Subcellular distribution of SERCA and calcium-activated ATPase in rabbit and human urinary bladder smooth muscle.

Previous studies have demonstrated that calcium storage and release from IP3-dependent sites in the sarcoplasmic reticulum play an important role in the contractile response of the rabbit urinary bladder to both field stimulation (mediated via neurotransmitter release) and bethanechol (direct muscarinic stimulation). In view of the importance of SERCA (see text) in urinary bladder smooth muscle function, we studied the distribution of SERCA by two methods; using Western blotting to quantitate the protein concentration and by enzyme analysis using thapsigargin to specifically inhibit SERCA. Rabbit and human samples of urinary bladder smooth muscle were homogenized and the homogenate separate into three particulate fractions by different centrifugation: the cell wall-nuclear, mitochondrial, and microsomal. The protein concentration of these three particulate fractions was determined and the SERCA protein level quantitated by Western blotting using SERCA-2 antibodies. The calcium ATPase activity was quantitated using standard enzymatic analysis and the thapsigargin sensitivity determined. The results demonstrated that (1) the concentration of SERCA was significantly greater in the microsomal fraction than in either of the other fractions for both rabbit and human bladder smooth muscle; (2) the enzymatic activities of both total calcium-activated ATPase and thapsigargin-sensitive calcium ATPase were evenly divided among the three fractions, and (3) the enzymatic activity of both total calcium-activated ATPase and thapsigargin-sensitive calcium ATPase of the rabbit exceeded that of the human. In conclusion, the distribution of SERCA and calcium ATPase of the rabbit bladder smooth muscle was similar to that in the human bladder smooth muscle, although activities in rabbit were significantly greater than those of human tissue.

Subcellular distribution of SERCA and calcium-activated ATPase in rabbit and human urinary bladder smooth muscle.

Previous studies have demonstrated that calcium storage and release from IP-3-dependent sites in the sarcoplasmic reticulum play an important role in the contractile response of the rabbit urinary bladder to both field stimulation (mediated via neurotransmitter release) and bethanechol (direct muscarinic stimulation). In view of the importance of SERCA in urinary bladder smooth muscle function, we studied the distribution of SERCA by two methods: using Western blotting to quantitate the protein concentration and by enzyme analysis using thapsigargin to specifically inhibit SERCA. Rabbit and human samples of urinary bladder smooth muscle were homogenized and the homogenate separated into three particulate fractions by differential centrifugation: nuclear-cell wall, mitochondrial, and microsomal. The protein concentration of these three particulate fractions was determined and the SERCA protein level quantitated by Western blotting using SERCA-2 antibodies. The calcium-ATPase activity was quantitated using standard enzymatic analysis and the thapsigargin sensitivity determined. The results demonstrated that: (1) the concentration of SERCA was significantly greater in the microsomal fraction than in either of the other fractions for both rabbit and human bladder smooth muscle; (2) the enzymatic activities of both total calcium-activated ATPase and thapsigargin-sensitive calcium ATPase were evenly divided among the three fractions, and (3) the enzymatic activity of both total calcium-activated ATPase and thapsigargin-sensitive calcium ATPase of the rabbit exceeded that of the human. In conclusion, the distribution of SERCA and calcium-ATPase of the rabbit bladder smooth muscle was similar to that in the human bladder smooth muscle, although activities in rabbit were significantly greater than those of human tissue.

Mini-laparotomy pelvic lymph node dissection minimizes morbidity, hospitalization and cost of pelvic lymph node dissection.

PURPOSE: We compared efficacy, morbidity and cost of mini-laparotomy pelvic lymph node dissection and laparoscopic pelvic lymph node dissection. MATERIALS AND METHODS: A total of 40 patients underwent mini-laparatomy, laparoscopic or standard pelvic lymph node dissection during a 24-month period. Nodal yield, complications, hospitalization and postoperative analgesic requirements were retrospectively evaluated. Operative expenses and the cost of postoperative hospitalization were standardized to a base cost for comparison. RESULTS: Mini-laparotomy pelvic lymph node dissection has an operative time (90 minutes) and nodal yield (9) similar to those of standard pelvic lymph node dissection, and morbidity and postoperative hospitalization (1.3 days) are comparable to those of laparoscopic pelvic lymph node dissection. The expense of the mini-laparotomy procedure is approximately 50% that of the laparoscopic procedure due primarily to the prolonged operative time (190 minutes) and disposable instrument costs ($665) of laparoscopic pelvic lymph node dissection. CONCLUSIONS: Mini-laparotomy pelvic lymph node dissection competes successfully with laparoscopic pelvic lymph node dissection in terms of efficacy and morbidity at significant cost savings.

Azlocillin and mezlocillin concentration in human prostatic tissue.

The concentrations of azlocillin and mezlocillin in human prostatic tissue obtained by transurethral resection or enucleation were measured after two 2.0-gram doses of either drug. The average plasma concentration of azlocillin and mezlocillin at time of tissue sampling was 64.9 and 36.3 micrograms/ml, respectively, and tissue concentration at the time of sampling was 22.9 micrograms/g for azlocillin and 9.4 micrograms/g for mezlocillin. The plasma/tissue concentration ratio for mezlocillin was 0.25 and for azlocillin 0.35. Concentrations of mezlocillin in tissue obtained by transurethral resection were similar to those obtained by enucleation. Azlocillin and mezlocillin in appropriate doses achieve a concentration in human prostatic tissue above the inhibitory concentration for common bacterial pathogens.

Single ectopic ureter: unusual presentation in adult male.

We present a case of an unusual and late presentation of a single ectopic ureter in an adult male. The patient presented with gross painless hematuria. The left ureter draining a dysplastic kidney terminated in the ipsilateral seminal vesicle.

Penile xenon (133Xe) washout: a rapid method of screening for vasculogenic impotence.

The radioactive inert gas xenon (133Xe) is a well-established isotopic indicator used to assess vascular status in many organ systems. We employed xenon-133 to evaluate male impotence. Xenon-133 was injected subcutaneously at the level of the coronal sulcus in the detumescent state. Using the gamma camera, sequential images were obtained and computer-generated curves calculated. The clearance time for 50 per cent washout of the injected 133Xe (T1/2) was then calculated for each patient, as well as a control group. Preliminary findings indicate a correlation with such established techniques of evaluating erectile impotence as history, physical examination, penile pulse Doppler tracings, and brachial-penile blood pressure index. The xenon-133 washout study was a rapid, minimally invasive, reproducible, and cost-effective method of screening those impotent patients for vasculogenic etiology of their erectile impotence. We recommend the addition of this method to the surgeon engaged in the care of impotent males.

Comparison of acid phosphatase isoenzymes of human seminal fluid, prostate, and leukocytes.

We used ion-exchange column chromatography and electrophoresis on polyacrylamide gel to compare the acid phosphatase isoenzymes of prostate and leukocytes. The major isoenzyme of the prostate is band 2A; only a trace of band 2B was observed. However, the major isoenzymes of leukocytes are band 4 and band 2B, and only a small amount of band 2A was observed. The three isoenzymes isolated from leukocytes or prostate gland react to the antiserum prepared against the aicd phosphatase isoenzyme of seminal fluid. Acid phosphatases of leukocytes other than the three isoenzymes mentioned above did not interact with the antiserum.

Concomitant germ cell tumors in monozygotic twins.

Simultaneously occurring embryonal cell tumors in a 38-year-old set of monozygotic twin brothers are reported. This is the eighth documented case of testicular cancer in twins and it is unique since it is the first report to describe bilateral testicular tumors in 1 of the twin brothers. The role of genetics in the development of certain cancers warrants further study. This documentation of testicular tumors in serologically proved identical twins is used as a basis for discussion of the role of heredity as a possible significant factor in germ cell tumor pathogenesis.