Ependymal stem cells divide asymmetrically and transfer progeny into the subventricular zone when activated by injury.

Evidence is presented to show that cells of the ependymal layer surrounding the ventricles of the mammalian (rat) forebrain act as neural stem cells (NSCs), and that these cells can be activated to divide by a combination of injury and growth factor stimulation. Several markers of asymmetric cell division (ACD), a characteristic of true stem cells, are expressed asymmetrically in the ependymal layer but not in the underlying subventricular zone (SVZ), and when the brain is treated with a combination of local 6-hydroxydopamine (6-OHDA) with systemic delivery of transforming growth factor-alpha (TGFalpha), ependymal cells divide asymmetrically and transfer progeny into the SVZ. The SVZ cells then divide as transit amplifying cells (TACs) and their progeny enter a differentiation pathway. The stem cells in the ependymal layer may have been missed in many previous studies because they are usually quiescent and divide only in response to strong stimuli.

Safety and side effects of immediate instillation of apaziquone following transurethral resection in patients with nonmuscle invasive bladder cancer.

PURPOSE: We studied the safety, tolerability and pharmacokinetics of a single immediate post-transurethral resection intravesical instillation of apaziquone for patients with nonmuscle invasive bladder cancer. MATERIALS AND METHODS: Patients with cTa-T1, G1-G2 urothelial cell carcinoma of the bladder underwent transurethral resection of bladder tumor(s) followed by a single intravesical instillation of apaziquone 4 mg/40 ml for 1 hour within 6 hours of transurethral bladder tumor resection. Adverse events and safety parameters were assessed on days 8 and 15 after transurethral bladder tumor resection. Blood samples were drawn before and during the instillation for pharmacokinetic analyses. The first 10 patients with pTa-T1, G1-G2 nonmuscle invasive bladder cancer were also evaluated by cystoscopy 3 months after treatment to determine mucosal healing. RESULTS: Of 20 patients receiving apaziquone 13 (65%) reported 35 adverse events, mostly grade 1 to 2. Eight patients (40%) reported 13 adverse events related to treatment, in particular dysuria, hematuria, bladder spasm, abdominal pain, asthenia and postoperative urinary retention. Three grade 3 and 1 grade 4 event(s) occurred, but these were considered unrelated to treatment. No other significant clinical changes were observed. Apaziquone and the active metabolite EO5a were not detected with pharmacokinetic analyses at any point of time. After 3 months no evidence of impaired mucosal healing was observed. CONCLUSIONS: A single immediate post-transurethral bladder tumor resection instillation of apaziquone was well tolerated with an expected good safety profile. Apaziquone and its metabolite EO5a were not detected systemically with pharmacokinetic analyses. These results have lead to further study of a single immediate instillation of apaziquone.

Human buccal absorption of flurbiprofen.

The buccal absorption of flurbiprofen was studied in normal men to quantify the transport from the oral cavity in humans and to evaluate the closed-perfusion cell apparatus as a means to study drug transport across externally accessible biologic membranes. Flurbiprofen was buccally absorbed by a passive diffusional mechanism and the rate of absorption was pH dependent. Membrane permeability coefficients for flurbiprofen were 4.3 x 10(-4) cm/sec at pH 5.5 and 2.1 x 10(-5) cm/sec at pH 7.0. These findings are in agreement with the pH relationship for buccal transport observed in dog experiments. Delineation of the effective permeability coefficients into components for the aqueous boundary layer and the lipoidal buccal membrane allowed for the prediction of the extent of absorption of the drug over a period of time. It was concluded that the buccal membranes of the human and dog were essentially lipoidal membranes with equivalent permeabilities and no evident aqueous pore pathways.

Incontinence associated with bilateral lesions of putamen.

From 1238 postmortem brain examinations, 44 subject cases with bilateral putamen lesions and 44 controls without bilateral putamen lesions were chosen and compared for the presence and severity of incontinence. The lesions were divided into three grades and the incontinence into five degrees of severity. Final results revealed that 39 of 44 subject cases had some degree of fecal incontinence compared with 12 of 44 controls. The subject cases included 14 with degree 4 incontinence while the controls had none. Forty-three of 44 subject cases exhibited urinary incontinence compared with 25 of 44 controls with the degree again being more severe in the subject cases. Using the chi 2 test, the probability that these differences were due to chance is minimal. Therefore, bilateral putamen involvement appears to be a significant factor in the occurrence of incontinence, and further study of this association is indicated.

Histologic grading of prostate cancer: a perspective.

The wide-ranging biologic malignancy of prostate cancer is strongly correlated with its extensive and diverse morphologic appearances. Histologic grading is a valuable research tool that could and should be used more extensively and systematically in patient care. It can improve clinical staging, as outlined by Oesterling et al (J Urol 138: 92-98, 1987), during selection of patients for possible prostatectomy by helping to identify the optimal treatment. Some of the recurrent practical problems with grading (reproducibility, "undergrading" of biopsies, and "lumping" of grades) are discussed and recommendations are made. The newer technologically sophisticated but single-parameter tumor measurements are compared with one important advantage of histologic grading: the ability to encompass the entire low to high range of malignancy. The predictive success of grading suggests that prostate cancers have more or less fixed degrees of malignancy and growth rates (a hypothesis of "biologic determinism") rather than a steady increase in malignancy with time. Most of the observed facts can be interpreted on that basis, including the interrelations of tumor size, grade, and malignancy. The increasing age-adjusted incidence of diagnosed prostate cancer is attributed to new diagnostic tools and increased diagnostic zeal.

Infant rat model of the shaken baby syndrome: preliminary characterization and evidence for the role of free radicals in cortical hemorrhaging and progressive neuronal degeneration.

Infants subjected to repeated episodes of violent shaking develop brain damage characterized by intracranial hemorrhage and progressive cortical atrophy. We have developed an animal model that mimics this pathological state and investigated its etiology and treatment. Anesthetized male rats, 6 days of age, were subjected to one episode of shaking per day for 3 consecutive days. Separate groups of rats were sacrificed 1 h postinjury on the third day of shaking for HPLC quantification of cortical .OH and vitamin E levels, and histological assessment of cortical hemorrhaging. Additional groups were sacrificed 7 or 14 days postinjury to demonstrate progressive neuronal degeneration via cortical wet weight comparisons. In comparison to noninjured shams, the results indicated that cortical vitamin E and .OH levels rose 53.7% (p < 0.005) and 457.1% (p < 0.001), respectively, in shaken infant rats. Brain histologies revealed a moderate-to-severe degree of cortical hemorrhaging in these animals 1 h postinjury. By 7 and 14 days postinjury, there was a 13.3% and 28.7% (p < 0.0001 vs. sham) loss of cortical tissue in shaken infants, respectively, indicating progressive neuronal degeneration. Treatment with 10 mg/kg (ip) of the 21-aminosteroid antioxidant, tirilazad mesylate, 10 min before and 2 h after each episode of shaking, resulted in a 53.1% attenuation of cortical .OH levels and a 34.9% decrease in brain hemorrhaging (p < 0.05 vs. vehicle). Tirilazad treatment did not, however, significantly effect cortical vitamin E concentrations at 1 h postinjury or the extent of progressive neuronal degeneration at either 7 or 14 days postinjury. The present animal model mimics the brain pathology seen in abused children. Our observation that tirilazad mesylate, an antioxidant-lipid peroxidation inhibitor, significantly reduces cortical .OH levels and brain hemorrhaging in shaken infant rats supports a role for oxygen radicals in the pathophysiology of this type of CNS injury. The failure of tirilazad to block progressive cortical degeneration suggests that mechanisms other than free radicals may be of prime importance in the mediation of this aspect of the pathology.

The external urethral barrier for stress incontinence: a multicenter trial of safety and efficacy. Miniguard Investigators Group.

OBJECTIVE: To assess the efficacy and safety of an external urethral barrier for the management of mild to moderate stress urinary incontinence in adult women. METHODS: Four hundred eleven women with the symptom of stress urinary incontinence in 12 United States centers participated. Additional inclusion and exclusion criteria were applied before protocol device use, and ultimately 390 subjects began device use. Outcome measures for efficacy and safety were assessed. Efficacy was evaluated by the number of leakage episodes using a voiding diary, subjective urinary leakage severity, incontinence impact scores, and pad testing. Safety was evaluated by symptom assessment, urinalysis, urine culture, measurement of postvoid residual urine volume, vulvar cytology, vaginal culture, and (n = 81) cystometric testing. RESULTS: Efficacy was indicated by statistically significant reductions in the number of leakage episodes, subjective leakage severity scores, incontinence impact scores, and pad-test loss during device use. The data also indicated that the device was safe, as evidenced by the lack of statistically significant changes in the percentage of subjects with urinary tract infections during device use or in postvoid residual urine volume and cystometric indices. Symptoms of vulvar irritation or lower urinary tract discomfort occurred in a small percentage of subjects but were generally transient, and only three women discontinued using the device. CONCLUSION: The external urethral barrier appears to be a safe nonsurgical alternative to absorbent products for the management of mild to moderate stress urinary incontinence in adult women.

Identifying types of female incontinence with retrograde urethrocystography.

The most specific radiographic findings characterizing stress incontinence (SI) on upright retrograde urethrocystography include replacement of a flat or rounded bladder base with a concave funnelled base; patency of the bladder neck with contrast material pooling in the proximal urethra; the descent of the intravesical Foley balloon beyond the internal meatus and into the proximal urethra. We found that neither a cystocele nor the dependent position of the urethra at the bottom of the bladder were diagnostic of SI if the above stigmata were absent. On the other hand the defect of urgency incontinence (UI) is functional. The bladder can usually be filled by retrograde urethral infusion (though in severe UI this may not be the case). An alert technician can frequently obtain a film when the patient is experiencing uninhibited voiding. The finding of contrast material throughout the urethra, in the distal urethra alone, or in the parameatal area is strongly suspicious for UI, especially when trabeculation is also seen. These findings in association with the stigmata of SI give warning of combined SI and UI.

Apical suspension test.

In stress incontinence, urinary leakage can be controlled by support of the anterior vaginal wall by applying pressure directly at the bladder neck (the Marshall test) or applying pressure remotely. In the latter maneuver, a cotton ball, gripped in the jaws of a sponge stick, engages the vaginal vault near the apex and rotates it posteriorly and horizontally. This tenses the anterior vaginal wall thereby controlling stress-induced leakage. The mechanism of action and some surgical implications are discussed.

Comparison of cystometrograms and urethral profiles with gas and water media.

Twenty-five female patients were studied using both gas and water media for custometrograms and urethral profiles. The results showed that gas and water did not generate equivalent data in either cystometrograms or urethral profiles. The cystometrogram data showed a high correlation of data between gas and water, with gas values consistently lower than water values. Urethral profile data with gas were very difficult to interpret in terms of water-generated data, and correlations were tenuous at best.

Evaluation of an implant that delivers leuprolide for 1 year for the palliative treatment of prostate cancer.

OBJECTIVES: To evaluate the Viadur implant, which delivers leuprolide acetate for the palliative treatment of advanced prostate cancer. METHODS: Inserted subcutaneously, the 4 x 45-mm implant uses osmotic pressure to deliver leuprolide continuously at a controlled rate for 1 year. This 19-center open-label study enrolled patients with prostate cancer who had had no prior therapy or showed biochemical evidence of treatment failure after prostatectomy or radiotherapy. Each patient received one implant. After 1 year, that implant was removed, another was inserted, and patients were followed up for 2 additional months. The primary efficacy measure was suppression of testosterone to less than the castrate threshold (50 ng/dL). RESULTS: Eighty patients were enrolled. The implant effectively suppressed testosterone in 79 patients (99%) within 2 to 4 weeks and maintained that suppression through the study period. In 1 patient, the testosterone was suppressed to less than 100 ng/dL within 4 weeks but was not less than 50 ng/dL until week 24. Prostate-specific antigen levels normalized (4 ng/mL or less) or a clinically significant decrease occurred in all patients. Leuprolide was rapidly absorbed, resulting in mean serum concentrations of 16.8 ng/mL 4 hours after implant insertion and 2.4 ng/mL at 24 hours; steady mean serum leuprolide concentrations were then maintained throughout the year, at approximately 0.9 ng/mL. Investigators were satisfied with the insertion and removal procedures. All patients reported satisfaction after 1 year of treatment. The safety profile of the implant was consistent with androgen ablation therapy. Most adverse events were mild, and the most common event was hot flashes. CONCLUSIONS: The leuprolide implant effectively suppressed testosterone concentrations to less than the castrate threshold and maintained that suppression throughout the study period.

Taste preconditioning augments odor-aversion learning.

On the basis of previous work that has shown a taste can potentiate odor-aversion conditioning in AX+ conditioning, 6 experiments used rats to examine the effects of pairing a preconditioned taste (A) with a novel odor cue (X) in an A+/AX+ aversion conditioning design. Experiments 1A and 1B demonstrated that a preconditioned taste produced a robust odor aversion that was significantly stronger than a potentiated odor aversion. The results of Experiment 2 showed that the robust odor aversion produced by A+/AX+ conditioning was not the result of the potentiated odor aversion summating with generalization from the taste aversion. The augmented odor aversion was produced only when the taste and odor stimuli were presented simultaneously (Experiment 3) and the preconditioned taste aversion was intact at compound conditioning (Experiment 4). Pairing a novel odor with a preconditioned taste was not sufficient to condition an aversion to odor (Experiment 5), although other results implicated a role for an association between odor and taste in the odor augmentation effect (Experiment 6). The present results have implications for current models of taste + odor interactions in flavor-aversion conditioning.

New techniques for the evaluation of bladder function.

The various techniques available for the dynamic assessment of bladder function, including uroflowmetry, cystometrography, urethral closure pressure profiles, and sphincteric electromyography are presented. All of these approaches are discussed in terms of the indications for their use and their functions in evaluation.

Immunocytochemical localization of an immunoconjugate (antibody IgG against prostatic acid phosphatase conjugated to 5-fluoro-2'-deoxyuridine) in human prostate tumors.

Current chemotherapeutic and/or endocrine treatments for adenocarcinoma of the prostate (CaP) do not selectively target neoplastic prostate cells. Therefore, new approaches are needed to improve treatment for prostate tumors. We hypothesized that because of the specific binding of antibody immunoglobulin G (IgG) against human prostatic acid phosphatase (PAcP), PAcP-IgG could function as a carrier protein for the conjugated chemotherapeutic drugs and that the immunoconjugate would then selectively localize (bind) to epithelial cells of human prostate tumors, but not to epithelial cells of other solid organs. Our objective was to test this hypothesis using human prostate, colon, and kidney tissue samples and human prostate pieces incubated in short-term organ culture. We used derivatives of 5-fluorouracil labeled with fluorescein isothiocyanate (FITC) and rabbit anti-PAcP-IgG tagged with CY3/rhodamine alone or as an immunoconjugate. Localization of PAcP-IgG alone and the immunoconjugate in prostate produced similar and specific immunostaining in prostate epithelial cells and their tumors, but not in epithelia of colon and kidney tissue sections or in prostate sections-treated with normal rabbit serum. Confocal microscopy showed co-localization of CY3 and FITC of the immunoconjugate in the same group of prostate epithelial cells and their tumors. Organ culture studies showed that human prostate tissue samples incubated with normal rabbit serum did not show any fluorescence whereas those cultured with PAcP-IgG immunoconjugate showed fluorescence in glandular epithelial cells. The later study also showed that in organ culture the immunoconjugate had penetrated and labeled prostate glands internal to the cut surfaces. Drug labeled with FITC did not localize specifically in the prostatic epithelium. Analysis of our data has shown that PAcP-IgG was needed for specific localization of the immunoconjugate in prostate glands. We conclude that PAcP-IgG was essential for delivery and binding of the drug in human prostate. This is the first report to show that PAcP-IgG-5-Fu-2'-d-based immunoconjugate was selective and specific to epithelial cells of human prostate and its tumors, as revealed by organ culture, immunocytochemical, and confocal microscopic techniques.

The relationship of cathepsin B and stefin A mRNA localization identifies a potentially aggressive variant of human prostate cancer within a Gleason histologic score.

Cathepsin B (CB) is involved in degradation of extracellular matrix proteins during tumor progression in human solid organ tumors (such as colorectal, bladder, and breast cancers), including human prostate cancer. Its activities are regulated by endogenous inhibitors (such as stefins or cystatins). Increased expression of cathepsin B message, protein, and membrane association have been linked to malignancy, but there are very few studies of their mRNA expression in prostate cancer using in situ hybridization techniques. Our objective was to determine the relationship of CB and stefin A (cystatin A) mRNA localization to the Gleason grading system for histologic scores in the hope of distinguishing aggressive and less aggressive variants of prostate cancer. We used a 25-base biotinylated oligonucleotide CB cDNA antisense probe to localize CB message and a 27-base biotinylated oligonucleotide stefin A cDNA antisense probe to localize stefin A message. Prostate samples from 41 prostatectomy patients were collected along with their pre-surgery serum PSA levels and clinical stage of the disease. Sections prepared from frozen prostate tissue samples were hybridized with the CB and stefin A and control pBR 322 probes using techniques reported by Sinha et al. [1] and their distribution quantitated by an image analysis system. Prostate sections treated with RNAse before hybridization or incubated with the pBR 322 control probe showed little or no reaction products, confirming that localization of CB and stefin A probes was specific. In prostate cancer, the reaction products were found in neoplastic and invasive cells and occasionally in stromal cells. The ratios of CB to stefin A were similar in normal prostate and benign prostatic hyperplasia (BPH) whereas they varied consistently within and between Gleason histologic scores for prostate cancer. These variations showed three localization patterns; namely, prostate cancers with higher levels of CB than stefin A, lower levels of CB than stefin A, and similar levels of CB and stefin A. All three patterns and ratios for CB and stefin A were found in prostate samples (22/41) represented by the Gleason histologic score 6 tumors. In these tumors, serum PSA levels ranged from 1 to 78 ng/ml and prostate cancers showed B, C, and D clinical stages. There was no correlation of CB/stefin A ratio and serum PSA values or clinical stage in a limited number of prostate cancer cases. Our data showed that there were prostate cancer cases within Gleason histologic scores which expressed high, similar, and low levels of CB when compared to stefin A. We postulate that prostate cancer cases showing higher levels of CB compared to stefin A probably represent an aggressive variant of this cancer within any one Gleason histologic score. If this is the case, aggressive variants of prostate cancer would occur within Gleason scores 3 to 10 even though higher scores are usually considered more aggressive forms of prostate cancers. Since our study is based upon a very limited number of frozen prostate samples, we emphasize that a larger series of archival prostate cancer samples along with their survival data should be analyzed to establish any relationship of CB/stefin A ratio and aggressive variants of this cancer. Therefore, our conclusion is tentative. Our study provides a partial explanation for differences in the clinical course of prostate cancer in patients. This is the first study to show that determination of CB and stefin A mRNA ratios may lead to identification of aggressive and less aggressive variants of prostate cancer within a Gleason histologic score.

Laboratory automation of high-pressure liquid chromatography.

An automated system for high-pressure liquid chromatography was developed. The system is built around commercial modules wherever possible, modified to varying degrees. An automatic sampler, a sample pump, a high-pressure sampling valve, a recorder with an integrator, and a high-pressure liquid chromatograph comprise the commercial instruments. Relays, solenoid valves, and timers control chromatographic events, i.e., duration of sampling and rinse, mobile phase pump refill, sample injection, and chromatographic time. The automated system is dependable over long periods of unattended operation. With the 40-sample capacity of the sample tray and the last sample stop capability, the automated system produces, for example, 40 20-min chromatograms in approximately 13 hr of unattended operation. Data demonstrate the reliability and utility of the system.

Combining sensory reinforcement and texture fading procedures to overcome chronic food refusal.

Previous research has demonstrated behavioral programs to be effective in treating children with selective food preferences. However, there are few examples of interventions for the child displaying almost total food refusal. The present program combined sensory reinforcement and texture fading procedures to treat a 4-year-old deaf, visually impaired child who only consumed milk and, occasionally, pureed baby food. Sensory reinforcement consisted of the contingent presentation of light and rocking motion following consummatory responses. Texture fading entailed gradually increasing food composition. Results indicated that treatment was associated with substantial increases in the consumption of solid foods. Effects were maintained following the withdrawal of sensory reinforcement and with meals presented outside of the original treatment settings. Various features of the program are highlighted and discussed.

Objective noninvasive evaluation of benign prostatic hypertrophy.

An objective noninvasive procedure has been developed to evaluate the urodynamics of benign prostatic hypertrophy. The test uses pneumatic occlusive cuffs similar to those used for blood pressure measurements and the electrical engineering concepts of open-circuit, short-circuit, and transient response measurements. The cuff is first inflated to measure pressure, then rapidly released, yielding the transient response and subsequent unimpeded flow. From the pressure and flowrate recordings as functions of time, objective evaluations of bladder strength and urethral obstruction are extracted.