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Development of a gas-phase oxygen biosensor using a blue copper-containing oxidase.

A gas-phase oxygen biosensor based on blue copper-containing oxidases was developed. Blue-oxidase enzymes, including laccase and ascorbate oxidase, have a blue chromophore prosthetic group, type 1 Cu+2, which can be reduced and decolorized with reducing substrates. When the enzyme is reoxidized with molecular oxygen, there is a concomitant return of the blue color. The oxygen biosensor consisted of the Rhus vernicifera laccase and ascorbate as substrate enclosed in pouches of low-density polyethylene under nitrogen gas. Operational stability of the biosensor was established by exposing it to different oxygen/nitrogen gas mixtures at 5 degrees C. Gas-phase oxygen concentrations were measured by keeping it under nitrogen gas and subsequently recording the rate of reappearance of the enzyme blue color, both visually and spectrophotometrically at 610 nm. The oxygen biosensor was able to detect a wide range of oxygen concentrations. The time required to recover the blue color, namely the biosensor response time, at the optimized assay conditions of 5 degrees C and a high-water activity level, was determined. This research describes the development of an oxygen biosensor with adequate activity and stability to measure gas-phase oxygen concentrations at 5 degrees C and high-water activity levels. The oxygen biosensor could be used to indicate oxygen concentrations above acceptable levels in headspace oxygen concentration which could affect the quality and safety of products packaged under initial low levels of oxygen concentration.

Ascorbate Oxidase↗

Efficacy of traditional herbal medicines in combination with acyclovir against herpes simplex virus type 1 infection in vitro and in vivo.

Traditional herbal medicines have been safely used for the treatment of various human diseases since ancient China. We selected 10 herbal extracts with therapeutic antiherpes simplex virus type 1 (HSV-1) activity. Among these, Geum japonicum Thunb., Rhus javanica L., Syzygium aromaticum (L.) Merr. et Perry, or Terminalia chebula Retzus showed a stronger anti-HSV-1 activity in combination with acyclovir than the other herbal extracts in vitro. When acyclovir and/or a herbal extract were orally administered at doses corresponding to human use, each of the 4 combinations significantly limited the development of skin lesions and/or prolonged the mean survival times of infected mice compared with both acyclovir and the herbal extract alone (P < 0.01 or 0.05). These combinations were not toxic to mice. They reduced virus yields in the brain and skin more strongly than acyclovir alone and exhibited stronger anti-HSV-1 activity in the brain than in the skin, in contrast to acyclovir treatment by itself. Combinations of acyclovir with historically used herbal medicines showed strong combined therapeutic anti-HSV-1 activity in mice, especially reduction of virus yield in the brain.

Acyclovir↗

Prophylactic treatment of cytomegalovirus infection with traditional herbs.

Hot water extracts of four traditional herbs, Geum japonicum, Syzygium aromaticum, Terminalia chebula and Rhus javanica, which have been shown to have anti-herpes simplex virus (HSV) activity in vivo, were examined for anti-cytomegalovirus (CMV) activity in vitro and in vivo in this study. They inhibited replication of human CMV and murine CMV (MCMV) in vitro. These anti-CMV activities in vivo were examined in an MCMV infection model using immunosuppressed mice. Mice were subcutaneously treated with various doses of cyclosporine, and immunosuppression and MCMV infection were monitored by suppression of antibody production and virus yield in the lung, respectively. Each herbal extract was orally administered to mice treated with 50 mg/kg of cyclosporine from a day before intraperitoneal infection, and the efficacy of herbs was evaluated by the reduction in the virus yield in the lung. Among them Geum japonicum, Syzygium aromaticum, and Terminalia chebula significantly suppressed MCMV yields in lungs of treated mice compared with water treatment. Efficacy of oral treatment with 750 mg/kg per day of Geum japonicum extract was similar to that of the intraperitoneal administration of 2 mg/kg per day of ganciclovir in increasing the body weight of infected mice and reducing the virus yield in the lungs. These herbs may be beneficial for the prophylaxis of CMV diseases in immunocompromised patients.

Animals↗

The metal site of stellacyanin: EXAFS studies of the Cu(II), Cu(I), Ni(II) and Co(II) derivatives.

Extended X-ray absorption fine structure (EXAFS) studies of Cu(II) (oxidized), Cu(I) (reduced), Ni(II) and Co(II) stellacyanin from Rhus vernicifera are reported. For Cu(II) stellacyanin, the coordination by three close ligands, viz. 2 N and 1 S, with the presence of smaller shells pointing to imidazole coordination, indicates similarities with the coordination in other so-called type 1 or 'blue'-copper proteins. Upon reduction, slightly longer ligand distances and an additional sulphur ligand are found. Ni(II) and Co(II) stellacyanin resemble Cu(I) and Cu(II) stellacyanin, respectively, in ligand distances, but have a tendency for three rather than two N (or O) ligands in the first shell. The results are compared with the three-dimensional model derived from 1H-NMR relaxation measurements for Co(II) stellacyanin, and are consistent with the proposal that apart from the three close ligands found in all blue-copper proteins, a sulphur from a disulphide bridge and the amide oxygen from an asparagine residue come to within coordinating distance of the metal in stellacyanin.

Binding Sites↗

Oxidant air pollution effects on plants of Joshua Tree National Monument.

Joshua Tree National Monument (JOTR) is located about 100 km east of the Los Angeles Basin, site of the heaviest concentration of photochemical oxidant (O(3)) air pollution in the US. This investigation was conducted to measure O(3) concentrations in JOTR and to determine the effects of O(3) on vegetation in the park. Potentially phytotoxic concentrations of O(3) were recorded in JOTR in 1984 and 1985, but peak concentration occurred at night, when most plant species would be less sensitive to O(3). No O(3) effects were observed on permanent vegetation observation plots in JOTR in 1984 or 1985. Controlled exposures of native summer annual and woody perennial species to O(3) showed that most did not develop visible O(3) injury symptoms except at concentrations higher than those expected in the park. However, Rhus trilobata Nutt. was injured at 0.10 ppm O(3), 4 h per day for 4 days. This species would be a useful bioindicator to assess the effects of O(3) on native desert plants.

Journal Article↗

Antibiotic screening of medicinal plants of the British Columbian native peoples.

One hundred methanolic plant extracts, 96 of which had documented medicinal uses by British Columbian native peoples, were screened for antibiotic activity against 11 bacterial strains. Eighty-five percent were found to have significant antibiotic activity against at least two of the bacteria tested. Ninety-five percent of the plants categorized as potential antibiotics based on their ethnobotanical usage were found to exhibit significant antibiotic activity. Seventy-five were found to be active against methicillin-resistant Staphylococcus aureus, 46 were active against an antibiotic supersusceptible strain of Pseudomonas aeruginosa and 18 of these were also active against a wild type strain. The extracts with the broadest spectra of activity were prepared from: Alnus rubra bark and catkins, Fragaria chiloensis leaves, Moneses uniflora aerial parts, and Rhus glabra branches.

Anti-Bacterial Agents↗

Antifungal screening of medicinal plants of British Columbian native peoples.

One hundred methanolic plant extracts were screened for antifungal activity against 9 fungal species. Eighty-one were found to have some antifungal activity and 30 extracts showed activity against 4 or more of the fungi assayed. The extracts with the greatest fungal inhibition were prepared from Alnus rubra catkins, Artemisia ludoviciana aerial parts, Artemisia tridentata aerial parts, Geum macrophyllum roots, Mahonia aquifolium roots and Moneses uniflora aerial parts. In addition to these, extracts prepared from the following plants also exhibited antifungal activity against all 9 fungi: Asarum caudatum whole plant, Balsamorhiza sagittata roots, Empetrum nigrum branches, Fragaria chiloensis leaves, Gilia aggregata aerial parts and roots, Glehnia littoralis roots, Heracleum lanatum roots, Heuchera cylindrica roots and Rhus glabra branches.

Antifungal Agents↗

Incidence of ozone symptoms on vegetation within a National Wildlife Refuge in New Jersey, USA.

During 1993-1996 and 2001-2003, we evaluated the percentage of plants (incidence) exhibiting ozone-induced foliar symptoms on vegetation within a National Wildlife Refuge located along the Atlantic Ocean coast of New Jersey, USA. Incidence varied among plant species and years. Bioindicator plants most sensitive to ozone, across all years, included native common milkweed (Asclepias syriaca) and wild grape (Vitis spp.), as well as introduced tree-of-heaven (Ailanthus altissima). Less sensitive bioindicators included Virginia creeper (Parthenocissus quinquefolia) and winged sumac (Rhus coppolina). Black cherry (Prunus serotina) and sassafras (Sassafras albidum) were least sensitive. The greatest incidence of ozone symptoms, across all plant species, occurred in 1996, followed by 2001>1995>1994>1993>2003>2002. A model was developed that showed a statistically significant relationship between incidence of ozone symptoms and the following parameters: plant species, Palmer Drought Severity Index, and the interaction of W126 x N100 measures of ambient ozone.

Air Pollutants↗

Homeopathic treatment of radiation-induced itching in breast cancer patients. A prospective observational study.

Following surgery for carcinoma of the breast, patients receive local radiotherapy. This can cause itching, which may be severe, in the radiation field. The affected skin usually is dry, rough and red. Twenty-five patients were treated homeopathically for radiation-induced itching. Fourteen patients developed itching during their course of post-operative radiation at 27 days median (range: 14-40). Eleven patients experienced itching in the radiation field after completion of treatment (median 21 days) after the end of their radiation treatment. A single dose of an individually selected homeopathic medicine in 30C dilution was given in the clinic, on the basis of repertorisation. Patients were asked to record a visual analogue scale (VAS) before prescription of the homeopathic medicine and at follow-up. Patients were evaluated at median 3 days (range: 1-27 days) after administration of the homeopathic medicine. In total, 14 of 25 patients (56%) responded to the first medicine. Nine patients had a second medicine, seven responded. Altogether 21 of 25 (84%) patients were successfully treated. The following medicines were employed successfully: Fl-ac 9/13, Rhus-t 3/5, Caust 2/3, Ign 2/2, Psor 2/2, gamma-ray 2/2 and Kali-bi 1/1. The VAS measurements before and after homeopathic treatment showed a reduction of the median value of 64mm (range: 20-100mm) to 34mm (median; range: 0-84mm). Homeopathic treatment of radiation-induced itching appears quite successful. The most frequently indicated and most frequently effective medicine was Fluoric acid. An approach that allows greater understanding of the patient as a whole in the short time available in a busy clinic may be required.

Adult↗

Hahnemann's experiments with 50 millesimal potencies: a further review of his casebooks.

BACKGROUND: A review of Hahnemann's clinical records at the Institute for History of Medicine of the Robert Bosch Foundation in Stuttgart shows that until the end of his life, Hahnemann continued to refine his clinical method, based on clinical cases. His "most perfected method" motivated him to write the sixth edition of the The Organon of the Healing Art, proposing solutions controlling the side effects he observed with repeated doses of homeopathic medicines. Unfortunately, this was published many years posthumously. The sixth edition of The Organon introduced the fifty-millesimal scale. OBJECTIVES: To identify the clinical cases treated with fifty-millesimal potencies and analyze Hahnemann's use of them. RESULTS: 1836 prescriptions of fifty-millesimal potencies were found, between 1837 and 1843 in three phases: initially sporadic; later compared regularly to centesimal dynamizations; and finally systematically. Thirty five medicines were identified in fifty-millesimal prescriptions, seven in potencies higher than 10 and only 3 (Sulphur, Mercurius solubilis and Rhus toxicodendron) used in the 30th degree. This accords with Haehl's information about the remedies in Hahnemann's case of fifty-millesimal potencies. CONCLUSIONS: Hahnemann probably decided to write the sixth edition, in 1840, to incorporate his latest experience with the repetition of potentized doses and periodically modified potencies. He must have revised it after February 1842 to include his latest findings with fifty-millesimal potencies in ascending degrees. Hahnemann's conception about the superiority of the fifty-millesimal in comparison with the centesimal dynamization was based on a significant number of experiments with the two scales.

Germany↗

Poison ivy: an underreported cause of erythema multiforme.

The relationship between herpes simplex virus (HSV) and erythema multiforme (EM) has been well described. Many authors contend that EM (excluding Stevens-Johnson syndrome and toxic epidermal necrolysis) occurs almost exclusively as a response to HSV infection. During the past year, however, we have observed several cases of EM complicating severe Rhus allergic contact dermatitis. Although this association has been previously documented, the paucity of cases in the literature, along with our experience, suggests that this is an underreported phenomenon. We describe 4 of our cases.

Adolescent↗

A systematic review of contact dermatitis treatment and prevention.

BACKGROUND: Contact dermatitis (CD) is a common occupational disease. There have been no systematic reviews of CD treatment or prevention. METHODS: Multiple databases were systematically searched. Using independent double review and published quality review criteria, articles were rated as good, fair, or poor. Treatment benefit data were tabulated and conclusions were based on the rated strength of published evidence. RESULTS: In all, 49 studies met inclusion criteria. Barrier creams containing dimethicone or perfluoropolyethers, cotton liners, and softened fabrics prevent irritant CD. Lipid-rich moisturizers both prevent and treat irritant CD. Topical skin protectant and quaternium 18 bentonite (organoclay) prevent rhus dermatitis. Diethylenetriamine pentaacetic acid (chelator) cream prevents nickel, chrome, and copper dermatitis. Potent or moderately potent steroids effectively treat allergic CD. There were no macrolide immunomodulator trials that met inclusion criteria. This review did not include studies of children, animals, or non-English language publications. CONCLUSIONS: A limited number of interventions effectively prevent or treat irritant and allergic CD, but well-controlled, outcome-blinded studies, particularly in the area of allergic CD prevention are needed.

Dermatitis, Occupational↗

Rangeland dynamics of southern Ethiopia: (2). Assessment of woody vegetation structure in relation to land use and distance from water in semi-arid Borana rangelands.

The structure and advancement of woody vegetation was studied in a semi-arid rangeland of southern Ethiopia under three land-use systems (communal land, a government ranch and a traditional grazing reserve enclosure) and along a distance gradient (near, middle and far) from water sources. A total of 54 woody plant species were identified. Based on the subjective opinions of the pastoralists, 85% of the identified woody plants have forage values to livestock. Cadaba farinsoa, Ormocarpum trichocarpum, Rhus natalensis, Acacia brevispica, Cordia gharaf and Hibiscus sparseaculeatus were reported to have fair to good palatability. Tree equivalent (TE) density of all woody plants combined was greater (P <0.01) in the government ranch (1) 188 TE ha(-1)) and the communal land (1083 TE ha(-1)) than in the traditional grazing reserve (419 TE ha(-1)), whereas this did not vary significantly (P >0.05) along the distance gradient from water. The most important encroaching woody plant species in the study areas were in descending order: Commiphora africana, Acacia drepanolobium, A. brevispica, Acacia. tortilis, Grewia tembensis and Lannea floccosa. The density of individual encroaching woody plant species along the distance gradient from water was not consistent. All encroaching woody species had the highest TE density in the communal land. The prevalence of these species followed the pattern of intensity of use within the communal grazing area. Most of the woody species had the highest abundance in the height class >0-2m regardless of land use and distance gradient from water. This study investigated the advance of severe woody encroachment in the communal and government sites as well as along the distance gradient from water. Some of the important contributing factors that can be suggested are heavy grazing pressure (in both the communal and government sites), expansion of cultivation and reduced mobility of livestock due to settlement of the pastoralists in the communal land.

Conservation of Natural Resources↗

Robustaflavone, a potential non-nucleoside anti-hepatitis B agent.

Robustaflavone, a naturally occurring biflavanoid isolated from Rhus succedanea, was found to be a potent inhibitor of hepatitis B virus (HBV) replication in 2.2.15 cells, with an effective concentration (EC50) of 0.25 microM, and a selectivity index (SI, IC50/EC90) of 153. Robustaflavone hexaacetate inhibited HBV replication with an EC50 of 0.73 microM, but exhibited no cytotoxicity at concentrations up to 1000 microM. Combinations of robustaflavone with penciclovir and lamivudine displayed synergistic anti-HBV activity, having the most pronounced effects when the combination ratios were similar to the ratio of EC50 potencies. Thus, a 1:1 combination of robustaflavone and penciclovir exhibited an EC50 of 0.11 microM and an SI of 684, while a 10:1 combination of robustaflavone and lamivudine exhibited an EC50 of 0.054 microM and an SI of 894. Statistical analyses of the combination data using the Combostat program confirmed that robustaflavone exhibited synergism with both penciclovir and lamivudine.

Acyclovir↗

Magnetic studies of the trinuclear center in laccase and ascorbate oxidase approached by EPR spectroscopy and magnetic susceptibility measurements.

The trinuclear centers in Rhus vernicifera laccase and Cucumis sativus ascorbate oxidase have been studied by EPR spectroscopy and magnetic susceptibility measurements over the wide range of 5 K to 300 K. The EPR spectra showed that type II copper receives increasing tetrahedral distortion with raising temperature. Magnetic susceptibilities of laccase showed that both of type I and type II coppers are almost fully paramagnetic since the antiferromagnetic interaction between type III coppers is extremely strong from 5 K to 300 K. On the other hand, the effective magnetic moment of ascorbate oxidase is contributed by ca. 1.7 Cu2+ even below ca. 100 K, since type II Cu is partly in the reduced form. The effective magnetic moment continuously increased with raising temperature because the antiferromagnetic interaction between type III coppers is not as strong as in the case of laccase. The simulation of the SQUID measurement results suggested that the conformational change of the ascorbate oxidase molecule caused the temperature dependence of the antiferromagnetic interaction. The type II Cu EPR signals in laccase and ascorbate oxidase were conspicuously broadened with raising temperature because of the increasing contribution of the triplet state by type III Cu's and/or of the rapid relaxation which finally led to only ca. 30% detection of the type II Cu signals at room temperature. The stepwise binding of azide to the trinuclear center made one of type III Cu's to be EPR detectable. SQUID measurements indicated that only one Cu in the trinuclear center is paramagnetic and other two Cu's are antiferromagnetically coupled for both of the one- and two-azide bound forms. The binding mode of azide to the trinuclear center was discussed based on some models.

Ascorbate Oxidase↗

Immunocytochemical studies on the origin and deposition sites of hydrolyzable tannins.

Two specific antibodies, directed toward hydrolyzable tannins, i.e. polygalloylated or oxidized derivatives of 1,2,3,4,6-penta-O-galloylglucose, and toward an acyltransferase from oak leaves that catalyzes the biosynthesis of this principal precursor, were used in immunocytochemical studies to determine the intracellular sites of origin and deposition of these polyphenolic plant constituents. Immunostaining of semi-thin sections from leaves and roots of young pedunculate oak (Quercus robur, syn. Quercus pedunculata) plants with marker enzymes and immunogold labeling of ultra-thin sections revealed immunoreactive sites for both enzyme and hydrolyzable tannins in chloroplasts, cell walls and intercellular spaces. The latter non-cytoplasmic (apoplast) compartments displayed characteristic aggregations of these two epitopes, thus indicating an intimate association of the biocatalyst and its products. Identical distribution patterns for hydrolyzable tannins were observed in leaves of Rhus typhina (sumac) and Tellima grandiflora (fringe cups) which, however, displayed no affinity toward the galloyltransferase antibody that had been raised against enzyme from oak. Controls with spinach leaves, known to be devoid of tannins, were inactive in all cases. The conclusion that, besides chloroplasts, cell walls and intercellular space serve as sites for the biosynthesis and deposition of hydrolyzable tannins was confirmed by analyzing extracts from these non-cytoplasmic compartments.

Journal Article↗

Antioxidant activity in medicinal plants associated with the symptoms of diabetes mellitus used by the indigenous peoples of the North American boreal forest.

Thirty-five plant species were selected from the published literature as traditionally used by the Indigenous Peoples of the boreal forest in Canada for three or more symptoms of diabetes or its complications. Antioxidant activities in methanolic extracts support the contribution of these traditional medicines in a lifestyle historically low in the incidence of diabetes. In a DPPH assay of free radical scavenging activity 89% of the methanol extracts had activity significantly greater than common modern dietary components, 14% were statistically equal to ascorbic acid and 23% had activities similar to green tea and a Trolox positive control. Superoxides produced with an NBT/xanthine oxidase assay found scavenging was significantly higher in 29% of the species as compared with the modern dietary components and Trolox. The methanol extracts of Rhus hirta, Quercus alba and Cornus stolonifera performed similarly to green tea's in this assay. Assessment of peroxyl radical scavenging using a DCF/AAPH assay showed 60% of the plant extracts statistically similar to Trolox while R. hirta and Solidago canadensis extracts were greater than green tea, ascorbic acid and Trolox. The majority of the species (63 and 97%, respectively) had scavenging activities similar to ascorbic acid in the superoxide and peroxyl radical scavenging assays.

Antioxidants↗

Digestibility, N balance and blood metabolite levels in Alpine goat wethers fed either water oak or shining sumac leaves.

Eight Alpine wethers (8-9 months of age, 27.5+/-1.10kg body weight (BW)) were randomly assigned to consume, free-choice, either shining sumac (Rhus copallina) or water oak (Quercus nigra) leaves as a sole diet. Leaves were collected and dried prior to feeding. A 14-day adaptation period was followed by a 4-day total fecal and urine collection. Chemical composition (%) of the fed water oak and shining sumac leaves revealed similar levels of OM (95.9 and 94.1) and N (1.54 and 1.42) but higher concentrations of cell wall fractions, NDF (54.8 and 31.2) and ADF (34.5 and 26.4), in water oak than shining sumac. Body weight of wethers differed between treatments, although this was not reflected in DM intake. Average daily intake values were 616g DM, 589g OM and 9.3g N. NDF intake was significantly lower (P=0.002) in goats-fed shining sumac than in those that consumed water oak (192 versus 330g). Daily fecal output of all components was higher (P<0.05) in water oak than shining sumac-fed goats. Apparent digestibilities (%) of all components were significantly lower in water oak than shining sumac-fed animals (DM 41 versus 63, OM 42 versus 64, NDF 24 versus 37, and N 27 versus 38). Urinary N excretion, N balance and concentrations of protein, plasma urea nitrogen (PUN) and glucose in the blood were similar between diets, averaging 2.6g N per day, 0.45g per day, 85.5mg/ml, and 14.1mg/dl, respectively. The results of this trial suggest that shining sumac is utilized better by goats than water oak. The use of dried leaves in this experiment may have led to possible negative effects on nutrient characteristics that animals consuming fresh leaves may not experience.

Journal Article↗