Long-term outcomes of urethral catheterisation injuries: a prospective multi-institutional study.

PURPOSE: There are no prospective data describing the incidence and spectrum of long-term complications associated with traumatic urethral catheterisation (UC). We prospectively monitored the long-term clinical outcomes and complications of patients with traumatic UC injuries. METHODS: A prospective study at two tertiary university hospitals was performed to record all referrals for iatrogenic urethral injuries caused by UC. Long-term follow-up was prospectively maintained by regular outpatient department visits and by monitoring all urological interventions and their outcomes from urinary catheter-related injuries. RESULTS: The incidence of traumatic UC was 13.4 per 1000 catheters inserted in male patients and 37 iatrogenic urethral injuries were recorded. The mean age was 74 ± 12 years and the mean length of follow-up was 37 ± 3.7 months. Urethral injuries were caused by inflating the catheter anchoring balloon in the urethra (n = 26) or by creating a false passage with the catheter tip (n = 11). In total, 29 patients (78%) developed urethral stricture disease during their follow-up; of which 11 have required at least one urethral dilation and two have required one urethrotomy. Three patients required long-term indwelling suprapubic catheter placement and seven patients opted for a long-term indwelling urethral catheter. There were eight patient mortalities; one of which was due to severe urosepsis resulting from catheter balloon inflation in the urethra. CONCLUSION: Catheter-related injuries are associated with significant long-term complications in this vulnerable patient cohort. In future, such injuries may be preventable if the safety profile of the urinary catheter is modified.

Characterisation of human urethral rupture thresholds for urinary catheter inflation related injuries.

Data on urethral catheter related injuries is sparse. In this study we aimed to characterise urethral diametric strain and urinary catheter inflation pressure thresholds that precede human urethral trauma during urethral catheterisation (UC). Human urethras were obtained from patients undergoing male to female gender reassignment surgery [(n = 9; age 40 ±â€¯13.13 (range: 18-58)) years]. 12Fr urinary catheters were secured in the bulbar urethra and the catheter's anchoring balloon was inflated with a syringe pump apparatus. Urethral diametric strain and balloon pressure were characterised with video extensometry and a pressure transducer respectively. Immunohistochemistry, Masson's trichrome and Verhoeff-Van Gieson stains evaluated urethral trauma microscopically. Morphological characterisation of the urethral lumen was performed by examining non-traumatised histological sections of urethra and recording luminal area, perimeter and major/minor axis length. Tearing (n = 3) and rupture (n = 3) of the urethra were observed following catheter balloon inflation. The threshold for human urethral rupture occurred at an external urethral diametric strain ≥ 27% and balloon inflation pressure ≥ 120kPa. Significant relationships were identified between urethral wall thickness and the level of trauma induced during catheter balloon inflation (p = 0.001) and between the pressure required to inflate the catheter balloon and the length of the major axis of the urethral lumen (p = 0.004). Ruptured urethras demonstrated complete transection of collagen, elastin and muscle fibres. In conclusion, urethral rupture occurs at an external urethral diametric strain ≥ 27% or with balloon inflation pressures ≥ 120 kPa. Incorporation of these parameters may be useful for designing a safety mechanism for preventing catheter inflation related urethral injuries.

Architecture of a transcribing-translating expressome.

DNA transcription is functionally coupled to messenger RNA (mRNA) translation in bacteria, but how this is achieved remains unclear. Here we show that RNA polymerase (RNAP) and the ribosome of Escherichia coli can form a defined transcribing and translating "expressome" complex. The cryo-electron microscopic structure of the expressome reveals continuous protection of ~30 nucleotides of mRNA extending from the RNAP active center to the ribosome decoding center. The RNAP-ribosome interface includes the RNAP subunit α carboxyl-terminal domain, which is required for RNAP-ribosome interaction in vitro and for pronounced cell growth defects upon translation inhibition in vivo, consistent with its function in transcription-translation coupling. The expressome structure can only form during transcription elongation and explains how translation can prevent transcriptional pausing, backtracking, and termination.

Internal temperature increase during photothermal tumour ablation in mice using gold nanorods.

Laser ablation (LA) is gaining large acceptance in the treatment of tumor. One of the main risks of this treatment is damaging the healthy tissue around the tumor. Among the solutions proposed to improve the selectivity of the LA and to localize heating to tumor tissue, the use of gold nanoparticles is one of the most promising. The aim of this work is threefold: i) to measure the temperature increase within the tumor during plasmonic photothermal therapy using gold nanorods; ii) to investigate the influence of nanorods concentration and laser settings on both the intra-tumoral temperature and the tumor surface temperature; iii) and to establish the nanorods concentrations able to cause tumor resorption at a defined laser settings. Two sets of trials were performed: i) 16 mice were divided in four groups with different treatment time (i.e., 5 min, 2 min, 1 min, and 30s), with constant gold nanorods amount (i.e., 12.5 µg) and laser power (i.e., 3 W·cm(-2)); ii) 16 mice were divided in four groups treated with different amount of gold nanorods (i.e., control, 12.5 µg, 25 µg, 50 µg) for 5 min at 2 W·cm(-2). Results show significant differences between internal and surface temperatures. We also demonstrate that this temperature difference increases with nanoparticle concentrations, decreases with laser power, and is not influenced by treatment time. This information is critical to improve the theoretical models that will guide future study designs in sensitive orthotopic tumor models.

Attitudes among junior doctors towards improving the transurethral catheterisation process.

OBJECTIVES: To evaluate the subjective opinions of junior doctors on their adequacy of training and confidence levels for performing transurethral catheterisation (TUC) and to investigate their subjective interest in a 'safety mechanism' that would eliminate the potential for urethral trauma during TUC. METHODS: An anonymous online survey was emailed to all interns that had a documented email address on the Royal College of Surgeons Ireland registry (2012-2013). The survey consisted of eight questions pertaining to TUC of male patients. RESULTS: The survey was delivered to 252 email addresses and the response rate was 52% (130/252). The vast majority (99%; n = 128) of interns felt confident inserting a transurethral catheter independently and 73% (n = 95) subjectively received appropriate training for catheterising male patients. The incidence of trauma after mistakenly inflating the catheter's anchoring balloon in the urethra was 3% (n = 4). The majority (90%; n = 116) of respondents were interested in a safety mechanism for preventing urethral trauma and 71% (n = 92) felt that a safety mechanism for urethral trauma prevention should be compulsory for all transurethral catheterisation among male patients. CONCLUSION: Despite pre-emptive training programmes, it appears that iatrogenic urethral trauma secondary to TUC remains a persistent morbidity in healthcare settings. Designing a safer transurethral catheter may be necessary to eliminate the risk of unnecessary urethral trauma in patients.

An analysis of radiation dose reduction in paediatric interventional cardiology by altering frame rate and use of the anti-scatter grid.

The purpose of this work is to investigate removal of the anti-scatter grid and alteration of the frame rate in paediatric interventional cardiology (IC) and assess the impact on radiation dose and image quality. Phantom based experimental studies were performed in a dedicated cardiac catheterisation suite to investigate variations in radiation dose and image quality, with various changes in imaging parameters. Phantom based experimental studies employing these variations in technique identified that radiation dose reductions of 28%-49% can be made to the patient with minimal loss of image quality in smaller sized patients. At present, there is no standard technique for carrying out paediatric IC in the UK or Ireland, resulting in the potential for a wide variation in radiation dose. Dose reductions to patients can be achieved with slight alterations to the imaging equipment with minimal compromise to the image quality. These simple modifications can be easily implemented in clinical practice in IC centres.

Case series evidence for changed interhemispheric relationships in cortical structure in some amputees.

Limb amputation and related changes in body feelings are associated with cortical functional reorganization that is reflected by increased interhemispheric asymmetry of body maps in the postcentral somatosensory cortex (PCS). As a pilot test to determine if limb amputation affects interhemispheric symmetry in PCS structure, we used MRI and computational morphometry to examine interhemispheric relationships of PCS thicknesses in a case series of eight lower limb amputees compared with 11 control subjects. As a further control, the same relationships were compared in the lateral occipital visual cortex (LOV) which, by nature of its visual connectivity, would be expected to be less related to amputation. The PCS thicknesses in the left and right hemispheres were positively related in control subjects, but not in amputees. The range of the PCS interhemispheric thickness differences (ID) in amputees was larger than the range in control subjects, and four of eight amputees had PCS ID that were at or above the maximal control subject ID. In contrast, LOV thicknesses in the two hemispheres were positively related and LOV ID ranges were similar in both amputees and control subjects. The results from this case series suggest the hypothesis that amputation alters PCS interhemispheric thickness relationships in some amputees. Further tests of this hypothesis would be useful to determine whether changes in structural symmetry contribute to known post-amputation alterations in PCS functional map symmetry and body feeling.

Use of a whole-slide imaging system to assess the presence and alteration of lymphatic vessels in joint sections of arthritic mice.

We investigated the presence and alteration of lymphatic vessels in joints of arthritic mice using a whole-slide imaging system. Joints and long bone sections were cut from paraffin blocks of two mouse models of arthritis: meniscal-ligamentous injury (MLI)-induced osteoarthritis (OA) and TNF transgene (TNF-Tg)-induced rheumatoid arthritis (RA). MLI-OA mice were fed a high fat diet to accelerate OA development. TNF-Tg mice were treated with lymphatic growth factor VEGF-C virus to stimulate lymphangiogenesis. Sections were double immunofluorescence stained with anti-podoplanin and alpha-smooth muscle actin antibodies. The area and number of lymphatic capillaries and mature lymphatic vessels were determined using a whole-slide imaging system and its associated software. Lymphatic vessels in joints were distributed in soft tissues mainly around the joint capsule, ligaments, fat pads and muscles. In long bones, enriched lymphatic vessels were present in the periosteal areas adjacent to the blood vessels. Occasionally, lymphatic vessels were observed in the cortical bone. Increased lymphatic capillaries, but decreased mature lymphatic vessels, were detected in both OA and RA joints. VEGF-C treatment increased lymphatic capillary and mature vessel formation in RA joints. Our findings suggest that the lymphatic system may play an important role in arthritis pathogenesis and treatment.

Ultra-sharp metal and nanotube-based probes for applications in scanning microscopy and neural recording.

This paper discusses several methods for manufacturing ultra-sharp probes, with applications geared toward, but not limited to, scanning microscopy (STM, AFM) and intra-cellular recordings of neural signals. We present recipes for making tungsten, platinum/iridium alloy, and nanotube fibril tips. Electrical isolation methods using Parylene-C or PMMA are described.

Sensory Constraints on Birdsong Syntax: Neural Responses to Swamp Sparrow Songs with Accelerated Trill Rates.

Both sensory and motor mechanisms can constrain behavioral performance. Sensory mechanisms may be especially important for constraining behaviors that depend on experience, such as learned birdsongs. Swamp sparrows learn to sing by imitating the song of a tutor, but sparrows fail to accurately imitate artificial tutor songs with abnormally accelerated trills, instead singing brief and rapid trills interrupted by silent gaps. This "broken syntax" has been proposed to arise from vocal-motor limitations. Here we consider whether sensory limitations exist that could also contribute to broken syntax. We tested this idea by recording auditory-evoked activity of sensorimotor neurons in the swamp sparrow's brain that are known to be important for the learning, performance and perception of song. In freely behaving adult sparrows that sang songs with normal syntax, neurons were detected that exhibited precisely time-locked activity to each repetition of the syllable in a trill when presented at a natural rate. Those cells failed to faithfully follow syllables presented at an accelerated rate, however, and their failure to respond to consecutive syllables increased as a function of trill rate. This "flickering" auditory representation in animals performing normal syntax reveals a central constraint on the sensory processing of rapid trills. Furthermore, because these neurons are implicated in both song learning and perception, and because auditory flickering began to occur at accelerated trill rates previously associated with the emergence of broken song syntax, these sensory constraints may contribute to the emergence of broken syntax.

Porcine extracellular matrix scaffolds in reconstructive urology: An ex vivo comparative study of their biomechanical properties.

Functional reconstruction of the human urinary bladder has been attempted by replacing defective bladder tissue with tissue-engineered xenogenic extracellular matrix (ECM) scaffolds. However, experimental studies that demonstrate the effects of implanted ECMs on important biomechanical properties such as total bladder capacity (TBC) and compliance (C) are lacking. In the current study, the effects of ECM scaffold surface area (SA) on TBC and C was assessed, ex vivo, in an ovine model (n=5). TBC and C were measured at pressures (P) of 5, 10, 15 and 20 mm Hg prior to performing a 3×3 cm (9 cm(2)) partial cystectomy defect. Equal-sized 3×3 cm (9 cm(2)) and larger 6×6 cm (36 cm(2)) urinary bladder matrix (UBM) scaffolds of porcine origin replaced the 3×3 cm cystectomy defect, and TBC and C were re-recorded for comparative analysis. The results showed that TBC decreased by 39.6%±0.005% (122.9 ml±15 ml, p<0.05) and C by 38.9%±0.51%, (ΔP=0-5mmHg, p<0.05) in ovine bladders reconstructed with 3×3 cm UBM scaffolds compared to their native values. It was also found that TBC increased by 25.6±0.64% (64.2 ml ± 8.8 ml, p>0.05) and C by 24.5±0.43% (ΔP=0-5mmHg, p>0.05) in the 6×6 cm UBM scaffold group compared to the 3×3 cm UBM scaffold group; however, these values were not statistically significant. The present work demonstrates that a fourfold increase in ECM scaffold SA relative to its intended defect does not lead to a significant improvement in TBC and C values.

In vivo vomeronasal stimulation reveals sensory encoding of conspecific and allospecific cues by the mouse accessory olfactory bulb.

The rodent vomeronasal system plays a critical role in mediating pheromone-evoked social and sexual behaviors. Recent studies of the anatomical and molecular architecture of the vomeronasal organ (VNO) and of its synaptic target, the accessory olfactory bulb (AOB), have suggested that unique features underlie vomeronasal sensory processing. However, the neuronal representation of pheromonal information leading to specific behavioral and endocrine responses has remained largely unexplored due to the experimental difficulty of precise stimulus delivery to the VNO. To determine the basic rules of information processing in the vomeronasal system, we developed a unique preparation that allows controlled and repeated stimulus delivery to the VNO and combined this approach with multisite recordings of neuronal activity in the AOB. We found that urine, a well-characterized pheromone source in mammals, as well as saliva, activates AOB neurons in a manner that reliably encodes the donor animal's sexual and genetic status. We also identified a significant fraction of AOB neurons that respond robustly and selectively to predator cues, suggesting an expanded role for the vomeronasal system in both conspecific and interspecific recognition. Further analysis reveals that mixed stimuli from distinct sources evoke synergistic responses in AOB neurons, thereby supporting the notion of integrative processing of chemosensory information.

Avian nucleus retroambigualis: cell types and projections to other respiratory-vocal nuclei in the brain of the zebra finch (Taeniopygia guttata).

In songbirds song production requires the intricate coordination of vocal and respiratory muscles under the executive influence of the telencephalon, as for speech in humans. In songbirds the site of this coordination is suspected to be the nucleus retroambigualis (RAm), because it contains premotor neurons projecting upon both vocal motoneurons and spinal motoneurons innervating expiratory muscles, and because it receives descending inputs from the telencephalic vocal control nucleus robustus archopallialis (RA). Here we used tract-tracing techniques to provide a more comprehensive account of the projections of RAm and to identify the different populations of RAm neurons. We found that RAm comprises diverse projection neuron types, including: 1) bulbospinal neurons that project, primarily contralaterally, upon expiratory motoneurons; 2) a separate group of neurons that project, primarily ipsilaterally, upon vocal motoneurons in the tracheosyringeal part of the hypoglossal nucleus (XIIts); 3) neurons that project throughout the ipsilateral and contralateral RAm; 4) another group that sends reciprocal, ascending projections to all the brainstem sources of afferents to RAm, namely, nucleus parambigualis, the ventrolateral nucleus of the rostral medulla, nucleus infra-olivarus superior, ventrolateral parabrachial nucleus, and dorsomedial nucleus of the intercollicular complex; and 5) a group of relatively large neurons that project their axons into the vagus nerve. Three morphological classes of RAm cells were identified by intracellular labeling, the dendritic arbors of which were confined to RAm, as defined by the terminal field of RA axons. Together the ascending and descending projections of RAm confirm its pivotal role in the mediation of respiratory-vocal control.

Precise auditory-vocal mirroring in neurons for learned vocal communication.

Brain mechanisms for communication must establish a correspondence between sensory and motor codes used to represent the signal. One idea is that this correspondence is established at the level of single neurons that are active when the individual performs a particular gesture or observes a similar gesture performed by another individual. Although neurons that display a precise auditory-vocal correspondence could facilitate vocal communication, they have yet to be identified. Here we report that a certain class of neurons in the swamp sparrow forebrain displays a precise auditory-vocal correspondence. We show that these neurons respond in a temporally precise fashion to auditory presentation of certain note sequences in this songbird's repertoire and to similar note sequences in other birds' songs. These neurons display nearly identical patterns of activity when the bird sings the same sequence, and disrupting auditory feedback does not alter this singing-related activity, indicating it is motor in nature. Furthermore, these neurons innervate striatal structures important for song learning, raising the possibility that singing-related activity in these cells is compared to auditory feedback to guide vocal learning.

A comparison of patient skin doses before and after replacement of a neurointerventional fluoroscopy unit.

AIM: After several embolization patients presented with radiation-induced skin injury in our neuroradiology centre, replacement of the centre's interventional fluoroscopy unit was prioritized. The aims of the present study were to compare the maximum skin dose delivered to each patient by the old and new units, to devise a method of estimating skin dose from the displayed dose-area product and to set local reference doses. MATERIALS AND METHODS: On the old unit, skin dose was measured using thermoluminescent dosimeters on 12 patients undergoing Gugliemi detachable coil embolization. Similar skin dose measurements were undertaken and the dose-area product was recorded for a further 12 patients on the new unit. RESULTS: The maximum skin dose measured on each patient on the old and new units had a mean of 2.2 Gy and 0.47 Gy, respectively, and a maximum of 4.1 Gy and 1.0 Gy, respectively. Maximum dose delivered to patients' skin by the new equipment was less than a quarter of the dose from the old equipment (p < 0.0001). CONCLUSION: The large reductions in skin dose reduced the risk of patients suffering radiation injury and confirmed the validity of replacing ageing interventional fluoroscopy equipment with modern equipment that incorporates dose management systems. As patient skin dose was correlated with dose-area product, local reference dose levels were set in terms of dose-area product; this enabled the operator to monitor the likely maximum patient skin dose during embolization procedures. Other centres could use a similar method to set their own reference doses.

Physiology of neuronal subtypes in the respiratory-vocal integration nucleus retroamigualis of the male zebra finch.

Learned vocalizations, such as bird song, require intricate coordination of vocal and respiratory muscles. Although the neural basis for this coordination remains poorly understood, it likely includes direct synaptic interactions between respiratory premotor neurons and vocal motor neurons. In birds, as in mammals, the medullary nucleus retroambigualis (RAm) receives synaptic input from higher level respiratory and vocal control centers and projects to a variety of targets. In birds, these include vocal motor neurons in the tracheosyringeal part of the hypoglossal motor nucleus (XIIts), other respiratory premotor neurons, and expiratory motor neurons in the spinal cord. Although various cell types in RAm are distinct in their anatomical projections, their electrophysiological properties remain unknown. Furthermore, although prior studies have shown that RAm provides both excitatory and inhibitory input onto XIIts motor neurons, the identity of the cells in RAm providing either of these inputs remains to be established. To characterize the different RAm neuron types electrophysiologically, we used intracellular recordings in a zebra finch brain stem slice preparation. Based on numerous differences in intrinsic electrophysiological properties and a principal components analysis, we identified two distinct RAm neuron types (types I and II). Antidromic stimulation methods and intracellular staining revealed that type II neurons, but not type I neurons, provide bilateral synaptic input to XIIts. Paired intracellular recordings in RAm and XIIts further indicated that type II neurons with a hyperpolarization-dependent bursting phenotype are a potential source of inhibitory input to XIIts motor neurons. These results indicate that electrically distinct cell types exist in RAm, affording physiological heterogeneity that may play an important role in respiratory-vocal signaling.

Synaptic transformations underlying highly selective auditory representations of learned birdsong.

Stimulus-specific neuronal responses are a striking characteristic of several sensory systems, although the synaptic mechanisms underlying their generation are not well understood. The songbird nucleus HVC (used here as a proper name) contains projection neurons (PNs) that fire temporally sparse bursts of action potentials to playback of the bird's own song (BOS) but are essentially silent when presented with other acoustical stimuli. To understand how such remarkable stimulus specificity emerges, it is necessary to compare the auditory-evoked responsiveness of the afferents of HVC with synaptic responses in identified HVC neurons. We found that inactivating the interfacial nucleus of the nidopallium (NIf) could eliminate all auditory-evoked subthreshold activity in both HVC PN types, consistent with NIf serving as the major auditory afferent of HVC. Simultaneous multiunit extracellular recordings in NIf and intracellular recordings in HVC revealed that NIf population activity and HVC subthreshold responses were similar in their selectivity for BOS and that NIf spikes preceded depolarizations in all HVC cell types. These results indicate that information about the BOS as well as other auditory stimuli is transmitted synaptically from NIf to HVC. Unlike HVC PNs, however, HVC-projecting NIf neurons fire throughout playback of BOS as well as non-BOS stimuli. Therefore, temporally sparse BOS-evoked firing and enhanced BOS selectivity, manifested as an absence of suprathreshold responsiveness to non-BOS stimuli, emerge in HVC. The transformation to a sparse auditory representation parallels differences in NIf and HVC activity patterns seen during singing, which may point to a common mechanism for encoding sensory and motor representations of song.

EGF-dependent cell cycle progression is controlled by density-dependent regulation of Akt activation.

The normal human breast epithelial cell line, MCF10A, was used to investigate the mechanism by which high-density inhibits EGF-dependent cell cycle progression. EGF-dependent Akt activation was found to be transient in high-density cells and sustained in low-density cells. High-density cells also showed decreased EGF receptor (EGFR) autophosphorylation, decreased retinoblastoma protein phosphorylation, and increased p27 protein expression. Although EGFR activation was decreased in the high-density cells, the activation was sufficient to stimulate EGFR substrates comparable to low-density cells. EGF-dependent activation of the Erk1/2 pathway and the upstream activators of Akt (Gab1, erbB3, PI3 kinase, and PDK1) showed no density dependency. Antagonists of Akt activity provided further evidence that regulation of Akt activation is the critical signal transduction step controlling EGF-dependent cell cycle progression. Both adenovirus-mediated expression of dominant-negative Akt and inhibition of PI3 kinase-mediated Akt activation with LY294002 blocked cell cycle progression of low-density cells. In summary, we report the novel finding that high-density blocks EGF-dependent cell cycle progression by inhibiting EGF signaling at the level of EGF-dependent Akt activation rather than at the level of EGFR activation.

Dose reduction to radiosensitive tissues in CT. Do commercially available shields meet the users' needs?

AIM: To assess the effectiveness and economy of routinely using commercially available in-plane bismuth shielding during CT scanning of the chest and brain. METHODS AND MATERIALS: Forty patients were scanned with thermoluminescent dosemeters (TLDs) in situ to measure the radiation dose to the thyroid and eye during CT scanning of the brain and chest. Half of the patients had the "AttenuRad" shield in place during scanning. RESULTS: Use of the shielding reduced the mean dose to the eye from 6.0 +/- 0.3 mGy to 4.9 mGy +/- 0.2 mGy and the thyroid dose from 16.4 mGy +/- 1.2 mGy to 7.1 mGy +/- 0.5 mGy. CONCLUSION: Use of the thyroid shield is recommended for all CT scanning of the chest. The eye shield does not produce as marked a reduction in radiation dose to the lens of the eye, when an angled gantry is used, since the eyes are not in the primary beam. Use of the eyeshield is justifiable where irradiation of the orbit is unavoidable, although whether artifacts would be a problem if the shield was used in this way was not assessed.

Monoamine oxidase inhibitors l-deprenyl and clorgyline protect nonmalignant human cells from ionising radiation and chemotherapy toxicity.

l-Deprenyl (R-(-)-deprenyl, selegiline) is an inhibitor of monoamine oxidase-B (MAO-B) that is known to protect nerve cells from a variety of chemical and physical insults. As apoptosis is a common mechanism of radiation-induced cell death, the effect of l-deprenyl on the survival of cultured cells and tissue explants was studied following exposure to gamma radiation. The results obtained were compared with the effects of the less-selective MAO-B inhibitor pargyline and the MAO-A inhibitor clorgyline. l-Deprenyl at a concentration of 10(-9) M protected the nontumorigenic cell line (HaCaT) and normal human urothelial explants from the effects of cobalt-60 gamma radiation, but did not protect tumorigenic human cell lines HaCaT-ras, HPV-transfected human keratinocytes (HPV-G cells), or PC3. Human bladder carcinoma explants were not protected. Clorgyline showed a smaller protective effect of normal cells, whereas pargyline had no effect. Radiation-induced delayed effects (genomic instability measured as delayed cell death) were prevented in normal cells by l-deprenyl but, interestingly, deprenyl appeared to increase the amount of delayed death in the tumorigenic cell lines. Studies using l-deprenyl prior to the exposure of nonmalignant cells to cisplatin showed that cell death due to this agent was also reduced. Treatment of cultures of nontumorigenic cells with l-deprenyl or clorgyline significantly increased the levels of the protein Bcl-2 following irradiation, but there was no such effect on the already-elevated levels of this protein in the tumour samples. Since the Bcl-2 has been shown to be an inhibitor of apoptosis or programmed cell death, this would imply that the protective effects of l-deprenyl and clorgyline involve activation of antiapoptotic pathways within the normal cell. This hypothesis is supported by data showing reduced levels of apoptosis in HaCAT cells and in normal bladder explant cultures following treatment with l-deprenyl.