Evaluation of the antimicrobial activity and materials compatibility of orthophthalaldehyde as a high-level disinfectant.

We tested the antimicrobial activity of orthophthalaldehyde (OPA) against 21 strains (16 species) of pathogenic microorganisms that cause hospital-associated infections. Changes in hepatitis B surface antigen (HBs-Ag) resulting from the addition of OPA to HBs-Ag-positive serum were measured using a radioimmunoassay. We also examined the effect of immersing medical instruments in OPA (0.55%) for 168 h at room temperature. OPA (0.5%, 0.37% and 0.25%) killed 11 strains of vegetative bacteria within 15 s, and it killed the test micro-organisms faster than 3.0% glutaraldehyde (GTA). Incubation with OPA or GTA caused levels of HBs-Ag to fall below a cut-off value within 30 s. OPA did not adversely affect instruments made from various materials. OPA demonstrated more effective antimicrobial activity than GTA against a range of microorganisms. We conclude that OPA should replace GTA as the first-choice high-level disinfectant for endoscopes, considering its antimicrobial efficacy and low inhalation toxicity.

Inhibitory activities of (-)-epigallocatechin-3-O-gallate against topoisomerases I and II.

The substitution of gallic acid at the 3 position of (-)-epigallocatechin-3-O-gallate (EGCG) increased the inhibition against topoisomerase I from calf thymus gland and topoisomerase II from human placenta, and the substitution of a hydroxyl group at the 3' position increased the inhibition against the topoisomerase I. These results suggested that the 3 and 3' positions of the EGCG molecule play important roles in the process of inhibition of topoisomerases I and II. EGCG showed strong inhibition against topoisomerases I from wheat germ, calf thymus gland and Vero cells, and showed weak or no inhibition against topoisomerases I from carcinoma cells such as A549, HeLa and COLO 201 cells. EGCG differentially inhibited the topoisomerases I from different sources.

Pentalenolactone I and hygromycin A, immunosuppressants produced by Streptomyces filipinensis and Streptomyces hygroscopicus.

Immunosuppressants isolated from Streptomyces filipinensis and S. hygroscopicus were identified with pentalenolactone I and hygromycin A, respectively. The compounds as well as cyclosporin A showed immunosuppressant activity in the mixed lymphocyte reaction, and pentalenolactone I and cyclosporin A suppressed IL-2 production, however, hygromycin A did not. Hygromycin A may have immunosuppressant activity by a different mechanism from pentalenolactone I, cyclosporin A and tacrolimus.

Isoaurostatin, a novel topoisomerase inhibitor produced by Thermomonospora alba.

A novel inhibitor of topoisomerase I designated as isoaurostatin (1) was isolated from the culture filtrate of Thermomonospora alba strain No. 1520. The structure of 1 was determined to be 6,4'-dihydroxyisoaurone on the basis of spectroscopic (NMR and MS) methods. Compound 1 inhibited the relaxation activity of calf thymus topoisomerase I in a noncompetitive manner and did not inhibit the relaxation and decatenation of human placenta topoisomerase II. Compound 1 is an inhibitor belonging to cleavable complex-nonforming type without DNA intercalation.

Biosynthesis of fattiviracin FV-8, an antiviral agent.

Streptomyces microflavus strain No. 2445 produces many derivatives of fattiviracin antibiotics. The major product of these derivatives is fattiviracin FV-8, which consists of four glucose and two trihydroxy fatty acid residues. We found that this strain has the ability to convert several sugars in the culture medium to glucose, and the glucose added to the medium is directly incorporated into the FV-8 molecule. Two trihydroxy fatty acid residues in the FV-8 molecule are derived from acetic acid, and production of FV-8 is inhibited by the addition of cerulenin, which is an inhibitor of fatty acid biosynthesis.

Antiviral activity of fattiviracin FV-8 against human immunodeficiency virus type 1 (HIV-1).

A novel antiviral agent, fattiviracin FV-8, purified from the culture broth of Streptomyces microflavus strain No. 2445, showed potent antiviral activities against human immunodeficiency virus type 1 (HIV-1), herpes simplex virus type 1 (HSV-1), varicella-zoster virus (VZV), and influenza A and B viruses. The action mechanism of fattiviracin FV-8 against HIV-1 was examined. As a result, the agent was thought to act on HIV-1 particles directly without lysis of the particles, and it affords the inhibition of viral entry into the host cells.

Inhibition of topoisomerases by fatty acids.

The inhibitory effects of various fatty acids on topoisomerases were examined, and their structure activity relationships and mechanism of action were studied. Saturated fatty acids (C6:0 to C22:0) did not inhibit topoisomerase I, but cis-unsaturated fatty acids (C16:1 to C22:1) with one double bond showed strong inhibition of the enzyme. The inhibitory potency depended on the carbon chain length and the position of the double bond in the fatty acid molecule. The trans-isomer, methyl ester and hydroxyl derivative of oleic acid had no or little inhibitory effect on topoisomerases I and II. Among the compounds studied petroselinic acid and vaccenic acid (C18:1) with a cis-double bond were the potent inhibitors. Petroselinic acid was a topoisomerase inhibitor of the cleavable complex-nonforming type and acted directly on the enzyme molecule in a noncompetitive manner without DNA intercalation.

Anti-herpes virus activity of fabaceous triterpenoidal saponins').

Anti-herpes simplex virus type 1 (HSV-1) activity of oleanane-type triterpenoidal saponins obtained from some fabaceous plants were examined. Among sophoradiol glycosides, the order of potency was kaikasaponins III>I>>sophoradiol monoglucuronide. It was suggested that the trisaccharide group showed greater action than the disaccharide group. Neither the monoglucuronide of sophoradiol nor that of soyasapogenol B showed activity. Among the trisaccharide group of soyasapogenol B, the order of activity was azukisaponin V>soyasaponin II>astragaloside VIII>>soyasaponin I. Therefore, the saponin having a glucosyl unit in the central sugar moiety seemed to show greater action. In comparison with the activities for a group having the same trisaccharide, the potency of the sapogenol moieties was found to be in the order of soyasapogenol E>sophoradiol>>soyasapogenol B. Hence, the carbonyl group at C-22 would be more effective than the hydroxyl group in anti-HSV-1 activity, while the hydroxyl group at C-24 could reduce the activity.

Anti-herpes virus activity of Solanum steroidal glycosides.

Since some Solanum-genus plants have traditionally been used for anti-cancer and anti-herpes agents from olden times, we examined anti-herpes simplex virus type 1 (HSV-1) activity of typical steroidal glycosides with the frameworks of spirostane (including nuatigenin glycoside), furostane, solasodane, tomatidane and ergostane (including dimer) obtained from Solanum plants. Among these steroidal glycosides, the spirostanol glycosides were most effective. An inclination was observed for the potency of activity to decrease in the order of spirostane, tomatidane, ergostane, solasodane, nuatigenin type, dimer of ergostane and furostane. It was also suggested that the activity depends on the kind of oligosacchride moiety.

Topostatin, a novel inhibitor of topoisomerases I and II produced by Thermomonospora alba strain No. 1520. III. Inhibitory properties.

A novel inhibitor of topoisomerases designated as topostatin was isolated from the culture filtrate of Thermomonospora alba strain No. 1520. The inhibitory activity of topostatin was shown to be pH- and temperature-dependent with a maximum around at pH 6 and 28 degrees C. The stability of topostatin decreased with decreasing pH and rising temperature. Topostatin inhibited topoisomerases I and II in a competitive manner with respect to DNA. The inhibitor also inhibited some restriction endonucleases such as Sca I, Hind III and Pst I, but not Alu I, Bam HI, Eco RI, RNase A, DNase I, DNase II and DNA ligase. Topostatin did not induce the nuclear accumulation of p53 protein by DNA damage in the normal human cells.

The Consumer Assessment of Health Plan Study (CAHPS) survey of children's health care.

BACKGROUND: Little is known about the experience of children and families with pediatric care. Asking parents about their experiences and the treatment of their children in health care plans can yield important information about selected aspects of medical care quality. Such data can be used to motivate, focus, and evaluate quality improvement efforts. METHODS: Development of the Child Core Survey followed the survey development principles of the Consumer Assessment of Health Plan Study (CAHPS) project, starting with assembly of existing instruments, consultation with experts, focus groups, and cognitive testing. A field test of the survey was conducted by mail among members enrolled in 1 of 25 plans originally identified as providing health care services to the public employees of the state of Washington (response rate, 52%). RESULTS: The 3,083 respondents rated personal doctors most highly, with overall care and specialty care rated nearly as well, and plan administration rated lowest. Parent-clinician and child-clinician communication, as well as spending sufficient time with the child were the strongest correlates of assessments of overall care and of personal doctors. Plans differed significantly in their performance along all the dimensions of child health care assessed in the survey except for aspects of access ("getting the care you need"). IMPLICATIONS: The Child Core Survey from the CAHPS provides a readily accessible method to assess the interpersonal care of children. Such data could be used to make plans accountable to the needs of children, to focus specific improvement initiatives, or both.

Inhibition of topoisomerases by antitumor prostaglandins.

Topoisomerase II from human placenta was strongly inhibited by prostaglandins such as delta 12,14-PGJ2, delta 12-PGJ2, PGA2, and PGA1, which have an antitumor activity. The topoisomerase II inhibitions by these prostaglandins were dose-dependent and IC50 values of delta 12,14-PGJ2, delta 12-PGJ2, PGA2, and PGA1 were 22 microM, 74 microM, 75 microM, and 98 microM, respectively. Topoisomerase I from calf thymus gland was inhibited by delta 12,14-PGJ2, delta 12-PGJ2, and PGA2, but not inhibited by other prostaglandins even at high concentrations. Only the prostaglandins having the antitumor activity inhibited topoisomerase II. The results suggest that the antitumor activity of prostaglandins may be related, at least in part, to inhibition of topoisomerase II in tumor cells.

Fattiviracin A1, a novel antiherpetic agent produced by Streptomyces microflavus Strain No. 2445. I. Taxonomy, fermentation, isolation, physico-chemical properties and structure elucidation.

A novel antiherpetic agent, fattiviracin A1, was isolated from the culture broth of strain No. 2445 identified as Streptomyces microflavus. It was purified through 1-butanol extraction, column chromatographies on Diaion HP-10 and silica gel and HPLC using a reverse phase column. The structure of fattiviracin A1 was determined by several spectroscopic experiments and chemical degradations. It is a new macrocyclic diester consisting of four D-glucose units and two (C24 and C33) hydroxy fatty acids. It is closely related to cycloviracins B1 and B2, but differs from these known compounds in both the length of its side chain and the sugar moiety.

1513-DMIa and 1513-DMIb, DNA methyltransferase inhibitors produced by Streptomyces sp. strain No. 1513.

Two new methyltransferase inhibitors were isolated from the culture filtrate of Streptomyces sp. strain No. 1513 and named 1513-DMIa and 1513-DMIb. 1513-DMIa and 1513-DMIb were distinguished in certain properties from DMI-1, DMI-2, DMI-3 and DMI-4 previously reported. The molecular weight of 1513-DMIa and 1513-DMIb were estimated to be 576 and 8400 from the results of FAB-MS and gel filtration, respectively. The inhibitory activities of 1513-DMIa and 1513-DMIb were shown to be pH- and temperature-dependent and both inhibited M. EcoRI in an uncompetitive manner with respect to DNA or S-adenosylmethionine (SAM).

Enhanced reporter gene expression in cells transfected in the presence of DMI-2, an acid nuclease inhibitor.

Cellular nuclease activity is a potential barrier to the successful delivery of foreign genes to mammalian cells. We tested the hypothesis that transfection in the presence of a specific DNase inhibitor can enhance the expression of foreign gene products. We have used DMI-2, a polyketide metabolite of Streptomyces sp. strain 560 to enhance the expression of bacterial chloramphenicol acetyltransferase (CAT) in the human lung adenocarcinoma cell line H441. DMI-2 has been shown previously to inhibit porcine DNase II, an acid pH nuclease contained in the endosomal/lysosomal compartment. Transfection of H441 cells in the presence of 0.1-1 microgram/ml DMI-2 caused: (1) 10-fold enhancement of CAT activity when the bacterial plasmid was complexed with either surfactant protein A-poly-lysine or transferrin-poly-lysine; (2) 1.5- to two-fold enhancement of CAT activity in cells exposed to lipofectin-DNA complexes: (3) no effect on transfection via calcium phosphate co-precipitation. DMI-2 alone showed no inherent transfection activity. In experiments using SP-A-poly-lysine and plasmid containing the beta-galactosidase reporter gene, DMI-2 increased the number of transfected cells. Methanolysis products of DMI-2 did not inhibit DNase II and did not enhance transfection efficiency. Taken together, the data support the hypothesis that nuclease action is a significant barrier to expression of foreign genes and inhibition of specific nucleases may facilitate transfection.

Inhibition of DNA topoisomerases by microbial inhibitors.

New inhibitors of DNA topoisomerase named 2280-DTI and 2890-DTI have been discovered in the culture filtrates of Micromonospora sp. strain No. 2280 and Streptomyces sp. strain No. 2890, respectively. Both inhibitors were purified from each culture filtrate by column chromatography on Diaion, Dowex and gel filtration. Both inhibitors were thermostable acidic substances with high molecular weight and inhibited topoisomerase I in a non-competitive manner. They differed from well-known inhibitors of topoisomerases such as camptothecin and doxorubicin, which inhibit the DNA rejoining reaction of the enzyme by intercalation into DNA strands or stabilizing the cleavable complex (enzyme-DNA reaction intermediate). 2280-DTI and 2890-DTI did not intercalate into DNA strands and also had no ability to stabilize the cleavable complex. It is suggested that 2280-DTI and 2890-DTI inhibit the DNA breaking and rejoining reactions of topoisomerase by direct action on the enzyme molecule.

Topostatin, a novel inhibitor of topoisomerases I and II produced by Thermomonospora alba strain No. 1520. I. Taxonomy, fermentation, isolation and biological activities.

A novel inhibitor of topoisomerases designated as topostatin was isolated from the culture filtrate of Thermomonospora alba strain No. 1520. Topostatin inhibited the relaxation of supercoiled pBR322 DNA by calf thymus topoisomerase I, and also inhibited the relaxation of supercoiled pBR322 DNA and decatenation of kinetoplast DNA by human placenta topoisomerase II. Topostatin had neither ability to stabilize the cleavable complex nor ability to intercalate into DNA strands. The inhibitor exhibited growth inhibitory activity against the tumor cells (SNB-75 and SNB-78) of central nervous system, but did not exhibit any antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts and fungi.

Topostatin, a novel inhibitor of Topoisomerases I and II produced by Thermomonospora alba strain No. 1520. II. Physico-chemical properties and structure elucidation.

Topostatin is a new topoisomerase inhibitor isolated from the culture filtrate of Thermononospora alba strain No. 1520. The inhibitor inhibits topoisomerases I and II, and it has neither ability to stabilize the cleavable complex nor ability to intercalate into DNA strands. The molecular formula of topostatin was determined as C36H58N4O11S based on the FAB-MS analyses, and the structure was elucidated to be a novel 14-membered ring containing peptide and terpenoid by various NMR spectroscopies.