Interaction between surface active drug (FK906:rennin inhibitor) and cyclodextrins in aqueous solution.

The aggregation behavior of FK906, which is a peptide like hypertensive agent, in aqueous solution was studied by static light scattering, 1H-nuclear magnetic resonance (NMR), and surface tension. These experiments showed a clear critical micelle concentration (cmc) at around 6.3 x 10(-3) to 1.3 x 10(-2) M of FK906 aqueous solution. The result of 1H NMR experiments revealed that FK906 aggregates primarily by hydrophobic interactions involving the benzyl moiety. The Debye plots from light-scattering studies showed that the apparent molecular weight of aggregated FK906 molecule is 1670 which corresponds to 2-3 molecules of FK906. The effect of alpha- and beta-cyclodextrins on the surface tension of FK906 aqueous solution was investigated. It appeared that the addition of alpha-cyclodextrin showed very small shift of cmc, but that of beta-cyclodextrin shifted the cmc to much higher concentration. The investigation on the surface tension of FK906 aqueous solution in the presence of beta-cyclodextrin indicated that FK906 forms a 1:1 complex with beta-cyclodextrin. On the basis of these experiments, it appears that beta-cyclodextrin has an ability to change the surface active property of FK906 in its aqueous solution. Therefore, it is expected that the addition of beta-cyclodextrin to FK906 aqueous solution may prevent the adsorption onto container walls and/or reduce the local irritancy.

Assay of magnesium in serum and urine with use of only one enzyme, isocitrate dehydrogenase (NADP+).

We report a method for assaying magnesium in serum and urine involving only one enzyme, isocitrate dehydrogenase (NADP+)(EC 1.1.1.42), which requires magnesium ion for activity. The enzymatic reduction of NADP+ by isocitrate increases in rate linearly up to at least 20 mmol/L magnesium in the presence of appropriate concentrations of the two metal-chelating reagents, EDTA and glycol ether diamine-N,N,N',N'-tetraacetate. Within-run (n = 20) CVs and day-to-day (n = 10) CVs for sera are < or = 1.5% and < or = 2.6%, respectively. Analytical recovery of magnesium in sera averages 96-100%. This method is not affected by bilirubin, hemoglobin, or lipemia. The method (y) gives the following results correlating with atomic absorption spectrophotometry (x): y = 1.03x + 0.06 mmol/L (n = 62, r = 0.995, Sylx = 0.03) for sera, and y = 1.03x - 0.10 mmol/L (n = 62, r = 0.989, Sylx = 0.19) for urines; with the calmagite method (x): y = 0.99x + 0.04 mmol/L (n = 62, r = 0.991, Sylx = 0.03) for sera, and y = 0.98x + 0.03 mmol/L (n = 62, r = 0.999, Sylx = 0.02) for urines.

Training intensities for aerobic exercise determined on untrained healthy men.

The purpose of this study was to determine the aerobic training intensity from the maximal and submaximal running exercise in 21 untrained adult men. To accomplish this, we evaluated the relationship between physiological (oxygen intake and heart rate) and physical parameters (running speed) of training intensity, and determined the training intensity at the submaximal exercise. Oxygen intake and heart rate were measured by a treadmill test. The maximal oxygen intake (VO2 max), and the aerobic threshold (AerT) and anaerobic threshold (AT) were measured to determine respiratory gas exchange. Running capacity was measured by a 12-min running and treadmill test. For the maximal exercise, there was a significant correlation (r = 0.88, P < 0.01) between VO2 max and 12-min running distance (speed). In addition, the oxygen intake and heart rate at AerT and AT in the submaximal exercise were linearly correlated with running speed. Three levels of training intensity at the submaximal exercise were termed: light, moderate, and heavy. Since AerT was the lower limit intensity and AT was the upper limit, we took the middle of their values as the moderate intensity. The end point for the determination of the training intensity at the submaximal exercise was estimated to be 85% VO2 max and 180 beats.min-1.

Inhibition of hepatic regeneration after 70% partial hepatectomy by simultaneous resection of the bowel in rats.

This study was aimed to evaluate bow simultaneous resection of the bowel influences hepatic regeneration after partial hepatectomy (HTX). Two hundred and sixty-four rats underwent 70% partial HTX, ileocecal resection (ICR), transverse colon resection (TR), colon amputation and simulatenous resection of the liver and the bowel (HTX+ICR, HTX+TR). Hepatic DNA synthesis was remarkably suppressed by simultaneous resection compared with the 70% HTX group (p < 0.01). In simultaneous resection groups, delayed enhanced hepatic protein synthesis (HPS) was observed after the operation as compared with the 70% HTX group, which showed an early postoperative peak of HPS. Postoperative anastomosis leakage occurred more frequently and survival rates were significantly lower in simultaneous resection groups. Higher plasma endotoxin levels of the portal and the peripheral veins were found in simultaneous resection groups as compared with other groups (p < 0.01-0.001). This study suggested that simultaneous resection of the bowel with partial HTX might inhibit hepatic regeneration and result in the increased risk of anastomosis leakage and high surgical mortality rate by increased plasma endotoxin levels and delayed enhanced HPS.

Significance of enzyme linked immunosorbent assay (ELISA) for antibodies to double stranded and single stranded DNA in patients with lupus nephritis: correlation with severity of renal histology.

The correlation between renal histology and class specific (IgG and IgM) antibodies to double stranded DNA (dsDNA) and single stranded DNA (ssDNA) was studied by enzyme linked immunosorbent assay (ELISA) in 40 untreated patients with systemic lupus erythematosus (SLE). The levels of IgG antibodies to dsDNA were significantly higher in patients with World Health Organisation class IV nephritis than in those with class I, class II, or class III nephritis. IgG antibodies to ssDNA were higher in patients with class IV than in those with class II nephritis. IgG antibodies to dsDNA showed a close correlation with the histological activity score and the amount of electron dense deposit. IgG antibodies to ssDNA showed only a weak correlation with the renal histological activity score. IgM antibodies to dsDNA and IgM antibodies to ssDNA were not correlated with renal histological features. Patients with moderate to severe nephritis had a lower ratio of IgM antibodies to dsDNA to IgG antibodies to dsDNA than those with mild nephritis. These results indicate that the measurement of IgG antibodies to dsDNA is predictive in evaluating renal histological activity in patients with SLE.

[One-year chronic oral toxicity study of propiverine hydrochloride in dogs followed by one-month recovery].

The chronic oral toxicity of propiverine hydrochloride (P-4), a new anti-pollakiuria agent, was studied in beagle dogs. Groups of 6 males and 6 females were treated with P-4 at doses of 0, 0.3, 1, 3, 9 mg/kg/day for one year and thereafter 2 animals of both sexes in each group were placed on withdrawal for one month. During administration and recovery period, no death occurred in any dosed animals. As a toxic sign, only the frequency of vomiting was increased in animals of 1, 3 and 9 mg/kg/day groups. Body weight, food and water consumption were not affected by the P-4 administration. In serum chemical examinations, gamma-GTP activity was increased in both sexes of 9 mg/kg/day group at 6 month of administration. Further decrease in total and free cholesterol, triglyceride and phospholipid, increase in GPT activity were detected in some animals of 9 mg/kg/day group at 12 month of administration. In addition decreasing tendency in levels of albumin was noted in males of 9 mg/kg/day group at 9 and 12 month of administration. And also, a gradual increase in total protein level and a gradual decrease in alkaline phosphatase activity were seen in control group, but in females or males of 9 mg/kg/day group, those changes were mild. Urine pH rised slightly in females of 3 mg/kg/day group and in both sexes of 9 mg/kg/day group. No specific findings attributable to P-4 treatment were detected in ECG, heart rate, funduscopy, hematology, fecal occult blood test and necropsy. The absolute and/or relative liver weight in males of 3 and 9 mg/kg/day groups were significantly increased. Light-microscopically, the hypertrophy of hepatocytes characterized by homogenization and enlargement of cytoplasmic space, and concentric inclusions in hepatocytic cytoplasm were detected in both sexes of 3 and 9 mg/kg/day groups. Corresponding to these microscopical findings, the following changes were observed electron-microscopically, the proliferation of smooth surfaced endoplasmic reticulum in hepatocytes in both sexes of 1, 3 and 9 mg/kg/day groups, lamellar bodies in hepatocytes in females of 3 mg/kg/day group, and in both sexes of 9 mg/kg/day group, and annulate lamellae in hepatocytes were detected in one female of 9 mg/kg/day group. After the recovery period, the above mentioned abnormalities were markedly attenuated or disappeared except the changes in hepatocytes. From these results, it seemed that 9 mg/kg/day of P-4 might be safety dose.(ABSTRACT TRUNCATED AT 400 WORDS)

Accumulation of pigment granules in lacrymal gland epithelium in practolol-treated beagle dogs.

A 6-month oral toxicity test of practolol was carried out in beagle dogs as a reference control for a newly developed beta blocker. No significant drug-induced changes were detected in any animals by various ophthalmological examinations such as ERG, tear flow, lysozymal activity in tears, etc. However, an unusual pathological change was detected in the lacrymal gland of all five dogs treated with practolol and not in control animals. Macroscopically, the lacrymal glands assumed a blackish brown to deep black colour on both the outside and the cut surface. Microscopically, fine, dark-brown pigment granules were present in the apical and supra-nuclear portion of the cytoplasm of predominantly serosal type epithelial cells. These pigments reacted positively to Schmorl's method for lipofuscin, but gave a negative PAS reaction for polysaccharide, Prussian blue for iron and Ziehl-Neelsen method for ceroid pigment. They were detected as membrane-bound electron-dense bodies by electron microscopy. Similar pigments were also deposited in the cytoplasm of the apocrine sweat gland. Although the mechanism of the accumulation of these granules is far from clear, concentration of practolol in the lacrymal gland is considered to be very closely related to the presence of these granules. A possible mechanism for ocular toxicity by practolol, involving this change, is discussed.

Clinical significance of antibodies to histones in systemic lupus erythematosus.

IgG antibodies to whole histones and to individual histones H1, H2A+H4, H2B and H3 were measured by enzyme linked immunosorbent assay in serum samples from 46 untreated patients with systemic lupus erythematosus (SLE) who had undergone renal biopsy. Patients with the WHO Class IV lupus nephritis had significantly higher levels of antibodies to histones (except H1) than those with milder forms of nephritis. Among antibodies to individual histones, the levels of antibodies to H2B histones best correlated with the renal histologic and the clinical activity of disease (rs = 0.759, rs = 0.577, p less than 0.001, respectively).

[Six-month chronic intravenous toxicity study of cefodizime sodium in dogs].

The chronic intravenous toxicity of cefodizime sodium (THR-221) was studied in beagle dogs. Groups of 6 males and 6 females were treated with THR-221 at doses of 0 (saline), 200, 400, 800 and 1600 mg/kg/day for 6 months. The THR-221 related symptoms were vomiting, excessive drinking behavior and salivation. The paleness of the visible mucosa and discoloration of vascular color by funduscopy due to systemic anemia were observed in one animal each of 800 and 1600 mg/kg/day groups. Body weight was depressed transiently or continuously in a few animals of 400-1600 mg/kg/day groups. The hematological, serum chemical and urinalysis findings in a few animals of 400-1600 mg/kg/day groups revealed decreases in RBC count, PCV and hemoglobin, an increase in reticulocyte count, a decrease in WBC count, a decrease in platelet count, slight increase in TP, and albumin, a decrease in AlP, and an increase in urinary Na. Light microscopically, deposition of hemosiderin and increased extramedullary hematopoiesis in the liver and spleen, and deposition of fibroid substance in the white pulp of the spleen and diffuse fibrosis in the bone marrows were detected in a few animals of 800 and 1600 mg/kg/day groups. Electron microscopically, no significant toxic changes were observed. The maximum nontoxic doses of THR-221 are estimated as 200 mg/kg/day in male and less than 200 mg/kg/day in female.

Preparation of phosphorylcholine derivatives of bovine serum albumin and their application to the affinity chromatography of C-reactive protein.

A simple method for the preparation of phosphorylcholine derivatives of bovine serum albumin (PC-BSA) by reductive alkylation of the amino groups of bovine serum albumin with choline phosphoryl glycoaldehyde is described. Choline phosphoryl glycoaldehyde was generated by periodate oxidation of glyceryl phosphorylcholine. PC-BSA was immobilized on SH-derivatized Toyopearl HW 65 by reacting the single SH group of PC-BSA with a bismaleimido reagent and then coupling maleimidated PC-BSA to the thiolated gel. The affinity purification of C-reactive protein (CRP), which is based on the Ca2+-dependent affinity of CRP for the phosphorylcholine residue of PC-BSA, was readily accomplished using the PC-BSA Toyopearl HW 65 column. The resulting CRP preparation from serum and pleural fluid was homogeneous as assessed by native polyacrylamide gel electrophoresis. PC-BSA derivatives were also shown to be reactive with Limulus polyphemus CRP.

Benign hemangioendothelioma of the pancreas with obstructive jaundice.

A 3-year-old boy with benign hemangioendothelioma of the pancreas is reported. He showed the presenting sign of obstructive jaundice. The boy was treated with temporary choledochojejunal bypass without additional treatment. In a 16-month follow-up the child has been well and asymptomatic. Laboratory data showed no abnormality. Biopsied specimen of pancreatic head showed hemangiomatous pattern histologically. Immunohistochemical studies of the biopsy revealed positive reaction to factor VIII-related antigen (FVIII-RAG), and endothelial cell marker, supporting an endothelial origin. Hemangioendothelioma has a favorable prognosis and spontaneous regression can be expected. So the first treatment of the tumor should be conservative.

Tumor-associated serum proteins in rats as detected by two-dimensional gel electrophoresis.

The sera from normal and tumor-bearing rats were separated by polyacrylamide gel disc isoelectric electrophoresis and then in a perpendicular dimension by polyacrylamide gel slab electrophoresis with a gradient from 4% to 20%. When the gel was stained with Coomassie brilliant blue, 100-150 components were separately observable with normal serum. With the sera from Rhodamine sarcoma-bearing rats, three more components were detected with a high reproducibility. They were of pI (isoelectric point) 4.5 and Rm (relative mobility) 0.63, of pI4.8 and Rm 0.63 and of pI 4.9 and Rm 0.75. The 4.5/0.63 (pI/Rm) and the 4.8/0.63 components appeared soon after transplanting the tumor, whereas the 4.9/0.75 component appeared at the late stage of its growth. Upon surgical removal of the grown tumor, the former two components disappeared rapidly, while the latter component disappeared gradually. Similar results were obtained with Yoshida sarcoma, and ascites hepatoma AH66 and AH130. A 3rd polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the 4.5/0.63 and the 4.8/0.63 components were each composed of a single polypeptide of approximately 64,000 daltons, and that the 4.9/0.75 component was composed of two different polypeptides of approximately 33,000 daltons and approximately 10,000 daltons. It was confirmed by an immunodiffusion experiment that none of these three proteins was alpha-fetoprotein.