Determination of propranolol enantiomers in human plasma and urine and in rat tissues using chiral stationary-phase liquid chromatography.
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Biomedical subjects
Publications and source records attributed to K Someya.
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With the purpose to use for therapeutic drug monitoring (TDM), blood concentrations of tobramycin (TOB) in each patient were measured by radioimmunoassay (RIA). A RIA kit of TOB (Clinical assay-Japan Travenol) was evaluated for precision and recovery, in that partial improvement of the method was made, in order to measure low level of TOB. The RIA was compared with high-performance-liquid-chromatography (HPLC), bioassay (BA) and 2 kinds of enzyme immunoassay (EIA) (EMIT and SLFIA). The RIA of TOB revealed high precision (1.8-2.4% in C.V.) and high reproducibility (5.0-6.9% in C.V.). It was found that this RIA kit can be used for measuring low level of serum TOB concentrations by a modification of the method. The total range of measurable blood level is from 0.1 to 16.0 micrograms/ml. The nearly one to one correspondence was observed between RIA and other 4 methods, when 154 samples obtained from 18 cases were measured. A representative case of TDM for TOB was demonstrated, in which predicted concentrations agreed fairly well with actual measured values at steady state. It was concluded that the RIA kit is useful for clinical application of TDM for the adequate dosage regimen of TOB. Modification of the method for rapid assay of a small number of samples will increase the clinical usefulness.
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The present studies examine the effect of starvation together with cold or hot exposure on thyroid hormone levels in rats. At 23 degrees C starved for 5 days, serum thyroid hormone levels decreased significantly compared with fed rats, averaging 3.6 +/- 0.5 micrograms/dl of thyroxine (T4), 47 +/- 11 ng/dl of triiodothyronine (T3), 1.4 +/- 0.3 ng/dl of free T4 and 39.6 +/- 5.1 pg/ml of reverse T3, respectively. At 15 degrees C rats starved for 5 days, serum free T4 level significantly more increased than that of 23 degrees C starved rats, while serum T4 level and T3 did not increase significantly. At 30 degrees C rats whether concomitant starvation or not, serum thyroid hormone levels of both group markedly more decreased than control rats. These experiment provide additional evidence that thyroid gland and the peripheral metabolism of thyroid hormone respond to variety situations such as cold or hot exposure together with starvation or not.
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Digestion of liver plasma membranes with trypsin and chymotrypsin prevented specific binding of 35-S or 131-I-bromosulphophthalein (BSP) to these membranes, in contrast concomitant appearance of BSP binding protein was observed in a high-speed (100,000 x g. 1 hr) supernatant of the extracts. On the other hand, BSP binding capacity of receptors was hardly destroyed by digestion with high concentration of phospholipase A, C. Gel filtration experiments on a column of Sephadex indicated that specific BSP binding to a protein in the high-speed supernatant was observed and this protein contained at least sialic acid and pentose suggesting a fragment of glycoprotein. In addition this molecular size was far smaller than 23,000, calibrated with bovine trypsin. Competitive inhibition was also observed between 131-I-BSP and BSP on a specific protein by gel filtration, while cholic acid did not affect its capacity to bind 131I-BSP. These results suggest that the specific protein on the liver plasma membrane is involved in the transport of organic anions across hepatocyte surface membranes.
Phagocytic ability of granulocytes obtained from 22 diabetic patients was studied by electron microscopy in order to explain their susceptibility to infection and was compared with that of granulocytes from 13 healthy controls. The main features of the procedure employed in this study were as follows; (1) morphologically quantitative assessment of phagocytosis, (2) no separate assay of engulfment and killing of bacteria, (3) granulocytes were mixed with 10 times their number of bacteria, (4) antibiotics were not used to kill extracellular bacteria, (5) use of a conic tube lined with collodion film for cell pellet formation, (6) each conic tube contained less than 1 X 10(6) granulocytes, (7) sections examined were the vertically cut surfaces of the thin buffy coat. Control subjects showed that most granulocytes did not contain intact intracellular bacteria. Assay of aliquots sampled after 30 minutes and 2 hours in controls showed no significant difference. However, in diabetics, eight cases showed similar results with controls and were thought to have normal function, and fourteen cases showed the increased number of granulocytes containing intact intracellular bacteria was much more marked in aliquots sampled after 2 hours and were considered as showing various degrees of impaired bactericidal ability.
Measurement of serum concentration of trypsin by RIA-Gnost Trypsin kit (Hoechst-Japan) was evaluated. The clinical usefulness of measuring serum trypsin level in diabetic patients was assessed. The measurement of trypsin using the radioimmunoassay (RIA) kit revealed good precision and reproducibility with intraassay error ranging from 3.6 to 5.5% in C.V. corresponding to mean trypsin concentration of 236.5-838.7 ng/ml and interassay error ranging from 8.1 to 11.1%. Tests for recovery and dilution were satisfactory for clinical use. Clinical materials included 35 normal subjects, 88 diabetics, 22 patients with liver diseases, 3 with acute pancreatitis, 7 with chronic pancreatitis and 3 with chronic renal failure. Serum trypsin concentration in normal controls was 157.6 +/- 59.9 ng/ml (m + 1 S.D.). Diabetic patients treated with diet therapy alone revealed serum trypsin level of 203.6 +/- 74.8 ng/ml (n = 50). In diabetics treated with sulfonil urea serum trypsin was 171.3 +/- 83.0 ng/ml (n = 25). In patients receiving insulin serum trypsin level was 90.5 +/- 49.0 ng/ml (n = 13). In patients with liver diseases, acute pancreatitis, chronic pancreatitis and chronic renal failure serum trypsin concentration were 236.9 +/- 88.0, 520.1 +/- 80.0, 113.0 +/- 75.6, and 2557 +/- 2771 respectively. Our results may indicate impaired pancreatic exocrine function in patients with severe diabetes mellitus. Increased serum trypsin level in diabetics treated with diet therapy may be due to stimulated excretion of trypsin resulted from restricted food intake. However, further study in larger number of patients is needed.