Preoperative assessment of endometrial carcinoma by three-dimensional power Doppler angiography.

OBJECTIVE: Preoperative evaluation of the depth of myometrial invasion in endometrial carcinoma is challenging. The objective of this study was to evaluate the usefulness of three-dimensional power Doppler angiography (3D-PDA) in this setting. METHODS: Sonographic and histological data on 100 consecutive cases of endometrial carcinoma were analyzed. The endometrial and myometrial vascular indices VI (vascularization index), FI (flow index) and VFI (vascularization flow index) were calculated by 3D-PDA. The results were compared with a complete surgical staging. RESULTS: The mean ( ± SD) age of patients was 67.1 ± 8.8 (range, 33-87) years. Forty-six patients had deep (≥ 50%) myometrial invasion. Eight patients had metastases, seven of them with deep invasion. Three patients were found to have carcinomas of non-uterine origin on histology, and these were excluded from further statistical analysis. The median endometrial and myometrial vascular indices were higher in the group with deep invasion than in the group without. Following multivariable analysis of the indices only the endometrial FI was independently associated with deep invasion (OR, 1.061; 95% CI, 1.023-1.099; P = 0.001). However, a greater endometrial volume was also an independent predictor of deep invasion (OR, 1.109; 95% CI, 1.011-1.215; P = 0.028). CONCLUSION: Our study suggests that endometrial and, to a lesser degree, myometrial vascular indices and endometrial volume correlate with the depth of myometrial invasion in endometrial carcinoma.

Tumor-associated trypsin participates in cancer cell-mediated degradation of extracellular matrix.

We have recently demonstrated that many cancer cell lines produce a novel trypsinogen isoenzyme called tumor-associated trypsinogen 2 (TAT-2). It was found during a search of the target protease for tumor-associated trypsin inhibitor (TATI). We now show that degradation of subendothelial cell extracellular matrix (ECM) by four different cell lines (COLO 205 colon carcinoma, K-562 erythroleukemia, CAPAN-1 pancreatic carcinoma, and HT 1080 fibrosarcoma) can be partially inhibited by TATI or neutralizing trypsin antibodies. When cells were cultured in serum-free medium on ECM, TATI and trypsin antibodies inhibited the release of immunoreactive fibronectin fragments from ECM by 47-54 and 40%, respectively. Degradation of isotopically labeled ([3H]serine, [3H]proline, and [35S]sulfate) ECM was also significantly prevented by TATI. At its maximum, it exerted a 57% inhibition on the degradation of [3H]serine-labeled ECM. Plasminogen added exogenously to the culture medium further potentiated the proteolysis of ECM. Interestingly, addition of enteropeptidase, an activator of TAT-2, also enhanced cell-mediated proteolysis as assessed by degradation of purified fibronectin coated onto the surface of wells. Immunoblot analysis showed that enteropeptidase-mediated proteolysis generated a pattern of fibronectin fragments similar to that obtained by digestion of purified fibronectin by TAT-2. These results demonstrate the existence of a proteolytic system in tumor cells which is dependent on the activation of TAT-2. We suggest that TAT-2 is involved in a protease cascade-stimulating tumor cell invasion and degradation of extracellular matrix.

Characterization of antigenic epitopes of potato virus Y.

Immunochemical analysis of overlapping synthetic hexapeptides covering the entire length of the coat protein of potato virus Y (PVY) revealed immunodominant regions both at the N-terminal and at the C-terminal end of the coat protein. Immunization of rabbits with synthetic peptides representing N- and C-terminal regions of the coat protein resulted in production of antibodies that reacted with PVY. Antigenicity of PVY peptides was found to correlate with predicted beta turns, with hydrophilicity and with predicted chain flexibility. Characterization of the immunochemical properties of PVY will facilitate the development of detection methods for potyviruses.

Effect of chemical modification of arginine and lysine residues of fibronectin on its antigenic and gelatin-binding activity.

The effect of chemical modification of arginine and lysine residues of fibronectin on its antigenic and gelatin-binding activity was studied by enzyme immunoassay techniques. Both modifications strongly reduced the gelatin-binding activity. Using conformation-specific antibodies it was shown that modification of lysines caused extensive conformational changes in the molecule. No such changes could be detected in arginine-modified fibronectin. The results suggest that arginine residues are directly involved in the binding of fibronectin to gelatin. Lysine residues seem to be important for maintaining a native conformation necessary for gelatin-binding.

Interaction of polyamines with proteins of human plasma: a preferential aggregation of fibrinogen and fibronectin (cold insoluble globulin).

The polyamines spermine and spermidine were found to aggregate proteins from human plasma and serum, apparently due to electrostatic interactions between these cations and anionic proteins. Fibrinogen and fibronectin (cold insoluble globulin) were identified as the major molecular components of aggregates, and fibronectin could be quantitatively removed from plasma by aggregation with spermine. The results indicate that fibrinogen and fibronectin have anionic groups which react with polyamines and which are essential for the solubility of these proteins.

Competitive enzyme immunoassay for human plasma fibronectin.

A competitive enzyme immunoassay for the determination of fibronectin in plasma is described. An enzyme conjugate prepared by coupling alkaline phosphatase to rabbit anti-human fibronectin antibodies by glutaraldehyde was used as principal reagent. The assay was performed by coating polystyrene tubes with purified fibronectin and reacting these coated tubes with a mixture of sample and enzyme-labeled antibodies. After overnight incubation, the amount of enzyme activity associated with the tube was determined. An assay range of 0.5-20 microgram/ml of fibronectin was obtained. The mean concentration of plasma fibronectin in female patients was found to be 270 microgram/ml (standard deviation 50 microgram/ml, n = 22). Denatured fibronectin had low activity in the assay. The presence of cross-reacting antigens in rat and guinea pig plasma was demonstrated by the enzyme immunoassay technique.

Affinity immunosensor for milk progesterone: identification of critical parameters.

The effects of several experimental parameters on the performance characteristics of a competitive-type immunochromatographic assay of milk progesterone were studied. Increasing the size of the colloidal gold particles used as a label increased both maximal signal obtained and sensitivity of the assay measured as slope of the progesterone standard curve. The concentration of the antibody used to prepare the detection zone was found to be critical factor, in that low concentrations of antibody resulted in a poor sensitivity. The compatibilities of various buffer systems with the assay were studied. The assay worked well with buffers having a broad pH range of 4.5-8.5.

A longitudinal study of screening for endometrial cancer by endometrial biopsy in diabetic females.

Diabetics are at high risk of developing endometrial cancer; the relative risk of endometrial cancer in diabetics is fourfold in comparison to non-diabetic controls. The purpose of this longitudinal study was to evaluate the effectiveness of screening asymptomatic diabetic females in terms of the premalignant and malignant endometrial findings, and to try to determine the optimal screening interval. In 1980-1981, a group of 462 diabetic females was identified and registered. One half of them (237) was invited to be screened. Endometrial samples were taken by using Vabra aspiration. The results of this first randomized screening in 1980-1981 have been published elsewhere. At that time 124 females participated. The remaining 225 females acted as an unscreened control group. Eight years later (1988-1989), both groups were invited to be screened. The Pistolet aspiration method was used. At this stage, group 1 (screened in 1980-1981) consisted of 78 females, and group 2 (not screened in 1980-1981) consisted of 148 females. In 85% (193/226) of the females, the uterine cavity was reached with the Pistolet instrument; 96% of the females found the pain acceptable. In the group screened twice (group 1), no pathologic lesions of the endometrium were found in the second screening. In the group screened for the first time (group 2), one female had endometrial adenocarcinoma (0.8%), one had complex hyperplasia without atypia (0.8%) and four had endometrial polyps (3.3%). In 165 cases of 193, both a cytologic and a histologic specimen were available. In 130 cases (79%) the cytology was of class I, including the one endometrial adenocarcinoma. In three cases (2%) it was of class II and in one case (1%) of class III. Endometrial biopsy by Pistolet aspiration was a method highly acceptable by the patients for examining the endometrium. However, cytologic examination was not able to show the existing endometrial adenocarcinoma. One endometrial sampling of asymptomatic diabetic females during early menopause could detect the bulk of the occult, slowly progressing lesions of the endometrium. Such screeening might be most efficient in terms of cost-benefit ratio.

Evaluation of three slide agglutination tests for rapid identification of Staphylococcus aureus.

Three slide agglutination tests for identification of Staphylococcus aureus were compared. The agglutination tests used for evaluation were Staphaurex (Wellcome Diagnostics), Staphyslide-Test (BioMerieux), and ANI S. aureus TEST (Ani Biotech Oy). A total of 347 isolates were analyzed, including 288 strains of S. aureus, 49 of S. epidermis, 11 of S. intermedius, 12 strains of other staphylococci and 14 non-staphylococcal strains. One hundred of the S. aureus strains were isolates from cases of food poisoning, 129 from mastitis and 59 from other clinical cases. The sensitivities of the tests were also compared using diluted suspensions of S. aureus strains and with purified Protein A dilutions. The results showed that the sensitivities of the tests were 98.6%, 97.9% and 99.0% for Staphaurex, Staphyslide-test and ANI S. aureus TEST, respectively. The specificities were 100% for the Staphyslide test and 98.8% for both the ANI S. aureus TEST and the Staphaurex test. The sensitivities measured with diluted S. aureus strain suspensions and Protein A solutions were equal with the Staphaurex and ANI S. aureus TEST. All the agglutination tests studied proved to be practical, easy to use and accurate for the rapid identification of S. aureus strains from culture isolates.

Reconstruction of the vulva with sensate gluteal fold flaps.

BACKGROUND AND AIMS: Soft-tissue reconstruction of the vulva following resection of malignancies is challenging. The function of perineal organs should be preserved and the reconstructed area should maintain an acceptable cosmetic appearance. Reconstruction with local flaps is usually sufficient in the primary phase after a radical vulvectomy. Numerous flaps have been designed for vulvar reconstruction usually based on circulation from the internal pudendal artery branches. In this paper we introduce our modification of the gluteal fold V-Y advancement flap as a primary reconstruction after a radical vulvectomy. PATIENTS AND METHODS: Twenty-two patients were operated with a radical vulvectomy because of vulvar malignancies. The operation was primary in eight and secondary in 14 patients. The reconstruction of the vulva was performed in the same operation for each patient. RESULTS: All flaps survived completely. Wound complications were registered in three patients. Late problems with urinary stream were corrected in two patients. A local recurrence of the malignancy was observed in six patients during the follow-up period. CONCLUSIONS: Gluteal fold flap is easy to perform, has a low rate of complications and gives good functional results. Even a large defect can be reconstructed reliably with this method. A gluteal fold V-Y advancement flap is sensate and our modification allows the flap to be transposed with lesser dissection as presented before.

Hemagglutinin activity of human plasma fibronectin.

Purified human plasma fibronectin at concentrations of about 30 microgram/ml was found to agglutinate trypsin-treated erythrocytes from certain species. The hemagglutination reaction was inhibited by specific antibodies to fibronectin, by relatively low concentrations of polyamines and by higher concentrations of basic amino acids and nonacetylated amino sugars. The divalent cations Ca2+ and Mg2+ and the chelating agent ethylenediaminetetraacetate did not affect the reaction. None of the neutral amino acids, neutral sugars or polyanions tested was inhibitory. The results imply that plasma fibronectin is capable of interacting with cell surfaces and support the idea of a similarity between cellular and plasma fibronectins.

Dissociation of fibronectin from gelatin-agarose by amino compounds.

Soluble fibronectin of human plasma was specifically dissociated at neutral pH from gelatin-agarose by several cationic amino compounds, notably the polyamines spermine, spermidine and putrescine, the basic amino acid arginine, and amino sugars. The neutral and acidic amino acids and the N-acetylated derivatives of amino sugars tested were ineffective. Gel-filtration experiments demonstrated that [14C]spermidine bound to fibronectin but not to gelatin.

Effect of lectins and chemical modification on the hemagglutinin activity of human plasma fibronectin.

Purified human plasma fibronectin has been shown to agglutinate protease-treated red cells [Vuento, Hoppe-Seyler's Z. Physiol. Chem. 360, 1327-1333, (1979)]. The present report shows that the activity is inhibited by low concentrations of lectins and by macromolecular serum factors. Chemical modification of carboxyl groups of fibronectin strongly inhibited the activity, but modification of amino groups of guanidinium groups had little effect on the activity. The results suggest that fibronectin receptors on erythrocyte surface are carbohydrate-containing molecules. Humoral macromolecular factors may control the interaction of fibronectin with cell surfaces. Chemical modification studies indicate that the parts of the fibronectin molecule responsible for the hemagglutinin activity are different from those mediating the binding of fibronectin to collagen.

Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions.

Fibronectin was purified from human plasma by affinity chromatography under nondenaturing conditions. The method was based on the previously known binding of fibronectin to gelatin. The novel features of our method are the use of arginine in the elution of fibronectin from immobilized gelatin [Vuento & Vaheri (1978) Biochem. J. 175, 333-336] and the use of arginine-agarose as second affinity step. The purified protein was homogeneous as judged by polyacrylamide-gel electrophoresis, analytical ultracentrifugation and two-dimensional immunoelectrophoresis. The yield was 60%. We propose that the method would be useful in preparation of fibronectin for studies on its biological activities, where it is important that the protein is obtained in a native state.

Identification of fibronectin fragments that bind to carboxy-group-modified proteins.

Limited proteolysis of human plasma fibronectin with chymotrypsin, trypsin or thermolysin has been used to localize binding sites responsible for binding [Vuento, Korkolainen & Stenman (1982) Biochem. J. 205, 303-311] of fibronectin to carboxy-group-modified proteins. These bindings sites are different from those mediating binding of fibronectin to gelatin or heparin. They are located close to the C-terminus of the polypeptide chains of fibronectin, and apparently overlap with the C-terminal fibrin binding site.

A monkey antigen crossreacting with carcinoembryonic antigen, CEA.

Normal monkey tissues were found to contain an antigen which crossreacts immunologically with the carcinoembryonic antigen (CEA) of the human digestive tract. The monkey antigen reacted with complete or partial identity to the normal crossreacting antigen (NCA) in humans when tested in immunodiffusion against anti-CEA or anti-NCA. Extracts of monkey tissues inhibited in radioimmunoassays measuring human NCA. It is possible that monkey foetuses and colonic tumours contain CEA.

Immunochromatographic assay for quantitation of milk progesterone.

We describe a rapid immunochromatographic method for the quantitation of progesterone in bovine milk. The method is based on a 'competitive' assay format using the monoclonal antibody to progesterone and a progesterone-protein conjugate labelled with colloidal gold particles. The monoclonal antibody to progesterone is immobilized as a narrow detection zone on a porous membrane. The sample is mixed with colloidal gold particles coated with progesterone-protein conjugate, and the mixture is allowed to migrate past the detection zone. Migration is facilitated by capillary forces. The amount of labelled progesterone-protein conjugate bound to the detection zone, as detected by photometric scanning, is inversely proportional to the amount of progesterone present in the sample. Analysis is complete in less than 10 min. The method has a practical detection limit of 5 ng of progesterone per ml of bovine milk.