F. De Negri

Microalbuminuria and endothelial dysfunction in essential hypertension

Microalbuminuria (urinary albumin excretion between 20 and 200 micrograms/min) and endothelial dysfunction coexist in patients with essential hypertension. To evaluate whether the two phenomena are related and the determinants of that association, we recruited 10 untreated males with essential hypertension and microalbuminuria without diabetes to be compared with an equal number of matched patients with essential hypertension excreting albumin in normal amounts and 10 normal controls. The status of endothelial function was inferred from circulating von Willebrand Factor antigen (vWF), a glycoprotein secreted in greater amounts when the vascular endothelium is damaged. vWF concentrations were higher in hypertensive patients with microalbuminuria than in hypertensive patients without and controls. Individual vWF and urine albumin-excretion values were correlated (r = 0.55, p < 0.002). Blood pressure correlated with both urinary albumin excretion and vWF. Left ventricular mass index and minimal forearm vascular resistances were comparable in patients with hypertension and higher than in controls; total and low-density lipoprotein cholesterol, triglycerides, lipoprotein-a, Factor VII, and plasminogen activator inhibitor-1 did not differ. Fibrinogen was higher and creatinine clearance lower in microalbuminurics. Albuminuria in essential hypertension may reflect systemic dysfunction of the vascular endothelium, a structure intimately involved in permeability, haemostasis, fibrinolysis, and blood pressure control. This abnormality may have important physiopathological implications and expose these patients to increased cardiovascular risk.

Modulation of hemostatic balance with antithrombin III replacement therapy in a case of liver cirrhosis associated with recurrent venous thrombosis

Patients with liver failure can present both thrombotic and hemorragic complications because of the deficiency in coagulation factors and inhibitors (protein C and S, antithrombin III) and impairment of fibrinolytic balance. Here we report the case of a 63-year-old man with liver cirrhosis, recurrent thrombosis, and features of low-grade consumption coagulopathy, showing severe antithrombin III deficiency (about 30% of normal values). Treatment with antithrombin III (2000 U/day) and low doses of heparin (5000 U b.i.d.) was successful in modulating the coagulation system toward an antithrombotic effect. After discharge from hospital the ambulatory treatment with antithrombin III concentrates (2000 U twice a week) allowed the attainment of antithrombin III activity of about 60% and prevented the patient from recurrence of venous thrombosis.

Interference of thyroperoxidase on immuno-cytochemical determination of steroid receptors in thyroid tissue

The presence of sexual steroid receptor proteins in thyroid tissue has been previously demonstrated by biochemical means. The aim of this study was to determine the estrogen (ER) and progesterone (PR) receptors in malignant (12 papillary and 1 follicular carcinoma) and nonmalignant (19 multinodular goiters, 1 Graves’ disease, 1 Hashimoto’s thyroiditis) thyroid diseases using immunocy-tochemical assay employing monoclonal anti-ER and anti-PR antibodies and the peroxidase-antiperoxi-dase technique. Positive results were obtained in 24/34 (70%) for ER (ER-ICA+) and 22/34 (64%) for PR (PR-ICA+). To evaluate the possible interference of thyroperoxidase in the immunostaining, in consecutive sections of a positive specimen, primary antibody or primary antibody plus bridging antibody or PAP complex was omitted. Using these modified procedures, staining distribution was similar to that obtained by the standard procedure: in contrast, no staining was found in the positive control, i.e. a breast cancer specimen. The inhibition of the endogenous peroxidase caused a loss of staining in both the standard and modified procedures on thyroid specimens; no staining modification was obtained in the positive control. These results suggest that the staining observed in thyroid tissue is not specific and related to the activity of thyroperoxidase on chromogen solution. The complete loss of staining after peroxidase inhibition appears to be in contrast with the results obtained by biochemical method, and different antigenicity of thyroid receptors in comparison with breast receptors may explain this discrepancy.

Combined factor VIII and IX inhibitors in a non-haemophilic patient: successful treatment with immunosuppressive drugs.

Summary.  A rare case of a patient with Sjogren syndrome and spontaneously acquired inhibitors of both factor VIII and factor IX is reported. Complete remission was obtained by means of immunosuppressive drugs.

Presence of endothelin-1 in the normal and pathological human thyroid

An immunohistochemical study with two rabbit polyclonal antibodies I-AR76 and CA-08-351 against Endothelin-1 (ET-1) was performed in 133 human thyroid specimens: 5 normal thyroids, 30 multinodular goiters (15 toxic and 15 nontoxic), 20 Graves’ diseases, 5 Hashimoto’s thyroiditis, 26 adenomas (6 Hürthle cell, 16 toxic and 4 nontoxic), 30 classic papillary carcinomas, 3 minimally invasive follicular carcinomas, 1 widely invasive follicular carcinoma, 3 undifferentiated carcinomas and 10 medullary carcinoma. All normal thyroids, non toxic multinodular goiters and non toxic adenomas, 4 (66%) Hürthle cell adenomas, 3 (15%) Graves’ diseases, 1 (33%) case of minimally invasive follicular carcinoma showed rare follicular cells with weak cytoplasmic immunoreactivity. Many immunoreactive follicular cells, with or without oxyphilic changes, were observed in all specimens of Hashimoto’s disease, while the lymphocytic infiltrate was always negative. Twenty-seven (90%) classic papillary carcinomas were positive. Immunoreactivity was intra-cytoplasmic, weak in 14 cases and intense in 13. The cells of toxic adenoma and toxic multinodular goiter were negative, whereas the acellular stroma was intensely positive in both cases. Medullary and undifferentiated carcinomas were negative. These results show ET-1 immunoreactivity in normal and pathological human thyroids. In particular, the high content of this peptide in the thyroid papillary carcinoma suggests that ET-1, whose mitogenic role has recently been emphasized, could be involved in the growth of this tumor.

Relationship between progesterone receptor and productive fibrosis as an index of tumor differentiation in breast cancer.

We studied the relationship between steroid receptors (SR) and various stromal parameters in 100 breast cancers. Each specimen was submitted to SR determination by the dextran-coated charcoal assay and to histologic examination. No relationship was found between the presence of SR and necrosis or fibroblastic cell content. There was an inverse correlation between SR positivity and the extent of lymphocyte infiltration (p < 0.05). ER + PR + status was strongly correlated with marked productive fibrosis (PF) (p < 0.005). When SR were separately assayed, only the presence of PR was correlated with PF (p < 0.005); similar results were obtained for PR levels (p < 0.01). PF was also associated with a higher nuclear grade (p < 0.001). In conclusion, in this study a strong correlation between PR positivity or concentration and extent of PF was demonstrated in breast cancer. Since PR synthesis is an expression of an intact regulatory pathway, our data suggest that stromal production in breast cancer is related to the degree of differentiation of malignant epithelial cells.

Activation of coagulation in smoking and non-smoking women using a third-generation oral contraceptive containing desogestrel

The concurret use of smoking and oral contraceptives affects the hemostatic balance, thereby inducing a thrombophilic state. In order to clarify the effects of this association on the hemostatic system, the possible changes in the markers of activation of coagulation (thrombin-antithrombin III complexes and prothrombin fragment F1+2) were evaluated in 35 women given a third-generation oral contraceptive for 6 months; 13 of these women (37.1%) were mild or moderate smokers. No differences were found in basal levels of the coagulation and fibrinolytic parameters between smokers and non-smokers. During oral contraceptive administration, both F1+2 fragment and thrombin-antithrombin III complex concentrations significantly increased both in smokers and in non-smokers (p < 0.01). Fibrinogen plasma levels increased in both groups (p < 0.01). Antithrombin III activity was reduced in both groups during treatment, but the difference was significant only in smokers (p < 0.05). Although the sample size of smokers was too small to draw definitive conclusions, present results appeared to confirm previous data about the effect of the concurrent use of smoking and oral contraceptives on antithrombin III levels, but did not demonstrate any additional effect of moderate smoking on the activation of the clotting system induced by this oral contraceptive preparation.