[Foam cell nephropathy in heterozygous female with Fabry's disease]. / Nefropatía de células espumosas en mujer heterozigota portadora de la enfermedad de Fabry.

We describe the clinical and pathological characteristics of a 49 year-old heterozygous female carrier of Anderson-Fabry's disease. Light microscopy and ultrastructural study of a renal biopsy showed the presence of foam cell nephropathy and galactosylceramide deposits affecting podocytes, the parietal epithelium of Bowman's capsule and the distal tubular cells, endothelial cells and medullary interstitial elements. Retrospectively, the presence of storage disease was confirmed in a hysterectomy specimen obtained two years previously. Our observation shows that heterozygotes for this disorder can not only carry and transmit the disease but may also develop pathological deposits in various organs. A renal biopsy from these carriers allows precise identification of the disease, facilitates adequate genetic counseling and gives the option of enzyme replacement therapy in patients who have pathological deposits.

[Immunohistochemical evaluation of the in vitro bromodeoxyuridine labeling index. 236 breast cancers studies by image analysis]. / Evaluation immunohistochimique de l'index de marquage après incorporation in vitro de bromodéoxyuridine. A propos de 236 cancers mammaires étudiés par analyse d'image.

We studied with computerized image analysis 236 breast cancer samplings after in vitro bromodeoxyuridine incorporation and immunohistochemical revelation. Labeling index values were compared with the usual prognostic factors and with the other studies in the literature. We established a positive correlation between labeling index and tumor size, histoprognostic grading, phase S and DNA index. A high labeling index was correlated with the absence of hormonal receptors but not correlated with the other prognostic factors. These results on tumor kinetics are similar to those obtained by flow cytometry and from other studies in the literature. However, this technic using optical microscopy allows for reliable selection of tumoral cells. Furthermore, the semi-automated image analysis provides an objective and reproducible evaluation of the labeling index.

[Acute intermittent porphyria: a possible cause of abdominal pain]. / Porfiria aguda intermitente: una posible causa de dolor abdominal.

Acute intermittent porphyria (AIP) is an inherited disorder of heme metabolism. It can produce a variety of symptoms including abdominal pain. In Spain it is an uncommon disease and consequently may not be included in the differential diagnosis of acute abdominal pain. Two cases of AIP are reported, both of which started with recurrent abdominal pain. A brief commentary of the main topics of the disease is made with special emphasis on the importance of an early diagnosis.

Detection of rare circulating breast cancer cells by filtration cytometry and identification by DNA content: sensitivity in an experimental model.

Current methods of detecting micrometastases in breast cancer fail in a large proportion of patients. Therefore an improved method for detection of metastases in blood samples could be of great clinical interest both for prognosis and selection of patients for adjuvant systemic therapy. We have developed a new non-invasive method which associates immuno-magnetic separation and filtration cytometry. The sensitivity of our procedure was evaluated in a model system using a mixture from a human breast cancer cell line (MCF-7) and a normal human blood sample. The identification of tumoral cells was achieved by measuring DNA content in comparison with standard cells. The lowest concentration of MCF-7 detected was 1 tumoral cell in 500,000 white blood cells. In addition, filtration cytometry provides a visual control of nuclei permitting the elimination of all doubtful cases and an automatic count of tumoral cells directly per ml of blood, which may be an independent predictor of early relapse. This new method may avoid unnecessary axillary lymph node dissection in patients with negative nodes. Our procedure seems suitable for the detection of rare circulating cells in routine laboratory testing and could be used in other applications.

Cell cycle expression of estrogen receptors determined by image analysis on human breast cancer cells in vitro and in vivo.

We have investigated, by image analysis, the cell cycle expression of estrogen receptors (ER) on MCF-7 cell line and on MCF-7 xenografts. The results demonstrate, in vitro as well as in vivo, an increase of ER concentration during the G0/G1-phase, followed by a decrease during the S-phase until the late S-phase where a rapid increase was noted. These results confirm that estrogens are involved in the DNA synthesis since ER is expressed in vivo at a maximal level in the late G1. In presence of saturating concentrations of 17 beta-estradiol, the mean ER concentration in G0/G1 phase is significantly decreased compared with the control cells cultured in estrogen-deprived medium. This indicates that 17 beta-estradiol down-regulates ER preferentially in the G0/G1 phase. These data suggest that ER in S and G2/M phases is unable to interact with its ligand. Consequently, estrogens may have no effects on the entry of cells in mitosis. Finally, after long-term tamoxifen treatment of MCF-7 xenografts, a tamoxifen-resistant tumor was developed which was characterized by a change in the profile of ER concentration during the G0/G1 phase. In conclusion, it is possible that the differences in cell cycle distribution of ER could be correlated with different phenotypes of breast cancer and also with different clinical phases of tumoral evolution. However, it remains to be known what is the clinical significance of the ER cell cycle expression in relation to tumor aggressiveness and survival.

Cell growth inhibitory effects of caulerpenyne, a sesquiterpenoid from the marine algae Caulerpa taxifolia.

Caulerpa taxifolia (Vahl) C. Agardh (Ulvophyceae, Caulerpales) is an algae of tropical origin that was accidentally introduced into the Mediterranean in 1984. Caulerpenyne (Cau) is the major metabolite present in Caulerpa taxifolia. This metabolite has previously been shown to be cytotoxic against cell lines in culture as in KB cells and fibroblasts from hamsters. Cau along with 6 other drugs representative of the major classes of anticancer products was tested against 8 cancer cell lines of human origin. Cau demonstrated growth-inhibitory effects in all cases with some variability between cell lines; this inter-cell variability was, however, less marked than that observed with the anticancer drug tested. Cells of colorectal cancer origin were the most sensitive to the presence of Cau with IC50 values of 6.1 and 7.7 microM. Increasing the duration of contact between Cau and the cells from 75 min to 29.5 hr did not improve the cytotoxic efficacy of this compound. When Cau was pre-incubated in the culture medium for from 7 to 83 min before being exposed to CAL 27 cells (head and neck cancer origin), there was a constant loss of cytostatic action of Cau as a function of Cau pre-incubation time. As the bovine serum albumin concentration increased in the culture medium, the concentration-response curves showed a constant shift towards the right, indicating a loss of cytostatic activity of Cau. In the presence of Cau, cells in culture clearly exhibited an early and marked shift into S phase followed by a blockade into the premitotic G2 M phase. Possible targets for CAU remain to be identified. Cau needs to be tested on tumor bearing animals to confirm this promising antiproliferative activity.

Effects of tamoxifen on potential doubling time of human breast cancer cell line determined by image cytometry of double fluorescent BrdU and DNA labeling.

Tamoxifen is extensively used for the treatment of human breast cancer. However, the mechanisms by which antiestrogens regulate the growth of estrogen receptor positive tumors have not been totally defined. A new methodology, using automated image analysis BIOCOM 500, was developed for determining potential doubling time (Tpot) of tumors. This new method was checked on three different human breast cancer cell lines (MCF-7, CAL 85-1, CAL 148) in comparison with flow cytometry and then applied to determine the effects of short-term tamoxifen treatment on Tpot of MCF-7 cells. Using the resulting bivariate contour plot of blue fluorescence (DNA content) versus green fluorescence (Bromodeoxyuridine content), a labeling index (LI) value of 0.39 +/- 0.05 and a Tpot value of 21 +/- 2.09 hours were determined for MCF-7 cells. As expected, data demonstrated that 72 hours of 1 microM tamoxifen treatment decreased the LI to 35% by increasing the proportion of G0/G1 cells. It increased the Tpot to 35% compared to untreated cells (Tpot = 31.8 + 4 hours) by a lengthening of G0/G1 phase without changing the length of S phase (Ts = 10.2 +/- 1 hours). At suprapharmacological concentrations (5, 10 microM), an approximately 50% increase in Tpot was observed without modification in Ts. These data suggested a specific cell cycle action of tamoxifen which was probably mediated by mechanisms other than estrogen inhibition, since these experiments were performed in estrogen-deprived medium. In addition, the automated imaging procedure appears to provide a rapid and quantitative approach to determine Tpot in fine needle biopsies which is useful for investigating alterations in cell growth after endocrine treatment or chemotherapy.

[Acute renal failure after the administration of enalapril as presentation form of Takayasu's arteritis]. / Insuficiencia renal aguda tras la administración de enalapril como forma de presentación de la arteritis de Takayasu.

The case of a 19 year old patient with Takayasu arteritis with exclusive involvement of the abdominal aorta and without involvement of the renal arteries who presented severe arterial hypertension during the acute phase of the disease is referred. The administration of a dose of 20 mg of enalapril maleate was followed in less than 24 hours by lameness of the lower limbs and acute oligoanuric renal failure, both of which were reversible after discontinuation of the drug. During the inflammatory phase of the disease, treatment with high doses of prednisone was effective in the control of the general symptoms and the biologic parameters of inflammation. Surgical revascularization was successfully carried out 5 months after diagnosis. Two years later the patient continues to be normotense and asymptomatic.

Oestrogen and progesterone receptor status in bone biopsy specimens from patients with breast cancer.

Of 16 breast cancer patients with histologically proven, tumour-infiltrating biopsy specimens most had low ER and PR values; the ER and PR contents varied between 0 and 135 and 0 and 44 fmol/mg protein, respectively. With the conventional clinical threshold of 10 fmol/mg protein, 8 specimens (50%) were ER-PR-, 4 (25%) ER-PR+, 3 (19%) ER+PR+ and 1 (6%) ER+PR-. ER levels were significantly lower in the tumoral bone lesion compared with the primary tumour. For 15 patients with negative biopsies and without endocrine treatment, ER and PR concentrations were quantifiable (2 fmol/mg protein or more) in 9 (60%) and 11 cases (73%), respectively. 8 of 9 patients over 55 (89%) were ER+ (2 fmol/mg protein or more). Conversely, for patients under 55, 1 of 6 (17%) was ER+ (P less than 0.001). Results for PR were similar. These data strongly suggest that steroid receptors are present in healthy bone tissue.

Factors affecting relapse in node-negative breast cancer. A multivariate analysis including the labeling index.

Between 1975 and 1982, 167 patients with carcinoma of the breast without axillary lymph node metastases were studied. The thymidine labeling index (LI), representing the percentage of cells in the DNA synthesis phase, was measured in all these patients. High LI values were more frequently encountered in young patients (P = 0.05), in low estrogen receptor (ER) tumor content (P = 0.007) and in high grade tumors (P = 0.0002). The overall 8-year relapse-free survival (RFS) was 68%. Univariate analysis demonstrated that RFS was influenced by histological grading (P = 0.03), ER (P = 0.03), PR (P = 0.02) and LI (P = 0.01). Multivariate analysis using the Cox regression model selected the LI as the single significant prognostic factor with regard to RFS (P = 0.037). These results emphasize the important role of cell proliferation kinetics in defining node-negative breast cancer patients with a high risk of relapse.

HPLC micromethod for simultaneous measurement of estradiol, progesterone, androgen and glucocorticoid receptor levels. Application to breast cancer biopsies.

Estradiol (ER), progesterone (PR), androgen (AR) and glucocorticoid receptor (GR) levels were assayed in 25 breast cancer tumors. The tissue was pulverized and homogenized in buffer, then divided into two parts: one was assayed by the standard dextran-coated charcoal method (DCC), with Scatchard plot analysis, the other was assayed by a micromethod developed in our laboratory, as described below: --incubation of the cytosol with several ligands (labelled and unlabelled) selected to avoid unwanted cross-reactions --DCC separation, followed by extraction of all receptor-bound steroids by precipitation of proteins with methanol/TCA --separation of these steroids on a high pressure liquid chromatography (HPLC) column using a methanol/water solvent --collection of the fractions of the column outlet and counting. Use of three labelled ligands and appropriate unlabelled ligands allowed assays of the four receptors. This micromethod was highly correlated with the standard method: ER = 0.985 (P less than 0.001); PR = 0.999 (P greater than 0.001); AR = 0.989 (P less than 0.001); GR = 0.867 (P less than 0.001). Thresholds of positivity were not modified. This micromethod allowed simultaneous measurement of several receptors in 40 mg biopsy specimens and can be applied to other hormone-dependent tissues.

Optimised analysis of tamoxifen and its main metabolites in the plasma and cytosol of mammary tumours.

Recent biochemical and pharmacological findings concerning tamoxifen (TMX) have proven that both the unchanged drug and the main metabolites, N-desmethyltamoxifen (NDT) and 4-hydroxytamoxifen (4OHT) are biologically active. An HPLC method based on on-line post-column UV irradiation with fluorescence detection is described. Optimized conditions allowed complete and rapid separation of TMX 4OHT, NDT and two other recently reported metabolites, Y and Z. This method was applied to plasma and cytosol drug and metabolite analyses. In plasma, from the moment of initial drug administration until the steady state (after 1 month or more of continuous oral TMX treatment), the values of NDT to TMX ratios were completely reversed: 22 to 215 in mean %, P less than 0.01. The presence of metabolites Y and Z is significant. 4OHT, hardly detectable at the first dose, was measured at the steady state with high interpatient variability. It is hypothesized that metabolite evolution with time may be due to auto-induction of drug metabolism. In cytosols, which were all obtained during continuous TMX treatment, the ratios between TMX and metabolites were comparable to those observed in plasma, but with greater interpatient variability. Metabolite Y was not detectable in cytosols. This variability was not linked to the levels of cytosolic oestradiol receptors before initiation of treatment.

Multiple hepatic adenomas after long-term therapy with testosterone enanthate. Review of the literature.

We report a case of multiple hepatic adenomas developed in a 32-year-old man with renal allograft after long-term therapy with testosterone enanthate. Histological assessment showed a benign hepatocellular adenoma, which has not regressed over a 6-month follow-up since androgen withdrawal. A review of primary hepatic tumors associated with androgenic-anabolic steroid therapy published in the English literature has been carried out, concentrating on the more relevant recent publications. No previous case of hepatic adenoma with possible relationship to the use of non-17-alpha alkylated compounds has been reported.

Simultaneous micro measurement of steroid receptors in breast cancer.

The oestradiol (RE) and progesterone (RP) receptor levels were analyzed in 26 tumour fragments (200-500 mg) from breast cancer patients. After pulverization of tissue, one part was analyzed by the routine dextran-coated charcoal (DCC) method and the other by a micromethod as follows: (i) cytosol incubation using the DCC method but in the simultaneous presence of [3H]oestradiol and [3H]R5020 (ii) extraction of the steroids bound to the receptor by precipitation with ethanol/TCA (iii) high pressure liquid chromatography (HPLC) on a modular system, with a C185 microns column and an elution by gradient mixture methanol/water. The fractions were collected and the radioactivity counted. The separation of oestradiol from R 5020 was rapid and complete. In addition dexamethasone was separated by this system making possible triple measures of RE, RP and glucocorticoid receptors. A highly significant correlation was obtained between the 2 methods: RE = 0.996, P less than 0.001; RP r = 0.975, P less than 0.001, implying that the thresholds of positivity, i.e. for therapeutic decisions, remain unchanged. Simultaneous measurement of RE and RP in a single needle biopsy is possible with this micromethod.

[Determination of beta2-microglobulin in breast, lung, colorectal and ENT carcinoma (author's transl)]. / Dosage de la beta2-microglobuline dans les cancers du sein, du poumon, du côlon et de la sphère ORL.

Significant increase of plasma level beta2-microglobulin in cancer patients (breast, bronchus, colorectal and ENT) occurs rarely. More, the highest levels observed are within the range of non malignant diseases. We cannot assume that beta2-microglobulin assay will be useful as tumor marker.