A solid-phase assay for the detection of anti-sperm antibodies.

PROBLEM: ELISA is an ideal assay method for a large-scale screening of anti-sperm antibodies among a large number of infertile males. However, conventional ELISA with whole spermatozoa needs time-consuming steps of centrifugation. METHOD: A solid-phase assay used for detecting anti-sperm antibodies was established. This assay is suitable not only for detecting circulating anti-sperm antibodies of IgG, IgM, and IgA subclass simultaneously but also for screening hybridomas secreting anti-sperm monoclonal antibodies (mAbs). The microtiter plates, on which solubilized sperm antigens are fixed, can be stored at -80 degrees C for up to six months without losing reactivity with anti-sperm antibodies. RESULTS: Using this assay, 53 sera (13 were proven positive and 40 were proven negative for sperm agglutination antibody) were tested. Although the false-negative rate was 0%, the false-positive rate was 32%. One thousand one hundred sixty-five supernatants from hybridomas constructed with splenocytes of mice who were hyperimmunized with human sperm and nonsecreting myeloma cells were tested by this solid-phase assay and two anti-sperm mAb secreting clones were selected and established. CONCLUSIONS: It is recommended that for research work this assay could be used for the first screening of the hybridoma secreting anti-sperm mAb, and for clinical use this assay might be suitable for the first screening of sera of infertile patients. However, conventional bioassays should follow to confirm the biological meaning of the positivity.

New Cavitron system (CUSA/CEM): its application for kidney surgery.

A new Cavitron Ultrasonic Surgical Aspirator system (CUSA/CEM), which contains electrocautery function at the hand piece, was used for partial nephrectomies in dogs without clamping the renal artery. The advantages of this new device (CUSA/CEM) over the original CUSA are: (1) it enabled more precise resection of the tissue in a dry field, (2) shorter operating time, and eventually (3) helpful for reduction of blood loss. The microscopic examination of the resected kidney revealed minimal reaction and damage to the adjacent kidney tissue as compared with the effect by electrocautery or microwave tissue coagulator.

Human prostatic cancer cell derived growth factor stimulating fibroblasts and its mechanisms.

A human prostatic cell line JCA-1 has been shown to release into culture medium an activity that promotes the proliferation of murine fibroblast 3T3 cells. The presence of additional serum-free conditioned culture supernatant from JCA-1 cells accelerates the 3T3 growth rate as determined by cell counts, [3H]thymidine uptake into DNA and colony formation in soft agar gel. The growth activity was determined not to derive from culture medium components. Treatment of JCA-1 cells with RNA synthesis inhibitor actinomycin D abrogated the activity, indicating that protein synthesis is required for the presence of this activity from JCA-1 cells. The molecular mass of this prostatic cell derived growth factor (PRGF) was larger than 10,000 KDa, and its interaction with fibroblasts resulted in an approximate 27 percent increase in replicating cell cycle phases S and G2M with a reciprocal decrease of G1 cells. These data indicated a mechanism causing a more rapid entry of cells into replication. The interaction between prostatic cancer cells and related tissues and their relation to pathogenesis is reviewed.

Application of microwave tissue coagulation in partial nephrectomy.

Microwave tissue coagulation was used during partial nephrectomy in 10 mongrel dogs, without clamping the renal artery. There were no major complications, such as retroperitoneal hematoma, abscess formation, or macroscopic infarction of the kidney tissue related to this new procedure. The advantages of microwave coagulation are reduced blood loss, shorter operative time, and minimal risk of vascular injury.

Maintenance intravesical bacillus calmette-guerin for superficial transitional cell carcinoma of bladder. A retrospective study.

Eighteen patients with recurrent and/or multifocal superficial transitional cell carcinoma of the bladder who were rendered tumor-free by transurethral resection and were then treated with either a single or second six-week course of induction Bacillus Calmette-Guerin (BCG) therapy, followed by maintenance therapy, were retrospectively reviewed. A 73 percent complete response rate was achieved in those patients treated prophylactically, while a 70 percent complete response rate was observed in patients treated for carcinoma in situ (CIS) with an average follow-up of twenty-nine months. Maintenance therapy may be warranted in those patients able to tolerate it without significant side effects.

Establishment of new human prostatic cancer cell line (JCA-1).

The establishment of a new human prostatic cancer cell line is described. This cell line was derived from a poorly to moderately differentiated prostatic adenocarcinoma. It has been maintained in tissue culture for fourteen months and has been passed fifty-two times. This cell line has an ability to form colonies in soft agar suspension cultures, and also is transplantable to nude mice. Tumors grown in nude mice revealed a poorly differentiated adenocarcinoma with positive PSA staining. Acid phosphatase activity was detected in freeze-thawed cells by enzymatic assay. A karyotype analysis demonstrated aneuploidy with a model chromosomal number of 69 and six marker chromosomes.

Expanding the role of the ureteroscope.

Although the rigid ureteroscope was designed exclusively for the diagnosis and treatment of pathological conditions of the ureter, its smaller diameter and extended length allow for its increased use in endoscopic procedures that cannot be performed successfully with a cystoscope. The indications can be divided into 3 categories: 1) difficult ureteral catheterizations, 2) augmented bladders and diverticula, and 3) narrow, strictured urethras and bladder necks. Several cases from each category are presented to illustrate the expanded role of the rigid ureteroscope in daily urological practice.

Benign soft-tissue neoplasm simulating sarcoma of spermatic cord.

A large, painless, slow-growing mass arising in the scrotum of a twenty-three-year-old man, which proved to be benign myxoma, simulated a primary sarcoma of the spermatic cord. Only 1 case of myxoma involving the spermatic cord has been reported previously. It is extremely important that multiple microscopic sections of the specimen be examined carefully so that the proper tissue diagnosis can be made. A false diagnosis of malignancy may lead to treatment that is not indicated and that has a high morbidity.

Immunosuppressive factor derived from renal cancer cells.

An immunosuppressive factor derived from conditioned medium of a human renal cancer cell line, KU2, was analyzed, using mitogen-activated normal peripheral blood lymphocytes. Addition of conditioned medium to lymphocyte cultures resulted in suppression of [3H]-thymidine incorporation to DNA of lymphocytes in a dose-dependent manner. This effect was cytostatic and reversible. This factor inhibited lymphocyte activation stimulated by mitogen at an early stage, and it also suppressed Interleukin-2 production by activated lymphocytes. This factor was non-dialyzable and heat sensitive at 56 degrees C. DNA replication of the cells was necessary for the production of this factor.

Pseudodiverticulum of the bladder: an unusual complication of pelvic trauma.

Investigation of frequent urinary tract infections 2 months after an accident in which a man sustained pelvic fractures and a head injury revealed a large pseudodiverticulum between the left bladder wall and pelvis, and extending inferiorly and medially through the separated symphysis pubis into the upper left thigh.

Fibroepithelial polyp associated with congenital ureteral diverticulum: report of 2 cases.

We report 2 cases of a single fibroepithelial polyp in conjunction with ureteral diverticula. Ureterorenoscopy is valuable in the identification of the benign nature of these lesions preoperatively to avert unnecessary nephroureterectomy. The etiology of fibroepithelial polyps remains unknown, although many theories have been proposed. Recent evidence has suggested that these lesions may be a developmental anomaly. Congenital ureteral diverticula also are thought to be of developmental origin. We hypothesize that fibroepithelial polyps and congenital diverticula of the ureter are part of a spectrum of the same developmental anomaly.

Value of proximal diversion and ureteral stenting in management of penetrating ureteral trauma.

Cases of penetrating ureteral trauma (17 gunshot wounds and 4 stab wounds) seen at two New York City hospitals over a nine-year period were reviewed. Early urologic complications (defined as urine drainage from the wound for greater than 2 weeks or need for a second operation) occurred in 50 percent of patients with a proximal ureteral injury but were less common when a nephrostomy and stent were used in the repair. A similar beneficial effect of stenting was seen in published cases, with a 91 percent complication rate for unstented repairs and a 15 percent rate when stenting and nephrostomy drainage were both used. The effect of stenting on midureteral repairs was less obvious, but the rate of urologic complications was lower in stented cases. None of the patients with distal ureteral injuries suffered a urologic complication. Stenting with and without a nephrostomy tube also produced good results in 2 patients with proximal ureteral injuries diagnosed late. We conclude that repair of penetrating ureteral injuries should include stenting and nephrostomy tube drainage in cases of proximal injuries, as well as generous debridement and water-tight closure. Midureteral injuries accompanied by gastrointestinal, pancreatic, and major vascular injuries should be stented and proximal diversion considered when prosthetic materials are used for vascular repairs.

Multiple penile horns: case report and review.

We report a case of unusually large multiple penile horns following removal of condylomata acuminata. Penile horns can grow rapidly, although malignant degeneration is uncommon. Wide excision with deep biopsy of skin at the base of the lesion probably is appropriate treatment.

A solid-phase immunoassay to screen anti-HLA monoclonal antibodies.

A solid-phase assay to detect anti-HLA monoclonal antibodies was developed. In this assay microtiter plates are coated with antigens solubilized from cultured lymphoid cells by sonication and then incubated with anti-HLA monoclonal antibodies. The antigen-antibody interaction is indicated by the development of color following the addition of peroxidase-conjugated anti-mouse Ig xenoantibodies and its substrate. The assay is rapid since it does not require centrifugations during the washing steps. Furthermore the assay is simple, reproducible and suitable to screen large numbers of samples and to detect antibodies recognizing determinants not exposed on the membrane of viable cells. The sensitivity of the assay is influenced by the pH of the buffer used to coat plates with antigens, by the number of cells used to prepare soluble antigens, by the incubation time of antigen preparations with plates and by the incubation time of antibody preparations with antigen-coated plates. Titration of anti-HLA monoclonal antibodies with known specificity and screening of hybridomas generated with splenocytes from mice immunized with cultured human lymphoid cells indicate that the sensitivity of the solid-phase assay is similar to that of the ELISA with lymphoid cells.

Bladder neck obstruction secondary to Brunn's cyst.

A large solitary Brunn's cyst caused a ball-valve type of obstruction of the bladder neck in a 29-year-old man without endoscopic or histological evidence of bladder inflammation. Unroofing and drainage restored normal voiding. The histopathological features of Brunn's nests are reviewed.

Ultrasonic aspiration of prostate, bladder tumors, and stones. Progress report.

The projected advantages of ultrasonic surgery on prostate, bladder tumors, and stones are: (1) precise histologic diagnosis without microscopic distortion as occurs with electrocautery; (2) continuous uninterrupted surgery; (3) improved hemostasis; (4) protection of external sphincter; (5) easy surgical technique to learn; (6) adaptability for video TUR; (7) can remove bladder stones and perform TURP at one sitting; (8) no fear of bladder wall injury during lithotripsy since stone is sucked to aspirator for dissolution; (9) no electrical burns or obturator reflex since electricity is not used to resect.

Correlation of heterogeneity index score (HIS) of DNA content in bladder cancer recurrence.

Automated flow cytometry (FCM) has been used to monitor the effects of therapy and progression of human bladder carcinoma. We have previously reported a computer-based model which has shown a correlation to relative mean DNA content in cell populations analyzed by FCM. Two patients with superficial transitional cell carcinoma of the bladder were followed for several months. Using FCM the patients' tissue samples were examined and heterogeneity index scores (HIS) were determined. Both cases had recurrence. The first patient has increased in grade with a respective increase in HIS within four months (grade I----II, HIS 27.2----80.8). The second patient also has recurrence in three months both times with a grade I tumor and a score of 106.2. High scores reflect large aneuploid populations which have shown to recur. Since such a correlation exists HIS may not only offer a more objective technique with quantitative results to monitor patients but possibly can distinguish the degree of tumor malignancy.

Monocyte cytolytic factor in promoting monocyte-mediated lysis of bladder cancer cells by bacillus Calmette-Guerin.

The role of monocytes in cell-mediated cytolysis of bladder cancer cells was investigated. Human peripheral monocytes released a cytolytic factor which lysed T24 bladder cancer cells and a number of human tumor cells, but not normal lymphocytes or fibroblasts. After incubation of monocytes with bacillus Calmette-Guerin for 48 hr. in vitro, cytolysis of T24 cells was increased up to 56.7 +/- 4.1%. Treatment of monocytes with actinomycin D (an inhibitor of RNA transcription) reduced release of cytolytic factor from 27.3 +/- 5.7% cytolysis to 4.5 +/- 1.4% (p less than 0.05). The response to mitomycin C was different between lymphokines and monocyte cytolytic factor. The mouse monoclonal antibody against human recombinant tumor necrosis factor did not neutralize monocyte cytolytic factor. These results show that this monocyte cytolytic factor is distinct from lymphokines and tumor necrosis factor. The evidence that bacillus Calmette-Guerin increases release of monocyte cytolytic factor may be associated with anti-tumor activity of bacillus Calmette-Guerin in intravesical therapy for treatment of bladder cancer.

Flow cytometric analysis of relative mean DNA content of urogenital cancer cells in fresh and paraffin-embedded materials.

The relative mean DNA content calculation was performed by flow cytometry on single cell suspensions prepared from fresh and paraffin-embedded specimens of 10 patients with surgically resected urogenital cancer. Samples were processed by a modified method of Hedley et al. including two hours of pepsinizing time, ribonuclease digestion, and propidium iodide staining. The mean DNA content which is a quantitative description of flow cytometric characteristics was significantly correlated between the fresh and paraffin-embedded materials (n = 10, r = 0.869, p less than 0.01). This method allows for the objective, retrospective analysis of DNA content in relation to diagnosis and prognosis of urogenital cancer.

Same-session endourological removal of upper and impacted lower urinary tract stones with the aid of prone flexible cystoscopy.

Calculi were removed from the upper urinary tracts and the distal ureter in single sessions in 2 patients with the aid of prone flexible cystoscopy and a through-and-through stone basket.