Cannabinoids in Urology. Which Benign Conditions Might They Be Appropriate to Treat: A Systematic Review.

There is growing evidence suggesting cannabinoids may provide suitable alternatives to conventional treatments in an increasing number of clinical settings. This review evaluates how cannabinoids are used to treat certain benign urological pathologies and to clarify the clinical value of this data. This review includes 62 papers and was undertaken per PRISMA's guidelines, it evidences the therapeutic potential of cannabinoids in the management of specific benign urological diseases, most notably neurogenic bladder dysfunction (clinical studies), renal disease (animal studies), and interstitial cystitis (animal studies). However, whilst cannabinoids are increasingly used, they cannot be considered reliable alternatives to more recognised treatments.

The impact of morphine treatment on bladder cancer cell proliferation and apoptosis: in vitro studies.

AIM: The aim of this study was to determine the effect of morphine on bladder cancer cell proliferation and apoptosis in vitro. MATERIALS AND METHODS: MTT assay was used to measure percentage growth of RT-112 human bladder cancer cells after 72 hours of morphine/morphine + naloxone treatment. Expression of µ-opioid receptors was assessed by Western blot and finally, apoptotic assay with CellEvent Caspase-3/7 Green Detection Reagent was carried out using confocal microscopy. RESULTS: The MTT assays showed that morphine increased RT-112 cell growth. Naloxone inhibited this growth enhancing effect. Western blot analysis regarding µ-opioid receptor expression in RT-112 cells remains inconclusive. Morphine was also found to decrease the rate of apoptosis of RT-112 cells, an effect which naloxone inhibited. CONCLUSIONS: This study provides evidence that morphine, at clinically relevant doses, causes RT-112 bladder cancer cell proliferation, possibly opioid receptor mediated and at least some of this effect might be due to decreased apoptosis. Clinically, this suggests that in patients with bladder cancer, managing pain with morphine might have detrimental consequences on patient outcomes and alternative pain relief should be considered if possible.

Health related quality of life (HRQoL) after cystectomy: comparison between orthotopic neobladder and ileal conduit diversion.

INTRODUCTION: Radical cystectomy and urinary diversion carries a high morbidity. Quality of life and body image are important considerations for urinary diversion (UD). We wanted to conduct a systematic review of literature to see which form UD offers a better quality of life (QoL). METHODS: We searched MEDLINE, Pubmed, EMBASE, CINAHL and the Cochrane library for studies using the following key words: 'quality of life' and 'ileal conduit', 'orthotopic neobladder', 'continent diversion' and 'urinary diversion'. All English language articles on UD surgery were included in the original search from 1990 to 2014. To improve the quality of evidence, we stratified our inclusion criteria into studies that report on QoL in both forms of UD using at least one validated questionnaire. RESULTS: Twenty-one studies (2285 patients) were included in our study all of which used at least one validated tool. The most frequently used tools were the SF-36, EORTC QLQ-C30 and FACT BL (10, 8, 5 studies respectively). None of the studies were randomised and only 4 studies were prospectively designed. Sixteen studies reported no difference in QoL between the two types of urinary diversion and four studies reported a better QoL with orthotopic neobladder of which 2 studies had younger and fitter patients. On the other hand, one study reported a better QoL in ileal conduit patients. CONCLUSION: Orthotopic neobladder urinary diversion shows a marginally better quality of life scores compared to ileal conduit diversion especially when considering younger and fitter patients.

Presentation patterns of invasive cancer of the cervix: results from Parirenyatwa Oncology and Radiotherapy Centre, Harare, Zimbabwe 1998-2010.

MAIN OBJECTIVE: The study sought to identify the presentation patterns of invasive uterine cancer of the cervix (CaCx) in Zimbabwe in terms of histology, stage of the disease, ages of patients and socio-economic status. DESIGN: Retrospective study from 1998 to 2010. SUBJECTS: All patients who registered for the first time with invasive CaCx over a systematically selected sample period of four years (1998, 2002, 2006 & 2010). SETTING: The main referral Radiotherapy and Oncology centre in Harare the capital city of Zimbabwe. RESULTS: Majority of patients (91.75%) presented with squamous cell carcinoma, 5.5% presented with adenocarcinomas and 2.75% presented with other types of histology. Late presentation was noted with the majority of the patients (89%) presenting with stage IIB and above. The common ages of patients at presentation were between 40 to 60 years. The majority of the patients (59.5%) were of low socio-economic status. CONCLUSION: In the developed countries CaCx is reducing in frequency, presentation tends to be early, treatment effective and there is decreasing mortality rate from this disease. However in developing countries the situation is not as positive and the disease remains a major concern. This is shown by the presentation pattern of patients with invasive CaCx in Zimbabwe. The patients are shown to present with late stage disease of the squamous cell type, primarily in the age ranges of 40 to 60 years and with the majority of the patients belonging to the low socio-economic status group.

Design of a continuous passive and active motion device for hand rehabilitation.

This paper presents the design of a novel, portable device for hand rehabilitation. The device provides for CPM (continuous passive motion) and CAM (continuous active motion) hand rehabilitation for patients recovering from damage such as flexor tendon repair and strokes. The device is capable of flexing/extending the MCP (metacarpophalangeal) and PIP (proximal interphalangeal) joints through a range of motion of 0 degrees to 90 degrees for both the joints independently. In this way, typical hand rehabilitation motions such as intrinsic plus, intrinsic minus, and a fist can be achieved without the need of any splints or attachments. The CPM mode is broken into two subgroups. The first mode is the use of preset waypoints for the device to cycle through. The second mode involves motion from a starting position to a final position, but senses the torque from the user during the cycle. Therefore the user can control the ROM by resisting when they are at the end of the desired motion. During the CPM modes the device utilizes a minimum jerk trajectory model under PD control, moving smoothly and accurately between preselected positions. CAM is the final mode where the device will actively resist the movement of the user. The user moves from a start to end position while the device produces a torque to resist the motion. This active resistance motion is a unique ability designed to mimic the benefits of a human therapist. Another unique feature of the device is its ability to independently act on both the MCP and PIP joints. The feedback sensing built into the device makes it capable of offering a wide and flexible range of rehabilitation programs for the hand.

Seminal tissue factor revisited.

Studies of seminal tissue factor (TF) are few and mostly based on small numbers. Due to the reported lack of factor (F) X in semen, it has been suggested that TF may not have a role in seminal coagulum formation. However, recent identification of a number of haemostatic factors in semen justifies a re-evaluation of its occurrence. Semen specimens were collected from sub-fertile (n = 19), normally fertile (n = 33), semen donors (n = 30) and vasectomized subjects (n = 62), some fractionated into sperm, a prostasome-rich fraction and seminal plasma. Functional and antigenic TF levels were measured and related to conventional fertility parameters. Semen contains high concentration of functional and antigenic TF. Most TF was found in seminal plasma prepared by low-speed centrifugation. When further fractionated by ultracentrifugation much of this may reside in the pellet (prostasomal fraction). It was also detectable on sperm. TF antigen levels were higher in vasectomized subjects than sub-fertile, normally fertile, donor (p = 0.02) and a 'pooled normal semen parameters' (PNSP) stratification (derived from a combination of measurements) (p = 0.06). The sub-fertile group showed a wider variation than normal, donor or the PNSP subjects. Seminal TF antigen levels correlated significantly with sperm agglutination (p = 0.03) and abnormal sperm morphology (p = 0.04). Subjects with anti-sperm antibodies also showed high TF antigen levels. In conclusion, semen contains functional and antigenic TF at high concentrations. A full complement of clotting factors probably exists in semen, so some pro-coagulant role for TF should not be excluded. Decreased seminal TF levels appear to be associated with seminal parameters that are known to favour male fertility.

Progesterone: a novel adjunct to intravesical chemotherapy.

OBJECTIVE: To investigate the effect of progesterone on multidrug-resistant urothelial cell lines, as the failure of intravesical chemotherapeutic drugs is often caused by multidrug resistance (MDR), mediated by the drug efflux pump P-glycoprotein (PGP), the function of which can be down-regulated by various compounds including steroid hormones. MATERIALS AND METHODS: Two urothelial cell lines (RT112S and MGH-U1S) and their MDR sublines (RT112R, to cisplatin; and MGH-U1R, a cell line expressing PGP) were used to assess the cytotoxic effects of progesterone, epirubicin and their combination. Cytotoxicity was assessed using a tetrazolium-based assay and in situ confocal microscopy. RESULTS: Cell lines sensitive to epirubicin (MGH-U1S, RT112S and RT112R) required a much lower dose of epirubicin to kill half the cells than did the MDR cell line. Progesterone was intrinsically cytotoxic to all cell lines with little difference among them. Combined therapy had no cumulative effect on epirubicin-sensitive cell lines, but reversed MDR in the MGHU1R cell line, both assessed by confocal microscopy and by the tetrazolium assay. CONCLUSIONS: Progesterone can reverse MDR in urothelial cells in vitro. This, combined with its effects on cell differentiation and apoptosis, together with its safety and tolerability compared to other MDR agents, suggests it may be a valuable adjunct to intravesical chemotherapy.

Intravesical meglumine gamma-linolenic acid in superficial bladder cancer: an efficacy study.

OBJECTIVES: Gamma-linolenic acid (GLA) is known to be cytotoxic to malignant cells. We assessed the efficacy of the novel intravesical formulation, meglumine gamma-linolenic acid (MeGLA), in a phase II trial, in patients with recurrent, superficial bladder cancer. PATIENTS AND METHODS: Thirty patients with recurrent, superficial transitional cell carcinoma (TCC) were recruited. The tumour pattern was recorded at flexible cystoscopy. Patients received a single intravesical instillation of 50ml of either 50mg (1mg/ml) (15 patients), or 125mg (2.5mg/ml) (15 patients) of MeGLA in water, retained for one hour. At subsequent cystoscopy, the tumour patterns were recorded, prior to undertaking routine cystodiathermy. Biopsies were obtained for histological assessment. Responses were divided into complete, partial or none. RESULTS: All 30 patients retained the drug for 1 hour without significant local or systemic side effects. There were 4 (13%) complete responses, 9 (30%) partial responses, and 17 (57%) non-responders. Histology showed no evidence of damage to surrounding urothelium. CONCLUSIONS: Our data confirms the safety and tolerability of MeGLA, which is consistent with findings from a previous phase I trial. A response rate of 43% also indicates that MeGLA has a significant cytotoxic effect against TCC and the results are similar to those obtained using standard, single-dose, intravesical regimens.

The effects of meglumine gamma linolenic acid (MeGLA) on an organ culture model of superficial bladder cancer.

The objective of this research was to assess the effects of the novel intravesical drug MeGLA in a physiologically representative model of superficial bladder cancer. Petri dishes were used to culture 5 mm square explants of rat bladder in minimal volumes of supplemented culture medium. Parental and resistant MGH-U1 urothelial cancer cells were transfected with a green fluorescent protein (GFP) vector. Transfectants were purified by flow cytometry. Cells were seeded onto the prepared organ cultures and imaging was performed using confocal microscopy. Confirmation of the tumour colonies was done using scanning electron microscopy. MeGLA was added at various concentrations to the colonies and its effects noted over several days. Results showed that colonies of GFP-MGH-U1 cells established themselves on the explants and could be identified by confocal microscopy. The colonies could then be followed over several days. The colonies were able to survive high concentrations of the drug of up to 1 mg/ml, 400 times the IC > 90% for monolayers and equivalent to doses in clinical use. We conclude that MeGLA is less effective in this system than on monolayer cell lines. However, it showed cytotoxic effects which were comparable to those seen with conventional agents in the same system.

Cellular resistance to mitomycin C is associated with overexpression of MDR-1 in a urothelial cancer cell line (MGH-U1).

OBJECTIVE: To compare multidrug resistance (MDR)-1 and MDR-3 gene expression in a new urothelial cancer cell line (MGHU-1, with resistance to mitomycin C) against controls and the established (epirubicin-resistant) MDR clone, and to correlate MDR with cytotoxicity data. MATERIALS AND METHODS: Resistance to mitomycin C was induced by the long-term exposure of wild-type MGHU-1 cells to increasing concentrations (20-400 nmol/L) of mitomycin C. The cytotoxicity of mitomycin C or epirubicin was then compared in MGHU-1, MGHU-MMC (mitomycin C-resistant) and MGHU-1R (established MDR) cells, using the tetrazolium biomass assay. The expression of MDR-1 and -3 was investigated by the reverse transcriptase-polymerase chain reaction, using cDNA-specific primers after titration, and compared with DNA and negative controls. RESULTS: MDR-1 and -3 were significantly and equally overexpressed in MGHU-1R, and associated with a dramatic increase in the 50% inhibitory drug concentration (P < 0.001) for mitomycin C and epirubicin against controls. In MGHU-MMC, the overexpression of MDR-1 was three times greater than that of MDR-3. The cytotoxicity profile for both agents was very similar to that of MGHU-1R. Trace amounts of MDR-1, but not MDR-3, were identified in the MGHU-1 wild-type. CONCLUSIONS: Urothelial cancer cell resistance to mitomycin C is associated with cross-resistance to epirubicin and overexpression of MDR-1, suggesting that mitomycin C falls within the MDR category. Clinical application of this methodology may allow patients to be identified who are unlikely to benefit from intravesical chemotherapy.

A model of superficial bladder cancer using fluorescent tumour cells in an organ-culture system.

OBJECTIVE: To develop a reproducible in vitro simulation of superficial bladder cancer for testing cytotoxic agents at clinically relevant concentrations. MATERIALS AND METHODS: Square explants (5 mm) of rat bladder were cultured in Petri dishes in minimal volumes of Waymouth's MB 752/1 medium supplemented with 10% fetal calf serum, antibiotics and glutamine. Parental and resistant MGH-U1 urothelial cancer cells were transfected with a green fluorescent protein (GFP) vector. Transfectants were purified by flow cytometry. Cells were seeded onto the prepared organ cultures and images obtained using confocal microscopy. The tumour colonies were confirmed using scanning electron microscopy. Conventional intravesical cytotoxic agents including epirubicin, mitomycin-C and estramustine were tested in the system. RESULTS: Colonies of GFP-MGH-U1 cells became established on the explants and were identified by confocal microscopy; the development of the colonies was then followed over several days. Staining the explant for viability allowed imaging of normal urothelium on the explant surface or surrounding skirt of urothelial cells. The conventional cytotoxic agents epirubicin, mitomycin C and estramustine showed the expected differential responses to parental and resistant cell types. The colonies were able to survive high concentrations of the drug, equivalent to those in clinical use. The colonies were imaged serially over a period of several days. CONCLUSION: This system provides a more realistic model for testing cytotoxic agents for use in intravesical therapy, by allowing clinically relevant concentrations of drugs to be tested. The differential properties of the parental and resistant cells are maintained. The model also enables the same tumour colony to be imaged over several days in culture. The model may also be adapted for use in testing the effects of drugs on normal urothelium and the study of the effects of growth factors.

Nonhomologous bioinjectable materials in urology: 'size matters'?

OBJECTIVE: To assess the factors that influence how particles might become fixed in tissues or migrate from them, by measuring the size of the injectable particles, their susceptibility to phagocytosis and their affinity for fibroblast attachment in culture. MATERIALS AND METHODS: The particle size of three types of particulate unphysiological bioinjectable material, i.e. Urocol (Genesis Medical, Ltd., London), Macroplastiquetrade mark (Uroplasty Ltd., Reading, UK) and Urethrin (Mentor Medical Systems, Wantage, UK) was analysed using phase-contrast light microscopy and confocal microscopy. Human monocytes from peripheral blood were incubated with the three materials in phagocytic studies, where ingestion was determined by confocal microscopy. A fibroblast cell line was used to ascertain the ability of the particles to act as a substrate for cell attachment in culture. RESULTS: The mean (SEM) maximum particle diameters of Macroplastique, Urethrin and Urocol were 209 (5.10) microm, 49 (1.52) microm and 14 (0.39) microm, respectively. Rat peritoneal macrophages and human peripheral blood monocytes commonly ingested Urocol particles; the phagocytosis of Urethrin was rare and that of Macroplastique was not detected. Fibroblasts adhered to Urocol paste and Urethrin particles, but not to Macroplastique. CONCLUSION: Published reports of particle size and phagocytosis are confusing, but a relationship clearly exists. Macroplastique is the largest particle and is least likely to be phagocytosed by human mononuclear phagocytes. Urocol paste is the slowest to dissipate in culture conditions; the flat surfaces of Urethrin, but not Macroplastique, can serve as a substrate for fibroblast anchorage.

Intracerebral clysis in a rat glioma model.

OBJECTIVE: Intracerebral clysis (ICC) is a new term we use to describe convection-enhanced microinfusion into the brain. This study establishes baseline parameters for preclinical, in vivo, drug investigations using ICC in a rat glioma model. METHODS: Intracranial pressure was measured, with an intraparenchymal fiber-optic catheter, in male Fischer rats 10, 15, 20, and 25 days after implantation of C6 glioma cells in the right frontal lobe (n = 80) and in control rats without tumor (n = 20), before and during ICC. A 25% albumin solution (100 microl) was infused through an intratumoral catheter at 0.5, 1.0, 2.0, 3.0, and 4.0 microl/min. Infusate distribution was assessed by infusion of fluorescein isothiocyanate-dextran (Mr 20,000), using the aforementioned parameters (n = 36). Brains were sectioned and photographed under ultraviolet light, and distribution was calculated by computer analysis (NIH Image for Macintosh). Safe effective drug distribution was demonstrated by measuring tumor sizes and apoptosis in animals treated with N,N'-bis(2-chloroethyl)-N-nitrosourea via ICC, compared with untreated controls. Magnetic resonance imaging noninvasively confirmed tumor growth before treatment. RESULTS: Intracranial pressure increased with tumor progression, from 5.5 mm Hg at baseline to 12.95 mm Hg on Day 25 after tumor cell implantation. Intracranial pressure during ICC ranged from 5 to 21 mm Hg and was correlated with increasing infusion volumes and increasing rates of infusion. No toxicity was observed, except at the higher ends of the tumor size and volume ranges. Fluorescein isothiocyanate-dextran distribution was greater with larger infusion volumes (30 microl versus 10 microl, n = 8, P < 0.05). No significant differences in distribution were observed when different infusion rates were compared while the volume was kept constant. At tolerated flow rates, the volumes of distribution were sufficient to promote adequate drug delivery to tumors. N,N'-Bis(2-chloroethyl)-N-nitrosourea treatment resulted in significant decreases in tumor size, compared with untreated controls. CONCLUSION: The C6 glioma model can be easily modified to study aspects of interstitial delivery via ICC and the application of ICC to the screening of potential antitumor agents for safety and efficacy.

Estramustine reversal of resistance to intravesical epirubicin chemotherapy.

OBJECTIVE: Failure of epirubicin treatment of superficial bladder cancer implies multidrug resistance (MDR) which is common. MDR is characterised by decreased cellular levels of drug. TCC cell lines sensitive to epirubicin and resistant to both epirubicin and mitomycin C were used to investigate augmented therapy by adding the MDR reversing agent estramustine to an in vitro model. METHODS: Cells were cultured as monolayers. Fluorescence analysis was performed by flow cytometry and confocal microscopy. Cells were exposed to epirubicin 20 microg/ml for 2 h and increasing amounts of estramustine. Cytotoxicity was determined under similar exposure conditions and MTT culture (dye reduction by live cells) allowed viable biomass to be read optically. RESULTS: Resistant cells accumulated eight times less epirubicin than sensitive cells. Confocal microscopy confirmed this for nuclear uptake. Accumulation in resistant cells can be increased to near-sensitive cell levels using 40 microg/ml estramustine. Image analysis of confocal fluorescence showed a shift from cytoplasm to nucleus. This correlated with increased cytotoxicity. CONCLUSION: Estramustine plus epirubicin chemotherapy can overcome MDR and may achieve more successful tumour killing in vivo. It may also prevent emergence of resistance. Primary TCC culture examination permits detection of sensitive and resistant cells and may predict outcome allowing a more logical treatment selection.

A prospective trial of flexible cystodiathermy for recurrent transitional cell carcinoma of the bladder.

PURPOSE: The flexible cystoscope has a proved role in the followup of patients with transitional cell carcinoma of the bladder but the full extent of its therapeutic role has yet to be defined. We analyzed 171 flexible cystodiathermies to assess patient tolerance and treatment success. Potential cost savings were also analyzed. MATERIALS AND METHODS: All patients with single or multiple small papillary (Ta) recurrences at followup flexible cystoscopy were treated with flexible cystodiathermy. Plain lubricating gel was used and no other analgesia was prescribed. A visual analog pain scale was completed by the patient after the procedure and an observer rating of discomfort was recorded. Followup for efficacy of treatment was performed. RESULTS: A total of 103 patients were treated with cystodiathermy during the last 3 years. Median followup was 21 months (range 12 to 42). Of the patients 52 (50.5%) had no recurrence of transitional cell carcinoma after treatment and 51 (49.5%) required treatment for recurrence. Only 13 patients (12.6%) had recurrences at or close to the original tumor site. Pain scales indicated that the procedure was well tolerated and all patients agreed to undergo it in the future if required. Estimated cost savings were approximately $66,500 per 100 patients. CONCLUSIONS: Flexible cystodiathermy is a well tolerated and efficacious treatment for recurrent small papillary (Ta) transitional cell carcinoma of the bladder. Since transitional cell carcinoma of the bladder frequently occurs in an elderly and often unfit population treatment that avoids general anesthetic offers considerable advantages.

Water as a tumoricidal agent in bladder cancer. In vitro studies in parental and resistant cell lines.

OBJECTIVE: Water is widely used as a tumoricidal agent during transurethral resection of bladder tumour (TURBT). Despite this, recurrences are common. One possible explanation may be that while the parental bladder cancer cells are sensitive to water, their multidrug resistant (MDR) clones which occur commonly, may be resistant to it. If so, is this a preserve of the MDR phenotype or do other mechanisms confer resistance to water? METHODS: The parental human urothelial cancer cell lines (MGHU-1 and RT112) and their drug-resistant variants (MDR and non-MDR) were exposed to short pulses of water. The MDR status of these cells was verified using confocal microscopy to detect intracellular accumulation of fluorescent drug (epirubicin). A clonogenic assay was used to establish tumoricidal efficacy on cells in suspension. A thiozolyl blue (MTT) assay was used on adherent cells to assess the residual viable biomass. RESULTS: In the MDR clone of cells, colony counts were reduced by 20% after treatment with water as compared to a 60% reduction in the parental cell line. There was a similar reduction in colony counts between the parental RT112 and its non-MDR cisplatin resistant variant. CONCLUSION: Water is less effective on MDR cells. Resistance to water may be a function of the MDR phenotype.

Intravesical chemotherapy with gamma linolenic acid becomes a realistic prospect in serum-free applications: in vitro cytotoxicity and systemic absorption studies.

PURPOSE: To assess the cytotoxicity of Meglumine gamma linolenic acid (MeGLA) in serum-free application on 2 urothelial cancer cell lines, to examine whether the instant kill action of MeGLA is retained in a serum free environment, and to study the pharmacokinetics of intravesical instillation of gamma linolenic acid (GLA). MATERIALS AND METHODS: The 2 human urothelial cancer cell lines (MGH-U1 & RT112) were utilized in classical cytotoxicity assays in which drug exposure lasted 2 hours in serum or in serum-free application. The thiozolyl blue (MTT) assay was used to quantify the residual viable biomass 5 days later. Immediate cytotoxicity was also compared in serum and serum-free application. Four Wistar rats were used to study the intravesical absorption profile of tritiated GLA (3H-GLA). RESULTS: There was a 10-fold enhancement of the lytic efficacy of MeGLA in serum-free application and this enhancement was also observed in experiments assessing instant kill. There was a similar enhancement of efficacy seen in the multi-drug resistant (MDR) clone of cells. The absorption profile showed < 2% of instilled counts were absorbed and the commonest destination for the absorbed GLA was the liver. CONCLUSIONS: The cytotoxic action of MeGLA was enhanced in serum free application. This enhancement was maintained when cells expressed the MDR phenotype. There was limited absorption from the bladder. MeGLA is a feasible intravesical agent for use in superficial bladder cancer.

Bladder cancer recurrence by implantation of exfoliated cells: is gamma-linolenic acid an effective tumoricidal agent?

OBJECTIVE: To compare the tumoricidal efficacy of meglumine gamma-linolenic acid (MeGLA), mitomycin C, epirubicin and water on two urothelial cell lines, and to establish the effect of serum protein levels derived from bladder cancer resection craters on the action of these agents. MATERIALS AND METHODS: The human urothelial cell lines MGHU-1 and RT112 and their drug-resistant variants were exposed to short pulses of aqueous MeGLA, mitomycin, epirubicin and water. Both adherent and suspended cells were exposed to these agents. The MTT viable biomass assay and a clonogenic assay were used to establish tumoricidal efficacy. These experiments were then repeated to assess the effect of added serum proteins on the test results. Estimates of protein in the waste irrigation fluid from 10 patients undergoing transurethral resection of bladder tumour (TURBT) were used to select the quantity of protein used in the study, to establish the clinical relevance. RESULTS: MeGLA caused > 95% reduction in the residual viable biomass of adherent cells, compared with < 50% reduction with any other agent. Both epirubicin and mitomycin were as effective as MeGLA in preventing colony formation from suspended drug-sensitive (parental) cells. However, using multidrug-resistant (MDR) cell lines, only MeGLA prevented any colony formation, although counts were greatly reduced by mitomycin and epirubicin. Water was least effective as a tumoricidal agent on both adherent and suspended cells. On the latter, water was markedly inactivated by adding 5% serum. TURBT waste irrigation fluid was found frequently to contain such quantities of serous fluid contamination, as shown by albumin estimates in waste fluid from 10 consecutive patients undergoing this procedure. CONCLUSION: MeGLA is an effective tumoricidal agent against both parental and MDR cell lines. Its efficacy is maintained in the presence of clinically relevant serum contamination.

Is gamma-linolenic acid an effective intravesical agent for superficial bladder cancer? In vitro cytotoxicity and in vivo tolerance studies.

The essential fatty acid gamma-linolenic acid (GLA) is an effective cytotoxic agent when applied topically and for prolonged periods to tumour cells. Topical application, by intravesical therapy, is firmly established in the treatment of superficial bladder cancer. However, this form of therapy is limited to a maximum duration of 2 h. At such a short drug exposure time, does GLA retain its cytotoxicity? We have examined this question by exposing the superficial bladder cancer cell lines MGH-U1 and RT112 to meglumine-GLA (MeGLA) for time intervals ranging from 30 min to 2 h, at drug concentrations ranging from 1000 to 1.95 microg/ml. The MTT viable biomass assay was used to assess cell kill. Greater than 90% inhibition was observed at a concentration of 125 microg/ml (IC > 90), at 2 h drug exposure. At shorter drug exposure times, higher drug concentrations were needed to induce the same effect. At 1 h drug exposure, the IC > 90 was recorded at 500 microg/ml. In vivo intravesical tolerance studies were conducted in rats. Rats exposed to 2.5 mg/ml MeGLA intravesically for 2 h or less remained well and bladder histology showed minimal changes. This study confirms that GLA retains its cytotoxicity at short drug exposure times and is well tolerated by normal bladder mucosa in vivo. Bladder mucosa tolerated > 10x the concentration required for the IC > 90 in vitro. MeGLA is therefore a feasible intravesical agent for superficial bladder cancer.

Surgical approaches to the thoracolumbar spine.

Surgical treatment of diseases involving the thoracolumbar spine offers a unique challenge to the spine surgeon. Although more familiar posterior techniques are useful for a variety of thoracolumbar problems, the management of disease in this region is often optimized by an anterior approach. Thoracolumbar surgery requires an understanding of the relevant surgical anatomy, the pathologic processes affecting this region, and the relative indications and contraindications for particular operative strategies. It is also essential to be familiar with the management of potential morbidity, methods for avoiding complications, and the selection of particular fusion techniques and instrumentation devices associated with specific operative strategies.