High-affinity folate receptor in human ovary, serous ovarian adenocarcinoma, and ascites: radioligand binding mechanism, molecular size, ionic properties, hydrophobic domain, and immunoreactivity.

High-affinity folate receptors are expressed in normal ovaries and ovarian carcinomas. Binding of [3H]folate in human ovary, serous ovarian carcinoma tissue, and ascites is a complex process that has not been well characterized. This study shows changes in binding affinity and mechanism of binding with decreasing receptor concentration, inhibition by folate derivatives, and a slow radioligand dissociation at pH 7.4 becoming rapid and complete at pH 3.5. The receptor seems to be positively charged since it elutes in the front effluent of a DEAE-Sepharose CL-6B ion-exchange column at pH 6.3. The gel filtration profile of Triton X-100-solubilized tissue and ascites contained two peaks of radioligand-bound receptor (25 and 100 kDa). Exposure of ascites to cleavage by phosphatidylinositol-specific phospholipase C resulted in a partial conversion of the 100-kDa peak to a 25-kDa peak. This suggests that the receptor may be anchored to the membrane by a glycosylphosphatidyl residue that inserts into Triton X-100 micelles, resulting in a large molecular size on gel filtration. The receptor in ovarian carcinoma tissue immunoreacts with antibodies against purified human milk folate receptor protein as shown by enzyme-linked immunosorbent assay, immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting (a single band of 45 kDa), and immunohistochemistry. In only three of seven ovarian carcinomas did expression of radioligand-bound receptors exceed levels found in five normal ovaries. However, only receptors in ovarian carcinoma specimens showed a high degree of immunoreactivity. Hence, even without elevations of the total receptor level, a folate receptor isoform homologous to human milk folate receptor protein seemed to prevail in serous ovarian carcinomas.

A folate binding protein in ascitic fluid, serum and ovarian tissue of patients with ovarian adenocarcinoma immunoreacts with antibodies against human milk folate binding protein.

The presence of a folate binding protein which immunoreacts with antibodies against human milk folate binding protein was demonstrated in ascitic fluids from seven patients with ovarian adenocarcinoma. Ascitic fluids collected from two patients with other malignancies contained non-immunoreactive FBP. Tumor tissue specimens from five patients with ovarian carcinoma contained immunoreactive FBP. By contrast to normal ovaries ovarian carcinoma tissue showed positive immunostaining on immunohistochemistry. Ascitic fluids from two patients with ovarian carcinoma exhibited single distinct bands on SDS-PAGE immunoblotting. The gel filtration profile of ovarian carcinoma tissue homogenate from two patients contained 25 and 1OOkDa peaks of radioligand-bound and immunoreactive folate binding protein, while ascitic fluid from one of the patients exhibited a large 100 kDa immunoreactive peak with no radioligand binding activity. The immunoreactive non-functional 100 kDa FBP could represent unprocessed precursor FBP. Future studies are necessary to evaluate whether determination of immunoreactive FBP in ovarian adenocarcinomatosis is of any diagnostic value.

Transvaginal ultrasound, endometrial cytology sampled by Gynoscann and histology obtained by Uterine Explora Curette compared to the histology of the uterine specimen. A prospective study in pre- and postmenopausal women undergoing elective hysterectomy.

BACKGROUND: The purpose of this study was to evaluate the diagnostic value of transvaginal ultrasound measurement of endometrial thickness, cytology obtained by Gynoscann, and histology of the endometrium sampled by Uterine Explora Curette compared with histology of the uterine specimen as the gold standard. METHODS: Consecutive patients admitted for hysterectomy had transvaginal ultrasound, sampling by Gynoscann, and Uterine Explora Curette done just before surgery, after informed consent. RESULTS: A total of 181 women entered the study. Sixteen had endometrial cancer, seven had atypical hyperplasia and nine had complex hyperplasia. A total of 168 patients had a transvaginal ultrasound done. At a cutoff limit of 4mm (endometrial thickness of 4mm or less indicating normal endometrium), the sensitivity was 90.3%, the specificity 24.8%, the positive predictive value 21.4% and the negative predictive value 91.9%. One endometrial cancer, one atypical and one complex hyperplasia were missed. The Gynoscann method showed a sensitivity of 62.5%, a specificity of 94.0%, a positive predictive value of 69.0% and a negative predictive value of 92.1%. Two cancers, three atypical and six complex hyperplasia were missed. The Uterine Explora Curette showed a sensitivity of 90.6%, a specificity of 100.0%, a positive predictive value of 100.0% and a negative predictive value of 98.0%. One endometrial cancer and two complex hyperplasia were missed. CONCLUSION: Transvaginal ultrasound is a reliable method in excluding endometrial pathology. The Uterine Explora Curette was superior to Gynoscann in diagnosing neoplasia of the endometrium. It was found to have the same diagnostic accuracy as conventional dilatation and curettage.

Folate receptors in malignant and benign tissues of human female genital tract.

We have characterized the folate receptor in malignant and benign tissues of human female genital tract (Fallopian tube and benign and malignant tissues of uterus). Radioligand binding displayed characteristics similar to those of other folate binding proteins. Those include a high-affinity type of binding (K = 10(10)M-1), apparent positive cooperativity, a slow dissociation at pH 7.4 becoming rapid at pH 3.5, and inhibition of binding by folate analogues. The gel filtration profile of Triton X-100 solubilized tissue contained two large peaks of 3H-folate labelled protein (> = 130 and 100 kDa) as well as a 25 kDa peak. Only a single band of 70 kDa was seen on SDS-PAGE immunoblotting. The large molecular size forms on gel filtration appear to represent folate receptors having a hydrophobic membrane anchor inserted into Triton X-100 micelles. The folate receptor of female genital tract showed cross-reactivity in ELISA and positive immunostaining with rabbit antibodies against human milk folate binding protein. Variations in the ratio of immunoresponse to total high affinity folic acid binding suggests the presence of multiple isoforms of the receptor in different types of malignant and benign tissues.

Characterization of the folate receptor in human molar placenta.

We have characterized a high-affinity folate receptor in human molar placenta tissue. Radioligand binding exhibited characteristics typical of other high-affinity folate binding proteins. Those included, positive cooperativity, a tendency to increased binding affinity with decreasing receptor concentration, a slow ligand dissociation at pH 7.4 becoming rapid at pH 3.5, and inhibition by folate analogues. The folate receptor cross-reacted with antibodies against human milk folate binding protein, e.g. the syncytothrophoblastic layer of molar placenta tissue sections showed strongly positive immunostaining. The gel filtration profile contained two radioligand-bound peaks (25 and 100 kDa), however, with considerable overlap. Only a single band of 70 kDa was seen on SDS-PAGE immunoblotting. The folate receptor in placental tissue may play a crucial role in the transfer of folate from maternal circulation to the fetus.

A high-affinity soluble folate receptor in fluids of non-neoplastic ovarian cysts: radioligand binding, molecular size, hydrophobic residue, and immunological properties.

The presence of a soluble folate receptor in fluids of non-neoplastic ovarian cysts was demonstrated. Radioligand binding exhibited characteristics typical of high-affinity folate-binding proteins. These included positive cooperativity, a tendency to increased binding affinity with decreasing receptor concentration, a slow ligand dissociation at pH 7.4 and inhibition by folate analogues. The folate receptor was probably synthesized in the lining epithelial cells of the cysts which showed positive immunostaining with antibodies against human milk folate-binding protein. The gel filtration profile of cystic fluid contained two radioligand-bound peaks, 25 and 100 kDa, whereas a single band of 70 kDa was seen on SDS-PAGE immunoblotting. Treatment with the enzyme phosphatidylinositol-specific phospholipase C resulted in a partial conversion of the 100 kDa peak to the 25 kDa peak. This suggests that insertion of a hydrophobic glycosylphosphatidylinositol tail into Triton X-100 micelles could give rise to large molecular size forms of the receptor on gel filtration.

Folate receptor in malignant effusions of ovarian carcinoma.

Binding of 3H-folate in human ovarian adenocarcinoma tissue was of a high-affinity type (K approximately 10(10) M-1) and displayed apparent positive cooperatively. A high-affinity folate receptor was also present in ascitic fluid and pleural effusion. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. The folate analogues methotrexate and in particular 5-formyltetrahydrofolate acted as inhibitors of 3H-folate binding in ascitic fluid. Ovarian adenocarcinoma tissue showed immunostaining with rabbit antibodies against human milk folate-binding protein. The gel filtration diagram contained two peaks of radiolabelled folate (at 25 and 100 kDa). The 25 kDa peak was predominant in ascitic fluid and pleural effusion. A single band of 70 kDa was seen on SDS-PAGE immunoblotting of tissue and malignant effusions. The concentration of folate receptor in tissue and fluid specimens could be determined by an immunochemical method (ELISA) utilizing antibodies against human milk folate-binding protein.

[Intraobserver variation in the classification of diseases]. / Intraobservatørvariationen ved klassifikation af sygdomme.

The aim of this study was to describe the variation within the observer in classification of diseases, with special regard to pregnancy, birth and maternity. Four observers A, B, C and D classified 150 random cases of pregnancy, birth and maternity on two different occasions with six months interval. The rate of agreement within the observer between the two different occasions was evaluated. The average agreement rate was 86% (71-92%) concerning diagnoses of pregnancy. 90% (84-95%) concerning maternity and 65% (53-75%) concerning delivery. The diagnostic spectrum of the observers and their criteria for the use of diagnoses were variable. The amount of variation correlated to the number of diagnoses and poorly defined diagnostic criteria. In conclusion, great variation was found within the observer in classifying diseases, especially concerning delivery, due to increased numbers of diagnoses and poorly defined diagnostic criteria. The setting-up of branch specialist committees is recommended, if we in the future want to use a public diagnostic register as a tool for research. These committees must achieve consensus concerning the need for classification within each specialty, define criteria for diagnosis and communicate recommendations and education in the system of classification.

[Interobserver variation in the classification of diseases]. / Interobservatørvariationen ved klassifikation af sygdomme.

The aim of this study was to describe the variation between observers in classification of diseases, with special regard to pregnancy, birth and maternity. Four observers A, B, C and D classified 150 random cases of pregnancy, birth and maternity. The rate of agreement between the observers and their use of diagnosis was evaluated. The total agreement-rate was 58% concerning diagnoses of pregnancy, 82% concerning maternity and 13% concerning delivery; with variation between the observers at 71-85%, 86-96% and 26-54%. The diagnostic spectrum of the observers and their criteria for the use of diagnoses were variable. In conclusion, great variation was found between observers in classifying diseases, due to different diagnostic criteria being used by the observers, and due to misunderstanding of the Classification of Diseases. The setting-up of committees of branch-specialists is recommended, if we in the future want to use a public diagnostic register as a tool for research. These committees must achieve consensus concerning the need for classification within each specialty, define criteria of diagnosis and effect recommendation or education in the system of classification.

[Ambulatory surgery and anesthesia. An inquiry study]. / Ambulant kirurgi og anaestesi. En enqueteundersøgelse.

Patients who had undergone surgical procedures as out-patients in either general or local anaesthesia were asked to complete a questionnaire with the aim of evaluating how they had experienced their operation, especially with respect to treatment and prevention of postoperative pain. Over a 12 month period, 851 patients (251 men and 600 women) who had undergone either abdominal, orthopaedic or gynaecological surgery as out-patients were given the questionnaire. Five hundred and fifteen patients (166 men and 349 women), i.e. 61%, answered. Nineteen percent had had their operation performed in local or regional anaesthesia, 30% in a combination of general anaesthesia with local infiltration, the remaining patients were operated under general anaesthesia. Eighty to ninety percent were satisfied with the preoperative information, and 88% were satisfied with the anaesthesia. Sixty-nine percent had had either no pain or almost no pain within the first 24 hours after the operation. Six percent found it difficult to stay home. Sixteen percent needed some kind of medical contact. Ninety-two percent said they would prefer out-patient surgery again as opposed to hospitalization, were they to need another operation. In conclusion, we found that these types of out-patient operations were acceptable to most of our patients (92%). We recommend widespread use of local anaesthetics in combination with general anaesthesia in order to minimize postoperative pain and facilitate the effect of postoperative analgetics.

Thermal artifacts and inter-gynecologist variation of laser cone biopsies of the cervix.

OBJECTIVE: To evaluate the extent of thermal artifacts and the inter-gynecologist variation in CO2 laser cone biopsies for cervical intraepithelial neoplasia (CIN) using high power density. METHODS: All 1180 histologic sections from 93 CO2 laser cone biopsies performed by nine gynecologists were evaluated by morphometry. Power density of 19,700 W/cm2 and spot size of 0.43 mm were used. The lengths of different epithelia, the distance from CIN to the excision margins, the thermal artifacts, and the extension of denudation were analyzed. Mean values and standard deviations were analyzed for all indices. The inter-gynecologist variation was analyzed using one-way analysis of variance, Tukey-Kramer Honestly Significant Difference test, and chi 2 contingency table analysis regarding cones classified as "good" or "not good". RESULTS: The mean length of analyzed surface per cone was 134 mm. In 3% of the sections, the sum of artifacts at the external excision margin exceeded the average "minimum free distance" from CIN to the margin (2.88 mm). At the endocervical excision margin, the free distance was exceeded in 17% of the sections. Eighteen percent of the cones had CIN at the excision margins, with no significant differences between gynecologists (P = .83). Differences existed between gynecologists regarding free minimum distance and minor thermal artifacts at the external margin, but when clinically relevant groups were applied, no significant differences were found (P > .05). CONCLUSION: We found that a high power density at CO2 laser cone biopsy resulted in specimens with minimal thermal artifacts compared to other authors who used low power density. No clinically relevant differences were found between the gynecologists regarding free margins or artifacts.

Outpatient CO2 laser excisional conization for cervical intraepithelial neoplasia under local anesthesia.

AIM AND OBJECTIVE: To evaluate the feasibility of CO2 laser excisional conization in patients with cervical intraepithelial neoplasia (CIN) in an outpatient setting under local anesthesia. DESIGN: A prospective study of women with cervical intraepithelial neoplasia over one year recording early and late complications, cure rate, and patients' compliance. The procedure was performed with a Coherent CO2 laser connected to the colposcope with a micromanipulator. The equipment attaining 19,700 W/cm2, spot diameter 0.43 mm, focus distance 300 mm, and continuous beam. RESULTS: The material included 81 women. Seventy-eight women were treated as outpatients, while three women were admitted for 24 hours observation due to surgical and technical complications during the treatment. Sixty-six percent of the women had bleeding less than 10 ml during the treatment. One woman was admitted 24 hours after surgery due to bleeding requiring suturing. Median operation-time was 11 minutes (range 3-60). All specimens contained CIN. In 76% of the cases the pre- and postoperative diagnosis were identical. Twelve specimens had CIN in one or both margins. Abnormal cytology after conization was found in three cases (4.2%). High patient compliance was found as 92% of the treated women would choose the same kind of treatment if it had to be repeated. CONCLUSION: High success rate, few complications and high patient compliance make the procedure adequate as a routine procedure in treatment and diagnosis of CIN.

[Acceptance of ambulatory laser conization under local anesthesia by Danish women]. / Danske kvinders accept af ambulant laserkonisation i lokalanaestesi.

The aim of the study was to evaluate the acceptability of CO2 laser excisional conization for cervical intraepithelial neoplasia under local anaesthesia in an outpatient setting. Seventy-seven patients, who underwent this procedure, were interviewed three months later concerning pain during treatment, pain and bleeding in the immediate postoperative period, and their general opinion about this procedure. The median duration of surgery was 11 minutes. The postoperative median observation time was 90 minutes. Seventy-one women experienced no discomfort during treatment. Thirty-one patients needed mild analgesics for an average of three days. Bleeding continued for a mean of 7.7 days after treatment and 73% experienced the bleeding as less, 8% as more as compared to menstrual bleeding. Ninety-two percent of the women would prefer this procedure if they had to undergo conization once again. In conclusion, we found a high acceptability in women treated with CO2 laser excisional conization under local anaesthesia in an outpatient setting.

Germ cell function and hormonal status in patients with testicular cancer.

Sperm counts, serum gonadotropins, and androgen levels were investigated in 39 seminoma patients and 58 patients with a nonseminomatous germ cell tumor of the testis after unilateral orchiectomy. In 58% of the patients, the total sperm count was below the lower reference value (80 million). A multiregression analysis demonstrated a correlation between a decreased total sperm count and the following three explanatory variables: (1) an elevated serum alpha-fetoprotein (AFP), (2) a history of chryptorchidism, or (3) a seminomatous tumor. In 42% of the patients, the sperm concentration and the sperm motility met criteria considered sufficient for cryopreservation. Serum follicle-stimulating hormone (FSH) was elevated in 33% of the patients. Androgens (serum testosterone [T] or urine 17-oxy-steroids [17-OS]) were subnormal in 5% of the patients, whereas serum luteinizing hormone (LH) was elevated in 14% of the patients without human chorionic gonadotropin beta-subunit (beta-HCG) in serum.

The long-term effect of cisplatin on renal function.

The long-term effect of cisplatin on renal function was studied in a follow-up investigation, 16 to 52 months after chemotherapy in 22 patients with disseminated nonseminomatous testicular cancer. The median cumulated cisplatin dose was 452 mg/m2 (range, 275-650 mg/m2). Prehydration with isotonic saline secured diuresis above 100 ml/hour. GFR (glomerular filtration rate: 51Cr-EDTA clearance) fell by 12.5% (median, P less than 0.01) compared with the pretreatment level. Effective renal plasma flow (125I-hippuran clearance) was estimated to be likewise reduced. The serum creatinine level rose 8 mumol/1 (median, P less than 0.05) during the treatment period with no further rise afterwards. Serum magnesium and urinary excretion of beta-2-microglobulin were normal. The results indicate a moderate and permanent reduction in GFR with no signs of long-term tubular defects in patients treated with cisplatin.

Maternal-serum-alphafetoprotein screening for fetal malformations in 28 062 pregnancies. A four-year experience from a low-risk area.

From 1st March, 1980 and up to 29th February 1984 a multicenter serum alpha-fetoprotein (S-AFP) screening project was carried out for the detection of severe fetal malformations. S-AFP was determined by a radio-immunoassay in 28 062 pregnant women between the 16th and 20th week of gestation. Patients with elevated S-AFP values, e.g. above 95 percentile, were examined further with a second S-AFP and by ultrasound scan. 244 amniocenteses (0.9%) were carried out to detect 62 malformations (21 anencephalies, 14 spina bifidas, 2 encephaloceles, 7 omphaloceles, 5 gastroschises, 4 chromosome abnormalities and 9 other malformations). Fifteen of the 16 cases of spina bifida could not be verified by ultrasound scan, whereas all other malformations except chromosome abnormalities were confirmed by ultrasonography. Two cases of spina bifida and one case of skin-covered encephalocele had normal S-AFP concentrations and were therefore not detected. There were no definitive false-positives, e.g. therapeutic abortion of a normal fetus. Our conclusion is that a nationwide S-AFP screening should be recommended.