A high-performance liquid chromatographic method for the determination of nalidixic acid (NA) in rat serum, brain and cerebrospinal fluid (CSF) was developed. NA in rat serum and brain homogenate was extracted and injected onto a reversed-phase column. CSF was directly analysed without extraction procedure. The limits of detection were 0.05 microgram ml-1 for serum, 0.07 microgram g-1 for brain and 0.02 microgram ml-1 for CSF, respectively. Calibration curves were linear over the concentration ranges 0.1-50 micrograms ml-1 for serum, 0.12-9 micrograms g-1 for brain and 0.05-10 micrograms ml-1 for CSF, respectively. The reproducibility of NA assay in rat biological media ranged from 1 to 4% relative standard deviations (RSD). The recoveries of NA added to serum and brain were higher than 96% with an RSD of less than 4%. The present method was found to be applicable to pharmacokinetic study of NA in rat serum, brain and CSF.

Animals↗

PubMed1993

Binding of an intrinsic ATPase inhibitor to the interface between alpha- and beta-subunits of F1FoATPase upon de-energization of mitochondria.

Yeast mitochondrial F1FoATPase has three regulatory subunit proteins: ATPase inhibitor, 9K protein, and 15K protein. Mutant yeasts lacking one or more of these protein factors were constructed by gene disruption [Ichikawa, N. et al. (1990) J. Biol. Chem. 265, 6274-6278; Yoshida, Y. et al. (1990) Eur. J. Biochem. 193, 49-53]. Dissipation of the electrochemical potential of protons of the mitochondrial inner membrane by an uncoupler or by a combination of valinomycin and potassium ions induced ATP-hydrolyzing activity of F1FoATPase in mitochondria of all the mutants, as in those of wild-type cells. However, the ATPase activity was inactivated within a minute in normal mitochondria, but was not suppressed in inhibitor-deficient mitochondria, and in mitochondria lacking either 9K or 15K protein, the inactivation of ATPase was slow and incomplete. Covalent binding of inhibitor protein to the enzyme was achieved with a zero length cross-linker, EEDQ, in uncoupled normal mitochondria, in which the inhibitor linked directly to both the alpha- and beta-subunits. This result strongly suggests that the binding site of the inhibitor protein is located at the interface between the two subunits.

Adenosine Triphosphate↗

PubMed1993

Aggressive granular lymphocyte leukemia of natural killer cell type in an elderly patient.

A 75-year-old man was admitted to our hospital because of intermittent fever. His peripheral blood picture showed granular lymphocyte (GL) proliferation. The GLs were immunologically and functionally phenotyped as natural killer cells. Chromosomal analysis of peripheral lymphocytes with interleukin-2 stimulation revealed an inversion of chromosome 9 with an unusual breakpoint, showing abnormal monoclonal proliferation of the GLs. Progressive increase of GL count and hepatosplenomegaly necessitated the start of combined chemotherapy. His condition was complicated by icterus and renal failure, and he died finally of respiratory failure. Autopsy revealed disseminated intravascular coagulation and infiltration of GLs in the bone marrow, spleen, and liver.

Aged↗

PubMed1993

[Myocardial revascularization with bovine internal thoracic artery graft (BIOFLOW): a case report].

Bovine internal thoracic artery graft (BIOFLOW) was used for coronary artery bypass grafting (CABG). A 64-year-old woman who had undergone triple CABG with saphenous vein five years ago was referred to our hospital again with anginal attack. Coronary angiogram showed the obstruction of the graft for LAD and new stenosis at right coronary artery (RCA). Gastroepiploic artery was not available because the patient had a history of gastric ulcer several times, and saphenous vein had harvested initial operation. We planned re-CABG for LAD with left internal thoracic artery and RCA with BIOFLOW. But at the operation, LAD was not graftable because of severe calcification. So that we performed single CABG to RCA with BIOFLOW. The BIOFLOW was patent at the 28 th postoperative day. From our experience, BIOFLOW can be expected as a graft for CABG when saphenous vein, internal thoracic artery and gastroepiploic artery are not available.

Aged↗

PubMed1993

[Adverse effect of prolonged formalin fixation on DNA ploidy in paraffin-embedded tissue].

From retrospective analysis of 174 formalin fixed, paraffin-embedded, HCC tissue samples, the possible correlation was investigated between the CV determined by FCM and the duration of formalin fixation. An adverse effect of prolonged formalin fixation on CV and fluorescence intensity in liver tissue was investigated prospectively in samples from 4 patients.

DNA, Neoplasm↗

PubMed1992

A procedure for the construction of linear restriction fragment maps of a 50- to 100-kb DNA.

A simple and effective procedure for the construction of linear restriction fragment maps was developed. Using a two-enzyme digestion, two-dimensional (2-D) electrophoresis procedure, all the restriction fragments in a 50- to 100-kb DNA can be individually resolved and displayed on a 2-D plane. This 2-D gel pattern, with appropriate markers, provides a fixed set of x, y coordinates for each fragment obtained from the single and double digestion as well as the relationship between the two steps. A matrix is constructed from the 2-D pattern. The vertical column shows all the singly digested individual fragments and their sizes obtained from each restriction enzyme treatment, and the dividing horizontal row shows all the doubly digested DNA fragments and their sizes after treatment with two enzymes. The order of arrangement is always from the smallest to the largest fragments. Using this matrix, two linear DNA restriction maps for these two enzymes can be simultaneously constructed in a self-reconfirming manner. As examples for this procedure, we describe the construction of two linear restriction fragment maps, a combination of EcoRI and BamHI digestion as well as a combination of EcoRI and HindIII digestion of lambda-phage DNA.

Bacteriophage lambda↗

PubMed1992

Airflow receptors in the lip and buccal mucosa.

Airflow receptor afferents in the oral mucosa responding to changes in intraoral air pressure during blowing were found to be innervated by the infraorbital nerve. They provided one response corresponding to the onset of blowing, a second related to an increase in air pressure, a third corresponding to the cessation of blowing, and a fourth that exhibited little change throughout sustained blowing. Intraoral air pressure in the cavity between the lips and the velopharyngeal portal may be monitored by these receptors, and the data they provide may contribute to the control of phonation.

Adult↗

PubMed1992

Antibacterial activity of crinitol and its potentiation.

An acyclic diterpene alcohol, crinitol [1], was identified in a marine brown alga, Sargassum tortile, as the principal antibacterial agent against Gram-positive bacteria, among which Propionibacterium acnes was most sensitive and, Staphylococcus aureus was least. To enhance its activity, crinitol was combined with several antioxidants, which presumably retard oxidative destruction of this molecule which possesses two easily oxidizable allylic alcohol groups. Two synthetic antioxidants, BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene), significantly enhanced the activity of crinitol, especially against Streptococcus mutans. Interestingly, crinitol also synergized BHT and BHA against this cariogenic bacterium.

Anti-Bacterial Agents↗

PubMed1992

Effect of fenbufen on the entry of new quinolones, norfloxacin and ofloxacin, into the central nervous system in rats.

The entry of two new quinolone antibacterial agents, norfloxacin and ofloxacin, into the central nervous system (CNS) of rats, and the effect of fenbufen on this was investigated. At various times after the administration of a bolus intravenous dose of norfloxacin or ofloxacin (10 mg kg-1) with or without fenbufen (20 mg kg-1), serum and cerebrospinal fluid (CSF) samples and whole brain were collected from the rats and the concentration of norfloxacin or ofloxacin in each sample was determined. Serum concentrations of both quinolones declined biexponentially with time and were significantly elevated by coadministration with fenbufen at the terminal phase. The fractions of these quinolones bound to serum protein were not altered by coadministration with fenbufen. Coadministered fenbufen raised the brain concentrations of both quinolones but did not affect their brain to serum unbound concentration ratios. In contrast, CSF to serum unbound concentration ratios as well as CSF concentrations of norfloxacin and ofloxacin were elevated by coadministration with fenbufen. Apparent diffusional clearances between blood and CSF of norfloxacin and ofloxacin estimated by the physiological model analysis increased by 1.9 and 2.6 times, respectively, after coadministration with fenbufen. These findings suggest that coadministered fenbufen may facilitate the entry of norfloxacin and ofloxacin into the CNS.

Animals↗

PubMed1992

Studies on single-dose toxicity of hydrophobically modified hydroxypropyl methylcellulose in rats.

Single-dose toxicological studies of hydrophobically modified hydroxypropyl methylcellulose (HM-HPMC, hydroxypropyl methylcellulose modified with stearylglycidylether) were conducted. A dispersion of HM-HPMC was administered to rats orally or by dermal application at doses up to 900 mg/kg. After the oral administration, the mean body weight of the 900 mg/kg group on the first day after administration was slightly but significantly lower (P less than 0.05) than that of the control group, and one rat had loose stools at 30 min. after the administration. No other abnormalities were noted. In the case of dermal application, no abnormalities were observed. No rats died, and no abnormalities in their organs were found by either route. In conclusion, there was no observed toxicity of HM-HMPC after oral or dermal administration at single dose up to 900 mg/kg under the conditions of these studies.

Administration, Cutaneous↗

PubMed1992

Primary dermal and eye irritability tests of hydrophobically modified hydroxypropyl methylcellulose in rabbits.

Primary dermal and eye irritation tests of hydrophobically modified hydroxypropyl methylcellulose (HM-HPMC, hydroxypropyl methylcellulose modified with stearylglycidylether), a new cellulose derivative used as a thickener for topical pharmaceuticals and cosmetics, were conducted in rabbits. A dispersion of HM-HPMC (3%) was applied to intact and abraded skins and reactions were observed. A very slight erythema was observed in both skins and this polymer was categorized as a "mild irritant". In the eye irritation test, with a dispersion of the same concentration, it was categorized as "marginal" in unrinsed eyes and "negative" in rinsed eyes.

Administration, Cutaneous↗

PubMed1991

Enhanced entry of ciprofloxacin into the rat central nervous system induced by fenbufen.

The effects of fenbufen on the transport of ciprofloxacin (CPFX) into the brain and cerebrospinal fluid (CSF) were investigated in rats. Periodically after a bolus i.v. dose of CPFX (10 mg/kg), alone or with fenbufen (20 mg/kg), to rats, aliquots of CSF and blood were collected and then the whole brain was readily excised from the animal after sacrifice by microwave irradiation. Serum levels of CPFX in the terminal phase were significantly elevated by the coadministration with fenbufen. However, the extent of CPFX binding to serum protein was not affected by fenbufen. Immediately after the coadministration with fenbufen, brain and CSF levels of CPFX were raised by about 15 to 70% and 70 to 100%, respectively. Both brain/serum unbound and CSF/serum unbound level ratios were increased by fenbufen at relatively early periods after drug injection. Analysis based on physiological models indicated that fenbufen significantly increased the apparent diffusion clearances of CPFX across blood-brain and blood-CSF barriers. These findings suggest that coadministered fenbufen may facilitate the entry of CPFX into the central nervous system not only by elevation of serum level but also by enhancement of permeability across the blood-brain or blood-CSF barrier.

Animals↗

PubMed1991

Electrolyte imbalance after canine pancreaticoduodenal allotransplantation using duodenocystostomy technique.

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Animals↗