conclusions TRPV4 is believed to be a mechanoreceptor in the bladder. This study demonstrates that TRPV4 is also involved in intercellular connectivity and structural integrity of the epithelium. [ABSTRACT FROM AUTHOR]

Prostate cancer (PCa) is the second most common diagnosed malignant disease in men worldwide. Although serum PSA test dramatically improved the early diagnosis of PCa, it also led to an overdiagnosis and as a consequence to an overtreatment of patients with an indolent disease. New biomarkers for diagnosis, prediction, and monitoring of the disease are needed. These biomarkers would enable the selection of patients with aggressive or progressive disease and, hence, would contribute to the implementation of individualized therapy of the cancer patient. Since the FDA approval of the long noncoding PCA3 RNA-based urine test for the diagnosis of PCa patients, many new noncoding RNAs (ncRNAs) associated with PCa have been discovered. According to their size and function, ncRNAs can be divided into small and long ncRNAs. NcRNAs are expressed in (tumor) tissue, butmany are also found in circulating tumor cells and in all body fluids as protein-bound or incorporated in extracellular vesicles. In these protected forms they are stable and so they can be easily analyzed, even in archival specimens. In this review, the authors will focus on ncRNAs as novel biomarker candidates for PCa diagnosis, prediction, prognosis, and monitoring of therapeutic response and discuss their potential for an implementation into clinical practice. [ABSTRACT FROM AUTHOR]

Background:Prostate cancer is the most commonly diagnosed cancer in men in Europe and the United States. Numerous studies have indicated genetics to have a major role in the aetiology of this disease; as much as 42% of the risk may be explained by heritable factors. Genome-wide association studies have detected an association between prostate cancer and chromosome 8p21-23. In this study, we analysed eight microsatellite (MS) markers in that region in order to confirm previous results and narrow down the location of candidate prostate cancer genes.Methods:292 cases and 278 controls were selected from the Netherlands Cohort Study (NLCS). The following MSs were used in the analyses: D8S136, D8S1734, D8S1742, D8S261, D8S262, D8S351, D8S511 and D8S520. Associations were evaluated using a χ2 test and logistic regression. We checked for any effects on the association by tumour stage.Results:Associations that were found confirmed previous research that pointed to the 8p21-23 region. Two MSs: D8S136 (odds ratio (OR), 0.69; P=4.00 × 10−28), and D8S520 (OR, 0.80; P=3.37 × 10−11), were consistently and strongly related with prostate cancer. Genotype analysis showed an additive effect for D8S136 (P-trend=6.22 × 10−03) and D8S520 (P-trend=2.62 × 10−22), suggesting an increased risk for people with a short number of repeats on both alleles at those markers.Conclusions:This study provides strong evidence that the 8p21-23 region is likely to harbour prostate cancer genes. [ABSTRACT FROM AUTHOR]

With ∼30 000 deaths annually in the United States, prostate cancer (PCa) is a major oncologic disease. Here we show that the microRNAs miR-130a, miR-203 and miR-205 jointly interfere with the two major oncogenic pathways in prostate carcinoma and are downregulated in cancer tissue. Using transcriptomics we show that the microRNAs repress several gene products known to be overexpressed in this cancer. Argonaute 2 (AGO2) co-immunoprecipitation, reporter assays and western blot analysis demonstrate that the microRNAs directly target several components of the mitogen-activated protein kinase (MAPK) and androgen receptor (AR) signaling pathways, among those several AR coregulators and HRAS (Harvey rat sarcoma viral oncogene homolog), and repress signaling activity. Both pathways are central for the development of the primary tumor and in particular the progression to its incurable castration-resistant form. Reconstitution of the microRNAs in LNCaP PCa cells induce morphological changes, which resemble the effect of androgen deprivation, and jointly impair tumor cell growth by induction of apoptosis and cell cycle arrest. We therefore propose that these microRNAs jointly act as tumor suppressors in prostate carcinoma and might interfere with progression to castration resistance. [ABSTRACT FROM AUTHOR]

One of the current challenges in the evaluation of novel agents for the treatment of advanced prostate cancer is the identification of a surrogate end point for overall survival (OS). Prostate-specific antigen (PSA) levels have been used as a screening tool and a biomarker of response to both hormonal and cytotoxic agents. However, PSA levels do not seem to be a suitable surrogate end point for OS in trials of targeted agents for castrate-resistant prostate cancer (CRPC). These findings suggest the need for adopting measures of efficacy that more accurately reflect the mechanisms of action of these agents in phase II trials, in order to realize improvements in OS in the phase III setting. The Prostate Cancer Clinical Trials Working Group (PCWG2) have recently made recommendations for the design of future trials and advised that PSA levels should not be the sole criterion on which to base clinical decisions. Here, we appraise the end points that have been used in phase II and III trials in patients with CRPC, and highlight the need for the adoption of the PCWG2 guidelines, the recommendations of which include radiographic imaging, in addition to bone scintigraphy, and symptomatic or radiographic disease progression criteria. [ABSTRACT FROM AUTHOR]

We investigated whether smoking is associated with mutations in the Von Hippel-Lindau (VHL) gene in 337 cases of sporadic renal cell carcinoma (RCC) among 120 852 people followed for 11.3 years; the findings suggest that smoking causes RCC independently of VHL gene mutations. [ABSTRACT FROM AUTHOR]

Presents abstracts of articles about oncology to be discussed at the Annual Scientific Meeting of the British Association of Urological Surgeons from June 23 to 27, 2003 in Manchester, England. `A high frequency of microsatellite instability is seen at tetranucleotide repeats in TCC of the bladder,' by J.W.F. Catto et al.

Benign prostatic hyperplasia (BPH) is one of the most common diseases affecting aging man. Attempts have been made to clarify the etiology and pathogenesis and, to that end, experimental models have been developed. To date, in vitro and in vivo models have been used, depending on the concept of the study. Spontaneous animal models are limited to the chimpanzee and the dog. Ethical and financial factors restrict the applicability of these models. The hormonal-induced canine BPH model is a good alternative that closely resembles human BPH in many aspects. The experimental models currently used for studying BPH are reviewed. Prostate Cancer and Prostatic Diseases (2000) 3, 28–33 [ABSTRACT FROM AUTHOR]

Finasteride is widely used for the treatment of benign prostatic hyperplasia (BPH). Its therapeutic efficacy is believed to be mediated through selective inhibition of prostatic 5α-reductase (type II). This prospective, controlled, randomized study examines various relationships between changes in tissue 5α-reductase isozyme activity, epithelial proliferative index and morphology in men with BPH treated with finasteride for 6 months, and correlates these with clinical response. Thirty-one patients presenting with symptomatic bladder outflow obstruction were prospectively randomized to receive either finasteride or placebo (2:1) for 24 weeks. Of these, 27 men aged 55–80 y (median 69 y) completed the study, including 18 patients on treatment. Symptom score determination, uroflow and prostate volume were assessed at baseline and end of study. Prostatic tissue was obtained by ultrasound-guided needle biopsy at baseline and by transurethral resection (TURP) at end of study for biochemical and morphometric analysis. Epithelial proliferation was examined quantitatively in the resected tissue using MIB-1 antibody to Ki-67 and counting the percentage of positively staining cells. Type II 5α-reductase activity was strongly inhibited by finasteride in resected BPH tissue, with over 100-fold decrease in Vmax (P=0.001), whereas the type I isozyme was inhibited 5-fold (P=0.005). Selective inhibition of type II 5α-reductase was demonstrated in all treated patients. No significant difference in epithelial proliferation was observed between the finasteride and placebo groups. Epithelial proliferation was, however, greater in prostatic tissue with histological manifestation of inflammation (2.02% vs 0.89%, P=0.001). Positive correlation between the total epithelial volume and the ratio of transition zone volume/total prostate volume was observed in the placebo group (r=0.834), whereas there was no such correlation in men taking finasteride... [ABSTRACT FROM AUTHOR]

E-cadherin is a Ca-dependent cell adhesion molecule involved in cell-cell interaction. In its normal physiological function it plays an important role in embryonic development and tissue morphogenesis. Recent studies have shown that in cancer development E-cadherin can act as a suppressor of invasion. Indeed, in several kinds of carcinomas allelic loss of the E-cadherin/Uvomorulin locus and decreased E-cadherin expression have been described. The importance of E-cadherin in human cancer development may be substantiated by molecular analysis of the E-cadherin transcript. Therefore, we isolated and characterized the human E-cadherin cDNA. Comparison of the nucleotide and deduced amino acid sequences revealed that the human E-cadherin is highly homologous to the mouse E-cadherin (uvomorulin) and to other members of the cadherin family. [ABSTRACT FROM AUTHOR]

Comparison of nucleotide sequence data of the 5′ region of a fes/fps viral oncogene with those of the v- fes/fps homologous regions of man and cat revealed the position of the 3′ portion of an as yet unidentified c- fes/fps exon. Comparative Southern blot and heteroduplex analysis of human and feline DNA immediately upstream of the v- fes/fps homologous regions showed extensive but discontinuous homology over a 9 kbp DNA stretch, which we have designated as fur. Northern blot analysis of mRNA from KG-1 myeloid cells with fes/fps-or fur-specific probes revealed a 3.0 kb fes/fps and a 4.5 kb fur transcript. Analysis of a number of tissues of an adult Wistar Lewis rat for the presence of fur transcripts revealed its differential expression pattern. An 0.95 kbp fes/fps-related and a 2.2 kbp fur-related cDNA recombinant clone were isolated from an oligo(dT)-primed KG-1 cDNA library. Comparative nucleotide sequence analysis of the fes/fps cDNA and its human genomic counterpart indicated that the cDNA contained genetic sequences that were identical to and colinear with exon 15-19 and , furthermore, that the poly(A) addition signal near the 3′ end of exon 19 was functional. Similar analysis of the 2.2 kbp fur cDNA indicated that the poly(A) addition signal of the fur transcript was in close proximity of the newly discovered fes/fps exon. The region in between contained a CATT sequence but no 'TATA' box. The fur transcript was characterized by a long noncoding region at its 3′ end. [ABSTRACT FROM AUTHOR]

The genetic and epigenetic changes that occur during cancer development result in apparent morphological changes. Light microscopic image analysis provides objective assessment of cellular and nuclear morphology. The complexity of changes reflects the basic nature of dedifferentitation: a multi-hit process. Image analysis methods proved valuable for the assessment of malignancy in bladder cancer. Clinically applicable systems have been developed to diagnose urothelial cell cancer and predict prognosis. What is the place of these systems in daily practice? [ABSTRACT FROM AUTHOR]

A cohort of 300 ACI rats was kept under standard laboratory conditions. After 30 months or upon natural death, complete autopsy was performed. In the genitourinary tract four kidney and five bladder tumors were found. Two of these bladder tumors, RBT323 and RBT157, are serially transplantable. In the fifth transplant generation the RBT323 tumor becomes metastatic to the lungs in more than 90% of animals. The metastatic ability of the RBT157 tumor changes from low to intermediate (50% of the rats have lung metastases) in the fourth passage. Histologically, the initial passages of the RBT323 and 157 tumors are grade II transitional cell carcinoma (TCC). The histological pattern of the RBT157 tumor remains essentially unchanged, whereas the RBT323 tumor progresses to a grade III tumor in the third passage. Electron microscopical studies reveal oblong elliptical and round vesicles lined by an asymmetrical unit membrane in the tumor cells, which stresses the urothelial origin of the tumors. Immunohistochemically both tumors show expression of cytokeratin 5, 7, 8 and 18. The progression of the tumors to a metastatic phenotype, however, is not associated with a specific change in the morphological characteristics. Cytogenetic analysis shows that both tumors are peridiploid with few marker chromosomes. Interestingly, both of these independently arising tumors exhibit a loss of chromosome 5. Rat chromosome 5 is syntenic to the major portion of human chromosome 9 (p23-qter). Loss of chromosome 9 is a cytogenetic trait of human superficial TCC, hence the RBT mocel is also in cytogenetic respect similar to human TCC. Two independently arising rat tumor lines that initially resemble superficial TCC both phenotypically and cytogenetically are described. Upon serial transplantation both lines progressed to a more aggressive tumor, albeit to a different extent (highly vs moderately metastatic). Thus this model system may be helpful in the identification of specific markers associated with the progression of superficial bladder cancer. [ABSTRACT FROM AUTHOR]

Transitional cell carcinoma of the bladder has a high recurrence rate after local treatment. Progression to a higher stage occurs in 10-30% of the recurrent tumors, and early detection of potentially progressive tumors is important. In the current study morphometric, densitometric, and chromatin textural features of nuclei of superficial bladder tumors (pTa-T1) were studied to determine the value of karyometric features in the prediction of tumor progression. Seventy-two histological samples from 36 patients, consisting of both the primary and the first recurrent superficial tumor, were analyzed. Patients were divided into two groups: those with tumor progression, defined as an increase in tumor stage or occurrence of metastatic disease, and those without. Discriminant analysis on four karyometric features resulted in correct prediction of prognosis of 78% and 97% in the primary and recurrent tumors, respectively ( P<0.001). Tumor grade and stage did not offer additional information concerning prognosis. Karyometric analysis of recurrent superficial transitional cell tumors can be useful in selecting patients who need a more aggressive therapy. However, tumor characteristics of recurrent tumors varied and continuous evaluation of the karyometric features is necessary for early detection of an increase in the malignant potential of the tumor. [ABSTRACT FROM AUTHOR]

The rod visual pigment, rhodopsin, and its illuminated form, opsin, were used to induce experimental autoimmune uveoretinitis in rats. Rhodopsin appears to be more pathogenic than opsin. A dose of 250 μg rhodopsin injected in Freund's complete adjuvant and pertussis adjuvant induces nongranulomatous inflammation with higher frequency, which starts earlier and is more severe than that induced by opsin. Two weeks postinjection, the mean score of rhodopsin-injected animals is more than twice as high as that of opsin-injected animals. The high pathogenicity of rhodopsin appears to be related to the biochemical integrity of the protein and depends on its state of illumination. The levels of the immune responses (both cellular and humoral) measured at day 10 postinjection do not account for the pronounced difference in pathogenicity between rhodopsin and opsin. The developmental patterns of severe uveoretinitis induced by rhodopsin or opsin were histologically evaluated and appear to be similar. In both cases we observed dense mononuclear and polymorphonuclear cell infiltrations in the retina and anterior uvea. Only in the severe stages does the choroid become involved. However, rhodopsin causes more pronounced involvement of the ciliary body, pars plana, and anterior chamber. The inflammation finally results in total elimination of the photoreceptor cell layer. [ABSTRACT FROM AUTHOR]

Over the past 10 years it has become apparent that invasion and metastasis are extremely complex processes; neoplastic cells must escape from the primary tumor, degrade the extracellular matrix, migrate to distant sites, arrest in the capillaries, and migrate through the basement membrane and underlying connective tissue to the metastatic site. Therefore, tumor cells must exhibit considerable flexibility in their adhesive interactions, and this is reflected in a complex and dynamic expression pattern of cell adhesion molecules, proteases, protease inhibitors, motility factors, and growth factors. Despite the recent explosion of information regarding adhesion-related molecules, questions as to their possible roles in normal tissue architecture and as to how alterations in their expression or structure may be responsible for the progression from a single malignant cell to a lethal metastatic disease need further investigation. Moreover, efforts should be made to use the obtained knowledge to contribute to improvements in the clinical management of cancer. In this review the different classes of cell adhesion molecules and proteases are summarized, with special emphasis being placed on molecules that have been shown to correlate with invasion and metastasis. Furthermore, the role of E-cadherin in cell adhesion and invasive processes is discussed in more detail, since E-cadherin may be considered promising as a candidate among cell-adhesion-regulating molecules to be used as a biomarker for malignancy. We also elaborate on the role of the catenins, which associate with and are important for the functioning of E-cadherin. [ABSTRACT FROM AUTHOR]

In this paper the predictive value of molecular prognostic parameters for bladder cancer is discussed. DNA ploidy has additional prognostic value for grade 2 tumors, irrespective of stage, with aneuploid tumors having a poor prognosis. Overexpression of the epidermal growth factor receptor (EGFR) can be used as a prognostic factor for the group of superficial tumors. Both abnormal E-cadherin and retinoblastoma (RB) expression have additional prognostic value for invasive tumors. The exact predictive value for the superficial tumors needs further study. The results with respect to p53 are conflicting and its exact role especially in the progression of pT1g3 tumors has to be clarified. In view of the discordance concerning its prognostic value, c-erbB-2 overexpression also needs further study. It appears that at this moment only a few molecular markers seem to have potential prognostic value, but their precise clinical relevance has to be studied more extensively. In particular the value of progression markers in the superficial TCC needs more attention. [ABSTRACT FROM AUTHOR]

This paper gives an overview of animal models available for the study of human renal cell carcinoma. Animal model systems can be divided into four categories: drug-induced tumors, virus-induced tumors, spontaneous tumors and human tumors transplanted into nude mice. Animal models have been used for improvement of diagnostic and therapeutic procedures. New markers for the diagnosis of more aggressive tumors can, for instance, be found by comparing metastatic with non-metastatic variants of the same primary tumor. Since advanced human renal cell carcinoma is refractory to chemotherapeutic agents, most therapeutic studies on animal models are immunotherapy studies. Combination treatment of biological-response modifiers such as interferon and tumor necrosis factor, with hyperthermia, bispecific monoclonal antibodies and high-energy shock waves, show promising results. Gene therapy will open new therapeutic possibilities. [ABSTRACT FROM AUTHOR]

An important molecular approach for the study of the complex processes implicated in prostatic carcinogenesis is the determination of steady-state mRNA levels of potentially relevant genes. One important category of these genes are the proto-oncogenes. Previous studies on the involvement of oncogenes in prostatic carcinogenesis, using primary human prostatic tumors, have shown that the ras and myc oncogenes might be related to the progression of this disease. We tested this hypothesis in the Dunning R-3327 rat prostatic cancer model system, which consists of a series of sublines that represent the different stages of tumor progression in prostatic cancer. We could not show a correlation between the expression of the ras and myc proto-oncogenes and the state of progression of the disease. The levels and patterns of expression of other proto-oncogenes, however, might be more useful as markers for malignancy and/or progression of prostatic cancer in rats. [ABSTRACT FROM AUTHOR]

The multitude of grading systems proposed for prostate adenocarcinoma illustrate the putative value of tumor grading as a predictor of prognosis. However, it also indicates the uncertainty and subjectivity of visual grading, as is confirmed by the results of several studies. Recent computer techniques enable fast image processing of microscoping images. Hence, a quantitative assessment of cellular and nuclear features can be obtained, eliminating subjectivity of visual grading. DNA-analysis and morphometric karyometry can be valuable for predicting prognosis in prostate tumors. For routine use, however, standardized preparatory techniques are mandatory. Up till now image analysis of microscopic images is a useful tool in addition to visual grading. Further standardization, inter- as well as intra-laboratory, is necessary to elaborate its value as a reproducible tool in tumor grading. [ABSTRACT FROM AUTHOR]

Immunocytochemistry with a rod-specific antiserum was used to study the post-hatch development (2 days-300 days) of photoreceptor elements within the pineal of the Japanese quail. At all ages staining was restricted to limited numbers of pinealocytes scattered throughout the gland. An enzyme-linked immunosorbent assay (ELISA), with the same rod-specific antibody, was then used to obtain a quantitative measure of rod opsin in total eye and pineal extracts in both the developing retina and pineal. The opsin content of both tissues shows a marked increase during the first 30 days after hatch and then plateaued to 0.84±0.02 nmoles opsin in the eye and 2.20±0.11 pmoles opsin equivalents in the pineal. The increase in opsin in the retina may be associated with continued post-hatch development of the photoreceptors. We then attempted to demonstrate the presence of the rhodopsin chromophore within pineal and retinal extracts using HPLC analysis. In both retinal and pineal extracts, 11-cis retinaldehyde was identified and a light-induced shift from the 11-cis to the all-trans isomer was clearly shown. This analysis also allowed us to calculate the total content of 11-cis and all-trans retinaldehyde (derived from both rod and non-rod photoreceptors) of the eye and pineal (eye: 1.7±0.2 nmoles; pineal: 4.6±0.5 pmoles). In the quail eye, the total amount of retinaldehyde is more than twice the amount of rod-like opsin. This probably reflects the large contribution of cones in the quail retina; the cone pigments will contribute to the retinaldehyde content but are not recognized by the rodspecific antibodies. In the pineal, we also found more than double the concentration of retinaldehyde than we would have predicted from the amount of rod-like opsin. These results, coupled with our immunocytochemical findings, suggest that the quail pineal contains at least two classes of photoreceptor, some 'rod-like', others 'non rod-like'. [ABSTRACT FROM AUTHOR]

A rod-specific antiserum was used to immunolabel elements within the retina and pineal of the adult Djungarian hamster and Welsh Mountain sheep. In the retina immunostaining was localized to the outer segments and perikarya of photoreceptor cells, while in the pineal limited numbers of labelled pinealocytes were scattered throughout the gland. An enzyme-linked immunosorbent assay (ELISA) was then used to obtain a quantitative measure of rod opsin in total eye and pineal extracts from the Djungarian hamster. Total rod opsin (±SEM) in the eye was measured by absorbance spectroscopy (1.88±0.10 nmoles opsin/eye) and by using the ELISA (1.75±0.02 nmoles opsin/eye). The opsin content from a total of 56 pineals gave a mean value of 0.34±0.01 pmoles opsin/pineal. Since a functional photopigment should be coupled in a 1∶1 ratio to a chromophore, we investigated whether we could identify 11 -cis and/or alltrans retinaldehydes in the pineal extracts by quantitative extraction and HPLC analysis as the oximes. No evidence of 11-cis or all-trans retinaloxime could be found, the chromatograms were indistinguishable from those produced by extracts of cortical brain tissue. We conclude that the opsin present within the adult hamster pineal is not coupled to the common vertebrate retinaldehyde chromophore, and as a result, is unlikely to be part of a functional photopigment. [ABSTRACT FROM AUTHOR]

Whereas cytokine therapy has proven efficacy in the treatment of metastatic renal cell carcinoma (RCC), many questions regarding the use of these drugs remain unanswered. In the present study we evaluated the antiproliferation effects of human recombinant alphainterferon (IFN), gamma-interferon and tumor necrosis factor-alpha (TNF) on eight human RCC xenografts. In particular, the importance of the administration route, dosage and tumor load was investigated. Response to the cytokines differed widely amongst the different tumors. Of three tested routes of administration (i.v., i.p. and s.c. peritumoral), only the s.c. peritumoral route was effective against tumor growth. After 6 weeks of therapy consisting of 150 or 1,500 units IFN/g given s.c. peritumorally three times a week or 30,000 units TNF/g given five times a week, alpha-IFN treatment resulted in 2%-100% growth inhibition; gamma-IFN, in 7%-80%; and TNF, in 35%-75% as compared with the untreated control. Growth of five of eight tumor lines could be inhibited completely by combinations of IFN and TNF, whereby the tumor dimensions at the beginning of therapy were decisive for the results. In some cases IFNs had optimal doses; however, the antitumor effects of TNF were always dose-dependent. Our studies indicate that the doses at which the optimal direct effects of cytokines are measured are critically dependent on the tumor treated. Although direct effects are only one part of the mode of action of cytokines, our results indicate that dosage of cytokines may need individualisation. [ABSTRACT FROM AUTHOR]

We have studied the effect of high-energy shock waves (HESW) alone or in combination with biological response modifiers (BRMs) or Adriamycin on the growth of the NU-1 human kidney cancer xenograft. When HESW are administered repeatedly (four sessions of 800 shock waves on days 0, 2, 4 and 6) a prolonged delay in tumor growth was found compared with that following a single administration. This effect was temporary, and several days after stopping the HESW administration the tumor regained its original growth potential (same doubling time). Tumor growth was suppressed for a longer period by the combination of 4 sessions of HESW and a single administration of Adriamycin, 5 mg/kg. Combination of HESW treatment with interferon alpha (5.0 ng/g body weight, three times/week) and tumor necrosis factor alpha (500 ng/g body weight, 5 days/week) s.c. around the tumor resulted in a complete cessation of tumor growth. While Adriamycin had an additive effect on HESW treatment, the combination with BRMs was highly synergistic. [ABSTRACT FROM AUTHOR]

High energy shock wave lithotripsy has proven to be an effective tool in the management of renal calculi. The effects of electrohydraulically generated high energy shock waves (HESW) on tumor cells were described only recently. Here we present data on the experimental design for treatment of tumor cells, using electromagnetically generated shock waves. The dertermination of the focal area, in which pressures are at least 50% of the maximum pressure, appeared to be essential. In vitro HESW treatment resulted in a dose dependant anti-proliferative effect on Dunning R-3327 PAT-2 rat prostate cancer subline, determined by temporal growth curve analysis after plating of treated cells in soft agar. Furthermore, it was shown that HESW treatment had a potentiating effect on Vinblastin treatment. The combination of HESW with Vinblastin appeared to have an additive in vitro antiproliferative effect on PAT-2 prostatic cancer cells. [ABSTRACT FROM AUTHOR]

The in vitro antitumor activity of recombinant alpha-and gamma-Interferon as well as recombinant Tumor Necrosis Factor alpha was tested on renal cell carcinoma xenografts using the double layer soft agar method. Using this assay, the effect of the drugs on the clonogenic potential of tumor cells in soft agar was determined and used as an indication for the antiproliferative capacity of these drugs. There appeared to be a differential response of the tested xenografts towards these drugs in a dose dependent way. However, when used in combination in most cases an additive or synergistic effect was observed. Even though again the response was differential, the combination of alpha-Interferon (10 ng/dish) and Tumor Necrosis Factor (100 ng/dish) resulted in a complete inhibition of colony formation in soft agar for both tumor lines. We conclude that there is a direct effect of alpha-Interferon, gamma-Interferon and Tumor Necrosis Factor on renal cell carcinoma xenografts. In combination drug tests the effect of alpha-Interferon and Tumor Necrosis Factor was strongly synergistic. The implication of these studies for in vivo use of these drugs remain to be established. [ABSTRACT FROM AUTHOR]

In a recent paper, we described the expression pattern of proto-oncogenes in primary human renal cell carcinoma [12]. To test the possibility of using xenografts as a useful alternative for such studies, we analyzed xenografts of a number of human renal cell carcinomas in nu/nu mice. Xenografts included RC2, RC14, RC21, RC43 and NC65. Northern blot analysis indicated that c- Ras was expressed in all these xenografts. The identify of the ras transcripts in the individual xenografts was further specified as c-Ha- ras, c-Ki- ras or N- ras. Expression of c- myc and the p53 gene was also found in a number of these tumors. Only RC21 failed to express the c- myc or the p53 gene. In all xenografts, a 3.0 kb c- fes/fps mRNA was present. In RC2, RC14, RC21 and RC43, low levels of the 4.8 kb ab 1 transcript were detectable. Transcripts of myb and sis could not be detected in any of the xenografts. The results indicated that the expression pattern of a variety of proto-oncogenes in xenografts of human renal cell carcinomas was similar to that in the primary tumors. [ABSTRACT FROM AUTHOR]

Renal cell tumors were screened for expression of the cellular oncogenes c- abl, c- fes, c- fms, c- myc, c- ras, and c- sis in dot blot hybridization analysis. Expression of c- ras and c- myc was clearly detectable in most of the 15 tumors that were studies. The c- fes oncogene appeared to be expressed in only two of them. Comparative Southern blot analysis of molecularly cloned human c- fes DNA and genomic DNA of the 15 renal tumors revealed no major genetic differences. Northern blot analysis of poly(A)-selected RNA from the fes-positive tumors with the complete viral v- fes oncogene of the Gardner-Arnstein strain of feline sarcoma virus as a molecular probe revealed hybridization of RNA species of 3.0 and 4.5 kb, respectively. The 3.0 kb c- fes transcript has also been reported in RNA from patients suffering from acute myelogenous leukemia [13]. The 4.5 kb transcript, however, has not been described before and represents either a c- fes-related splicing intermediate or, more likely, a completely processed transcript. The results of this study could imply that human c- fes coding sequences are more extensive than was previously assumed. [ABSTRACT FROM AUTHOR]

The retinal proteins opsin, α-transducin, S-antigen and interstitial retinol-binding protein (IRBP) are essential for the processes of vision. By use of immunocyto-chemistry we have employed antibodies directed against these 'photoreceptor proteins' in an attempt to identify the photoreceptor systems (retina, pineal and deep brain) of the Japanese quail. Opsin immunostaining was identified within many outer (basal portion) and inner segments of retinal photoreceptor cells and limited numbers of photoreceptor perikarya. Opsin immunostaining was also demonstrated in limited numbers of pinealocytes with all parts of these cells being immunoreactive. These results differ from previous observations. In contrast to the results obtained with the antibody against opsin, S-antigen and α-transducin immunostaining was seen throughout the entire outer segments and many photoreceptor perikarya of the retina. In the pineal organ immunostaining was seen in numerous pinealocytes in all follicles. These results conform to previous findings in birds. In addition, IRBP has been demonstrated for the first time in the avian retina and pineal organ. These findings underline the structural and functional similarities between the retina and pineal organ and provide additional support for a photoreceptive role of the avian pineal. No specific staining was detected in any other region of the brain in the Japanese quail; the hypothalamic photoreceptors of birds remain unidentified. [ABSTRACT FROM AUTHOR]

We present the first evidence that purified rhodopsin can induce experimental autoimmune uveoretinitis (EAU) in monkeys. Injection of a highly purified lipid-free rhodopsin preparation provokes severe chorioretinitis with concomitant anterior uveitis. The onset of disease is earlier, its frequency is higher, and the inflammation is considerably more severe than in EAU induced under similar conditions by opsin. The first inflammatory cells are observed in the ciliary body and pars plana. Within a few days the inflammation extends into the anterior chamber, choroid, and retina. Retinitis predominates in the central area, while chorioretinitis is observed in the periphery, both accompanied by damage to and elimination of the photoreceptor cells. The monkeys develop high cellular and humoral immune responses against rhodopsin and opsin. The cellular response maximum just precedes the onset of EAU. This may indicate that cellular immunity has an important role in the pathogenesis of rhodopsin-induced EAU. [ABSTRACT FROM PUBLISHER]