Monoclonal antibodies to Kaiso: a novel transcription factor and p120ctn-binding protein.

The POZ-zinc finger protein Kaiso belongs to a rapidly growing superfamily of BTB/POZ zinc finger transcription factors implicated in embryonic development and cancer. Kaiso interacts with the catenin p120(ctn), but the significance of the interaction remains unknown. Although p120(ctn) is normally found in association with E-cadherin at cell-cell junctions, it can translocate to the nucleus under certain circumstances. Thus, the p120(ctn)-Kaiso interaction may regulate transcriptional events, as has been described previously for the classical catenin, beta-catenin and the LEF1/TCF transcription factor. To facilitate further study of Kaiso and to determine the physiological relevance of its interaction with p120(ctn), we have generated and characterized a panel of five Kaiso-specific monoclonal antibodies (MAbs) that function in immunoblotting, immunoprecipitation, and immunofluorescence analyses.

Successful treatment of severe hemorrhagic cystitis with cystectomy following matched donor allogeneic hematopoietic cell transplantation.

We report a case of severe hemorrhagic cystitis complicating high-dose cyclophosphamide (CY), total body irradiation and allogeneic hematopoietic stem cell transplantation (HSCT). Supportive care with i.v. hydration, platelet transfusions, continuous bladder irrigation and aluminum irrigation of the bladder was ineffective and the patient developed multiple complications from hemorrhagic cystitis. His condition became critical with persistent bleeding, pulmonary edema and acute renal failure. These problems resolved following successful simple cystectomy and ileal conduit. The patient did not develop any acute or chronic complications following surgery. He is alive with a good quality of life and in third remission after receiving salvage chemotherapy 14 months after his transplantation.

Selective uncoupling of p120(ctn) from E-cadherin disrupts strong adhesion.

p120(ctn) is a catenin whose direct binding to the juxtamembrane domain of classical cadherins suggests a role in regulating cell-cell adhesion. The juxtamembrane domain has been implicated in a variety of roles including cadherin clustering, cell motility, and neuronal outgrowth, raising the possibility that p120 mediates these activities. We have generated minimal mutations in this region that uncouple the E-cadherin-p120 interaction, but do not affect interactions with other catenins. By stable transfection into E-cadherin-deficient cell lines, we show that cadherins are both necessary and sufficient for recruitment of p120 to junctions. Detergent-free subcellular fractionation studies indicated that, in contrast to previous reports, the stoichiometry of the interaction is extremely high. Unlike alpha- and beta-catenins, p120 was metabolically stable in cadherin-deficient cells, and was present at high levels in the cytoplasm. Analysis of cells expressing E-cadherin mutant constructs indicated that p120 is required for the E-cadherin-mediated transition from weak to strong adhesion. In aggregation assays, cells expressing p120-uncoupled E-cadherin formed only weak cell aggregates, which immediately dispersed into single cells upon pipetting. As an apparent consequence, the actin cytoskeleton failed to insert properly into peripheral E-cadherin plaques, resulting in the inability to form a continuous circumferential ring around cell colonies. Our data suggest that p120 directly or indirectly regulates the E-cadherin-mediated transition to tight cell-cell adhesion, possibly blocking subsequent events necessary for reorganization of the actin cytoskeleton and compaction.

Infusion of dendritic cells pulsed with HLA-A2-specific prostate-specific membrane antigen peptides: a phase II prostate cancer vaccine trial involving patients with hormone-refractory metastatic disease.

BACKGROUND: A phase II trial was conducted to assess the efficacy of infusions of dendritic cells (DC) and two HLA-A2-specific PSMA peptides (PSM-P1 and -P2). This report describes thirty three subjects with hormone-refractory metastatic prostate cancer without prior vaccine therapy history who were evaluated and reported as a group. METHODS: All subjects received six infusions of DC pulsed with PSM-P1 and -P2 at six week intervals. Clinical monitoring was conducted pre-, during, and post- phase II study. Data collected include: complete blood count, bone and total alkaline phosphatase, prostate markers, physical examination, performance status, bone scan, ProstaScint scan, chest x-ray, as well as assays to monitor cellular immune responses. RESULTS: Six partial and two complete responders were identified in the phase II study based on NPCP criteria, plus 50% reduction of prostate-specific antigen (PSA), or resolution in previously measurable lesions on ProstaScint scan. CONCLUSIONS: Over 30% of study participants in this group showed a positive response at the conclusion of the trial. This study suggested that DC-based cancer vaccines may provide an alternative therapy for prostate cancer patients whose disease no longer responds to hormone therapy.

Evaluation of phase I/II clinical trials in prostate cancer with dendritic cells and PSMA peptides.

BACKGROUND: A phase I trial involving patients with advanced prostate cancer was conducted to assess the safe administration of dendritic cells (DC) and HLA-A0201-specific prostate-specific membrane antigen (PSMA) peptides (PSM-P1 or -P2). Thirty-three of the phase I participants were subsequently enrolled in a phase II trial, which involved six infusions of DC pulsed with PSM-P1 and -P2 peptides. METHODS: Clinical monitoring was conducted up to 770 days from the start of the phase I study. Data collected included: complete blood count, bone and total alkaline phosphatase, prostate markers, physical examination, performance status, bone scan, ProstaScint scan, and chest X-ray, as well as assays to monitor cellular immune responses. RESULTS: Nine partial responders were identified in the phase II study based on National Prostate Cancer Project (NPCP) criteria, plus 50% reduction of prostate-specific antigen. Four of the partial responders were also responders in the phase I study, with an average response duration of 225 days. Their combined average total response period was over 370 days. Five other responders were nonresponders in the phase I study. Their average partial response period was 196 days. CONCLUSIONS: The responses observed in the phase I and II clinical trials were significant and of long duration. The partial-responder group included patients who continued to respond from phase I, as well as those who started to respond during the phase II trial.

Follow-up evaluation of prostate cancer patients infused with autologous dendritic cells pulsed with PSMA peptides.

BACKGROUND: We recently conducted a phase I clinical trial administering autologous dendritic cells pulsed with prostate-specific membrane antigen (PSMA) peptides to advanced prostate cancer patients. Participants were divided into 5 groups receiving 4 or 5 infusions of peptides alone (PSM-P1 or -P2; groups 1 and 2, respectively), autologous DC (group 3), or DC pulsed with PSM-P1 or -P2 (groups 4 and 5, respectively). Seven partial responders were observed. Follow-up evaluation of these responders is presented in this report. METHODS: Clinical monitoring for hematological studies and prostate markers was conducted up to 370 days from the start of the phase I study. Data collected include: lymphocyte, hematocrit, alkaline phosphatase, prostate-specific antigen (PSA), free PSA, and PSMA levels. RESULTS: Groups 4 and 5 (patients infused with DC pulsed with PSM-P1 or -P2) represented 5/7 responders. The length of response was between 100 days (1 patient) to 200 days or above (6 patients). Four patients still remained responsive at the end of the period of observation. CONCLUSIONS: The responses observed in this phase I clinical trial are significant and of long duration. Most of the responders were in treatment groups infused with DC pulsed with PSM-P1 or -P2, suggesting the requirement of both components for effective immunotherapy.

Bladder volume determination using a dedicated, portable ultrasound scanner.

Urethral catheterization, the standard method for measurement of bladder volume, is associated with patient discomfort plus the risks of urethral trauma and urinary tract infection. A portable ultrasound instrument that automatically determines bladder volume was used for 164 determinations and the digital readout correlated with the catheterized volume (r2 equals 0.79). This instrument is a noninvasive alternative to urethral catheterization for the determination of bladder volume in most patients.

Percutaneous nephrostomy tract incision using modified Otis urethrotome.

An Otis urethrotome has been modified to permit passage into the kidney over an 0.038-inch guidewire by drilling a 3/16-inch hole through the tip. I have used this instrument to aid in the performance of 36 nephrostomy tract dilations without significant complications. When compared with 20 Amplatz tract nephrostomy dilations, the new method was quicker, requiring 4.3 minutes versus 6.5 minutes. This instrument is especially useful for tracts where significant renal fascial scarring makes Amplatz dilation difficult. Ease of tract dilation, as well as decreased time to dilate, make this technique a useful addition to standard nephrostomy tract dilation techniques.

Intrathecal methadone and morphine for postoperative analgesia: a comparison of the efficacy, duration, and side effects.

A double-blind study of patients selected at random compared the analgesic and adverse effects of intrathecal methadone (1 mg) with those of intrathecal morphine (0.5 and 1 mg). The study was conducted on 30 patients who underwent major orthopedic or urologic surgery. The intrathecal opioid was administered at the end of surgery, and assessments began 1 h thereafter and continued for 20 h. Pain measurements, supplementary analgesia requirements, and adverse effects were recorded. Intrathecal morphine (0.5 and 1 mg) provided effective and prolonged analgesia. Methadone, however, was unable to ensure the same degree of analgesia; consequently, the median pain scores were consistently higher following methadone than morphine (0.5 and 1 mg) (P less than 0.05). The time to the onset of discomfort severe enough to require supplemental morphine was longer after intrathecal morphine than that following methadone (24 and 29 h with morphine 0.5 and 1 mg; 6.5 h with methadone; P less than 0.05). Respiratory depression (increases PaCO2) was not associated with methadone and morphine 0.5 mg but was common following morphine 1 mg (P less than 0.05). Facial pruritus was unique to intrathecal morphine. Urinary retention requiring bladder catheterization was more frequent following morphine than methadone, although this was not statistically significant. Nausea and vomiting were common to all groups. Intrathecal morphine (0.5 and 1 mg) provides superior postoperative analgesia to 1 mg methadone. Various explanations for the observed differences between the drugs are discussed, including the possibility that the dose of methadone used in the subarachnoid space was inadequate and that a larger dose might have produced an effect equal to that of morphine.

Case-control study of men with suspected chronic idiopathic prostatitis.

We studied prospectively 50 asymptomatic men (24 men from infertile couples and 26 normal volunteers) with no history of genitourinary infection and 34 men referred for symptoms of chronic prostatitis. Both groups were evaluated by urethral and prostatic secretion cultures for Chlamydia trachomatis, 4-glass prostatic localization cultures for facultative aerobic bacteria (Ureaplasma urealyticum, Mycoplasma hominis and selected fungi) and counts of prostatic fluid leukocytes. The men with symptoms of prostatitis had more than 1,000 leukocytes per mm. in prostatic secretions more often than did controls (11 of 27 versus 0 of 44, p less than 0.001). The concentration of Ureaplasma urealyticum in prostatic secretions was 1 log higher in prostatic fluid localization cultures than in first voided urine in 0 of 30 patients versus 13 of 50 controls (p equals 0.0014). Chlamydia trachomatis was not isolated from any patient or control. No other significant differences were found between patients and controls. We did not identify an infectious cause of chronic nonbacterial prostatitis.

Treatment of congenital penile curvature due to disparate corpora cavernosa by the Nesbit technique: a rule of thumb for the number of wedges of tunica required to achieve correction.

Treatment of pure congenital penile curvature due solely to asymmetry of the corpora cavernosa as described by Nesbit lacks precision. Intraoperative verification of Nesbit correction by artificial erection after wedge excision may be impossible because leakage through the sutured corporeal incisions prevents full erection. We present a rule of thumb to calculate the number of 1 cm. wedges of tunica albuginea to be excised to correct an individual deformity. The number of such wedges required is equal to the measured difference in centimeters between the lengths of the asymmetric, concave and convex surfaces of the affected penis, between the pubis and mid glans. Results of application of this principle in 6 patients are presented.

Transitional cell carcinoma of the urinary bladder: histologic clearance with combined 5-FU chemotherapy and radiation therapy. Preliminary results of a bladder-preservation study.

Fourteen patients with transitional cell carcinoma of the urinary bladder were treated with 4,000 cGy of pelvic irradiation concurrent with two 96-hour infusions of 5-fluorouracil (5-FU). Three weeks after completion of this regimen, patients underwent repeat cystoscopy and deep-muscle biopsy at the site of their original neoplasms. Eight of 14 (57%) had no tumor left in the biopsy specimen, and they received an additional course of chemotherapy and radiation therapy to a total dose of 4,400 cGy to the pelvis and 6,000 cGy to the bladder. Five of the 14 had residual tumor in the biopsy specimen (one did not undergo biopsy) and went on to planned cystectomy. Two of the five had no tumor in the cystectomy specimen. Overall, ten of the 14 patients (71%) have been downstaged to a condition of P0 (no tumor) following 4,000 cGy and two courses of 5-FU. Of eight patients with retained bladders, seven remain well at a median follow-up of 7 months. At a range of follow-up of 3-21 months and a median of 7 months, 13 of 14 patients remain tumor-free. This regimen results in a greater percentage of downstaging than conventional irradiation alone, and may allow bladder preservation for those with radiation therapy- and chemotherapy-responsive tumors.

Diagnosis of bacteriuria in men: specimen collection and culture interpretation.

Bacteriologic criteria for diagnosing urinary tract infections in men have not been well defined. For determination of the bacterial colony count in voided specimens that most accurately reflects bladder bacteriuria, culture results were compared for voided urine and bladder urine (obtained by suprapubic aspiration and urethral catheterization) from men with various genitourinary problems. Bladder bacteriuria was found in 36 (47.3%) of 76 sets of specimens from 66 individuals. Culture results of bladder specimens showed excellent agreement with those of clean-catch midstream-void and uncleansed first-void specimens (weighted kappa = 0.924 and 0.906, respectively). The criterion for clean-catch midstream-void specimens that best differentiated sterile from infected bladder urine was growth of greater than or equal to 10(3) cfu of one predominant species/ml; this definition had a sensitivity of 0.97 and a specificity of 0.97. Uncleansed first-void specimens were equally sensitive (0.97) but less specific (0.91-0.92) in detection of bacteriuria. Pyuria (greater than or equal to leukocytes/mm3) and irritative genitourinary symptoms showed modest correlations with bladder bacteriuria.

Diuretic renography in evaluating dilated upper urinary tract in children.

The diuretic renogram provides a previously unavailable noninvasive method for assessing and following urinary tract dilation of diverse cause, and defining true obstruction in children. Technetium-99m-DTPA (diethylenetriaminepentaacetic acid) is injected intravenously and a posteriorly placed gamma scintillation camera used to image the kidneys, ureters, and bladder. Furosemide is subsequently injected to stimulate a diuresis, and the washout pattern of isotope is monitored (time-activity histogram). Nonobstructive, obstructive, and poor renal function patterns were seen. Forty patients with varying degrees of hydroureteronephrosis were evaluated. Tracings were performed serially and compared with the clinical course in time, as well as with pressure flow studies and operative findings. All patients with a nonobstructed diuretic renogram did well, except for 1 boy in whom ureteropelvic obstruction developed later. Difficulty in evaluating the obstructive renogram pattern occurred only in some children with severe (grade V) hydronephrosis. Further standardization of the diuretic renogram with regard to time of diuretic injection, state of patient hydration, and regions of imaging may improve diagnostic accuracy.

Urinary calculous disease in Southeast Asian immigrants.

Recent immigrants from Southeast Asia accounted for 39 of 149 hospital admissions for treatment of urinary tract stones. Presumptive diagnosis of a urinary calculus was possible in only 19 per cent of the refugees compared with 60 per cent of other patients treated at the same hospital (p less than 0.005). Calculi in Southeast Asian immigrants were larger (p less than 0.001), and surgical procedures were required more often (p less than 0.05) than for other patients with calculi. Urinary stones should be considered a likely cause of abdominal or urinary tract complaints in recent immigrants from Southeast Asia.

Emphysematous pyelonephritis caused by Candida albicans.

We report a case of emphysematous pyelonephritis that was proved by histological examination and culture to be caused by Candida albicans. The fungal infection caused ureteral obstruction. Nephrectomy alone resulted in complete recovery.

A modification of the Otis urethrotome as an aid to fascial dilation of percutaneous nephrostomy tracts.

An Otis urethrotome has been modified by drilling a 3/64-inch hole through the tip to permit passage into the kidney over a 0.038-inch guide wire. This instrument has been used to aid in the performance of 12 nephrostomy tract dilations without significant complications. When compared to 10 Amplatz tract dilations, the modified Otis method required an average of 3.6 minutes versus 8.7 minutes to dilate the nephrostomy tract to 24F. Ease of tract dilation as well as decreased time to dilate make this technique a useful adjunct to standard nephrostomy tract dilation techniques.

Prostatitis and epididymitis.

Ascending spread of urethral pathogens may be the mechanisms of infection of the prostate and epididymis. Sexually transmitted organisms cause most epididymitis in young men. Although evidence is mounting to suggest that sexually transmitted organisms may cause prostatitis, data are insufficient at this time to base a therapeutic approach on this concept. The authors present approaches to the management of these two disorders.