N-Oxidation of benzydamine (BZY) mediated by flavin-containing monooxygenase (FMO) was evaluated by microdialysis in vivo in different regions of rat brain and liver. The probe was implanted into local regions of the brain, such as the olfactory bulb, cerebral cortex, corpus striatum, hippocampus and cerebellum, or the hepatic lobe. By perfusing BZY via the probe, BZY N-oxide was identified in the dialysate. The estimated concentrations of BZY N-oxide in extracellular fluid were almost the same as those in the olfactory bulb, hippocampus and cerebral cortex, half the concentration in the hepatic lobe; however, the concentration in the corpus striatum was lower and that in the cerebellum was higher than in the other regions. These results demonstrate that the extracellular concentration of BZY N-oxide formed in vivo was unexpectedly high in every brain region.
Routine analysis of the carbohydrate moieties of a glycoprotein is critical for ensuring the consistene quality of biopharmaceutical products. Fluorophore-assisted carbohydrate electrophoresis (FACE) is a recently introduced method for the separation, by polyacrylamide gel electrophoresis, of oligosaccharides labeled with 8-amino-naphthalene-1, 3, 6-trisulphonic acid (ANTS).In this study, we have evaluated the applicability of the FACE method to analysis of the carbohydrate moieties of three different recombinant human erythropoietins (rHuEPOs), two Chinese hamster ovary (CHO) cell-derived rHuEPOs, and one baby hamster kidney (BHK) cell-derived rHuEPO. The N-linked oligosaccharides released from the rHuEPOs were labeled with ANTS. Enzymic sequence analysis of the N-linked oligosaccharides was performed by the FACE method. The results showed that the FACE method was useful for the analysis of sialo-, asialo-, and agalacto-oligosaccharides in the same gel. Its usefulness for rapidly and reliably revealing the oligosaccharide profiles of given glycoproteins, and its reproducibility were also confirmed in this study. In conclusion, the method can be used for evaluating the quality consistency of recombinant glycoprotein products in terms of the carbohydrate moiety.
Agglutination of fifty bacterial strains isolated from gastric-intestinal tract of chickens with yeasts treated with glutaraldehyde and glycine was assayed to study possible adhesion mechanisms to epithelia. It was found tha Lactobacillus fermentum, Lactobacillus fermentum subsp. cellobiosus and Lactobacillus animalis agglutinated yeasts massively and this agglutination was inhibited in each strain by different representative sugaars : agglutination was inhibited in three selected strains with 0.2 M mannose. The three strains agglutinated chicken red blood cells, but did not agglutinate human red blood cells, whereas the agglutinating capacity for other animal erythrocytes was variable.The lectins were partially purified on a Sephadex G-75 column and affinity chromatography. The molecular weight of each of the protein subunits was estimated as about 50-67 kDa using electrophoretic techniques. After treatment with lipase and metaperiodate modifications bacterial superficies and/or liberation of some structures produced an unspecific interaction with increase in the yeast agglutination and hemagglutination. The optimal conditions of work were : pH 6-7, Ca2+ 1mM, Mg2+ 2mM and 37-42°C.
A growth-supperssing factor for bovine artery endothelial cells (BAEGSF) was purified from the conditioned medium of a mouse lymphoma P388D1 cell culture in the presence of 100 μg/ml carboxymethylated curdlan. The purification steps included, in order, ammonium sulfate fractionation and eight stafes of column chromatography on Macro-Prep Ceramic Hydroxyapatite, Q-Sepharose, Sephacryl S-300 HR, Matrex PBA-30, CHT II, Resource-Q, anti-bovine serum albumin (BSA) agarose, and Superdex 200HR columns. The purified BAEGSF showed two bands with silver staining on a sodium dodecyl sulfate polyacrylamide gel under reducing conditions (SDS-PAGE) and their molecular weights were estimated as approximately 55 and 63 kDa, while the molecular weight of the purified BAEGSF was estimated as about 65 kDa by gel filtration using Superdex 200HR. This result shows that BAEGSF obtained from Superdex 200HR chromatography is a partialy purified preparation and suggests that one of the two bands on SDS-PAGE corresponds to BAEGSF. BAEGSF was shown not to have a lethal effect on endothelial cells, but had an inhibitory action on the proliferation of these cells. Furthermore, the growth-suppressing activity of BAEGSF for bovine artery endothelial cells (BAE) was not inhibited by anti-transforming growth factor-β (TGF-β), anti-tumor necrosis factor-α (TNF-α), and anti-interleukin-1 (IL-1) antibodies. These results suggest that BAEGSF is different from TGF-β, TNF-α, and IL-1 wich have been reported to inhibit BAE growth.
S-(1, 2-Dicarboxyethyl)glutathione (DCE-GS) was found in Saccharomyces cerevisiae, but not in bacterial species nor in a unicellular alga (Acetabularia acetabulm). The enzyme that catalyzes condensation of L-malate and glutathione (GSH) to form DCE-GS was partially purified from baker's yeast. It had a molecular mass of 49 kDa and was monomeric and the Km values were 2.2 and 1.4 mM for L-malate and GSH, respectively. The enzyme had a pH optimum of 7.5. DCE-GS levels in yeast cells were significantly higher in aerobic cultures than in anaerobic ones. DCE-GS was synthesized in cells cultured between 20 and 35°C.
We investigated the inhibitory effect of the glycoprotein fraction (fraction 2) extracted from Miscanthus sinensis ANDERSSON (M. sinensis) on biphasic cutaneous reactions in mice passively sensitized with IgE. Biphasic skin reactions with peak responses at 1 (IPR, immediate phase reaction) and 24 h (LPR, late phase reaction) were caused by passive sensitization with an anti-dinitrophenol IgE monoclonal antibody (anti-DNP IgE mAb) followed by an epicutaneous challenge of 0.1% dinitrofluorobenzene (DNFB) in 100% ethanol. Intraperitoneal injection of fraction 2 before the DNFB challenge significantly inhibited the biphasic ear swelling response in passively sensitized mice in a dose-dependent manner (1-30 mg/kg). We also found that fraction 2 was effective at inhibiting the vascular permeability in mouse ear induced by an injection of compound 48/80, histamine or serotonin. In addition, fraction 2 inhibited scratching behavior as weel as ear edema observed within 2 h after DNFB challenge. Marked inhibition was observed in both passively sensitized and non-sensitized mice. The locomotor activity of mice was also reduced by the administration of fraction 2 as well as by diphenhydramine. These results suggest that the inhibitory effect of glycoprotein fraction 2 of M. sinesis on an IgE-mediated allergic inflammatory reaction is due to the protection of mediator-induced vascular permeability and that in addition to the inhibition of an inflammatory reaction, a sedative action is responsible for the inhibition of allergy-induced scaratching responses.
Slow channel blockers play a major role in the treatment of cardiovascular disease. The intention of this study was to investigate the electrophysiological properties of MM 4 (1-[N-[2-(3, 4-dimethoxy-phenyl)ethyl]-N-methylaminoacetyl]-1, 2, 3, 4-tetrahydropyrido[2, 3-b][1, 4]thiazepine fumarate) and MM 6 (1-[N-[2-'3, 4-dimethoxy-phenyl)ethyl]-N-methylaminopropionyl]-1, 2, 3, 4-tetrahydropyrido[2, 3-b][1, 4]thiazepine fumarate), two newly synthesized compounds structurally related to KT-362 (5-[3-[[2-(3, 4-dimethoxy-phenyl)ethyl]-amino]-1-oxopropyl]-2, 3, 4, 5-tetra-hydro-1, 5-benzothiazepine fumarate), by means of the conventional intracellular microelectrode technique. In various guinea pig heart muscle preparations, MM 4 and MM 6 exerted very similar effects though the action of MM 6 was more pronounced. In a concentration range from 3 to 100 μmol/l the compounds did not produce any significant change in transmembrane action potential parameters of papillary muscle and left atria, whereas the action potential duration at 20% and 50% time to repolarization in spontaneously beating Purkinje fibers was significantly shortened. In sinoatrial nodes action potential amplitude, Vmax, rate of activity and slope of slow diastolic depolarization were decreased, whereas the time to 50% and 90% repolarization was significantly prolonged. A decrease in the slow calcium inward current may account for the observed effects. In contrast to KT-362, MM 4 and MM 6 do not seem to affect the fast sodium inward current. It was concluded that replacement of the 1, 5-benzothiazepine nucleus by a 1, 4-pyridothiazepine structure and/or methylation of the side chain may weaken or even eliminate sodium channel blocking ability while calcium antagonistic characteristics are preserved. Shortening of the side chain might result in a general loss of activity.
Diabetic cataracts are though to be caused by hyperglycemia associated with disturbed glucose metabolism.Diabetes mellitus often involves abnormal lipid metabolism in addition to abnormal glucose metabolism. To date, however, very few studies have counted hyperlipidemia as a risk factor for diabetic cataracts. The present study was undertaken to determine whether this abnormal lipid metabolism is a risk factor for diabetic cataracts in rats. Cataracts were caused by streptozotocin (STZ) administration in the ordinary diet or cholesterol rich diet fed rats. When rats with STZ (65 mg/kg)-induced diabetes mellitus were fed an ordinary diet, cataracts became evident at 9 weeks in 26.7% of animals, and increased to an incidence of 53.3% after 10 weeks of STZ treatment.However, in rats with STZ-induced diabetes mellitus that were fed a cholesterol rich diet to induce severe hyperlipidemia and low high density lipoprotein (HDL) cholesterol, cataracts were observed one week earlier, after 8 weeks of treatment, in 40.0% of animals, with an increase to a 53.3% incidence and an 86.7% incidence after 9 and 10 weeks of STZ treatment, respectively. Plasma glucose levels did not differ between the goups. These results suggest that hyperlipidemia and low HDL cholesterol are associated with an earlier onset and an elevated incidence of diabetic cataracts. We then investigated the relationship between plasma lipids and cataracts by STZ (45-85 mg/kg) administration. The results showed that the onset of cataracts correlated positively with plasma total cholesterol, triglyceride, non-HDL cholesterol and glucose levels, and negatively with HDL cholesterol levels. The results of this study suggest that hyperlipidemia and low HDL cholesterol levels may be risk factors for the onset of diabetic cataracts and that diabetic cataracts may be accelerated by hyperlipidemia and low HDL cholesterol in rats.
Providone-iodine (PVP-I), an antibacterial medicine, was infiltrated in sepiolite (SPL). The available iodine content in this new pharmaceutical product, a sepiolite preparation containing povidone-iodine (PVP-I-SPL), was retained at 98.9 and 98.3% during storage at 40°C for 3 and 6 months, respectively. The effective removal of various gasses, including ammonia, hydrogen sulfide, ethylmercaptan and acetaldehyde, was achieved by use of PVP-I-SPL. Especially, the concentration of ammonia gas was reduced more tha half after 30 min of exposure, suggesting that PVP-I-SPL has excellent ability to adsorb ammonia gas. The satisfactory antibacterial effect of PVP-I-SPL was also obtained by testing its minimum bactericidal concentration (MBC). No irritation reactions to the rabbit auricle or ophthalmic mucosa or to human skin were observed by the skin irritation test.The PVP-I-SPL preparation has bactericidal activity and gas-adsorbing ability; therefore, this pharmaceutical product should be useful for the prevention of infections and deodorization in hospital rooms and houses, as well as in nursing homes for elderly people.
We studied here the effects of 8 kinds of kampo-hozais clinically used to treat atopic dermatitis (Shofu-san, Toki-inshi, Unsei-in, Oren-gedoku-to, Ji-zuso-ippo, Jumi-haidoku-to, Juzen-taiho-to, Hochu-ekki-to) on delayed-type hypersensitivity (DTH), using three types of murine models such as picryl chloride (PC)-induced (contact hypersensitivity), sheep red blood cell (SRBC)-induced (Jones-Mote's reaction) and tuberculin-induced DTH response, in order to clarify and to compare the immunopharmacological action of Kampo-hozais. Most of the kampo-hozais investigated here suppressed PC-induced contact hypersensitivity, especially at the inductive phase. Comparing the efficacies of these kampo-hozais on the three types of DTH responses in mice, they were generally divided into 4 groups. Shofu-san significantly reduced PC-induced and tuberculin-induced DTH responses but not a SRBC-induced DTH response. On the other hand, Toki-inshi reduced contact hypersensitivity, tuberculin type DTH response and Jones-Mote's reaction. Ji-zuso-ippo and Juzen-taiho-to suppressed mainly Jones-Mote's reaction, and Unsei-in, Oren-gedoku-to and Jumi-naidoku-to intensively suppressed contact hypersensitivity. We thought that these findings could help us understand how to use these kampo-hozais properly.
We prepared a new liquid preparation for eradication of Helicobacter pylori (HP), and examined drug release in vitro and in vivo. The liquid preparation mainly consisted of a sodium alginate (AG) aqueous solution containing ampicillin (ABPC), an antibiotic drug, or methylene blue, a dye. Drug release was retarded by Ca pre-treatment (0.10 M, 20 s) of the AG preparation in in vitro drug release studies due to gel-formation at the liquid surface. In in vivo experiments, the AG preparations were administered orally to rats. THe rats were divided into two groups, with or without pre-administration of ranitidine hydrochloride (RH, an H2-blocker). The total remaining % of ABPC in the stomach was high in the rats administered the AG preparation compared to the ABPC solution. The AG preparation might float in the stomach without adhering to the gastric wall in the rats without pre-administration of RH. THe total remaining % of ABPC at 30 min was almost 100% in the RH pre-administration rats administered the AG preparation, and about 80% of the drug existed in fraction 2 (implying adhesion of the preparation on the gastric mucus). At 60 min, the total remaining % in the AG preparation plus Ca (mean 87%) increased about 2-fold compared to that in the AG preparation alone (mean 44%). In this case, a large portion of the remaining ABPC also existed in fraction 2. This preparation may be usuful for eradication of HP.
Cytochrome P-450 3A (CYP3A) is a drug-metabolizing enzyme dominant in the human liver. We have designed a useful method for evaluation of induction of CYP3A mRNA by various drugs using HepG2 cells known to retain liver-cellular functions. Using semi-quantitative reverse transcription-PCR (RT-PCR), we demonstrated that cultured HepG2 cells constitutively expressed CYP3A mRNA. This mRNA was expressed at high levels in culture for several days and was further induced by several drugs (e.g. rifampicin (RFP), dexamethasone). Treatment of HepG2 cells with RFP induced CYP3A mRNA in a dose- and time-dependent manner. Cells in culture for 48 h with 1 and 50 μmol/l RFP increased 2.7- and 5.0-fold in CYP3A mRNA expression in comparison with untreated controls, respectively. In contrast, no change in the amount of CYP3A mRNA was observed when the cells were treated with cimetidine which has been shown to inhibit CYP3A activity. Our method using a combination of HepG2 cells and RT-PCR allowed evaluation of the degree of induction of CYP3A mRNA both easily and rapidly.
The enhancing effects of dimethyl-β-cyclodextrin (DM-β-CyD) on the absorption of cyclosporin A (CsA) after oral administration to rats under bile duct-cannulated and -noncannulated conditions were investigated.The dissolution rate of CsA was markedly augmented by complexation with DM-β-CyD. In a closed loop in situ study, DM-β-CyD considerably increased the cumulative amounts of CsA in the mesenteric venous blood after injection of the aqueous CsA suspension into the small intestinal sac of rats. In addition, the cumulative amount ratio of M1, the dominant metabolite of CsA in rats, to CsA in the mesenteric venous blood for up to 40 min after the injection of the CsA-DM-β-CyD suspension into the sac was lower than that of the CsA suspention alone.DM-β-CyD inhibited the bioconversion of CsA in the small intestinal microsomes of rats. THese results indicate that the bioconversion of CsA was abated by complexation with DM-β-CyD. An in vivo study revealed that DM-β-CyD incerased the transfer of CsA to blood, not lymph, with low variability in the absorption after oral administration of the CsA suspension to rats. The variability of bioavailability of DM-β-CyD complex was lower than tha of Sandimmune, although the extent of bioavailability of DM-β-CyD was only a little higher than that of Sandimmune. The bioavailability of CsA or its DM-β-CyD complex was appreciably decreased by the cannulation of the bile duct of rats, and the extent of the lowering in the bioavailability in the presence of DM-β-CyD was much less serious than that of CsA alone. The present results suggest that DM-β-CyD is particularly useful in designing oral preparations of CsA with an enhanced bioavailability and a reduced variability in abdorption.
MSI-78 is a peptide analog of naturally occurring magainin 2 isolated from the skin of Xenopus laevis. The peptide is known to have one of the strongest antibacterial activities in magainin 2 analogs against methicillin-resistant Staphylococcus aureus (MRSA). To find novel compounds superior to MSI-78, we have further designed, synthesizing 1, 1-di(4-aminobutyl)-6-benzylindane (PM4) and 1, 1-dibenzyl-6-(4-aminobutyl) indane (PM5), and tested their inhibitory ability of the growth of S. aureus. In an in vitro assay, PM4 showed the same antibacterial activity against the bacterium as MSI-78, and non-hemolytic activity against human red blood cells (RBCs) at the MIC (minimum inhibitory concentration )value, in contrast to the latter. On the other hand, PM 5 showed stronger antibacterial activity than MSI-78, but being still accompanied with hemolysis at the MIC value. Otherwise, stronger decarboxylase activity for oxaloacetate was observed in PM5, rather than magainin 2 analogs or Oxaldie 1 as a control peptide, but not in PM4.
A rapid and simple cytochrome P450 (CYP) 2C19 genotyping system was established by making several modifications in previously reported procedures. PCR conditions were modified to be capable of simultaneous amplification of CYP2C19ml and CYP2C19m2 regions. Intensive bands of 169 bp for the CYP2C19ml region and 329 bp for the CYP2C19m2 region with low background were obtained by using PCR condition involving an initial denaturation of 5 min at 94°C, 35 cycles of 1 min at 94°C; 1 min at 53°C; 1 min at 72°C, and final extension of 5 min at 72°C. Next, the optimal restriction enzyme digestion conditions were determined by using PCR products from a subject of wt/wt genotype. Both products were completely digested with 5 U of the corresponding enzyme (Sma I for CYP2C19ml and Bam HI for CYP2C19m2) for 1 h incubation at an optimal temperature. The incidence (16%; 5/32) of subjects homozygous for mutant alleles determined by an established assay system agreed well with the incidence of the poor metabolizer (PM) phenotype in the Japanese population. The established genotyping system would, therefore, be applicable to the clinical laboratory testing of patients with a PM phenotype of CYP2C19 to select appropriate and effective medication.
Glycyrrhetic acid mono-glucuronide (GAMG), 1-(18β-glycyrrhet-3-yl)-β-D-glucopyranuroic acid, was hydrolyzed to glycyrrhetic acid (GA) by GAMG β-D-glucuronidase in Eubacterium sp. GLH from human intestinal bacteria. The enzyme had an optimum pH of 5.0 and was purified from a crude extract by Butyl Toyopearl 650 S, Toyopearl HW-55 S, Hydroxyapatite and DEAE-Toyopearl 650 M column chromatography. the purified enzyme showed a specific activity of 495 nmol/min/mg protein and a single band on Coomassie brilliant blue staining and a molecular weight of about 43 kDa on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis.The apparent molecular weight was 49.5 kDa, as estimated by Toyopearl HW-55 S column chromatography. Also, the enzyme seemed to have a sulfhydryl group(s) in its active site with a K, m value of 77×10-3 M.
Deoxycytidine kinase from Ehrlich carcinoma cells was purified 10400-fold by ammonium sulfate fractionation and affinity chromatography using Sepharose 4B coupled to 2'-C-cyano-2'-deoxy-1-β-D-arabinofuranosyl-N4-palmitoylcytosine, with a yield of 45%. The purified enzyme preparation showed a single major band with a molecular weight of 32000 on SDS-PAGE. The enzyme phosphorylated deoxyadenosine, deoxyguanosine, cytidine, and several deoxycytidine analogues as well as deoxycytidine. Also, the kinetic parameters of the enzyme for the substrates were estimated.
The present study was conducted to determine the characteristics of the effects of Keishi-ka-shakuyaku-to (Gui-Zhi-Jia-Shao-Yao-Tang; TJ-60) on diarrhea. Significant repression was noted by TJ-60 at 1000 mg/kg, p.o.for diarrhea induced by pilocarpine, barium chloride or castor oil. Under normal conditions, TJ-60 did not influence small intestinal transit by its oral treatment even at 1000 mg-kg, however, it dose-dependently improved the acceleration of such transit caused by neostigmine. TJ-60 did not influence the resting tonus in isolated small intestine, but did selectively inhibit low frequency electrostimulated contractions.These results indicate that the antidiarrheal effects of TJ-60 may be due to the inhibition of excessively accelerated small intestinal movement, and that the inhibition of acetylcholine release by parasympathetic nerves is partly involved in the mechanism of this antidiarrheal action.
Nitro reduction is a critical step in the mutagenic activation of nitroarene. Nitroarene and quinone are known to be reduced by common enzymes, and thus, naphthoquinone (NQ) was studied for its effects on the mutagenicity of nitroarene in the Ames test using Salmonella typhimurium TA98 without S9. The mutagenicity of 1, 3-dinitropyrene in TA98 was found to increase 9- and 6-fold as much in the presence of 70 nmol/plate of 2-methyl-1, 4-NQ and 2-hydroxy-1, 4-NQ, respectively. Mutagenicity also became greater in 1, 3, 5-trinitronaphthalene, 1-nitropyrene and 3-nitrofluoranthene. Seventy nmol/plate of 2-methyl-1, 4-NQ incerased the mutagenicity of 1-nitropyrene by 10.5-fold as much.
The effects of orally administered dimethyl-4, 4'-dimethoxy-5, 6, 5', 6'-dimethylene-dioxybiphenyl-2, 2'-dicarboxylate (DDB) on the hepatotoxicity induced by carbon tetrachloride, acetaminophen or ethanlo were investigated in rats and mice. Either single or repeated DDB pretreatment (50 or 200 mg/kg) did not alter the hepatotoxicity induced by carbon tetrachloride (0.2 or 1.0 ml/kg, i.p.) in female rats as indicated by increases in the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and sorbitol dehydrogenase (SDH) in serum. The hepatotoxicity of acetaminophen (350 mg/kg, i.p.) was also unaffected in male mice pretreated with DDB (50 mg/kg/d) for a week. However, DDB administration (50 mg/kg/d for 7d) decreased the hepatic fatty degeneration induced by repeated ethanol treatment (0.75 g/kg. i.p., ×2 times a day for a week) in rats as shown by the accumulation of triglycerides and cholesterol in the liver. Malondialdehyde (MDA) formation in liver homogenates was inhibited by DDB treatment. The significance of the action of DDB on alcoholic fatty liver generation in clinical sttings is discussed.
The complete amino acid sequence of [2Fe-2S] ferredoxin from Nicotiana tabacum has been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by trypsin and Asp-N endoproteinase digestions. This ferredoxin exhibited 9, 10, 8, and 10 differences respectively in its amino acid sequence, when compared with the ferredoxins of Datura stramonium, D. metel, D. adborea, and Physalis alkekengi var.francheti but 17-28 differences for other angiosperms, and 34-37 differences for fern and horsetails. These results are in agreement with the taxonomic position for these plants.
1-(β-D-Arabinofuranosyl)-2-thiocytosine (araSC), a 2-substituted derivative of cytarabine (araC), has been investigated for its cytotoxicity, enzymatic stability, plasma concentration-time profile in mice, and cytokinetics.This derivative showed strong cytotoxicity in several mammalian cell lines, although activity (IC50s) was weaker than araC. Greater stability to mouse cytidine deaminase was observed; the half-line in the presence of the enzyme was about 4-times longer than that of araC. The plasma concentration-time profile in mice in vivo showed prolonged retention of araSC when compared with araC. Cytokinetic study using flow cytometry indicated a non-S-phase specific effect of this compound.
The effects of various fatty acid glycerol esters (FAGE) on the transport of ceftibuten in rat intestinal brushborder membrane vesicles (BBMV) were investigated. The HLB numbers of FAGE used were in the range of 6.5 to 15.7. The ceftibuten uptake by BBMV was examined by a rapid filtration method in the presence or absence of FAGE of a non-solubilizing concentration (0.03%, w/v). Tetraglycerol monostearate, hexaglycerol monostearate, hexaglycerol sesquistearate, hexaglycerol tristearate and decaglycerol tristearate were found significantly to enhance the ceftibuten uptake under an inward H+-gradient condition, while FAGE with oleate or laurate did not affect it. The extent of ceftibuten uptake with FAGE correlated with the HLB of the esters (r=-0.90, p<0.001).To elucidate the mechanism of its enhancing action, the effects of FAGE on the membrane lipid fluidity were examined using the fluorescence probes diphenylhexatriene (DPH) and trimethylammonium diphenylhexatriene(TMA-DPH) which distribute in the hydrophobic inner and outer regions of the lipid layer, respectively. The increase in ceftibuten uptake induced by FAGE proved to correlate with an increase in fluidity of the outer lipid layer (r=-0.83, p<0.003) but not the inner lipid layer of BBMV. Hence, the enhancing effect of FAGE with the lower HLB on the ceftibuten transport in the presence of the H+-gradient should, at least in part, be explainded by the interaction with the outer lipid regions of BBMV.
Streptomyces sp. KM1-30 was isolated from soil as a producer of antimutagens by screening with a modified Ames test. The chemical structure of the antimutagenic metabolite was identified as streptovaricin C, which is known to inhibit DNA dependent RNA polymerase from E. coli and RNA dependent DNA polymerase from RNA tumor viruses, by MS and 1H-, 13C-NMR analyses. Addition of streptovaricin C to the cultures of UV treated Salmonella typhimurium TA100 or Trp-P-2-treated S. typhimurium TA98 decreased the frequency of mutation without a decrease in viable cell counts. The effect of streptovaricin C to the mutation induced by UV and Trp-P-2 was not desmutagenic, but antimutagenic.