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Aims: To determine several aspects of the epidemiology of chronic inflammatory bowel disease (IBD), including distribution of the various forms of IBD, sex, age at diagnosis considering lag-time to diagnosis, criteria used in the diagnosis, the relationship between educational level and activity, familial aggregation, phenotype (site and clinical type), number of admissions and mean hospital stay/year, surgical requirements and mortality., Patients and Methods: We carried out a retrospective (1954-1993) and prospective (1994-97) descriptive epidemiologic population study, in the fifth health district of Gijón in Asturias (Spain), with 225,798 inhabitants. A total of 595 patients diagnosed with chronic IBD according to the diagnostic criteria described by Lennard-Jones and Truelove for Crohn's disease (CD) and ulcerative colitis (UC) and according to Ashley B. Price's criteria for indeterminate colitis (IC) were studied. In all patients a complete clinical follow-up was performed., Results: During the study period, we diagnosed 595 patients with IBD [305 patients with UC (51.3%), 272 (45.7%) patients with CD and 18 (3%) with IC]. Sex distribution was 287 females (48.2%) and 308 males (51.8%), with a male/female ratio of 1.07. Mean age at presentation was 38.79 +/- 17.44 years (UC: 43.37 +/- 17.55; CD: 33.98 +/- 16.16; IC: 33.73 +/- 13.48), p = 0.000. Age at onset prior to diagnosis was as follows: UC: 42.03; CD: 30.47; IC: 30.99 (p = 0.000). Diagnostic criteria used in UC was symptomatic in 99.01% (p = ns), endoscopic in 95.04% (p = 0.000), and pathologic in 87.21% (p = 0.000); in CD diagnostic criteria used was radiologic in 85.29% (p = 0.000). A total of 29.1% of patients with UC and 66.7% of those with CD had higher education (p = 0.0005). Family history was found in 9.8%. Anatomical site was as follows: in UC: rectum 21%, 28.2% rectum and sigmoid, 22.3% left colitis, 4.2% distal to hepatic flexure and 24% pancolitis; in CD: 32.72% terminal ileum, 19.11% colon, 37.13% ileo-colon, 11.02% extensive intestinal and 3.67% gastro-duodenal. A total of 8.37% of patients with UC and 14.51% of those with CD had been hospitalized during the previous 4 years; mean hospital stay was 1.63 days in UC and 2.27 days in CD. The mean surgical requirements were 0.54 +/- 1.08 (31.59%); UC: 0.11 +/- 0.36 (10.2%); CD: 1.04 +/- 1.38 (56.25%), p = 0.000. The mortality rate was 48.73 deaths/1,000 inhabitants (UC: T = 65.57; CD: T = 33.08; IC:T= 0) p = ns. The standardized mortality ratio was 4.83 (UC: 6.51; CD: 3.28)., Conclusions: We highlight the uniformity of the distribution of IBD in relation to types of disease and sex. Patients with CD had a higher level of education. Genetic components play an important role in these diseases and familial aggregation was high, especially in CD. Complicated situations are infrequent in this group of patients. Morbidity was higher in patients with CD as reflected by surgical requirements and hospital stay. Mortality was lower in CD than in UC.

Aim: To determine the incidence and prevalence of inflammatory bowel disease in our area and to compare our results with those of other series from Spain and other parts of the world., Patients and Methods: Descriptive epidemiologic population study, retrospective (1954-1993) and prospective (1994-1997) in health district V of Gijón in Asturias (Spain) with 225,798 inhabitants. Diagnostic criteria used were those described by Lennard-Jones and Truelove for Crohn's disease and ulcerative colitis and those described by Ashley B. Price for indeterminate colitis. Annual incidence was expressed per 100,000 inhabitants. Prevalence was calculated excluding cured patients: proctocolectomized in ulcerative colitis (10 cases) and deaths (29 cases)., Results: During the period studied, 595 patients were diagnosed with bowel disease (305 patients with ulcerative colitis, 272 with Crohn's disease and 18 with indeterminate colitis). Mean annual incidence (1954-1997) was 6.128 (95% CI: 2.90-9.36). In the 4-year prospective study the incidence was 15.49 (95% CI: 11.19-21.79), 9.36 for ulcerative colitis, 6.08 for Crohn's disease and 0.77 for indeterminate colitis (UC/CD: 1.58). Prevalence was 246.23 (95% CI: 225.6-226.70, 212.79 for ulcerative colitis, 116.47 for Crohn's disease and 7.97 for indeterminate colitis., Conclusions: Incidence and prevalence obtained in our environment were higher than those described in other areas of Spain and were similar to those found in areas of Europe and other parts of the world with a higher incidence. Incidence and prevalence have increased since 1980, probably due to the widespread use of endoscopy as a diagnostic technique. Rates were higher in the prospective study than in the retrospective one.

Objective: Apolipoprotein D is a protein component of the human plasma lipid transport system but is also associated with a more favorable prognosis in women with breast cancer. This retrospective study was undertaken to examine the tumoral expression of apolipoprotein D in epithelial ovarian cancer and to analyze the possible correlation with tumor and patient characteristics as well as androgen receptors and their prognostic significance., Methods: Immunohistochemical evaluation was used to examine apolipoprotein D expression in paraffin blocks from 68 epithelial ovarian carcinomas., Results: A total of 18 (26.4%) tumors stained positively. No significant correlation was found between apolipoprotein D expression and patient or tumor characteristics and androgen receptor status. However, apolipoprotein D expression was significantly associated with prognosis in patients with residual tumor greater than 1 cm. Thus, patients with apolipoprotein-D-negative tumors had a poorer overall survival than those with apolipoprotein-D-positive tumors (P = 0.039). In addition, multivariate analysis demonstrated that apolipoprotein D expression was an independent prognostic factor with initial tumor size in this group of patients (P = 0.005)., Conclusions: Our results led us to consider the existence of apolipoprotein D expression by a significative percentage of ovarian carcinomas, and this protein expression might be of clinical usefulness for identifying lesions with different evolution., (Copyright 2000 Academic Press.)

Background: Male breast cancer (MBC) is a rare tumor in comparison to the same disease in the female population (FBC). Classical prognostic factors (tumor size, node status, estrogen receptor positivity, histological grade) have a similar prognostic value in both tumors, but MBC seems to have a worse outcome. Several markers under estrogen control have been shown with a similar incidence in breast tumors of both sexes, while androgen-induced markers have been detected in a higher percentage of breast tumors in males., Aims and Methods: Our purpose was to compare in 68 MBC and in 68 FBC the expression of Pepsinogen C (Pep C) and Apolipoprotein D (Apo D), two proteins under androgen control, by immunohistochemical methods., Results: Pep C was expressed by 52 of 68 (76.4%) MBC patients, whereas 34 of 68 (50%) FBC showed positive staining. Apo D was expressed by 57 of 68 (83.8%) MBC patients, while 40 of 68 (41.2%) FBC stained positively. Differences between percentages of positive expression were significant (p<0.005 for Pep C; p<0.0001 for Apo D). Moreover, differences between expression levels of Pep C and Apo D in both populations of patients were significant. The mean value of Pep C was significantly higher in male breast tumors (HSCORE = 141.3) than in the females (HSCORE = 80.3) (p<0.0001). Similarly, Apo D mean value was significantly higher in MBC (HSCORE = 161.5) than in FBC (HSCORE = 102.3) (p=0.006)., Conclusion: These differences can open the field of a more selective hormonal therapy that should not be based on estrogen receptor status only, but also on androgen receptor status.

Background: c-erbB-2 is a transmembrane signaling molecule closely related in structure to the epidermal-growth-factor receptor (EGFR) but biologically distinct from it. c-erbB-2 has been implicated in cell transformation and tumor pathogenesis, but very little is known about its content and clinical significance in colorectal cancer., Aims: To evaluate the c-erbB-2 content in colorectal cancer and its possible relationship with clinicopathologic parameters from tumors and prognostic significance., Methods: Membranous and cytologic c-erbB-2 oncoprotein contents were examined by an immunoenzymatic assay in tumors and paired normal surrounding mucosa samples from 131 colorectal cancer patients. In addition, survival analysis were prospectively performed in a subgroup of 69 consecutive patients with resectable colorectal carcinomas, who underwent a mean follow-up period of 28 months., Results: In the overall group of patients, c-erbB-2 levels were significantly higher in membranous than in cytosolic samples, in neoplastic tissues (5,830.4 +/- 1085.3 vs. 934.2 +/- 107.5 NHU/mg protein; p < 0.0001) and in surrounding normal mucosa samples (5,257.8 +/- 646.3 vs. 837.4 +/- 187.4 NHU/mg protein; p < 0.0001). Nevertheless, a significant positive relation was found between membranous and cytosolic oncoprotein levels in these two paired sets (p < 0.0001, for both). There were no significant differences in membranous or cytosolic c-erbB-2 protein levels between neoplastic tissues and surrounding mucosa samples in this overall group of patients. In addition, the results did not show significant correlations of these oncoprotein contents with clinicopathologic parameters from tumors such as location, stage, histologic grade, and DNA content or S-phase fraction. However, the results indicated that low membranous c-erbB-2 content (< 4,500 NHU/mg protein) in tumors predict shorter relapse-free survival and overall survival (p < 0.05, for both) in resectable colorectal cancer patients., Conclusions: There are a wide variability of both membranous and cytologic c-erbB-2 contents in colorectal carcinomas, which seems to correspond to the biological heterogeneity of these tumors. In addition, our results also demonstrate that high membranous c-erbB-2 levels are associated with lesions of favorable evolution in resectable colorectal cancer patients.

Background: Apolipoprotein D (ApoD), a protein component of the human plasma lipid transport system, is an androgen regulated protein in both breast and prostate cancer cell lines., Methods: Here we examined, by immunohistochemical assay, the ApoD expression in normal, benign and malignant prostate tissues., Results: ApoD was detected in the two analyzed normal prostate tissues, in 16 (94.1%) of 17 benign prostatic hyperplasias, and in 36 (63.1%) of 57 prostate carcinomas. In prostate carcinomas, ApoD immunostaining values did not show significant relationship neither with age of the patients, stage and histologic grade of the tumors, nor with preoperative serum levels of prostatic specific antigen (PSA). However, ApoD-negative immunostaining predicted shorter survival duration in the subgroup of 46 patients with nonresectable prostate cancer (p = 0.02)., Conclusions: Our results led us to consider that Apo D expression by prostate carcinomas migth be of clinical usefulness for identificating lesions with different evolution.

Background: Apolipoprotein D is a glycoprotein of the human plasma whose functional role remains unclear. On the other hand, this protein is also produced by breast carcinomas and is positively associated with a favorable outcome of patients. However, none study has focused on metastasic lesions., Aim: To analyze apolipoprotein D expression in breast cancer patients and their synchronous metastasic axillary lymph nodes., Methods: We analyzed by immunohistochemical assay both, the tumoral expression of apolipoprotein D in primary tumors and in their synchronous metastasic axillary lymph nodes of 30 node-positive breast cancer patients., Results: Of the primary tumors, 28 (93.3%) showed a positive immunostaining for apolipoprotein D, although there was wide variability immunostaining values. On the other hand, 16 (53.3%) patients showed a positive immunostaining for the protein in their tumoral lymph nodes. In addition, there was a significant positive relationship between the tumoral expression of apolipoprotein D in primary tumors and metastasic lymph nodes (P < 0.05). However, only immunostaining values of the protein in primary tumors achieve statistical signification (P < 0.05) as prognostic factor of favorable evolution to predict overall survival from patients., Conclusions: Apolipoprotein D is also expressed in metastasic lymph nodes of breast carcinomas, but with a different pattern of immunostaining and less clinical significance than in primary tumors.

Background: Human glandular kallikrein (hK2) belongs to the serine protease family of enzymes and has high sequence homology with prostate-specific antigen (PSA). The physiological role of hK2 has not as yet been determined, but there is evidence that it can regulate the proteolytic activity of PSA through processing and activating pro-PSA, an inactive precursor. Thus, it is conceivable that these two secreted proteins may coexist in biological fluids. Currently, hK2 is considered an androgen-regulated and prostate-specific protein. Recently, it has been demonstrated that hK2 is expressed in the breast cancer cell line T-47D after stimulation by steroid hormones, and we reported that hK2 can be detected in a subset of breast tumor extracts. These data suggest that hK2 may be expressed in tissues other than the prostate, such as those in which PSA has already been detected. Because hK2 is a secreted protein, it may be present in various biological fluids., Methods: We analyzed milk samples from lactating women, amniotic fluid from pregnant women, and breast cyst fluid from patients with gross breast cystic disease, using a highly sensitive and specific immunoassay for hK2., Results: hK2 was present in all three biological fluids. We suggest that the female breast may produce hK2 and provide evidence that hK2 may have value as an additional marker for the discrimination between type I and type II breast cysts., Conclusions: The female breast produces hK2 in addition to PSA. More studies are necessary to establish the role of this kallikrein in nondiseased breast, gross breast cystic disease, and breast cancer.

Pepsinogen C is a proteolytic enzyme involved in the digestion of proteins in the stomach; it is also synthesized by a significant percentage of female breast carcinomas. In addition, it has been demonstrated that pepsinogen C is one of the few proteins induced by androgens in breast carcinoma cells. Here we evaluate the expression of pepsinogen C by immunoperoxidase staining in normal breast tissue from 3 male patients, 15 gynecomastia tissues, 2 male in situ breast carcinomas, and 68 male invasive breast carcinomas. Pepsinogen C immunostaining values were quantified in male breast tumors using the HSCORE system, which considers both the intensity and the percentage of cells staining at each intensity. The results indicated positive immunohistochemical staining for pepsinogen C in all gynecomastia tissues, the two in situ ductal carcinomas, and 52 of 68 invasive breast carcinomas (76.4%). The three normal breast tissues analyzed showed negative staining for pepsinogen C, whereas invasive tumors showed clear differences among them with regard to the intensity and percentage of staining cells. In addition, pepsinogen C scores were significantly higher in well-differentiated (grade I, 188.7) and moderately differentiated (grade II, 145.8) tumors than in poorly differentiated (grade III, 98.5) tumors (p = 0. 032). Similarly, significant differences in pepsinogen C content were found between estrogen receptor (ER)-positive tumors and ER-negative tumors (158.5 vs. 44.3, respectively; p = 0.009). Patients with pepsinogen C-positive tumors reached longer relapse-free and overall survival periods than did those with tumors with negative staining, but no statistical differences were observed between survival curves calculated for these two groups of patients. This results demonstrate expression of pepsinogen C by gynecomastias and by a high percentage of male breast carcinomas and may indicate an important role of pepsinogen C in the pathophysiology of male breast diseases.

The expression CA195 in serum, defined by monoclonal antibody CC3C195 (IgM), was studied in 67 patients with pancreatic cancer and in 138 patients with biliary or pancreatic benign disease. The results were compared with carcinoembryonic antigen (CEA) expression. The overall sensitivity of the CA195 assay (> 12 U/ml) was higher than that for CEA (89.5% vs. 53.7%) (p < 0.001). Sensitivity was increased to 92.5% with the simultaneous use of the two antigens, but the difference was statistically significant only with CEA (p < 0.001). The specificity of CA195 calculated from all patients with benign diseases was lower than that of CEA (73.1% vs. 89.8%). However, using a cutoff value of 100 U/ml for CA195, the specificity of this antigen (82%) was higher than that of CEA. These results demonstrate that marked elevations of tumor antigen CA195 are relatively specific for pancreatic carcinoma, and that this antigen is superior to CEA for diagnosing pancreatic cancer by virtue of its higher sensitivity.

Collagenase-3 (MMP-13) is a matrix metalloproteinase recently identified on the basis of differential expression in normal breast tissues and in breast carcinoma. To date, collagenase-3 expression has been reported only in breast carcinomas and in articular cartilage of arthritic patients; the presence and possible implication of this enzyme in the progression of other malignant tumours are unknown. In this study collagenase-3 mRNA expression has been analysed by northern blot in a series of 35 matched squamous cell carcinomas of the larynx and the corresponding adjacent non-neoplastic tissues. In addition, mRNA expression of membrane type 1-matrix metalloproteinase (MT1-MMP) and gelatinase A, two matrix metalloproteinases which have the ability to activate collagenase-3 in vitro, was also examined in the same cases. No collagenase-3 expression was detected in any of the 35 normal mucosae, but collagenase-3 mRNA was observed in 20 of the 35 carcinomas (57 per cent). Western blot analysis revealed the presence of collagenase-3 protein in those carcinomas with high levels of mRNA expression, whereas no protein was detected in the carcinomas with negative mRNA expression, or in any of the normal tissues. The protein was localized predominantly in tumour epithelial cells. Collagenase-3 expression correlated significantly with better histological differentiation of the tumours (p = 0.026), as well as with advanced local invasion (p = 0.026). Collagenase-3 upregulation was also significantly associated with MT1-MMP and gelatinase A overexpression. These findings suggest that collagenase-3 expression may contribute to the progression of a significant subset of squamous cell carcinomas of the larynx and that its coordinate overexpression with MT1-MMP and gelatinase A may have a cooperative effect in the progression of the tumours.

Human collagenase-3 (MMP-13) is a member of the matrix metalloproteinase family of enzymes that was originally identified in breast carcinomas and subsequently detected during fetal ossification and in arthritic processes. In this work, we have found that collagenase-3 is produced by HCS-2/8 human chondrosarcoma cells. An analysis of the ability of different cytokines and growth factors to induce the expression of collagenase-3 in these cells revealed that basic fibroblast growth factor (bFGF or FGF-2) strongly up-regulated the expression of this gene. By contrast, other factors, including interleukin-1beta and transforming growth factor-beta, previously found to induce collagenase-3 expression in other cell types, did not exhibit any effect on the expression of this gene in chondrosarcoma cells. Further analysis of the bFGF-induced expression of collagenase-3 in human chondrosarcoma cells revealed that its effect was time and dose dependent, but independent of the de novo synthesis of proteins. Western blot analysis revealed that the up-regulatory effect of bFGF on collagenase-3 was also reflected at the protein level as demonstrated by the increase of immunoreactive protein in the conditioned medium of HCS-2/8 cells treated with bFGF. Immunohistochemical analysis of the presence of collagenase-3 in a series of 8 benign and 16 malignant cartilage-forming neoplasms revealed that all analyzed malignant chondrosarcomas stained positively for collagenase-3, whereas only 2 of 8 benign lesions produced this protease. In addition, the finding that bFGF was detected in all analyzed chondrosarcomas, together with the above in vitro studies on HCS-2/8 cells, suggest that this growth factor may be an in vivo modulator of collagenase-3 expression in these malignant tumors. These results extend the pattern of tumor types with ability to produce this matrix metalloproteinase and suggest that collagenase-3 upregulation may contribute to the progression of human chondrosarcomas.

Background: In populational studies the standardized questionnaires of multidimensional functional evaluation, particularly the Older Americans Resources and Services Program-Multidimensional Functional Assessment Questionnaire (OARS), have shown to be the most adequate tools to evaluate the elderly persons living within the community., Methods: A health care survey was performed by personal interview in the homes of a stratified randomized sample of people over the age of 65 years in Vigo, Galicia, Spain obtained from the Municipal Inhabitants Patronage (n = 841) using complete validated versions of OARS in the Spanish and Gallician languages., Results: The prevalence of severe/absolute handicap in the basic functional areas among this population over 65 years was 7.7% in physical health, 6.2% in mental health, 5.2% in economic resources, 6.7% in social work, and 9.5% in daily life activities (DLA). Twenty-two percent of the population presented severe/absolute handicaps in at least one of the dimensions. One point nine percent of the population studied required institutionalization: 2.6% required economic aid, 1.4% required home help for the instrumental DLA and 1.1% required help for both the basic and the instrumental DLA although these were not being provided. One point three percent were found to be ideal candidates for a telealarm/teleaid program., Conclusions: The use of the OARS questionnaire in Spain is useful to obtain information to be applied in the planning of health care services directed at elderly people.

The human p27kip1 gene encodes a cyclin-dependent kinase inhibitor implicated in the negative regulation of the cell cycle. In order to elucidate the possible role of p27 mutations in the development or progression of human breast cancer, we have studied the occurrence of genetic abnormalities in this gene in a series of 30 primary breast carcinomas. Direct sequence analysis of the polymerase chain reaction amplified human p27 gene revealed the occurrence of two sequence variations with respect to the reported sequence; both variants were also present in the lymphocyte DNA from the same patients. First, a silent G to A change at codon 142 (Thr) was detected in a single case. Second, a T to G transversion at codon 109, resulting in a Val to Gly change, was observed in eight tumour DNA samples (26%) and in 31 out of 80 unrelated normal individuals (39%). This latter change creates a Bg/I restriction site that might be useful for genetic analysis of human tumours. Despite the occurrence of these polymorphisms, we did not however find any evidence of somatic mutations in the coding region of the p27 gene. On the basis of these results, we suggest that alterations in the integrity of the human p27 gene are not common events in human breast carcinomas.

Apolipoprotein D (apo D) is a glycoprotein involved in the human plasma lipid transport system and present at large amounts in cyst fluid from women with gross cystic disease of the breast. Apo D expression in breast carcinomas was examined by immunoperoxidase staining of a series of 163 tumors. A total of 60 (36.8%) tumors were negative for apo D immunostaining, 28 (17.2%) carcinomas were weakly positive, 33 (20.2%) were moderately stained, whereas the remaining 42 (25.8%) tumors were strongly stained with the specific antibodies. No significant correlation was found between apo D content and tumor size, lymph node involvement, or biochemical parameters such as estrogen receptors, cathepsin D, or pS2 protein. However, the finding of a significant association between apo D and menopausal status of patients or differentiation grade of tumors, with apo D values being lower in tumors from premenopausal women or in poorly differentiated carcinomas, suggested a potential value of this glycoprotein as a prognostic factor in breast cancer. Preliminary analysis of relapse-free survival and overall survival in a subgroup of 152 women with a mean follow-up of 42 months confirmed that low apo D values were significantly associated to a shorter relapse-free survival and poorer survival. According to these data, we propose that apo D in combination with other well-established prognostic factors may contribute to more accurately identify subgroups of breast cancer patients with low or high risk for relapse and death.

The authors present a prospective double blind test aimed at objectively determining the acceptance of flavoured orally administered enteric diets specific to patients with glucose metabolism alterations (Glucerna and Precitene Diabet), in the light of the taste disorders described in such patients. Sixty-two patients were studied, 32 diabetics and 32 control patients, who were given a sample of each product; the level of acceptance was quantified on a modified wine-tasting scale. The average taste of the diabetics was 12.1 points (SD = 4.34) for Glucerna, and 10.1 (SD = 3.91) for the Precitene Diabet. In the control patients, the scores were 12.7 (SD = 3.78) and 13.2 (SD = 3.23) respectively. Multiple regression analysis did not reveal significant differences in taste according to age, sex or place of origin. Average taste among the diabetic patients as a whole with both products was 11.2 points and, for the nondiabetics, 12.92. The model detected significant differences (p = 0.01) between the two groups. The average taste of the diabetics was less than that of the non-diabetics, irrespective of all the remaining variables examined, including the type of preparation. These results confirm the lower oral acceptance in diabetic patients, possibly associated with disorders in the sense of taste, showing the utility of the modified wine-tasting scale as a test in evaluating the acceptance of enteric diets which must be administered orally.

Zn-alpha 2-glycoprotein (Zn-alpha 2-gp), a protein present at high levels in breast cyst fluid, has been measured in 104 breast tumour cytosols by using an immunoenzymatic assay. Concentrations of Zn-alpha 2-gp ranged from 0 to 23.5 micrograms/mg of total soluble protein, with an average value of 2.4 micrograms/mg. There was no significant correlation between Zn-alpha 2-gp and menopausal status, tumour size or lymph node involvement, or between this protein and biochemical parameters such as oestrogen receptor, cathepsin D or pS2 levels. However, there was a significant association between Zn-alpha 2-gp and histological grade of tumours, with higher Zn-alpha 2-gp levels in well-differentiated tumours (mean 4.6 micrograms/mg) than in moderately (1.8 micrograms/mg) or poorly (0.9 micrograms/mg) differentiated tumours. On the basis of these results, we propose that Zn-alpha 2-gp may be considered as a biochemical marker of differentiation in breast cancer.

The protein composition of breast secretions from 99 premenopausal women with benign or malignant breast diseases and from 70 control women without breast pathologies has been studied by using polyacrylamide gel electrophoresis. These fluids have been classified into two types according to their major polypeptide components. Type I fluids are defined by three major distinctive bands at Mr 44,000, 24,000, and 17,000, while those designated Type II present distinctive bands at Mr 80,000, 15,000, and 14,000. Amino acid sequencing and immunoblotting analysis demonstrated that proteins in Type I secretions correspond to Zn-alpha 2-glycoprotein, apolipoprotein D, and gross cystic disease fluid protein-15, while those from Type II fluids have been identified as lactoferrin, lysozyme, and alpha-lactalbumin. Most women (93%) without breast pathology and most patients (88%) with benign diseases had secretions with a Type I polypeptide pattern. By contrast, a large percentage (57%) of secretions from women with breast carcinoma presented a Type II protein pattern. Further studies with a large number of women will be useful for corroborating the potential clinical interest of breast fluid protein analysis.

Breast secretions can be classified into two types according to their major protein components. Type I fluids contain Zn-alpha 2-glycoprotein, apolipoprotein D, and gross cystic disease fluid protein-15, while Type II fluids are characterized by the presence of lactoferrin, lysozyme, and alpha-lactalbumin. In this study, the polypeptide composition of breast secretions from 719 nonlactating women was evaluated by using polyacrylamide gel electrophoresis. The required amount for the analysis (1 microliter) was obtained from 50% of control women and from 75% of women with mammary disease. There were more secretors in premenopausal than in postmenopausal women, as well as in parous than in nulliparous women. Evaluation of factors affecting protein composition of breast secretions revealed that Type II fluids were found in the majority of women who had given birth in the last four years and in a high proportion of oral contraceptive users. After excluding both of these groups, Type II fluids were detected in 47% of patients with breast cancer, but only in 8% of control women and in 16% of women with benign breast diseases. Taken together, these results suggest that protein analysis of breast secretions could be an useful tool for the study of breast pathologies.

Cyst fluids from 55 premenopausal women with gross cystic breast disease were classified by K+/Na+ ratio: 19 with ratio over 1 (type I) and 36 with ratio less than 1 (type II). Immunoradiometric assay of cathepsin D in both types of cyst fluids revealed the presence of large amounts of this proteinase. The average concentration of cathepsin D in type I cyst fluids was 63.3 nmol/l, which was significantly higher than that corresponding to type II cyst fluids (35.1 nmol/l). Immunoprecipitation analysis of intracystic cathepsin D demonstrated that this protein was present as the 52 kD non-processed precursor form of the molecule. Since procathepsin D is a useful prognostic marker in breast carcinoma, we suggest that cyst fluid quantification of cathepsin D could aid to detect patients affecting of gross cystic disease with higher risk for developing breast cancer.

We used sodium dodecyl sulfate-polyacrylamide gel electrophoresis to study cyst fluids from women with breast gross cystic disease. The subjects could be classified into two categories according to the concentrations of protein GCDFP-70 in the cyst fluid: those with Type I cysts had a very low content of this protein; those with Type II cysts had very high concentrations. Analysis of the amino acid sequence of GCDFP-70 from both cyst fluid types confirmed that this protein is human plasma albumin. The average concentration of albumin found in Type I cyst fluids was 0.32 g/L and that corresponding to Type II was 10.16 g/L. Thus, albumin quantification from cyst fluids or analysis by either polyacrylamide or agarose gel electrophoresis provides a simple procedure for classifying these fluids, yielding results that correlate well with previous classifications based on other measurements such as sodium, potassium, and chloride concentrations. This albumin-based quantification method may improve the classification of breast cysts and might be useful in further studies on functional changes in the cysts and their relationship to breast cancer.

Breast cyst fluids were aspirated during the mild luteal phase of the menstrual cycle from 88 patients with gross cystic disease. According to intracystic Na+/K+ ratio three categories of cyst were identified: Type I with low Na+/K+ ratio, low levels of chloride, glucose, and pH, and high concentrations of dehydroepiandrosterone-sulphate (DHAS) and apocrine epithelium; Type II with high Na+/K+ ratio presenting levels of chloride, glucose, pH, and DHAS similar to those found in plasma, and flattened epithelium; Type III with intermediate values for the above parameters. The findings suggest that the cysts could correspond to the different functional stages of the epithelium lining the cysts. Cyst classification was evaluated using a discriminant analysis, which allows breast cyst fluids to be assigned within a category defined by the Na+/K+ ratio. The apparent correct rate for each category was more than 87%. The variables that better discriminate among groups of cysts were chloride, glucose, intracystic prolactin, and pH, respectively. These discriminant functions could be used as criteria for an optimal classification of breast cysts, which might help study of the epidemiology of breast cancer and its relationship to gross cystic disease.