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Isolation of a 14 kDa antigen from Taenia solium cyst fluid by HPLC and its evaluation in enzyme linked immunosorbent assay for diagnosis of porcine cysticercosis.

A fraction with a major band of 14kDa was obtained from crude cyst fluid of Taenia solium cysticerci by 2-step chromatography. A first fraction isolated by gel filtration (Sephacryl S-300 high resolution) was purified using an anion exchange column (Mono Q HR 5/5) on high performance liquid chromatography. Evaluation of the analytic sensitivity of this fraction (F3) was carried out in an antibody enzyme linked immunosorbent assay (Ab-ELISA-F3) using serum samples from pigs experimentally infected with different doses of T. solium eggs. The cross-reactivity of F3 was evaluated with serum samples from pigs that were naturally or experimentally infected with Taenia hydatigena, Taenia saginata asiatica, Fasciola hepatica, Trichinella spiralis, Metastrongylus apri, Trypanosoma congolense and Sarcoptes scabiei, and with serum samples of rabbits hyper-immunised with metacestode cyst fluid of T. hydatigena and T. solium. Antibody titres of lightly or heavily infected pigs differed in their kinetics. However, the increase in F3-specific antibodies could not be related to the infection level. Analysis of the specificity of the F3 showed that serum samples of pigs infected with other parasites did not recognise this antigen. Cross-reaction with T. hydatigena occurred in ELISA using cyst fluid as antigen, but the F3 antigen fraction was not recognized by rabbit hyper-immune serum samples to T. hydatigena. Evaluation of the diagnostic sensitivity and specificity of the Ab-ELISA-F3 was done by a non-parametric receiver operating characteristic (ROC) analysis using 66 serum samples from Zambian village pigs. The total number of cysticerci of these pigs was determined by dissection (28 pigs harboured T. solium cysticerci and 38 were negative at dissection). In addition, 58 serum samples from Cameroonian pigs (28 pigs from cysticercosis-free farms and 30 pigs with cysticerci at tongue inspection) were used in a separate ROC analysis. The results from the ROC analysis yielded a low diagnostic value (area under ROC curve=0.48) with the sera from the Zambian pigs while a relatively high diagnostic value was obtained with the sera from Cameroonian pigs (area under ROC curve=0.78). The main factor contributing to a low diagnostic value based on the Zambian serum samples seemed to be the false-positive reactions that were likely caused by the occurrence of transient antibodies in the non-infected animals.

Animals↗

An androgen binding protein in the cyst fluid of patients with polycystic ovary syndrome.

Androgen binding proteins (ABP) have not been described in the ovary. We have studied cyst fluids from three patients with the Polycystic Ovary Syndrome and one patient with choriocarcinoma for the presence of ABP. All contained a high affinity, low capacity binding protein for dihydrotestosterone and 17beta-estradiol. Comparison of this protein with the plasma androgen binding protein (TeBG) in polyacrylamide gel electrophoresis and equilibrium dialysis did not reveal any significant differences in molecular size (Kg) net charge (Yo), radius R, molecular weight (mol wt), heat stability, steroid specificity, or binding affinity (Ka). Cyst fluid and plasma could not be distinguished from each other on the basis of protein composition as measured by cellulose acetate electrophoresis. In the one patient studied, the binding capacity for DHT in cyst fluid and plasma was 58.1 and 16.1 ng/Gm protein, respectively.

17-Hydroxycorticosteroids↗

Structures and concentrations of fifteen different steroid sulfates in human breast cyst fluids.

By applying capillary gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS), a simultaneous quantitation of all important steroid sulfates present in a number of breast cyst fluids, has been obtained. The fact that prevailing androgen sulfate structures in the cyst fluids are different from those in blood suggests at least intracystic metabolism of blood-born precursors. Particularly greater amounts of 5 alpha-reduced steroids are found in breast cysts. 5 alpha-Androstane-3 alpha,17 beta-diol is a major androgen sulfate of breast cyst fluids, its concentration being some 2000-fold that of blood. After prolonged topical application of progesterone on the breast, an accumulation of the sulfates of several pregnanediol isomers could be observed.

Adult↗

Radioimmunoassay of human epidermal growth factor in human breast cyst fluid.

Epidermal growth factor (EGF) was assayed radioimmunologically in three secretions of the human mammary gland: milk (n = 16), colostrum (n = 8) and breast cyst fluid (n = 139). The immunoreactivity (immunoreactive EGF) detected in these three fluids corresponds directly to monomeric EGF, as can be shown by the complete cross-reaction with the standard and the identical behaviour in chromatography on Sephadex G50. The mean concentration of EGF is significantly greater in breast cyst fluid (241 +/- 143 ng/ml) and in colostrum (197 +/- 56 ng/ml) than in milk (107 +/- 22 ng/ml). The range of individual values in cyst fluid is large, 5-945 ng/ml. A possible role of EGF as a paracrine or autocrine factor in the pathology of cystic dysplasia of the mammary gland and in the increased risk of malignant transformation is hypothesised.

Binding, Competitive↗

Growth factor profiles in breast cyst fluid identify women with increased breast cancer risk.

Gross cystic disease and breast cancer are hormonally induced diseases that may share a common biochemical environment conducive to abnormal proliferative responses. Breast cyst fluid samples were analyzed for specific growth factors and levels were compared with breast cancer risk. Growth factor profiles identified both women at increased breast cancer risk and subgroups of women with distinct clinical manifestations of gross cystic disease. Women at increased risk for breast cancer demonstrated in their breast cyst fluid lower levels of platelet-derived growth factor and transforming growth factor (TGF)-beta, compared to women at lower risk. The presence of multiple cysts was associated with increased mitogenic activity, increased epidermal growth factor (EGF) and TGF-beta breast cyst fluid levels, and recurrent cysts were associated with higher levels of EGF. Unique growth factor profiles were associated with each risk group or clinical state, suggesting that distinct proliferative environments, associated with different clinical outcomes, are present in the breast tissue of women with gross cystic disease.

Adult↗

Isolation and partial characterization of species-specific and cross-reactive antigens of Echinococcus granulosus cyst fluid.

Two parasite antigens have been isolated from Echinococcus granulosus hydatid cyst fluid using hydrophobic interaction chromatography, anion exchange chromatography and gel filtration chromatography. Initial characterization of the antigens indicates that both are glycoproteins, of approximately 20 and 48 kDa (Eg20 and Eg48). When the two antigens were tested with a battery of antisera from patients with heterologous parasitic infections, only Eg20 was found to be specific for E. granulosus. The Eg48 antigen cross-reacted with the sera of 33% of E. multilocularis patients. In both antigens, some of the epitopes recognized by antibodies in the sera of hydatid patients were periodate-sensitive. This suggests the involvement of carbohydrates in at least some of the antigenic determinants. Due to the abundance of the Eg48 antigen in the hydatid cyst fluid, it would be the more practically useful antigen for disease diagnosis, especially in countries where only E. granulosus is endemic.

Animals↗

Transforming growth factor alpha, beta 1 and beta 2 in breast cyst fluid.

Women who have palpable breast cysts lined by apocrine metaplastic epithelium (intracystic Na/K < 3) may have a higher risk of breast cancer than those who have breast cysts lined by flattened epithelium (intracystic Na/K > 3). Transforming growth factor-alpha (TGF-alpha) exerts a proliferative effect on many cell types and may play a role in mammary carcinogenesis. Transforming growth factor-beta (TGF-beta) stimulates the proliferation of mesenchymal cells but inhibits mammary epithelial cell proliferation and may exert a protective effect against breast cancer development. The aim of this study was to measure the distribution of TGF-alpha, TGF-beta 1 and TGF beta 2 in the two sub-groups of breast cysts. TGF-alpha was undetectable in all but 2 of the 46 samples of breast cyst fluid tested. TGF-beta 1 ranged from undetectable up to 25.4 ng/ml (n = 46) while the intracystic concentrations of TGF-beta 2 were generally higher, ranging from 0.30 ng/ml to 125 ng/ml (n = 42). There was no significant difference between the distribution of TGF-beta 1 levels in the 2 cyst groups, but TGF-beta 2 levels were significantly higher (P < 0.001) in the high electrolyte ratio group (Na/K > 3). A negative correlation was found between TGF-beta 1 and TGF-beta 2 levels in the high electrolyte ratio group (rs = 0.605, p = 0.006, n = 19). TGF-alpha is unlikely to play an important role in breast cancer development in women with palpable breast cysts. The significantly higher concentrations of TGF-beta 2 in the high electrolyte ratio group may explain, in part, the lower risk of breast cancer in this group of women.

Biopsy, Needle↗

Alpha 1-antichymotrypsin complexes in human breast cyst fluids.

Alpha 1-antichymotrypsin, a serum protease inhibitor, was found in 72 breast cyst fluids aspirated from women affected by gross cystic breast disease. When fractionated by gel chromatography, the presence of protein complexes or aggregates was demonstrated. A different distribution of the alpha 1-antichymotrypsin appeared to be related to the ionic composition of the breast cyst fluid; when compared with metabolically active apocrine cysts, a statistically significant increase of alpha 1 protease inhibitor values in flattened epithelial cysts was revealed (P < 0.001). Two-dimensional immunoelectrophoresis showed in apocrine cysts (Na/K ratio < 3) a characteristic double peak of alpha 1-antichymotrypsin immunoprecipitin curve. The relationship between this alpha 1 protease inhibitor and electrolyte profiles may provide further knowledge about the imbalance between proteases and their inhibitors on functional changes of gross cysts and might be useful in studies on their mechanism of formation and relationship to subsequent breast cancer.

Exudates and Transudates↗

Liesegang rings in renal cyst fluid.

Peculiar ring-like structures identified as Liesegang rings (LRs) were found in renal cyst fluid from three patients with benign renal cysts. They ranged in size from 5 to 820 mu. Most had a double-layer outer wall with equally spaced radial cross-striations and an amorphous central nidus. Special stains were performed in one case, and the results are discussed. Reports of LRs in cystic or inflamed tissues have recently appeared in the literature. Some LRs have been mistaken for eggs or mature components of the giant kidney worm, Dioctophyma renale. We propose that cytologic assessment of renal cyst fluid in conjunction with histologic examination decreases the likelihood of misdiagnosis of LRs.

Aged↗

A giant cystic mesenchymal hamartoma of the liver: diagnosis, management, and study of cyst fluid.

A 21-yr-old West Indian woman with a giant hepatic cyst is reported. Preoperative investigations revealed that it was not a simple congenital cyst but that it was highly vascular. Studies of the cyst fluid obtained by percutaneous catheter drainage suggested that the epithelial lining was of bile duct origin. Fluid output was increased by secretin administration, but remained unchanged after amiloride. Bromsulphthalein did not enter the cyst fluid after intravenous injection. A hemihepatectomy was performed and the histologic appearances of the lesion were consistent with cystic mesenchymal hamartoma, a rare lesion in this age group. The patient is well 4.5 yr later.

Adult↗

Detection of hydatid antigen in human cyst fluid by reverse latex agglutination test.

Diagnosis of cystic echinococcosis (CE) has, sometimes, proved to be of some difficulties whether by serological, radiological, or even by direct microscopic examination of cyst fluid. Therefore, with an aim to apply and assess another method of diagnosis, of pre - intra - or post-operative application, the present work had studied the use of reverse latex agglutination (RLA) test in the diagnosis of CE, as a test that could directly detect hydatid antigen in human cyst fluid samples. The results were reliable when compared with the direct microscopic examination of cyst fluid samples. Also, it was more reliable than indirect haemagglutination test. It showed 100% sensitivity. No false positive reaction was observed with samples from cystic tumours, with a resultant specificity of 100%. Moreover, the test is easy to perform with a visually interpreted results within 2-3 minutes. The reagents are inexpensive, stable and easily available. These merits make the RLA test suitable for the diagnosis of CE, particularly in suspected cases with seronegative results or cases with sterile cysts.

Animals↗

Intracytoplasmic eosinophilic inclusion bodies in benign breast cyst fluids.

Intracytoplasmic eosinophilic inclusion bodies (EIBs) observed in cytologic preparations of benign breast cyst fluid aspirates were morphologically similar to the well-known EIBs in urine specimens. EIBs have not been previously described in breast cyst fluids. A chi-square test proved a statistically significant correlation between the presence of intracellular EIBs and cellular degeneration. The possible significance of this finding with regard to the origin of EIBs is discussed.

Breast Diseases↗

Simultaneous Assay of ErbB-2 Protein and Carcinoembryonic Antigen in Cyst Fluid as an Aid in Diagnosing Cystic Lesions of the Breast.

C-erbB-2 oncoprotein (ErbB-2 protein)and carcinoembryonic antigen (CEA)were simultaneously evaluated in the breast cyst fluid of 6 patients with intracystic cancer, 6 patients with intracystic papilloma, and 42 patients with gross cystic disease. A combination test, using 12 ng of ErbB-2 protein and 150 ng of CEA/ml of cyst fluid, respectively, as cutoff values resulted in a positive detection rate of 83%(5/6 cases) for intracystic breast cancer. Two of the 6 patients with intracystic papilloma and 2 of the 42 patients with gross cystic disease were positive. In addition, elevated levels of these markers in cyst fluid correlated well with the expression of these antigens in the corresponding tumor. Thus, it can be concluded that measurements of ErbB-2 protein and CEA aid in the diagnosis of intracystic breast cancer, and the assay of ErbB-2 protein is valuable for detecting tumors in which this protein is overexpressed.

Journal Article↗

Evidence for a potent lipid secretagogue in the cyst fluids of patients with autosomal dominant polycystic kidney disease.

Transepithelial fluid secretion appears to be an important factor in the progressive enlargement of cysts in autosomal dominant polycystic kidney disease. Evidence indicates that the fluid within cysts harbors an autocrine, paracrine, or endocrine secretagogue with the capacity to modulate the rate of cyst expansion. Fluids from five patients with autosomal dominant polycystic kidney disease were studied to determine the chemical nature and the physiologic function of the putative secretagogue. The secretory activity of cyst fluid assayed with polarized monolayers of Madin Darby canine kidney cells could be ascribed to a lipophilic substance of molecular weight < 3,500 d that was not destroyed by freezing, boiling, or proteolytic digestion. This lipid stimulated the production of intracellular cAMP and increased the rate of fluid secretion when added to either surface of cultured renal epithelial cells. Anion exchange chromatography revealed biologic secretory activity to a greater extent in the neutral lipid than in the fatty acid and phospholipid fractions separated from cyst fluid. More extensive chromatographic separation showed preferential appearance of the secretagogue in a fraction of neutral lipids enriched in monoglycerides. Among several candidate lipids, 1-mono-arachidonyl glyceride and arachidonic acid were found to mimic the effect of the cyst fluid to stimulate fluid secretion by Madin Darby canine kidney cells; however, their abundance in cyst fluid was insufficient to account for the degree to which secretion was stimulated by cyst fluid. Moreover, the effect of the arachidonic acid species to stimulate fluid secretion was inhibited by treatment with indomethacin, whereas the effect of the cyst fluid was not. On the basis of this study, it was concluded that human autosomal dominant polycystic kidney disease cyst fluid contains an anonymous lipid with the capacity to stimulate fluid secretion in renal epithelia. This potent endogenous modulator of fluid transport may have an important role in determining the rate at which cysts expand in autosomal dominant polycystic disease.

Animals↗

CA 125 and carcinoembryonic antigen levels in cyst fluid, ascites and serum of patients with ovarian neoplasms.

The levels of CA 125 and carcinoembryonic antigen (CEA) in tumour cyst fluid, ascites and serum of patients with epithelial ovarian neoplasms were estimated. In addition, the serum concentration of both antigens was evaluated 10 days after operation. Independently of histological type of ovarian tumour, CA 125 and CEA values were significantly higher in cyst and ascitic fluid than in corresponding patients' serum. The higher values of both markers were also found in malignant than benign ovarian cysts. CEA occurred at higher concentrations in mucinous carcinomas whereas CA 125 values were comparable both in mucinous and serous cyst fluid. The levels of these markers in cyst fluid did not correlate with the serum concentrations. The significant individual differences existed in the ability of tumour cells to produce and release both studied antigens. Post-operative CEA levels returned to normal more slow than CA 125 values.

Adenocarcinoma, Mucinous↗

Biochemical analysis of breast cyst fluid as a possible predictor of breast carcinoma development.

The occurrence of breast cancer in patients with gross cystic disease is 2-5 times higher as compared to control group of women. During 3 years, 183 cyst fluid samples were analyzed in 129 females, in 30 patients of them the samples were analysed repeatedly. The distribution of the Na+/K+ ratio, considered as the measure of cancer risk, was found to be bimodal. In repeated analyses the type I cyst fluid markedly predominated (Na+/K+ < or = 4.0). A direct dependence on this ratio was found in the concentration of glucose, albumin, carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA) and its specific form, TPS; an indirect dependence was found for the level of uric acid, phosphates, alkaline phosphatase (ALP) and alpha-amylase (AMS). The predominance of apocrine metaplasia cells released into the cyst fluid is characteristic of type I cysts. A definitive assessment of significance of these parameters will be enabled by a long-term follow-up of the disease in the respective patients.

Adult↗

Estradiol, gonadotropins, and tumor markers in ovarian cyst fluid.

BACKGROUND: The study was designed to improve the discrimination between functional and neoplastic ovarian cysts in order to avoid unnecessary surgery. METHODS: Concentrations of tumor markers (CA 125, CEA, CASA, CA 72-4) and hormones (estradiol, FSH, LH) in cyst fluid were detected by enzyme immuno- or immunoradiometric assays. Wilcoxon test was used to evaluate the correlation of cyst fluid markers and histology. RESULTS: One hundred and thirty-eight ovarian cyst aspirates were investigated. Seventy-one patients (51.5%) had functional cysts whereas 67 (48.5%) had benign (n=59) or malignant (n=8) cystic tumors. Statistically significant correlations of CA 125 (p<0.0005) and CASA (p<0.02) with neoplastic histology were found. No significant correlation could be detected between CA 72-4, CEA, or hormone values and histology. Elevated estradiol concentrations are suspicious for functional cysts in premenopausal age. Low FSH and LH levels seem to be an indicator for functional cysts in peri- and postmenopausal age. CONCLUSIONS: The assessed analytes could not reliably distinguish between functional and neoplastic ovarian cysts. Our results indicate that CA 125 is a marker for neoplastic histology in a proportion of ovarian cysts. The use of FSH and LH in the diagnosis of postmenopausal blastomas needs further investigation.

Adolescent↗