Intermediate-term survival of robot-assisted versus open radical cystectomy for muscle-invasive and high-risk non-muscle invasive bladder cancer in The Netherlands.

BACKGROUND: Radical cystectomy with pelvic lymph node dissection is the recommended treatment in non-metastatic muscle-invasive bladder cancer (MIBC). In randomised trials, robot-assisted radical cystectomy (RARC) showed non-inferior short-term oncological outcomes compared with open radical cystectomy (ORC). Data on intermediate and long-term oncological outcomes of RARC are limited. OBJECTIVE: To assess the intermediate-term overall survival (OS) and recurrence-free survival (RFS) of patients with MIBC and high-risk non-MIBC (NMIBC) who underwent ORC versus RARC in clinical practice. METHODS AND MATERIALS: A nationwide retrospective study in 19 Dutch hospitals including patients with MIBC and high-risk NMIBC treated by ORC (n = 1086) or RARC (n = 386) between January 1, 2012 and December 31, 2015. Primary and secondary outcome measures were median OS and RFS, respectively. Survival outcomes were estimated using Kaplan-Meier curves. A multivariable Cox regression model was developed to adjust for possible confounders and to assess prognostic factors for survival including clinical variables, clinical and pathological disease stage, neoadjuvant therapy and surgical margin status. RESULTS: The median follow-up was 5.1 years (95% confidence interval ([95%CI] 5.0-5.2). The median OS after ORC was 5.0 years (95%CI 4.3-5.6) versus 5.8 years after RARC (95%CI 5.1-6.5). The median RFS was 3.8 years (95%CI 3.1-4.5) after ORC versus 5.0 years after RARC (95%CI 3.9-6.0). After multivariable adjustment, the hazard ratio for OS was 1.00 (95%CI 0.84-1.20) and for RFS 1.08 (95%CI 0.91-1.27) of ORC versus RARC. Patients who underwent ORC were older, had higher preoperative serum creatinine levels and more advanced clinical and pathological disease stage. CONCLUSION: ORC and RARC resulted in similar intermediate-term OS and RFS in a cohort of almost 1500 MIBC and high-risk NMIBC.

Evaluation of chemotherapy with magnetic resonance imaging in patients with regionally metastatic or unresectable bladder cancer.

OBJECTIVES: To determine whether the failure of chemotherapy in patients with regionally metastatic or unresectable transitional cell carcinoma (TCC) of the bladder can be predicted early in the course of chemotherapy with magnetic resonance (MR) imaging. METHODS: In this prospective study, 36 patients with regionally metastatic or unresectable TCC of the urinary bladder underwent MR imaging before and after two, four, and six cycles of chemotherapy with Methotrexate, Vinblastine, Adriamycin (doxorubicin) and Cisplatin (MVAC). The response after two cycles of MVAC was evaluated by using conventional tumour size parameters with unenhanced MR imaging and with changes in the time to the start of tumour or lymph node enhanced at fast dynamic contrast-enhanced MR imaging. The results obtained with these techniques were compared with the findings at histopathology in cystectomy or transurethral resection specimens that were obtained after chemotherapy. Duration of survival was defined as the time from the start of chemotherapy until disease-specific death. Kaplan-Meier curves were drawn to determine the difference in prognosis between responders and nonresponders. RESULTS: After two cycles of chemotherapy, the accuracy, sensitivity, and specificity in distinguishing responders from nonresponders with conventional MR imaging were 69%, 81%, and 50%, respectively. With the fast dynamic contrast-enhanced technique, accuracy, sensitivity, and specificity were 92%, 91%, and 93% respectively. The median bladder cancer specific survival was 28 months for all patients studied. Responders to chemotherapy at fast dynamic contrast-enhanced MR had better median disease-specific survival than nonresponders (42 months vs. 12 months [p<0.0001]). CONCLUSION: We can predict whether a patient will respond to chemotherapy after two cycles of chemotherapy with fast dynamic contrast-enhanced MR imaging.

Comparison of orthotopic sigmoid and ileal neobladders: continence and urodynamic parameters.

OBJECTIVES: The objective this of the study was to compare continence rates and urodynamic parameters among patients who had undergone orthotopic bladder substitution with sigmoid or ileal segments. METHODS: Continent urinary reservoirs were constructed in 112 patients. Fifty patients received a sigmoid neobladder (SN) and 62 patients an ileal neobladder (IN). Thirty-four patients with an SN (mean age 64.4 years), and 20 with an IN (mean age 57.8 years) agreed to postoperative urodynamic evaluation at a median time after surgery of 18 and 37 months, respectively. Continence and urodynamic parameters were compared in both groups. RESULTS: The average reservoir capacity of the SN (296 ml) was lower than the IN (546 ml). The majority of patients voided by the Valsalva maneuver and achieved good peak flow rates [SN group 16.6 (range 7-32) ml/s, IN group 25.5 (range 5-35) ml/s]. Of the patients with an SN 26 (76%) and with an IN 15 (75%) emptied to near completion with a post-void residual (PVR) of less than 100 ml. Daytime continence was achieved in 90% of IN patients and 85% of SN patients. Only 9% of patients with an SN and 60% of patients with an IN were continent at night. CONCLUSION: A neobladder constructed from detubularized ileum or sigmoid achieves adequate capacity with a satisfactory daytime continence rate. Nighttime incontinence in patients with IN can be at least partly explained by periods of high pressure due to neobladder contractions in combination with a relaxed sphincter during sleep. The low nighttime continence rate of the SN is probably related to its small functional capacity.

The effect of N-acetyl-L-cysteine on the viscosity of ileal neobladder mucus.

N-acetyl-L-cysteine (NAC) proved to be an effective mucolytic in pulmonary secretions. Our goal was to investigate the in vitro effect of NAC on viscosity of ileal neobladder mucus. The urine of a patient with an ileal neobladder was collected during the first 7 days postoperatively and stored in a refrigerator. After precipitation, the urine was decanted. The residue was stirred to a homogeneous suspension. To samples of 4.5 ml mucus, 0.5 ml NAC 10% was added. To the control sample, 0.5 ml water was added. The samples were incubated in a water bath at 37 degrees C for 5, 30 and 60 min. Viscosity was measured in the Bohlin VOR Rheometer. The viscosity of the ileal neobladder mucus decreased quickly after incubating with NAC 10%. Viscosity increased slightly after I h of incubation. The viscosity in the control sample was higher than in the other incubated samples. NAC was found to decrease the viscosity of ileal neobladder mucus, supporting the in vivo experience that NAC can be useful in patients with an ileal neobladder to facilitate the evacuation of mucus by decreasing viscosity.

Return to fertility after extended chemical castration with a GnRH antagonist.

BACKGROUND: Antagonistic analogues of GnRH for the treatment of prostate cancer may be used clinically in persons for whom return to fertility after such treatment is important or desirable. The purpose of this study was, therefore, to evaluate the effects of a long term treatment with orntide, a GnRH antagonist, on testosterone levels and fertility in male rats. METHODS: Two groups of male rats received either 120-day orntide microspheres (8.8 mg orntide/kg/120 days) or vehicle alone (control group). Serum orntide and testosterone levels in both groups were monitored at certain intervals for 9 months from the initiation of treatment. After recovery of normal serum testosterone levels in the treated animals, each rat was housed with two proven breeder, but drug-naive, females. RESULTS: All mates of treated rats achieved pregnancy as rapidly as the mates of control rats although two of the control rats did not sire a litter with either female and one sired only one litter. The mean size of the litters of treated (12.3 offspring per litter) and control (10.6 offspring per litter) were similar. All offspring were grossly normal morphologically and behaviorally during the time to weaning. CONCLUSIONS: These results suggest that lack of fertility due to testosterone suppression is reversible after cessation of treatment with this GnRH antagonist.

Effect of the concurrent LHRH antagonist administration with a LHRH superagonist in rats.

PURPOSE: The purpose of this study was to investigate the effect of a novel LHRH antagonist, Orntide acetate, on the initial testosterone elevation in rats during treatment with a LHRH superagonist, Leuprolide acetate. METHODS: Thirteen groups of a rat animal model were administered either liquid Orntide or Orntide PLGA microspheres before or simultaneously with Leuprolide injections. Serum levels of testosterone were monitored during the time course of the study using a radioimmunoassay method. RESULTS: Administration of a single daily dose of liquid Orntide resulted in testosterone suppression within 6 h to levels below 0.5 ng/ml (castration level). However, combined administration of liquid Orntide and liquid Leuprolide did not have a significant effect on the initial testosterone elevation in studied rats. Similarly, there was no effect when liquid Orntide was co-administered with Leuprolide microspheres. Administration of Orntide microspheres 48 h before Leuprolide microspheres suppressed testosterone levels below the castration level within 24 h, however, did not prevent a rise in testosterone serum concentration upon administration of Leuprolide microspheres. Also, a second testosterone peak was observed between days 3 and 15 in the animals which were simultaneously treated with Orntide microspheres and Leuprolide microspheres. CONCLUSIONS: Orntide acetate was found to be an effective LHRH antagonist with a rapid onset of pharmacological action and a short biological half-life. Administration of a single dose of liquid Orntide or Orntide microspheres, resulted in rapid testosterone suppression without an initial elevation, as seen with LHRH superagonists. However, combined administration of Orntide and Leuprolide did not have an effect on the initial testosterone elevation in rats.

Transurethral marsupialization of a medial prostatic cyst in patients with prostatitis-like symptoms.

PURPOSE: The results of transurethral marsupialization as treatment for medial prostatic cysts were assessed. MATERIALS AND METHODS: Between June 1992 and August 1994 we performed transrectal ultrasound on 704 patients with symptoms of bladder outlet obstruction or lower urinary tract symptoms and a medical prostatic cyst was found in 34 (5%). Transurethral marsupialization of the cyst via incision of the prostatic floor under transrectal ultrasound guidance was performed in 18 patients. Followup was 12 to 25 months (mean 18). RESULTS: Patients with a medial prostatic cyst complained of prostatitis-like symptoms (77%), scrotal pain (62%), impaired micturition (32%), small volume ejaculation (35%), painful ejaculation (24%), hemospermia (24%) and infertility (12%). After marsupialization of the cyst, symptoms resolved completely in 14 patients (78%), improved in 17 (94%) and did not improve in only 1 (6%). No complications of this procedure were noted. The 16 patients who did not undergo surgery still complain of prostatitis-like symptoms without evidence of bacterial prostatitis. CONCLUSIONS: We believe that a medial prostatic cyst can cause prostatitis-like symptoms and that marsupialization of the cyst can provide symptom relief in the majority of patients.

Hydra head activator peptide has trophic activity for eukaryotic neurons.

The synthetic undecameric peptide, pGlu-Pro-Pro-Gly-Gly-Ser-Lys-Val-Ile-Leu-Phe, known as the hydra head activator peptide, present in high concentrations in mammalian hypothalamus and intestine, was tested for neurotrophic activity in a survival assay using cultured chick embryonic sympathetic and dorsal root ganglion cells, and for morphological differentiation activity on neuroblastoma cells. Hydra head activator peptide supported neuron survival. The optimal active concentration, 1 pM, was very similar to the concentration that causes bud and head formation in hydra. Maximal neuron survival obtained with hydra head activator peptide was close to that obtained with nerve growth factor: both substances enhanced survival up to 3 times that of control cultures. Bradykinin, which has some amino acid sequence homology with hydra head activator, was inactive as a neurotrophic factor. Hydra head activator induced rapid morphological differentiation of the mouse neuroblastoma cell line Neuro-2A. Neuro-2A responded to the peptide by process extension, 4 h after its addition to the culture medium. Neurotrophic factors isolated to date have been characterized by their ability to maintain cell viability and enhance neurite outgrowth. Hydra head activator peptide met these two criteria when tested in 3 different neuron culture systems. Our results suggest that the head activator peptide may act as a neurotrophic factor for neurons in other species, including mammals.

Expression of neurotrophic activity in Xenopus oocytes injected with mRNA from wounded rat cerebral cortex.

Injury to the cerebral cortex of the rat brain has been shown to induce the expression of neurotrophic factors for dissociated peripheral and central neurons in culture. We confirm this phenomenon and report that Xenopus laevis oocytes injected with mRNA extracted from wounded rat cortex expressed similar neurotrophic activity. To detect the low amounts of neurotrophic factors that could be expected from the oocyte translation system, a miniaturization of the assay for neurotrophic and cell-surviving activity was developed using Terasaki microtiter plates for culture of chicken embryo sympathetic ganglion cells. Messenger RNA (mRNA) was size-fractionated on a sucrose gradient and RNAs from each fraction were injected into oocytes. Neurotrophic activity was recovered from the homogenates and from the incubation media of oocytes injected with mRNA from 7 day post-lesion cortex. Messenger RNAs in the active fractions ranged in size from 0.8 to 1.8 kb. As much as 20% of the activity was secreted by the oocytes. No significant neurotrophic activity was detected from oocytes injected with mRNA fractions extracted from the cortex of control rats or from other gradient fractions from post-lesion cortex.

1,1,3 tricyano-2-amino-1-propene (Triap): a small molecule which mimics or potentiates nerve growth factor.

1,1,3 Tricyano-2-amino-1-propene (Triap) is a small molecular weight compound which increases the rate of nerve and tissue regeneration in several experimental systems. Early experiments with this compound showed that, like nerve growth factor (NGF), Triap induced neurite formation in chick spinal ganglia. To assess the similarity between NGF and Triap, we compared the effects of Triap and NGF on a rat pheochromocytoma cell line (PC12) and on cell survival in a primary chick neuronal culture. In the latter, Triap at less than 0.01 nM preserved neurons and caused them to extend neurites as did 1 nM NGF. Triap induced neurite outgrowth in the PC12 cell line giving a maximal response (40-50% of the maximal response of NGF) at a concentration of 20 micrograms/ml (151 microM). Triap's morphological effects were not inhibited by antibodies directed against NGF or the NGF receptor. Low concentrations of Triap also potentiated the morphological effects of NGF. Triap induced an increase in cell-substratum adhesion and cellular hypertrophy in PC12 cells and also potentiated the adhesive actions of NGF. Triap had no effect on ornithine decarboxylase activity even though it potentiated NGF's effects on this enzyme. These data indicate that Triap induces neurotrophic effects and does not seem to act through the same mechanisms as NGF but can potentiate many of NGF's morphological and biochemical actions.

Toxic effect of phenytoin on developing cortical neurons in culture.

Studies were undertaken to determine the effect of chronic phenytoin exposure on developing neurons. Cerebral cortex from 16-day fetal mice was utilized to prepare primary dissociated cell cultures. Phenytoin was added to the cultures 10 days after plating and the cultures were harvested on day 17. Cortical cultures were assayed for neuronal cell number by phase microscopy and for high-affinity uptake of 3H-labeled gamma-aminobutyric acid (GABA) by both radioautography and scintillation spectrometry. Neuronal cell counts demonstrated a highly significant decrement in the number of neurons in cultures exposed to phenytoin at 15, 25, and 50 micrograms/ml. 3H-GABA-labeled neurons constituted 13% of the neurons present in both control and phenytoin-exposed cultures. These data indicate that phenytoin is toxic to cortical neurons in culture and that GABAergic neurons are affected to the same extent as the total neuronal population.

Benzodiazepine receptor development in cultures of fetal mouse cerebral cortex mimics its development in vivo.

Using a recently developed assay method which maintains cell integrity, we have performed a developmental study of the benzodiazepine (BDZ) receptor on cells of fetal mouse cortex in cultures which were either 'neuron-poor' or 'neuron-rich'. The developmental profile of the BDZ receptor in culture, when assayed on the intact cell, closely mimicked its development in vivo, reaching maximum values at 35-42 days after conception. Further, the time course of development paralleled that reported for GABA neurons both in vivo and in vitro. We have also shown that the relative delay in development of the clonazepam-displaceable portion of BDZ binding approximated a similar delay in the reported development of glutamic acid decarboxylase activity, both parameters presumably reflecting specific neuronal development. Thus, the ontogenesis of specific BDZ receptor binding in culture bears marked similarity to the time course of development of the GABAergic system.

An in situ assay for determination of benzodiazepine binding.

Benzodiazepine (BDZ) receptors have been demonstrated recently in a variety of mammalian tissues. However, assay methods for such receptors have required that disrupted tissues be used. We have developed an in situ assay for this receptor utilizing intact cells cultured from the cerebral cortices of fetal mice which is more sensitive and physiologic than those used previously. Results obtained with this assay differ in the following ways from those in which disrupted tissues are used: (1) total and specific BDZ binding was as much as 10-fold higher in the in situ assays; (2) Scatchard analysis of the binding data is consistently nonlinear, revealing at least two binding sites with KD values of 5.5 and 303 nM, and (3) presumed nonneuronal receptors were found in abundance.

Adverse effects of phenobarbital on morphological and biochemical development of fetal mouse spinal cord neurons in culture.

The effects of chronic phenobarbital treatment on neuronal development were assessed in cell cultures of fetal rodent spinal cord. The activity of the principal enzyme involved in acetylcholine synthesis, choline acetyltransferase (CAT), and counts of large spinal cord neurons were used as indicators of neuronal development Phenobarbital (30 to 120 micrograms/ml) added to cultures for two- or six-week periods produced dose-dependent decreases in both CAT activity and neurons counts. Barbituric acid at doses equimolar to those at which phenobarbital produced these decreases had no effect.

Neuronal maturation in mammalian cell culture is dependent on spontaneous electrical activity.

Fetal mouse spinal cord (SC) and dorsal root ganglion (DRG) neurons undergo a process of maturation in cell culture lasting a month or more. We have investigated the role of electrical activity in this maturational process with the use of tetrodotoxin (TTX), the specific blocker of the voltage-sensitive sodium channel responsible for action potential generation. This agent completely eliminates the spikes and related synaptic activity which occur abundantly in untreated cultures. Such blockade of electrical activity in the cultures, when begun early (day 1 or day 8 in vitro), results in a 85-95% reduction in the number of large SC neurons, without affecting DRG neuron numbers. TTX treatment initiated when cultures are mature (day 70) has no significant effect on either DRG or SC neurons. Intermediate effects are obtained when treatment is initiated at day 35 in vitro. The activity of the nerve-specific enzyme choline acetyltransferase, is significantly decreased by early TTX treatment, while DNA and protein content of the cultures (primarily contributed by glial and fibroblastic cells) is not affected.

Neuroblastoma differentiation involves both the disappearance of old and the appearance of new poly(A)+ messenger RNA sequences in polyribosomes.

The cholinergic mouse neuroblastoma cell line NS20Y was adapted to undifferentiated growth in suspension culture. When suspension cells were transferred to surface culture and treated with dibutyryl cyclic AMP, the cells underwent differentiation as assessed by biochemical, morphological, and physiological criteria. Differentiated NS20Y cells in co-culture with mouse muscle cells had the capacity to form functional neuromuscular junctions with the muscle cells. The sequence complexities of the poly(A)-containing messenger RNA (poly(A)+ mRNA) of the differentiated, process-forming cells (P-cells) and undifferentiated cells in suspension culture (S-cells) were measured by analysis of the kinetics of hybridization of the mRNAs with their complementary DNAs (cDNAs). There were less than 100 high abundance and approximately 8000 low abundance poly(A)+ mRNAs in both differentiation states. Heterologous hybridization reactions and recycling of the cDNA probes revealed that 9.7% and 6.8% of the messages in P- and S-cells, respectively, were specific to those differentiation states. The P-cell-specific sequences included approximately 3 high abundance and 320 low abundance poly(A)+ mRNAs. The S-cell-specific sequences included approximately 3 high abundance and 250 low abundance poly(A)+ mRNAs. We conclude that the increment in NS20Y differentiation results in both the disappearance of old, and the appearance of new mRNAs in polyribosomes.