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A method has been developed for the direct determination of 7-hydroxycoumarin (7HC) and 7-hydroxycoumarin-glucuronide (7HCG) in urine without sample clean-up. Separation was carried out in 90% 100 mmol l-1 phosphate buffer, 11 mmol l-1 deoxycholic acid (sodium salt) and 10% acetonitrile, on a 47 cm uncoated silica capillary at 20 kV with detection of the analytes at 320 nm. The linear detection range for concentration versus peak area for the assay is from 0 to 100 micrograms ml-1 for both analytes, with a limit of quantitation of 2 micrograms ml-1 for 7HC and 5 micrograms ml-1 for 7HCG in urine. Inter- and intra-assay results showed sr values in peak areas of between 0.5 and 13%. The method was applied to the direct determination of 7HC and the glucuronide conjugate in urine from two volunteers administered with 250 mg of coumarin. The samples were also analysed by another CE method and by using HPLC. There was no statical difference between the results determined by each of the methods. Up to 83% of the coumarin administered was excreted as 7HC or 7HCG. The majority of the 7HC excreted was in the glucuronide form (98%) with 2% occurring as free 7HC.

Both coumarin (1,2-benzopyrone) and warfarin (4-hydroxycoumarin) have been shown to prevent the recurrence of malignant melanoma. Their action is macrophage-dependent and the dosage is critical. In 1984 a multicentre, prospective, randomised, double-blind trial of coumarin, given as a daily 50-mg dose for 2 years after surgery in patients with high-risk melanoma, was started. the patients had lesions greater than 1.70 mm thick and TNM stage IB or stage II disease. To date there are 4 recurrences in the coumarin-treated group of 13 patients, and 10 recurrences in the placebo-treated group of 14 patients (P < 0.01). There were no toxic effects.

The measurement of serum sialic acid in sera from individuals with leukemia, breast, gastrointestinal, lung and bladder cancers, brucellosis and manic depression is described. The serum sialic acid levels were also examined in mice bearing Landschutz ascites tumours. The presence of sialic acid-enriched glycopeptides in serum was shown. The results indicate that while sialic acid may be significantly elevated in sera of some tumour-bearing individuals, it is not solely tumour-derived. Its application as a tumour marker is limited and requires critical analysis.

The T lymphocytes populations of 22 young healthy adults, 21 healthy middle aged and older blood donors, 35 non-pregnant women of child bearing age and 14 patients with advanced malignant disease were assessed and compared. It was found that the mean T cell counts in the middle aged and older controls were significantly lower than in the healthy young adults and were further reduced in the patients with malignant disease. The addition of the proteolytic agent brinase (protease 1 obtained from Aspergillus oryzae) to the rosetting test increased the T cell counts signficantly in all groups. This was mot marked in the older age groups and the patients with malignant disease. The proteolytic agent is shown to exert its effect on the lymphocytes in the test. Slow intravenous infusion of either brinase or streptokinase into patients with malignant disease is shown to result in increased T lymphocyte counts pari passu with a restoration of skin allergy. The significance of these findings and possible mode of action of the proteolytic agents in increasing T cell activity are discussed.

Thein vitro effects of the proteolytic enzyme brinase on ascites tumour cells were studied histochemically. Dissociation of protein from phospholipid attachment in surface and cytoplasmic membranes was seen, accompanied by activation of lysosomes in the cytoplasm. These changes are similar to those found by other authors in ascites cells exposed to specific antibody and indicate that changes in membrane lipid function should be envisaged in assessing the effects of proteolytic treatment of cells. Lysosomal activation also resulted from exposure of the ascites cells to tragacanth, a metaphase-blocking polysaccharide.Two patients with melanoma showed substantially increased lysosomal activity in their lymphocytes afterin vitro treatment with brinase.

The value of the oral anticoagulant warfarin sodium and fibrinolytic agents is discussed in relation to cancer surgery. A controlled trial of 128 patients showed that in a variety of recurrent cases the addition of warfarin to chemotherapy doubled the 2-year survival rate. The best results were obtained in postmenopausal patients with breast cancer. Warfarin depresses cellular immune responses which might militate against its use for cases undergoing "curative" surgery. Instead, induction of fibrinolysis by streptokinase or Brinase is suggested, because it increases the activity of the cellular immune mechanism. The results to date of an ongoing controlled randomized trial of streptokinase with surgery of tumors of the large bowel are presented, showing that the trends are in favor of streptokinase therapy; however, insufficient time has elapsed to make it, as yet, statistically significant. The action of streptokinase-induced plasmin and Brinase on lymphocytes is described.

Dysfunction of the cellular immune mechanism was found in 23 patients with persistent symptoms of brucellosis of more than one year's duration when skin tested for delayed hypersensitivity to dinitrochlorobenzene (DNCB). Eighteen patients had low normal or depressed E rosetting capacity of their lymphocytes. Anti-anergic treatment with levamisole 150 mg daily for one month and for two consecutive days each week for six months produced an exacerbation of symptoms after one to four weeks followed by complete relief from symptoms in 15 patients and partial relief in four. Vitamin A was used to potentiate the effect of levamisole in seven patients.

1. Two thousand, one hundred and seventy-three patients with cancer or chronic infections were treated with coumarin in a clinical trial. 2. 0.37% of these patients developed elevated liver enzyme levels during therapy with coumarin. 3. This hepatitis was probably a form of idiosyncratic hepatotoxicity and may have been immune in origin.