Relationship between interleukin-8 levels and myeloperoxidase activity in human gastric mucosa.

Although interleukin (IL)-8 is well known as a chemotactic agent for neutrophil migration in vitro, the relationship between IL-8 activity and the degree of neutrophil infiltration in gastric mucosa is still unclear. In the present study, we investigated IL-8 and myeloperoxidase activity, a marker of neutrophil infiltration, in gastric antral mucosa using biopsy samples in 23 patients with no gastric lesions. The results indicate that there is a good correlation between IL-8 and myeloperoxidase activity (y = 0.173x + 13.9; r = 0.49, P < 0.01). Furthermore, IL-8 and myeloperoxidase activity are significantly higher in Helicobacter pylori-positive patients than in H. pylori-negative patients. In conclusion, an increase of IL-8 activity in the gastric mucosa causes increased neutrophil infiltration in human gastric mucosa and H. pylori infection accelerates these reactions in the mucosa.

Insulin antibodies and management of diabetes after total pancreatectomy.

For 18 patients after total pancreatectomy, serial determinations of plasma immunoreactive insulin bound with insulin antibody (binding IRI) were performed to ascertain the relationship between insulin antibody formation and diabetic stability. As a result, seven patients treated with human DNA insulin experienced neither unstable diabetic control (UDC) nor binding IRI elevation. On the other hand, six (58%) of 11 patients treated with beef and porcine insulins began to experience UDC with elevation of binding IRI. For these six patients, the binding IRI level was as low as 242 +/- 100 microU/ml at the onset of UDC and continued to increase (maximum binding IRI, 2048 +/- 1707 microU/ml) without spontaneous recovery from UDC. However, three patients recovered from UDC by the transfer to human DNA insulin, with a small decrease in binding IRI (470 to 4400 microU/ml) at the end of UDC. An equilibrium binding assay and the kinetic analysis of insulin antibodies showed that high-affinity antibody capacity (Ab1) for beef insulin was 28 +/- 6 ng/ml immediately before UDC, 306 +/- 120 ng/ml (115 to 578 ng/ml) during UDC, and 28 +/- 12 ng/ml immediately after UDC. Low-affinity antibody capacity was not correlated with diabetic stability. Therefore, in the totally pancreatectomized patients, it was concluded (1) that Ab1 was associated with the UDC onset or the recovery and (2) that UDC developed with far lower levels of Ab1, in contrast with patients with insulin-dependent diabetes. This is a reason why serial determination of insulin antibody, especially Ab1, is necessary for control of diabetes after total pancreatectomy.

Relationships between red cell cation contents and blood pressure level.

Red cell sodium (R-Na), potassium (R-K) and sodium to potassium ratio (R-Na/K) were studied in 2,542 adults participating in periodic medical examinations. The mean values of R-Na and R-Na/K in untreated borderline (n = 406) and definite hypertensives (n = 485) were higher than those in normotensives (n = 1,651), but that of R-K was not different among the groups. The increased R-Na and R-Na/K in the hypertensives were observed only in those under 50 years, but not in those 50 years and over. In the normotensive subjects, age and sex affected the intraerythrocyte cation contents, but a positive family history of hypertension did not. Although a few characteristics correlated with both R-Na and mean blood pressure, and with both R-Na/K and mean blood pressure, multiple regression analyses revealed that R-Na and R-Na/K independently and significantly contributed to the variation in mean blood pressure. It was also indicated that these relations of R-Na and R-Na/K to mean blood pressure were significant in those under 50 years, but not in those 50 years and over. While several characteristics confound the relationships between red cell cation contents and blood pressure, these results suggest that the contributions of R-Na and R-Na/K to the mechanisms of human hypertension still persist in young to middle-aged people.

High-performance liquid chromatography of coenzyme Q-related compounds and its application to biological materials.

A convenient and precise method for the separation and determination of coenzyme Q (CoQ)-related compounds (CoQ homologues, plastoquinone-9, ubichromenol-9, etc.) was developed using high-performance liquid chromatography (HPLC). All compounds tested were separated using a reverse-phase column with a suitable mobile phase and detected at a wavelength of 275 nm. CoQ extracts in plasma and erythrocytes were purified by thin-layer chromatography prior to HPLC analysis, but such purification was not necessary when determining CoQ in urine and tissues. Hydroquinone forms of CoQ existing in animal tissues were oxidized to the corresponding quinone forms with potassium hexacyanoferrate(III). This HPLC method was applied satisfactorily to the determination of the contents of CoQ homologues in human and animal samples. CoQ10 was the only homologue detected in human samples, and CoQ8, CoQ9 and CoQ10 were native homologues of CoQ in rat tissues. Ubichromenol-9 and plastoquinone-9 were not detected in these samples.

Abnormal relationship between dietary sodium intake and red cell sodium transport in salt-sensitive patients with essential hypertension.

The effects of high sodium intake on erythrocyte 22Na efflux rate constants were studied in 25 patients with essential hypertension and 9 normal subjects. With changes in sodium intake from 100 mEq to 300 mEq/day, both total and ouabain sensitive 22Na efflux rate constants decreased significantly (p less than 0.001) in "salt-sensitive" patients (-0.031 +/- 0.005 and -0.035 +/- 0.006 /hr, respectively), but these responses were variable in "nonsalt-sensitive" patients and in normal subjects. The "salt-sensitive" patients showed a significant increase in their body weight, while intraerythrocyte sodium contents remained unchanged in the both groups. These results suggest that the abnormal change in membrane Na-K-ATPase activity may, at least in part, be involved in the mechanism of sodium susceptibility in patients with essential hypertension.

Studies on bile acids: The microquantitative separation of cellular bile acids by gas-liquid chromatography.

1. A method is described for the quantitative isolation of bile acids from cellular material. Homogenates of rat liver are freeze-dried and extracted exhaustively with 95% (v/v) ethanol containing 0.1% (v/v) of aq. ammonia (sp.gr. 0.88) and purified by anion-exchange chromatography on Amberlyst A-26. 2. The extracted bile acid conjugates are subjected to either of two hydrolytic procedures, one involving chemical and the other enzymic agents. A unique feature in this study is the introduction of an enzyme, a clostridial peptide-bond hydrolase, for the rapid cleavage of bile acid conjugates, replacing the classical drastic chemical hydrolysis with strong alkali. 3. After hydrolysis, free bile acids are methylated and converted into their trifluoroacetates for final determination by gas-liquid chromatography on a triple component column, FS-1265-SE30-NGS. 4. For the purpose of identification of peaks, bile acid methyl esters are converted into their trimethylsilyl ethers by allowing the methyl esters to react with a new and potent silyl donor, bis(trimethylsilyl)acetamide. 5. The technique affords us a means of studying the metabolism of bile acids at the cellular and subcellular levels in tissues.