Production of active bovine cathepsin C (dipeptidyl aminopeptidase I) in the methylotrophic yeast Candida boidinii.

The heterologous production of active bovine cathepsin C (CTC; dipeptidyl aminopeptidase I) was investigated. Attempts to express CTC in Escherichia coli were hampered by formation of inclusion bodies that were partially degraded. To overcome this impediment, secretion of recombinant CTC was attempted in the methylotrophic yeast Candida boidinii. A DNA fragment encoding bovine procathepsin C was synthesized based on preferred codon usage in C. boidinii and placed downstream of the C. boidinii proteinase A signal sequence resulting in secretion of active CTC into the culture medium. The gene was expressed under the control of the methanol-inducible formate dehydrogenase gene promoter. Production levels were significantly improved by using a protease-deficient strain, changing medium composition, and by lowering the temperature of induction. When the recombinant C. boidinii was grown for 90 h in a jar-fermenter, active CTC was secreted with a yield of up to approximately 12 mg/l.

Synthetic studies of vitamin D analogs. XXII. Synthesis and antiproliferation activity of putative metabolites of 1 alpha, 25-dihydroxy-22-oxavitamin D3.

As putative metabolites of 1 alpha,25-dihydroxy-22-oxavitamin D3 (OCT), 24-hydroxylated OCT in 24(R) and 24(S) forms, 24-ketoOCT, 26-hydroxylated OCT in 25(S) and 25(R) forms, pentanorOCT and pentanor-ketoOCT were synthesized from the steroidal 20(S)-alcohol. Their antiproliferation activities towards human promyelocytic leukemia cells (HL-60 cells) are also reported. Oxidized derivatives at the C-24 position, 24-ketoOCT, 24(R)-hydroxylated OCT and 24(S)-hydroxylated OCT, showed activities comparable to or slightly weaker than that of OCT, while 26-hydroxylated OCT was less active than OCT. Truncated OCT, pentanor OCT and pentanor-ketoOCT, were inactive at 10(-7)-10(-10) M.

Adhesion of peripheral blood lymphocytes of children with arthritis to human umbilical vein endothelial cells.

To determine whether adhesion of peripheral blood lymphocytes (PBL) of patients with juvenile rheumatoid arthritis (JRA) may be enhanced, adhesion of PBL of children with JRA, children with seronegative spondyloarthropathies (SSA), age-appropriate and adult controls, to human umbilical vein endothelial cells (HUVEC) was assessed in vitro. B and CD4 T lymphocytes in initial, adherent, and non-adherent cell fraction were identified by flow cytometry. B lymphocytes of all the younger subjects combined had a higher adherence to activated HUVEC compared with B lymphocytes of the adult donors. Except for greater adherence of HLA-DR+ CD4 T cells, lymphocytes of children with JRA showed no enhanced adhesion to either unactivated or activated HUVEC. The percentage of B cells adherent to activated HUVEC in each of the subject groups was 1.5-3.6-fold higher than adherent CD4 T lymphocytes. Surface analyses indicated higher percentages of CD49d (alpha 4)+ and CD29 (beta 1)+ CD4 T lymphocytes in adherent cells, but less of a differential in CD49 (alpha 4)+ and no difference in CD29 (beta 1)+ B lymphocytes. There were fewer Leu-8 (L-selectin)+ B and Leu-8+ CD4 T cells among adherent cells. The data suggest a greater adhesive capacity of B lymphocytes compared with CD4 T lymphocytes which is unrelated to disease, and the possibility that B lymphocytes may utilize adhesion molecules distinct from those of CD4 T lymphocytes. Only a small subset of T cells of patients with JRA may have an enhanced capacity for adhesion to endothelium.

[A case of sudden death from massive hematemesis due to rupture of an atherosclerotic aneurysm into the esophagus].

A 86-year-old woman was admitted to our hospital following vomiting of a small amount of blood. On the 8th hospital day, the patient suddenly died of massive hematemesis. Autopsy disclosed thoracic aortic aneurysm with esophageal fistula.

Serum dopamine beta-hydroxylase activity in essential hypertension.

1. Serum dopamine beta-hydroxylase activity was determined in normotensive control subjects and patients with labile or established essential hypertension. The enzyme activity was 25-9 +/- 1-9 (SEM) 29-6 +/- 2-5 and 25-1 +/- 1-9 micronmol min-1 1-1, for control, labile and established hypertensive subjects respectively. 2. Neither blood pressure nor serum dopamine beta-hydroxylase activity was changed in normotensive control subjects by administration of phentolamine; however, in patients with essential hypertension blood pressure was significantly decreased (P is less than 0-01) and serum dopamine beta-hydroxylase activity was slightly increased. With propranolol administration, blood pressure and the serum enzyme activity were not significantly changed in normotensive or hypertensive subjects. 3. Our results suggest that there is no correlation between serum dopamine beta-hydroxylase activity and blood pressure.

Hypotensive action and increased plasma renin activity by Ca2+ antagonist (Nifedipine) in hypertensive patients.

The Ca2+ antagonistic coronary vasodilator, Nifedipine, was sublingually administered by a dose of 30 mg to 19 patients with hypertension. Blood pressure of patients with with essential hypertension (n=14) decreased from 177 +/- 24 to 123 +/- 13 mmHg systolic and from 108 +/- 12 to 80 +/- 11 mmHg diastolic (mean +/- SD) (p less than 0.01). Plasma renin activity (PRA) increased significantly from 0.73 +/- 0.62 to 1.50 +/- 1.02 ng/ml/h (p less than 0.05). The same tendency was observed in malignant and renovascular hypertension. In primary aldosteronism (n = 2), blood pressure decreased but PRA did not increase. Hypotensive action and increased plasma renin activity by Ca2+ antagonist, Nifedipine, were clearly demonstrated in patients with hypertension.

Serum dopamine-beta-hydroxylase activity in essential hypertension and in chronic renal failure with hypertension.

Elevated serum DBH (dopamine-beta-hydroxylase) activity was found in essential hypertension. The elevated level was not reduced when blood pressure was brought to normotensive level by administration of thiazide or rauwolfia. In contrast, serum DBH activity was low in both normotensive and hypertensive patients treated on prolonged hemodialysis. However, there was no correlation between serum DBH activity and blood pressure level. It was suggested that the pathogenesis of high blood pressure might be different between essential hypertension and hypertension with chronic renal failure, and that measurement of serum DBH activity might help for clinical differentiation of essential hypertension from certain forms of secondary hypertension.