Detection and clearance of prostate cells subsequent to ultrasound-guided needle biopsy as determined by multiplex nested reverse transcription polymerase chain reaction assay.

OBJECTIVES: To determine if circulating prostate cells are detectable subsequent to transrectal ultrasound (TRUS)-guided biopsy, and if so, whether cells remain in circulation for up to 4 weeks. METHODS: Blood samples were drawn from 90 patients with elevated serum prostate-specific antigen (PSA) levels and/or abnormal digital rectal examination. Two samples were drawn from all patients immediately prior to TRUS and 30 minutes postbiopsy. Blood samples were also obtained 1 week postbiopsy from 83 patients, and 1 month postbiopsy from 61 patients. Multiplex nested reverse transcription polymerase chain reaction assay (RT-PCR) for PSA and prostate-specific membrane antigen (PSM) was performed on total ribonucleic acid (RNA) from each sample. Results were reported as positive if at least one of the targets was detected. RESULTS: Of 45 patients with biopsy-proven adenocarcinoma, 22 were RT-PCR positive prebiopsy and all remained positive 30 minutes postbiopsy. Of 23 patients with adenocarcinoma who were RT-PCR negative prebiopsy, 5 (22%) converted to positive 30 minutes postbiopsy (P < 0.001). Four of these 5 patients returned to negative after 1 week or 1 month. Of 45 patients without cancer at biopsy, 32 were RT-PCR negative prebiopsy and 6 (19%) converted to positive 30 minutes postbiopsy (P < 0.001). Although four of six available samples were still positive at 1 week, all four samples available 1 month postbiopsy were negative. CONCLUSIONS: Detection of circulating prostate cells subsequent to biopsy occurred in 11 of 55 (20%) previously RT-PCR negative patients, a proportion twice that reported in the literature. We attribute this higher proportion to the simultaneous detection of PSA and PSM mRNA in our multiplex assay. Conversion rates were similar in patients regardless of biopsy result. Testing of serial postbiopsy blood demonstrates clearing of these cells by 4 weeks in most patients.

Toxicity of ultrasound in mice: neonatal studies.

Pregnant mice were exposed to ultrasound (continuous wave, 2 MHz) on Day 8 of gestation to determine effects on the progeny. The most significant finding was a decrease in mean uterine weight of the female progeny. The thresholds for this effect were 140 s at 0.5 W/cm2 and 60 s at 1 W/cm2, which were below the thresholds previously reported for other effects in mice. We suggest that this indicates a delay or impairment of maturation of the mice exposed in utero. Exposure of the dams to spatial average intensity of 1 W/cm2 for 40 and 60 s had no effect on body weight of the progeny, compared with sham-treated controls. In this experiment the body weights of progeny from sham-treated controls were significantly lower than those from untreated controls on Days 10, 17, and 25 of age. After exposure in utero to 0.5 W/cm2 for 180 s, statistically significant decreases in mean body weights of the neonates were observed, but only on Day 25 of age, in both sexes compared with sham-treated controls. At necropsy at Day 25 of age, neonatal organ weights relative to body weights were not significantly affected for the thymus in either sex or for the seminal vesicles and tests ion comparison with sham-treated controls.

Mutagenicity of 2- and 3-carbon halogenated compounds in the Salmonella/mammalian-microsome test.

Short-chain, 2- and 3- carbon halogenated hydrocarbons were tested for mutagenicity for Salmonella typhimurium strain TA 100 both with and without the presence of S-9. Without exception, all brominated derivatives were more mutagenic than the chlorinated derivatives, usually by a substantial order of magnitude. 2-Fluoroethanol, the only fluorinated compound tested, showed little or no mutagenic activity up to 100 micromole per plate concentration. Two highly purified propane derivatives containing a halogen atom on each of the three carbons showed little or no direct mutagenic activity. A third trihalogenated compound with a halogen atom on each carbon atom showed some direct mutagenic activity, probably due to impurities. However, all three trihalogenated compounds were highly active mutagens following S-9 activation. The presence of a double bond in the case of 1, 2, 3-trichloropropene resulted in a higher level of direct mutagenic activity than 1, 2, 3-trichloropropane, but activation with S-9 resulted in a further increase in mutagenic activity with the former compound. On the other hand, S-9 caused a substantial decrease in mutagenic activity of most compounds containing a double bond. With the presence of an alcoholic group in a compound, the addition of S-9 caused variable responses, increasing the number of his+ revertant colonies due to 2, 3-dibromopropanol but had little or no effect with five other compounds containing an alcoholic group. Evidence is also presented that the position of a double bond in relation to the halogen atoms may influence mutagenic activity.

Effects of ultrasound on the mouse exposed at different stages of gestation: acute studies.

At five stages of gestation, mice were exposed dorsally to ultrasonic irradiation at 1 W/cm2 (spatial and temporal average), 2 MHz, and 34 and 37 degrees C (temperature at beginning of exposure). The results indicated that ultrasonic exposure up to at least 100 s is not hazardous to a pregnant mouse or its offspring in utero under the conditions of our experiment. Thresholds for both undesirable and lethal effects on the dam and its embryos or fetuses appeared between 100 and 200 s. Temperature rises in the uterus to more than 40 degrees C and similarity of results obtained with continuous-wave and burst modes of irradiation suggest a thermal mechanism as one cause of these effects. Preliminary conclusions are that thresholds for both tissue damage to the dam and deleterious effects on pregnancy were similar; however, they occurred at spatial average intensity and exposure levels far in excess of those used in clinical diagnostic medicine.

Mutagenicity of halogenated and oxygenated three-carbon compounds.

Four structurally related three-carbon compounds, known for their antifertility activity in the male, and the brominated derivatives of two of these compounds were tested for mutagenic activity by the Salmonella typhimurium test of Ames et al. In the presence of strain TA-100, a base-pair substituion detector strain, 1,2-dibromo-3-chloropropane (DBCP), was the most active compound tested but required enzymatic conversion by 59 microsomal preparation to an active mutagen. Three of these compounds containing an epoxide group-epichlorohydrin, epibromohydrin, and glycidol-were highly active direct mutagens, not requiring 59 for activation, alpha-Chlorohydrin was the least active compound tested; alpha-bromohydrin was 40 times more active than its chlorinated analog. Epibromohydrin was only slightly more active than epichlorohydrin, but both were highly active. With both of the halogenated epoxides, 59 preparation caused a substantial decrease in mutagenic activity at every concentration tested. All six compounds showed dose-related responsiveness for the base-pair substitution detector strains used. However, they were relatively inactive against the frameshift detector strain of S. typhimurium, TA-98. Glycerol, propylene glycol, and n-propanol, which are also three-carbon compounds containing one or more hydroxy groups, were inactive when trested at high concentrations with strain TA-100.

Comparison of segmented polyether urethane with polyethylene IUDs in rabbits.

A segmented polyether urethane IUD was compared with a polyethylene IUD in rabbits. The contraceptive efficacy of urethane IUDs was excellent. Moreover, the purulent slippery deposit present with the polyethylene IUDs and observed by Davis et al. (1) was absent. Our studies indicate that the polyether urethane IUDs have a high degree of antifertility activity in rabbits and these IUDs have reduced inflammatory response, based on leucocytic infiltration and tissue debris in the uterine lumen. It is suggested that polyether urethane IUDs not requiring copper or other medication can be designed for high contraceptive efficacy, intrauterine compatibility and with the necessary rigidity for proper uterine retention in humans.

Postpartum survival of mice exposed in utero to ultrasound.

We found no credible enhancement of neonatal mortality in mice exposed in utero on Day 8 of gestation to ultrasound at 0.44 W/cm2 (+/- 20%) spatial average intensity, 2 MHz, continuous wave, for 60--180 s in a 37 degrees C water bath. This result is compared with the adverse effect on neonatal survival reported by Curto [in Ultrasound in Medicine (Plenum, New York, 1976), Vol. 2, pp. 535--536] for exposure on Day 14 of gestation at about 0.125 to 0.5 W/cm2, 1MHz, continuous wave, for 180 s in a 30 degrees C bath. It is concluded that a temporary reduction in fetal viability, lasting from 4 days to less than 10 days after exposure, could account for both sets of data. It is also evident from these data that improved methods of determining intensities within mammalian tissues are highly desirable.

Studies with timed-pregnant squirrel monkeys (Saimiri sciureus).

Timed-pregnant monkeys were produced in a large nonhabituated colony of Saimiri sciureus of Bolivian origin. In a colony of 373 females and 40 males, 277 females (74%) were considered to be inseminated, based on microscopic observation of sperm and/or detection of a coagulum (plug) in the vagina. Forty-six full-term progeny were delivered. The mean gestational period was 152.5 days (SD = 3.9 days). For continuous cohabitation, the median time to insemination was five days, with 75% of inseminations occurring within eight days. Pregnancy evaluation (mouse bioassay) indicated a high level of resorptions within the first 50 days of gestation. This may help explain the low birth rates reported for other nonhaibuated colonies.

Investigation of the occurrence of a contraceptive tetrapeptide in hamster embryos.

The synthetic tetrapeptide H-Thr-Pro-Lys-OH, the extract of two-cell hamster embryos, and the oviductal contents devoid of embryos failed to show any in vivo antifertility activity. In addition, the tetrapeptide was not found in the biologic preparations of embryo extract and remaining oviductal contents. These findings are not in agreement with those of previous reports.

Pregnancy diagnosis in squirrel monkeys: hemagglutination test, radioimmunoassay, and bioassay of chorionic gonadotropin.

The Nonhuman Primate Pregnancy Test may be useful for diagnosis of pregnancy in squirrel monkeys between 40 and 60 days in pregnancy. However, single determinations have an inherent 10% risk of false negative responses caused by low chorionic gonadotropin levels; thus, initial negative test responses should be followed within one week by an independent confirmatory test. Preliminary results with this hemagglutination inhibition test compare favorably with bioassay and may be useful, in conjunction with conventional uterine palpation, for diagnosis of pregnancy in squirrel monkeys.