Prognostic significance of preoperative DNA flow cytometry in surgically-treated cervical cancer.

OBJECTIVE: To evaluate the prognostic significance of preoperative DNA flow cytometry compared with other clinical and histologic variables in cervical carcinoma. STUDY DESIGN: Sixty-four patients with FIGO Stage Ib-II cervical cancer treated with radical abdominal hysterectomy and systematic pelvic lymphadenectomy were analyzed. The mean follow-up was 3.4 (range 0.3-9.8) years. DNA flow cytometry was performed with fresh tumor tissue. Four biopsies were recut from the surgical specimen within 30 minutes of the operation. The ectocervix was divided into four quadrants and a specimen obtained from each. DNA-low-grade tumors (diploid, near-diploid, tetraploid and near-tetraploid) were distinguished from DNA-high-grade tumors (aneuploid and hypoploid). Carcinomas with more than one non-diploid stem line were considered heterogeneous. An S phase fraction >7% was classified as low, 7% - < 14% as moderate, and > or = 14 as high. DNA ploidy, DNA heterogeneity, S phase fraction and various clinical and histological variables were related to disease-free survival. RESULTS: In the univariate analysis patients with DNA-low-grade carcinomas had significantly better disease-free survival than patients with DNA-high-grade tumors (82% vs 45%, p = 0.021). Carcinomas with an S-phase fraction < 7% were associated with better disease-free survival (0.8) than those with an S-phase fraction 7% - > 14% (0.62) and those with > or = 14% (0.64), but this was not statistically significant. Cox stepwise regression analysis showed DNA-heterogeneity, age, grade, parametrial involvement and extrapelvic metastasis to be independent prognostic factors. CONCLUSION: DNA ploidy and DNA heterogeneity are of prognostic importance in cervical cancer. DNA flow cytometry may be used preoperatively to identify low-risk and high-risk patients within a given stage.

Self-regulation of the endothelin receptor system in choriocarcinoma cells.

The human trophoblast secretes endothelin-1 (ET-1) and expresses ET receptors. The present study tested whether the transformed BeWo, JAR and JEG-3 choriocarcinoma cells: (1) secrete endothelin-1 (ET-1); (2) express both ET-A and ET-B receptor subtypes; and (3) have the potential to allow for autologous regulation of ET-receptor proteins. The cells were cultured for 24/48 h with or without 10% FCS and, in experiments on receptor regulation, with ET-1 (5-20 nM and 10 microM). ET-1 secretion was measured by RIA and receptor levels by immunoblotting. All cell types secreted ET-1 albeit at different levels and sensitivity to FCS. All cell lines expressed both ET-A (JEG-3>BeWo=JAR) and ET-B (JEG-3=JAR>BeWo) receptor subtypes, which could be up- and downregulated depending on ET-1 concentration, culture time and FCS presence. It is concluded that BeWo, JAR and JEG-3 choriocarcinoma cells secrete ET-1 and express both ET-A and ET-B receptor subtypes. The receptor levels can be regulated by ET-1. This provides the molecular basis for an autocrine system with the potential of autologous regulation of yet unidentified ET-1-induced functions.

Perioperative myocardial cell injury: the relationship between troponin T and cortisol.

STUDY OBJECTIVE: To investigate whether there is an association between Troponin T (TnT), reflecting myocardial cell injury, and cortisol, reflecting the degree of surgical trauma and associated stress, in light of our recent evaluation of TnT as a marker of perioperative myocardial cell injury. DESIGN: Prospective, cohort study. PATIENTS: 70 patients (67.4 +/- 8.7 yrs) with definite or at-risk coronary artery disease (CAD) undergoing elective noncardiac surgery (vascular n = 38, abdominal n = 21, orthopedic n = 8) with general (n = 63) or regional (n = 4) anesthesia with postoperative on-demand analgesia. MEASUREMENTS AND MAIN RESULTS: Morning blood samples for TnT (upper limit of normal: <0.2 ng/mL), CK-MB (reference range

Numerical chromosomal aberrations of chromosome 1 and 7 in dysplastic cervical smears.

Cervical smears with Papanicolaou's staining (PAP) reveal only morphological characteristics of epithelial cells of the cervix uteri. Since chromosomal aberrations are known to play a role in malignant transition, we analyzed cervical smears for numerical changes of the chromosomes 1 and 7 with fluorescence in-situ hybridization to probe for a diagnostic value of these chromosomes in the characterization of cervical dysplasia. Cervical smears were collected from 21 patients with suspect histology of curettage or biopsy specimen, 14 of them having been subsequently graded as cervical intraepithelial neoplasia (CIN) III and 5 as CIN II. Nineteen normal cervical smears (PAP I-II) served as controls. Smears were hybridized with chromosomal enumeration probes for chromosome 1 and 7. Disomic cells (2 copies of chromosome 1 and 7) were decreased in the CIN II (63%) and CIN III group (57%) with respect to the control group (77%). Cells with 3 signals for chromosome 7 were significantly more frequent in the CIN III and the CIN II group than in the control group (6.7, 6.4 and 0.7%, respectively). Only the CIN II group (10%), but not CIN II (6%), showed a significant trisomy for chromosome 1 as compared with the controls (3.8%). A close correlation between the incidence of trisomy 1 or 7 and PAP grading was observed. PAP III-IIID smears with high trisomy 1 counts corresponded to CIN III histology, while all CIN II patients were PAP III-IIID with low incidence of trisomy 1. We conclude that trisomy of chromosome 7 is a feature of cervical dysplasia and seems to be an early event in dysplastic transition. In contrast, trisomy of chromosome 1 is observed only in high grade dysplasia and may be a marker for pre-malignant lesions.

Exfoliative cytology of a lymphoepithelioma-like carcinoma in a cervical smear. A case report.

BACKGROUND: Lymphoepithelioma-like carcinoma of the cervix (LELC) is cytologically identical to its counterparts at other sites, such as the nasopharynx. LELC can be suspected on a cervical cytologic smear. The differential diagnosis includes nonkeratinizing squamous cell carcinoma with prominent stromal inflammation, carcinoma with intense stromal eosinophilia, glassy cell carcinoma, malignant lymphoma (especially lymphoepitheloid-Lennerts lymphoma) and metastatic Schmincke-Regaud tumor. CASE: A 55-year-old female presented with an ulcerated endophytic tumor in the cervix. Exfoliative cytology showed uniform, large tumor cells, often associated with inflammatory cells, with round or oval nuclei and one or more prominent nucleoli. The cytoplasm was finely granular to flocculent, and the nuclei were uniformly vesicular. The chromatin was peripherally marginated. The cell borders were indistinct. There was no evidence of dyskeratotic or keratinized cells, koilocytes or glandlike formations. These findings were highly suspicious for LELC and were confirmed by biopsy. Flow cytometry showed DNA aneuploidy, with a DNA index of 1.08. In situ hybridization was negative for human papillomavirus 16 and 18. CONCLUSION: LELC of the uterine cervix has cytologic features that are sufficiently characteristic for a specific cytologic diagnosis. The diagnosis, nevertheless, has to be proven by histology.

Amniotic fluid insulin levels in nondiabetic pregnant women: an update.

Elevated amniotic fluid insulin levels in diabetes are frequently described but there are few systematic data on metabolically healthy women to define normal ranges. Previous studies had too high normal ranges because they were based on unspecific insulin binding radioimmunoassays. The aim of the study was to update normal amniotic fluid insulin data and to define a reliable normal range in the course of a nondiabetic pregnancy. Amniotic fluid insulin levels were measured in 841 amniotic fluid samples of 707 nondiabetic women undergoing amniocentesis for hydramnios, suspected malformation, determination of lung maturation, Rhesus antibodies and cordocentesis. Mean (+/- SD) of amniotic fluid insulin level was 3.6 (+/- 2.1) microU/mL at 31.5 (+/- 4.9) weeks of pregnancy. The 97th percentile was 8.2 microU/mL. Insulin levels show a biphasic course between 16th and 42nd weeks of pregnancy with a zenith at 30th week. Only two cases (0.3%) had unexplicably elevated amniotic fluid insulin levels > or = 10 microU/mL. Thus, in nondiabetic women amniotic fluid insulin levels > 10 microU/mL are unlikely.

Circadian rhythm of the soluble p75 tumor necrosis factor (sTNF-R75) receptor in humans--a possible explanation for the circadian kinetics of TNR-alpha effects.

Circadian alterations of several immune functions in vivo are well established, and may have important physiological and clinical implications. In line with this, tumor necrosis factor (TNF)-alpha has been implicated in the circadian regulation of body temperature. As soluble TNF receptors (TNF-R) act as naturally occurring competitive inhibitors of this cytokine, we investigated plasma levels of the soluble sTNF-R55 and sTNF-R75 in comparison with plasma cortisol and body temperature in nine healthy male volunteers during a defined 16 h light/8 h dark cycle. It was found that sTNF-R75, but not sTNF-R55, exhibited a clear-cut circadian rhythm with a significant (P < 0.01) peak at 7:51 a.m. +/- 58 min. The phase of the sTNF-R75 rhythm preceded that of cortisol by approximately 1 h and inversely corresponded to the circadian rhythm of body temperature. Moreover, the individual amplitudes of sTNF-R75 and body temperature exhibited a significant (P < 0.01) positive correlation. These results suggest that (i) the two sTNF-R are regulated independently, (ii) the sTNF-R75 rhythm is not primarily due to the cortisol rhythm and (iii) the fluctuation of the sTNF-R may contribute to the regulation of body temperature by modulating the availability of free TNF.

Toward universal criteria for gestational diabetes: relationships between seventy-five and one hundred gram glucose loads and between capillary and venous glucose concentrations.

OBJECTIVES: Replacement of the two-step, 100 gm, 3-hour National Diabetes Data Group procedure by the one-step, 75 gm, 2-hour World Health Organization oral glucose tolerance test has been hindered by a paucity of data comparing the two tests during pregnancy. The current series compared 100 gm and 75 gm glucose loads and glucose measurements in venous plasma or capillary blood. STUDY DESIGN: After a 75 gm oral glucose tolerance test 30 gestational diabetics and 30 metabolically healthy pregnant women were randomly assigned to a second 75 or 100 gm test within 3+/-1.3 (mean+/-SD) days. Glucose levels at both tests was measured in capillary blood and venous plasma, as were insulin and C peptide. RESULTS: In controls 1-hour maternal glucose levels (112 vs 128 mg/dl) and 2-hour levels (104 vs 113 mg/dl) differed significantly after a 75 or 100 gm load (paired t test). In gestational diabetes mellitus, however, there was no difference (176 vs 178 mg/dl) but a low insulin/glucose quotient at 1 hour. Only 2-hour levels differed significantly (133 vs 149 mg/dl). In controls glucose measurement in capillary blood and venous plasma differed significantly at 1 hour (126 vs 115 mg/dl) and 2 hours (111 vs 104 mg/dl) independently of the glucose load. In gestational diabetes mellitus, however, glucose measurement in capillary blood and venous plasma differed neither in 1-hour levels (179 vs 174 mg/dl) nor in 2-hour levels (142 vs 139 mg/dl). CONCLUSION: In metabolically healthy women both different loading and different blood fractions lead to statistically different blood glucose levels at 1 and 2 hours. In gestational diabetes mellitus, however, 1-hour glucose levels do not differ after a 75 or 100 gm load or after glucose measurement in capillary blood or venous plasma. This is due to elevated insulin resistance shown by a low insulin/glucose quotient at 1 hour. For comparison of tests in gestational diabetes mellitus only, 2-hour values must be adjusted by 16 mg/dl after different loading.

Anti-insulin antibodies and birth weight in pregnancies complicated by diabetes.

Free insulin cannot cross the placenta but insulin complexed to anti-insulin antibodies has been demonstrated in cord blood. We studied whether antibody-bound insulin in diabetic patients can evoke fetal macrosomia independently of maternal metabolic control. In 457 non insulin-treated controls and 173 insulin-treated diabetic patients we measured 1187 anti-insulin antibody levels and maternal blood glucose, maternal fructosamine, cord blood insulin, cord blood C-peptide, cord blood fructosamine and amniotic fluid insulin. Mean anti-insulin antibody levels in maternal blood and cord blood were significantly higher in insulin treated diabetic patients (4.6 and 5.4 U/ml) than in controls (1.8 and 1.7 U/ml) with maxima of 89.2 in maternal and 120.0 U/ml in cord blood, respectively. In insulin treated diabetic patients 16.6% (maternal blood) and 22% (cord blood) anti-insulin antibody levels were above the 97th percentile. There was a high significant correlation between maternal and cord blood anti-insulin antibodies (R = 0.987, P = < 0.0001), but no correlation of anti-insulin antibodies with maternal (glucose, fructosamine) or fetal (insulin, C-peptide, and fructosamine in cord blood, amniotic fluid insulin) metabolic parameters. While maternal and fetal metabolic parameters correlated with birth weight neither maternal nor cord blood anti-insulin antibody levels correlated with birth weight. These findings do not support the hypothesis that maternal anti-insulin antibodies independently influence fetal weight.

Comparative evaluation of three assay systems for automated determination of hemoglobin A1c.

We evaluated three newly introduced systems for automated determinations of hemoglobin (Hb) A1c, which allow the processing of large amounts of samples in a routine clinical laboratory. We compared these methods--the Variant HPLC, the Hi-Auto A1c analyzer system, and the Roche immunoassay--with the Diamat HPLC system. All showed good precision and good concordance with the Diamat HPLC. The reference range for Hb A1c has to be determined by the laboratory for each assay system. Interference study showed no statistically significant influence of anemia, polycythemia, rheumatoid factor, or chronic hemodialysis, although individual Hb A1c values can be influenced by polycythemia (when measured with the Hi-Auto A1c analyzer) and by chronic hemodialysis (when measured with the Variant HPLC). HPLC was not suitable for measuring Hb A1c in the examined cases of hemoglobin variants; assaying fructosamine seems to be better for monitoring these patients.

Exfoliative cytology of invasive neuroendocrine small cell carcinoma in a cervical cytologic smear. A case report.

BACKGROUND: Neuroendocrine small cell carcinoma of the cervix (NSC) is cytologically identical to its counterparts at other sites, such as the lung, and can be suspected on a cervical cytologic smear. It has to be distinguished from poorly differentiated nonkeratinizing squamous small cell carcinoma, adenocarcinoma with carcinoid features, embryonal neuroblastoma, embryonal rhabdomyosarcoma, stromal sarcoma and non-Hodgkin's lymphoma. CASE: An 18-year-old woman had invasive NSC. Exfoliative cytology of the cervix showed tumor cells, single or in small clusters and files, with darkly staining nuclei. The chromatin pattern was coarse, with small, prominent chromocenters, and the nuclei were often invisible. The cytoplasm was so reduced as to be barely discernible. Mutual molding of adjacent nuclei was frequent. Immunohistochemistry showed positive staining for pancytokeratin and neuron-specific enolase. Flow cytometry showed aneuploidy, with a DNA index of 1.93. CONCLUSION: The cytologic appearance of NSC in a cervical cytologic smear is characteristic. The diagnosis, nevertheless, has to be proven by the identification of neuroendocrine differentiation.

Multicentre clinical evaluation of the COBAS CORE CEA, CA 125 II and PSA tumor marker assays.

The aim of our study was to evaluate the clinical usefulness of the tumor markers CEA, CA 125 and PSA using the COBAS CORE system from Roche Diagnostic Systems. Our results demonstrate that determination of these markers on the COBAS CORE immunoassay analyser provides the performance required for routine use in clinical practice. The results obtained in this clinical trial validate the correlation with disease extension, a characteristic that defines and determines the clinical utility of the tumor markers. We also conclude that learning to operate the COBAS CORE system is simple, as is management of the system through the user-friendly software.

Diagnostic value of gonadal steroid hormones and gonadotropins in ovarian cystic lesions.

During a 12-mo period the fluid aspirated from 32 ovarian cystic lesions was assayed for 17 beta-estradiol (E2), progesterone (P), testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH). All cysts were subsequently subjected to histologic or cytologic examination. In 84.6% (11/13) of the proven follicular cysts, the E2 content was greater than 100 pg/ml, in 76.9% (10/13) the P content was greater than 13 ng/ml, and in 69% (9/13) the T content was greater than 0.5 ng/ml. In 94.7% (18/19) of the ovarian cysts of other types, the E2 content was less than 100 pg/ml and the P content was less than 13 ng/ml. The FSH and LH contents of both types of cysts were variable, and no consistent pattern was discerned. In contrast, cytologic examination identified 69.2% (9/13) of the follicular cysts. In conclusion, consideration of the E2 and P contents with cytologic examination allowed the identification of all follicular cysts. Accordingly we recommend the estimation of E2 and P as part of the assessment of all ovarian cyst aspirates obtained from premenopausal patients.

Evaluation of the Cobas Core system for use in routine diagnosis of thyroid disorders.

The aim of this clinical study was to evaluate the TSH, FT4, FT3, T3, and T4 assays designed for use on the Cobas Core Immunoassay Analyzer. Performance of these enzyme immunoassays (EIAs) was compared with well established and routinely used radioimmunoassays (RIA) or immunoradiometric assay (IRMA). In the first part of the study, the between-run and within-run data as well as the linearity of the Cobas Core assays was carried out, showing a very high precision; most coefficients of variation (CVs) were below 8%. In the second part of the study, serum samples from patients with different thyroid diseases were evaluated, and the results compared with those obtained by RIA or by IRMA. All results were highly comparable with the clinical aspects of all investigated thyroid disorders, and the data of TSH, FT4, T3 and T4 were completely in concordance with those assessed by RIA or IRMA methods; however, only the FT3 assays had a moderately correlation. In contrast to RIA methods, Cobas Core EIAs provided precise results in patients with thyroid hormone transport protein anomalies or T4/T3 autoantibodies; moreover, elevated serum concentrations of thyroxine binding globulin (TBG) did not affect measurement of total thyroid hormone concentrations.

[A rapid method for diabetes screening in pregnancy]. / Schnellmethode zum Diabetesscreening in der Schwangerschaft.

Gestational diabetes affects about 3% of pregnancies in German-speaking countries. Roughly one third of these pregnancies develop a requirement of insulin. In unrecognised and hence untreated pregnancies, perinatal morbidity and mortality are increased 20-fold. Gestational diabetes is asymptomatic and only 60% of patients have risk factors for the condition; thus, general screening with oral glucose tolerance testing is necessary to detect all cases. Most antenatal medical care is provided by gynaecologists in practice and by general practitioners who do not have a sophisticated laboratory at their immediate disposal. General screening requires a test that is simple, inexpensive and quick but nonetheless meets high quality standards. A new microcuvette rapid test fulfills these requirements; the results are ready within a few minutes' time. We performed parallel blood glucose measurements with a standard enzymatic method and with the rapid test in 500 unselected pregnant women undergoing an oral glucose tolerance test at our obstetric clinic. The mean fasting, 1-hr and 2-hr values were 77,128 and 106 mg/dl, respectively, as measured by the enzymatic test and 75,129 and 107 mg/dl as measured by the rapid test. The results of the reference enzymatic method and the rapid test agreed at a high level of significance (r = 0.98; p < 0.0001).

[Premature labor and fetal fibronectin in cervical and vaginal secretions]. / Frühgeburt und fetales Fibronectin im Zervikal- und Vaginalsekret.

This study aimed to determine whether quantitative measurements of fetal fibronectin in cervical vaginal secretions reflect the prognosis for imminent premature delivery. 166 patients, who were pregnant from between 25 and 36 weeks, were included in the study over a six months' period. Group A included 60 women with premature contractions, which ceased after tocolytic therapy. Group B consisted of 25 women, who delivered prematurely within 1 week. Group C contained 22 patients, with confirmed PROM and group D contained 24 patients in whom PROM was suspected, but not confirmed. The control group (group E) consisted of 35 patients with uneventful pregnancies. Fetal fibronectin levels in groups B and C differed significantly from those in the other groups. Therefore, the positive predictive value of elevated fetal fibronectin levels for premature delivery was 86.1%.

Serum immunosuppressive acid protein predicts lymph node metastases in patients with renal cell carcinoma.

OBJECTIVE: To evaluate the usefulness of serum immunosuppressive acid protein (IAP) as a marker for lymph node metastases in patients with renal cell carcinoma. PATIENTS AND METHODS: Serum IAP levels were studied in 73 patients with renal cell carcinoma, prior to therapy. Serum IAP levels were correlated with the clinical and the post-operative histopathological stage. A threshold value of 800 micrograms/ml was applied. RESULTS: Statistical analysis revealed a better correlation between IAP concentration and disease than with any other variable. Sensitivity and specificity were 94% and 84%, respectively, thus providing good positive (65%) and negative (98%) predictive values regarding lymph node involvement. CONCLUSION: Measuring serum IAP levels can be a valuable contribution to lymph node staging in patients with renal cell cancer. The choice of cut-off value of 800 micrograms/ml has yet to be supported by a larger series.

Detection and evaluation of persisting stress-induced hormonal disturbances by a post stress provocation test in humans.

Eighteen healthy army officers were subjected after prolonged rest to exhaustive ergometric work for about 15 minutes. Before and afterwards blood was taken from the cannulated antecubital vein for determination of free and sulfoconjugated catecholamines, cortisol, glucose, and white blood cell count. One week later, the same procedure was repeated with the same subjects with the difference that the probands underwent about 2.5 hours of difficult mountain climbing and a subsequent rest of 1.5 hours before ergometry. The most important results were: 1) total and bound fractions of catecholamines showed some significant differences between the first and second ergometry due to the previous mountain climbing stress; 2) serum cortisol did not increase after the first ergometry but did so significantly after the second ergometry due to the previous stress; low cortisol is not always indicative of the absence of stress; 3) the absolute number of white blood cells increased in both situations, correlated significantly with the severity of the stress and the individual increases were more person than situation specific and; 4) blood glucose remained unaffected in both situations. We conclude that a previous stress experience can affect a second stress response and that such a post stress provocation test can uncover persistent hormonal alterations. This procedure may be useful for the evaluation of inaccessible stress situations from subsequent stress measures.

[Mucoviscidosis screening with immunoreactive trypsin. Initial experiences in Austria]. / Mukoviszidose-Screening mit immunreaktivem Trypsin (IRT). Erste Erfahrungen in Osterreich.

Up to now, 16.338 IRT-measurements have been carried out on dried blood spot specimens; 15.505 of them were taken in the first week of life. Related to a provisionally chosen cut-off point of 750 ng/ml, 134 newborns (= 0.86%) showed an elevated IRT-value and subsequently were recalled between the fourth and sixth week of life for a second IRT-determination. Twenty-five out of 116 reinvestigated children again showed an elevated value, as based on likewise provisional, age-dependent reference values. Four of these children subsequently were identified as CF patients by sweat testing. So far, we did not encounter any false-negative IRT values. We also commenced to establish a profile of reference values for the first twelve weeks of life; as yet, there are not enough data for definitely defining these limits of normality. In conclusion, IRT-screening appears to be a reliable method for identifying CF patients in the newborn period. Our preliminary results indicate an incidence of CF of 1 to 3880 in the southeast of Austria.