Coffee consumption in hypertensive men in older middle-age and the risk of stroke: the Honolulu Heart Program.

OBJECTIVE: To examine the association between coffee consumption and the development of stroke in men at high risk for cardiovascular disease. METHODS: Coffee intake was observed from 1965 to 1968 in a cohort of men enrolled in the Honolulu Heart Program with follow-up for incident stroke over a 25-year period. Subjects were 499 hypertensive men (having systolic or diastolic blood pressures at or above 140 and 90 mm Hg, respectively) in older middle-age (55 to 68 years) when follow-up began. Past and current cigarette smokers were excluded from follow-up. RESULTS: In the course of follow-up, 76 men developed a stroke. After age-adjustment, risk of thromboembolic stroke increased significantly with increases in coffee consumption (P = 0.002). No relationships were observed with hemorrhagic stroke. When adjusted for other factors, the risk of thromboembolic stroke was more than doubled for men who consumed three cups of coffee per day as compared to nondrinkers of coffee (RR = 2.1; 95% CI = 1.2-3.7). CONCLUSIONS: Although in need of further confirmation, consumption of coffee appears to be positively associated with an increased risk of thromboembolic stroke in hypertensive men in older middle-age. Findings suggest that it may be prudent to advise older middle-aged men with hypertension who consume large amounts of coffee to consider reducing their coffee intake.

Action of adenosine triphosphate on human skin and gingival fibroblasts. Protective action of fibronectin.

If cultured in media supplemented with adenosine triphosphate (ATP), EDTA, trypsin, thrombin, or incubated at 0-15 degrees C, human skin fibroblasts (HSF) and human gingival fibroblasts (HGF) change from long attached elliptical to round floating cell cultures. Also, if treated with ATP, EDTA, trypsin, thrombin, or incubated at 0-15 degrees C, the attached HFS or HGF monolayers detach from plastic substratum and form floating round cells that progressively aggregate together and die. The described experiments examined the role of cellular and extracellular ATP on HSF and HGF attachment. These two types of fibroblasts differed in their cellular ATP levels and their response to metabolic inhibitors. ATP causes destruction of microtubules as monitored by colcemid uptake and cellular detachment. Fibronectin protects both HSF and HGF from the effects of extracellular ATP.

Correlation between tyrosine hydroxylase activity, melanogenesis, and estradiol binding in human melanoma cells.

Cytosols fractions from human mammary carcinoma, or malignant melanoma cells, and tyrosine hydroxylase (TH) bind 3H-estradiol (3H.E) with strong affinity. Monoclonal antibodies secreted by cloned cell hybrids which were formed by the fusion of murine myeloma cells with spleen cells from BALB/c mice pre-immunized with purified TH were used to differentiate between 3H.E-binding protein in the cytosols of various neoplastic cells. These monoclonal TH-antibodies inhibited the enzymic activity and the binding of 3H.e to TH or to cytosol fractions of melanotic melanoma cells, but had no effect on 3H.E binding to cytosol fractions from various mammary epithelial or carcinoma cell lines. Cultivation of estradiol-responsive melanotic melanoma cells in media supplemented with the TH-antibodies, caused loss of pigmentation, the cells became amelanotic and non-responsive to 17 beta-estradiol. The results indicate that the estrogen does not bind to melanoma cells via a receptor as for mammary carcinoma cells but to tyrosine hydroxylase.

Monoclonal anti-tyrosine hydroxylase antibodies.

Tyrosine hydroxylase (TH) and cytosol fractions from human mammary carcinoma and malignant melanoma cells bind 17 beta-[3H]estradiol with strong affinity. Sucrose density gradients suggested differences between the binding of 17 beta-[3H]estradiol to TH and melanoma cell cytosol compared with the binding of the estrogen to mammary carcinoma. Monoclonal antibodies secreted by cloned cell hybrids which were formed by the fusion of murine myeloma cells with spleen cells from BALB/c mice immunized with purified TH inhibited the enzymatic activity and the binding of 17 beta-[3H]estradiol to TH or to cytosol fractions of melanotic melanoma cells, but had no effect on 17 beta-[3H]estradiol binding to cytosol fractions from various mammary epithelial or carcinoma cell lines. Cultivation of estradiol-responsive melanotic melanoma cells in media supplemented with the TH antibodies produced loss of pigmentation, and the cells became amelanotic and nonresponsive to 17 beta-estradiol. The results indicate that tyrosine hydroxylase of melanotic melanoma cells is an estrogen-binding protein.

Long-term cultivation of human mammary carcionoma: proliferation and differential biochemical properties of the cultured cells.

Human mammary carcinoma cell cultures proliferated from primary explants in Eagle's essential medium (MEM) supplemented with insulin, fetal calf serum (FCS) and/or human alpha-a1-antitrypsin. Human mammary carcinoma cells differed from normal mammary epithelial cells by the following catalytic activities: a. Thymidine uptake into the carcinoma cells was 6 to 10 fold greater, whereas thymidine conversion to CO2 was half to one fifth that of normal cells. b. The nucleolytic activity patterns of the mammary carcinoma cells preferred polycytydylic acid and double helical polynucleotides, whereas those of the normal mammary cells preferred polyuridylic acid and had no effect on double helical polynucleotides. c. The polymerase activity most evident in mammary carcinoma cells is a hybrid-dependent DNA polymerase which is guided by the ribo-strand of the template poly (rA) . poly(dT). In contrast the all-ribo template poly (rA) . poly(rU) showed little activity. d. There was slight or statistically non-significant difference between the amino acid composition of material cleaved from mammary carcinoma cells prepared from tumor tissues and from cells cultivated 10 months in vitro. e. There was no difference between the molar proportions of the carbohydrate components of the cell membrane from fresh tumor tissue and long term in vitro cultivated cells. f. The granules from long term in vitro cultured mammary carcinoma cells contained high collagenolytic, caseinolytic, fibrinolytic and esterolytic activities.