Detection of thyroid peroxidase mRNA in peripheral blood of patients with malignant and benign thyroid diseases.

The aim of this study was to evaluate thyroid peroxidase (TPO) mRNA expression in peripheral blood of patients with benign and malignant thyroid disease. Included were 120 thyroid cancer patients, 85 patients with goitre or Graves' disease (GD) and 54 healthy volunteers. TPO mRNA expression was analysed in peripheral blood by nested RT-PCR. In cancer patients, RT-PCR results were compared with staging, grading and serum thyroglobulin (TG) measurement. TPO transcripts were detected in 7/10 (70%) patients with known metastases of thyroid cancer and in 39 of 110 (36%) patients without metastases (P<0.05), in 15/44 (34%) patients with goitre, in 17/41 (41%) cases with GD and in 4/54 (7.4%) subjects in the control group (P<0.05, controls vs all patients with thyroid disease). Among cancer patients without metastatic disease, RT-PCR results correlated positively with lymph node status (P=0.05), grading (P=0.01) and elevated serum thyroglobulin levels (P=0.03). This is the largest study investigating the use of the TPO-RT-PCR assay. Positivity in TPO-RT-PCR correlates significantly with metastatic disease in cancer patients and with the presence of thyroid disease in general. To date, TPO-RT-PCR cannot substitute for standard techniques in the diagnosis of local recurrence or metastatic spread in thyroid cancer patients. However, as results of TPO-RT-PCR correlate significantly with lymph node status, grading and serum TG measurements in patients with non-metastatic disease, TPO seems to be an interesting molecular marker to look at in follow-up studies.

3D interstitial HDR brachytherapy combined with 3D external beam radiotherapy and androgen deprivation for prostate cancer. Preliminary results.

BACKGROUND: Evaluation of feasibility, tolerance and efficiency for a new 3D interstitial HDR brachytherapy technique combined with 3D external beam radiotherapy and androgen deprivation for prostate cancer. PATIENTS AND METHODS: Between January 1997 and August 1998 we treated 35 patients with stage cT1-3 N0 M0 prostate cancer. Thirty-two patients with a follow-up of 12 to 28 months (median: 18 months) were evaluated. After ultrasound-guided transrectal implantation of 4 non-parallel needles, CT based 3D brachytherapy treatment planning ("Offenbach system") was performed. All patients received 4 fractions brachytherapy using a fractional dose of 5 or 7 Gy. Time between each fraction was 14 days. After brachytherapy 3D external irradiation followed up to 39.6 or 45.0 Gy. All patients received androgen deprivation, starting 2 to 19 months before brachytherapy, ending 3 months after 3D external radiotherapy. RESULTS: Posttreatment PSA levels dropped to < 1.5 ng/ml in 29/32 patients (91%). In 25 patients PSA levels were < 0.5 ng/ml, in 4 patients 0.5 to 1.5 ng/ml. In 2 patients we noted biochemical relapse. Transrectal implantation was very well tolerated. Grade 3 acute urinary toxicity occurred in 1 patient. We noted no Grade 2 or higher acute gastrointestinal toxicity. One patient developed a Grade 3 late urinary toxicity. No patient showed late gastrointestinal side effects. All 140 dose-volume histograms for 3D HDR brachytherapy were analyzed. CONCLUSIONS: The new 3D HDR brachytherapy technique, combined with 3D external irradiation and androgen deprivation, is a feasible, so far well-tolerated and effective treatment in the short-time follow-up of median 18 months.

Molecular detection of thyroglobulin mRNA transcripts in peripheral blood of patients with thyroid disease by RT-PCR.

The sensitive detection of circulating tumour cells in patients with differentiated thyroid cancer may precede the detection of relapse by other diagnostic studies--such as serum thyroglobulin-and thus may have important therapeutic and prognostic implications. We performed reverse transcription-polymerase chain reaction (RT-PCR) on blood samples from patients diagnosed with thyroid disease using two different RT-PCR sensitivities. Additionally, tissue specificity of TG mRNA-expression was determined using RNA extracts from 27 different human tissues. The lower limit of detection was 50-100 TG mRNA producing cells/ml blood using a 'normal' RT-PCR sensitivity and 10-20 cells/ml blood using a 'high' sensitivity. With the normal sensitivity TG mRNA was detected in 9/13 patients with thyroid cancer and metastasis, 63/137 patients with a history of thyroid cancer and no metastasis, 21/85 with non-malignant thyroid disease and 9/50 controls. With the high sensitivity TG mRNA was detected in 11/13 patients with thyroid cancer and metastasis, 111/137 patients with a history of thyroid cancer and no metastasis, 61/85 with non-malignant thyroid disease and 41/50 controls. Interestingly, using the normal RT-PCR sensitivity TG mRNA transcripts are specific for thyroid tissue and detectable in the peripheral blood of controls and patients with thyroid disease, which correlates with a diagnosis of metastasized thyroid cancer. However, with a high RT-PCR sensitivity, TG mRNA expression was found not to be specific for thyroid tissue and was not correlated with a diagnosis of thyroid cancer in patients. As a consequence, to date TG mRNA detected by RT-PCR in the peripheral blood cannot be recommended as a tumour marker superior to TG serum-level.

Elevated placental cytokine release, a process associated with preterm labor in the absence of intrauterine infection.

OBJECTIVE: To investigate the role of cytokines in normal term and preterm labor in the absence of intrauterine infection. METHODS: Cytokine (interleukin [IL]-1 beta, IL-6 and tumor necrosis factor-alpha [TNF-alpha]) release was estimated from placental and decidual cell cultures from 22 nonlaboring women at term with cesarean deliveries, 18 women with spontaneous labor at term, and 21 women with preterm labor (19-36 weeks gestation) who delivered vaginally or by cesarean, according to gestational age. Eight of 21 women delivering preterm had clinical evidence of intrauterine infection, and 13 were not infected. RESULTS: Placental cell cultures obtained from women with spontaneous term labor released significantly larger amounts of cytokines (median: IL-1 beta 6450 pg/mL, IL-6 1821 ng/mL, and TNF-alpha 13,506 pg/mL) compared with placental cell cultures from nonlaboring women at term (median: IL-1 beta 2602 pg/mL, IL-6 993 ng/mL, TNF-alpha 3475 pg/mL; P < .02). Placental cells from women delivering preterm with intrauterine infection did not produce significantly different amounts of cytokines (median: IL-1 beta 3929 pg/mL, IL-6 1084 ng/mL, TNF-alpha 2847 pg/mL) when compared with those of nonlaboring women at term, whereas placental cells from uninfected women delivering preterm produced significantly larger amounts of cytokines (median: IL-1 beta 22,903 pg/mL, IL-6 1899 ng/mL, TNF-alpha 15,005 pg/mL; P < .01) than cells from nonlaboring women at term. Cytokine release from decidual cell cultures was similar in all groups tested. CONCLUSION: In the absence of intrauterine infection, preterm labor was associated with elevated placental cytokine release.

[Prenatal chromosome analysis using the FISH technique allows fetal aneuploidy detection within a few hours]. / Die pränatale Chromosomenanalyse mit Hilfe der FISH-Technik ermöglicht das Auffinden fetaler Aneuploidien innerhalb weniger Stunden.

Last years evaluation of fluorescence-in-situ-hybridization (FISH) allowed detection of chromosomal abnormalities by using DNA probes, binding to chromosomes in the nucleus. Because it is possible to directly examine interphase nuclei, FISH-technique, in contrast to traditional cytogenetic analysis has the advantage of small loss of time in case of urgent decisions on perinatal management. Karyotyping was performed on fetal cells, obtained from 72 pregnancies after amniocentesis, by both classical cytogenetics and fluorescence in situ hybridization using commercially available kits which utilise the alpha satellite probes for chromosomes 13 + 21 and 18. The classical cytogenetics demonstrated that the fetal karyotype was normal in 67 cases and abnormal in 5 cases (four with trisomy 21 and one with a translocation trisomy 18). With the FISH-technique it was possible to obtain accurate diagnosis of trisomy 21 within 24 hours of sampling. The distribution of the number of signals in the chromosomally normal and abnormal fetuses was significantly different, but we were not able to identify the fetus with translocation trisomy 18. We conclude that in the investigation of fetuses with ultrasonographic diagnosed malformations, FISH provides a rapid technique for detection of numerical chromosomal aberrations, but replacement of classical cytogenetics is not possible because of its limitations for identification of subtle structural chromosomal abnormalities.